bluuebunny/crossref_metadata_2025_split huggingface dataset:
Transduction of donor hematopoietic stem-progenitor cells with Fas ligand enhanced short-term engraftment in a murine model of allogeneic bone marrow transplantation
Authors: Whartenby Katharine A. , Straley Erin E. , Kim Heeje , Racke Frederick , Tanavde Vivek , Gorski Kevin S. , Cheng Linzhao , Pardoll Drew M. , Civin Curt I. 
Published: October 2002

AbstractFas-mediated apoptosis is a major physiologic mechanism by which activated T cells are eliminated after antigen-stimulated clonal expansion generates a specific cellular immune response. Because activated T cells are the major effectors of allograft rejection, we hypothesized that genetically modifying allogeneic bone marrow (BM) cells prior to transplantation could provide some protection from host T-cell attack, thus enhancing donor cell engraftment in bone marrow transplantation (BMT). We undertook studies to determine the outcome of lentiviral vector-mediated transduction of Fas ligand (FasL) into lineage antigen-negative (lin−) mouse BM cells (lin− BMs), in an allogeneic BMT model. FasL-modified lin− BMs killed Fas-expressing T cells in vitro. Mice that received transplants of allogeneic FasL+ lin−BMs had enhanced short-term engraftment, after nonmyeloablative conditioning, as compared to controls. We observed no major hepatic toxicity or hematopoietic or immune impairment in recipient mice at these time points. These results suggest potential therapeutic approaches by manipulating lymphohematopoietic stem-progenitor cells to express FasL or other immune-modulating genes in the context of BMT.
Activation of Rac-1, Rac-2, and Cdc42 by hemopoietic growth factors or cross-linking of the B-lymphocyte receptor for antigen
Authors: Grill Brock , Schrader John W. 
Published: October 2002

AbstractInterleukin-3 (IL-3)–induced activation of endogenous Rac-1, Rac-2, and Cdc42. Rac-1 was also activated by colony-stimulating factor-1 (CSF-1), Steel locus factor (SLF), granulocyte-macrophage colony-stimulating factor (GM-CSF), and IL-5 or by cross-linking the B-lymphocyte receptor for antigen (BCR). The activation of Rac-1 induced by cross-linking the BCR or by IL-3 stimulation was blocked only partially by Ly294002, with about 25% to 30% of Rac-1 activation still occurring in the absence of detectable increases in phosphatidyl-inositol-3 kinase (PI-3K) activity. Overexpression of constitutively active mutants of H-Ras, N-Ras, or M-Ras resulted in activation of coexpressed Rac-1 through an Ly29402-resistant, PI-3K–independent mechanism. Overexpression of constitutively active mutants of p21 Ras, or Rac-1, but not of PI-3K, was sufficient for activation of p38 mitogen-activated protein kinase (MAPK) in cells of hemopoietic origin. Inhibition of increases in PI-3K activity by Ly294002 had no effect on the IL-3–induced activation of p38 MAPK. In contrast, Ly294002 partially inhibited the activation of p38 MAPK induced by cross-linking of the BCR, although some p38 MAPK activation occurred in the absence of increases in the activity of Rac-1 or PI-3K. The activation of Rac-1, Rac-2, and Cdc42 by IL-3 and other hemopoietic growth factors is likely to be an important component of their actions in promoting growth, survival, and function.
Genomic and proteomic analysis of the myeloid differentiation program: global analysis of gene expression during induced differentiation in the MPRO cell line
Authors: Lian Zheng , Kluger Yuval , Greenbaum Dov S. , Tuck David , Gerstein Mark , Berliner Nancy , Weissman Sherman M. , Newburger Peter E. 
Published: October 2002

Abstract
We have used an approach using 2-dimensional gel electrophoresis with mass spectrometry analysis combined with oligonucleotide chip hybridization for a comprehensive and quantitative study of the temporal patterns of protein and mRNA expression during myeloid development in the MPRO murine cell line. This global analysis detected 123 known proteins and 29 “new” proteins out of 220 protein spots identified by tandem mass spectroscopy, including proteins in 12 functional categories such as transcription factors and cytokines. Bioinformatic analysis of these proteins revealed clusters with functional importance to myeloid differentiation. Previous analyses have found that for a substantial number of genes the absolute amount of protein in the cell is not strongly correlated to the amount of mRNA. These conclusions were based on simultaneous measurement of mRNA and protein at just a single time point. Here, however, we are able to investigate the relationship between mRNA and protein in terms of simultaneous changes in their levels over multiple time points. This is the first time such a relationship has been studied, and we find that it gives a much stronger correlation, consistent with the hypothesis that a substantial proportion of protein change is a consequence of changed mRNA levels, rather than posttranscriptional effects. Cycloheximide inhibition also showed that most of the proteins detected by gel electrophoresis were relatively stable. Specific investigation of transcription factor mRNA representation showed considerable similarity to those of mature human neutrophils and highlighted several transcription factors and other functional nuclear proteins whose mRNA levels change prominently during MPRO differentiation but which have not been investigated previously in the context of myeloid development. Data are available online athttp://bioinfo.mbb.yale.edu/expression/myelopoiesis.
Phorbol ester stimulates a protein kinase C–mediated agatoxin-TK–sensitive calcium permeability pathway in human red blood cells
Authors: Andrews Dina A. , Yang Lu , Low Philip S. 
Published: October 2002

AbstractCalcium entry into mature erythrocytes (red blood cells; RBCs) is associated with multiple changes in cell properties. At low intracellular Ca2+, efflux of potassium and water predominates, leading to changes in erythrocyte rheology. At higher Ca2+ content, activation of kinases and phosphatases, rupture of membrane-to-skeleton bridges, stimulation of a phospholipid scramblase and phospholipase C, and induction of transglutaminase-mediated protein cross-linking are also observed. Because the physiologic relevance of these latter responses depends partially on whether Ca2+ entry involves a regulated channel or nonspecific leak, we explored mechanisms that initiate controlled Ca2+ influx. Protein kinase C (PKC) was considered a prime candidate for the pathway regulator, and phorbol-12 myristate-13 acetate (PMA), a stimulator of PKC, was examined for its influence on erythrocyte Ca2+. PMA was found to stimulate a rapid, dose-dependent influx of calcium, as demonstrated by the increased fluorescence of an entrapped Ca2+-sensitive dye, Fluo-3/am. The PMA-induced entry was inhibited by staurosporine and the PKC-selective inhibitor chelerythrine chloride, but was activated by the phosphatase inhibitors okadaic acid and calyculin A. The PMA-promoted calcium influx was also inhibited by ω-agatoxin-TK, a calcium channel blocker specific for Cav2.1 channels. To confirm that a Cav2.1-like calcium channel exists in the mature erythrocyte membrane, RBC membrane preparations were immunoblotted with antiserum against the α1A subunit of the channel. A polypeptide of the expected molecular weight (190 kDa) was visualized. These studies indicate that an ω-agatoxin-TK–sensitive, Cav2.1-like calcium permeability pathway is present in the RBC membrane and that it may function under the control of kinases and phosphatases.
Dioxygen Activation by Copper Complexes. Mechanistic Insights into Copper Monooxygenases and Copper Oxidases
Authors: Itoh Shinobu , Fukuzumi Shunichi 
Published: October 2002

Abstract
Reactions of copper(I) complexes with molecular oxygen have been examined using a series of N-alkyl-bis[2-(2-pyridyl)ethyl]amine tridentate ligands (R′Py2R) and N,N-dialkyl-2-(2-pyridyl)ethylamine didentate ligands (R′Py1R1,R2) at low temperature. The tridentate ligands predominantly provide (μ-η2 : η2-peroxo)dicopper(II) complexes (side-on type peroxo complex), while the didentate ligands enhance O–O bond homolysis of the peroxo species to produce bis(μ-oxo)dicopper(III) complexes. With the (μ-η2 : η2-peroxo)dicopper(II) complexes supported by the tridentate ligand, efficient oxygenation of phenolates to the corresponding catechols has been accomplished to provide a good model reaction of tyrosinase. The bis(μ-oxo)dicopper(III) complexes, on the other hand, undergo aliphatic ligand hydroxylation as well as oxygen atom transfer to sulfides to give the corresponding sulfoxides. In the reaction of bis(μ-oxo)dicopper(III) complex with 10-methyl-9,10-dihydroacridine (AcrH2) and 1,4-cyclohexadiene (CHD), a new active oxygen intermediate such as a (μ-oxo)(μ-oxyl radical)dicopper(III) or a tetranuclear copper-oxygen complex has been suggested to be involved as the real active oxygen species for the C–H bond activation of the external substrates. A mixed valence bis(μ3-oxo) trinuclear copper(II,II,III) complex has also been assessed using the didentate ligand with the smallest N-alkyl substituent (methyl). Mechanistic details of the above reactions as well as ligand effects on the copper(I)-dioxygen reactivity are discussed systematically.
Simple high-performance liquid chromatographic method to monitor vigabatrin, and preliminary review of concentrations determined in epileptic patients
Authors: George S , Gill L , Braithwaite R A 
Published: October 2002

SUMMARY. A simple and rapid high-performance liquid chromatographic method has been developed for the determination of vigabatrin concentrations in plasma or serum. The assay uses only 100 µL of specimen and has been found to be linear over a concentration range of 1 to 50 mg/L. The limit of detection has been determined as 1 mg/L, and the between-batch coefficient of variation for the two internal quality controls routinely analysed ( n=33) has been found to be less than 5%. There was no evidence of interferences in the assay from other commonly prescribed anti-epileptic drugs. This method has been applied to routine clinical specimens to determine the concentration of vigabatrin in 47 patient specimens over a 12-month period. It was found that only 63% of the male group and 53% of the female group were within the proposed target concentration of 5 to 35 mg/L. In addition, it was found that 26% of the male group and 36% of the female group were found to have concentrations below 5 mg/L, which may indicate lack of compliance and/or lack of therapeutic efficacy of treatment.
Non-fasting reference intervals for the Abbott IMxTM homocysteine and AxSYMTM plasma folate assays: influence of the methylenetetrahydrofolate reductase 677 C→T mutation on homocysteine
Authors: Donnelly James G , Isotalo Phillip A 
Published: October 2002

Plasma homocysteine comes under both genetic and nutritional control. B vitamins and particularly folate are important factors in homocysteine metabolism. We have obtained reference intervals for total plasma homocysteine and plasma folate. We have also determined the influence of methylenetetrahydrofolate reductase (MTHFR) genotype on plasma homocysteine concentrations in healthy individuals. Reference intervals for Abbott IMxTM homocysteine and AxSYMTM plasma folate assays were established using 116 volunteers recruited from hospital staff. Exclusion criteria included cardiac, hepatic or renal disorders, and use of over-the-counter prescription medications. An exception was the inclusion of three women using oral contraceptives and one woman receiving post-menopausal oestrogen supplementation. Methylenetetrahydrofolate reductase 677C-T genotyping was performed on 101 of the volunteers to determine whether the MTHFR 677T allele influences homocysteine concentrations in healthy individuals. Reference intervals for homocysteine and folate were determined using the mean±2 standard deviations of the data. Folate/homocysteine ratios were sorted by MTHFR C677T genotype. Homocysteine correlated negatively with plasma folate. Mean male homocysteine concentrations were significantly higher (9· μmol/L; P<0·05) than the mean value (7·1 μmol/L) obtained for females. Mean homocysteine values were significantly higher in subjects who were homozygous for the MTHFR 677T allele when compared with the 677CC genotype ( P<0·05). Ratios of folate/homocysteine were 20% and 7·44% lower in the male and female 677TT group than in the 677CC group, respectively. The mean homocysteine value of 43 volunteers who were taking multivitamins was not significantly different from that of 73 who were not vitamin supplemented. Conversely, the mean folate value was slightly greater, and statistically significant, in the group taking vitamin supplements. The mean folate values and reference intervals were not significantly different when grouped by sex or age. MTHFR 677C T mutations influenced homocysteine values observed in our study of healthy volunteers, even though we did not observe outright folate-deficient individuals. Our random homocysteine values were similar to the fasting homocysteine values obtained in other studies.
Immune complex transfer enzyme immunoassay for anti-ovalbumin IgA in serum
Authors: Hirota Kouichi , Kamashima Tomomi , Hashida Seiichi , Totani Masayuki 
Published: October 2002

Background: An immune complex transfer enzyme immunoassay for antiovalbumin immunoglobulin A (IgA) was developed. Methods: Serum-specific antibody was reacted simultaneously with 2,4-dinitrophenyl-bovine serum albumin-ovalbumin conjugate and ovalbumin- β-D-galactosidase conjugate. The complex formed from the three components was trapped onto polystyrene balls coated with anti-2,4-dinitrophenyl group immunoglobulin G, eluted with &epsi; N-2,4-dinitrophenyl-L-lysine and transferred to polystyrene balls coated with anti-human IgA α-chain. Bound β-D-galactosidase activity was determined by fluorometry. Results & Conclusions: The detection limit of this method for the measurement of specific anti-ovalbumin IgA was 9 fmol/tube, which was 20-fold lower than that of the enzyme-linked immunosorbent assay (ELISA). Because serum interference with this method was lower than that with the ELISA, the detection limit of this method was 300-fold lower than that by the ELISA. Anti-ovalbumin IgA was detected in 100% of healthy subjects, which was confirmed by pre-incubation with an excess amount of ovalbumin.
Interference in immunoassay is an underestimated problem
Authors: Ismail AAA , Walker PL , Cawood ML , Barth JH 
Published: October 2002

The presence of antibodies in some patients' serum has long been known to be a potential source of interference in immunoassays, as shown by numerous case reports. These often appear after the introduction of a new analyte (e.g. troponin) and then decrease in number as the topic becomes exhausted. This highlights the persistent and intrinsic nature of this problem, despite attempts by the manufacturers to compensate for this source of error. However, an explanation of the immunoanalytical basis underpinning these assays could be more effective in raising awareness than intermittent case reports. In this review we have outlined the use of antibodies as reagents, the factors determining how they bind to antigen(s), and the nature of the immune response in order to explain the insidious and unpredictable nature of this form of interference. Studies on the prevalence of interference have yielded values ranging from 0.05 to more than 2%. However, these figures are analyte- and assay-specific, influenced by the study design, and are not therefore generally applicable. It is also highly likely that figures on prevalence and incidence will worsen in the future because of the wider use of monoclonal antibodies as diagnostic and therapeutic tools. Clinical laboratories should be alert to assay interference from antibodies irrespective of its nature, as immunoassays will remain an indispensable analytical tool, unlikely to be replaced in the foreseeable future by a practical alternative.
Vagus nerve stimulation: quality control in thyroid and parathyroid surgery
Authors: Lambert A. W. , Cosgrove C. , Barwell J. , Oxenham S. , Wilkins D. C. 
Published: October 2002

This paper describes the use of the Neurosign 100 Nerve Monitor and vagus nerve stimulation in the identification and assessment of the integrity of the recurrent laryngeal nerve (RLN) during thyroid and parathyroid surgery.Vocal fold function was assessed pre- and post-operatively in all patients undergoing thyroid and parathyroid surgery. The nerve monitor, used in association with endotracheal electrodes, was used to confirm correct RLN identification and demonstrate its integrity at the completion of surgery.There were 21 unilateral and 19 bilateral neck explorations. In these 40 patients, 57 of 59 RLNs were identified. The nerve monitor demonstrated RLN continuity in all but one case (equipment failure: electrode misplacement) after initial identification. Vagus nerve stimulation was performed in 21 patients without adverse sequelae. Damage to the RLN was identified in one of these patients, in whom direct RLN stimulation close to the larynx had failed to indicate discontinuity. Post-operatively this patient had a transient unilateral vocal fold palsy.The use of the Neurosign 100 Nerve Monitor is no substitute for meticulous surgery. Stimulation of the vagus nerve may be a more sensitive means of assessing RLN integrity during thyroid and parathyroid surgery than stimulation of the RLN itself. Confirmation of RLN integrity allows the surgeon to proceed with confidence to the contralateral side of the neck during hazardous bilateral explorations.
Laryngeal granuloma: characteristics of the covering epithelium
Authors: Luzar Boŝtjan , Gale Nina , Klopĉiĉ Ulrika , Fischinger Janez 
Published: October 2002

The purpose of the present study was to evaluate the biological behaviour of the marginal epithelium, that proliferates and eventually covers laryngeal granulomas, and to reveal the applicability of the recently re-introduced Ljubljana classification when reporting reactive epithelial hyperplastic lesions.A retrospective clinical and histomorphological analysis was performed on 149 laryngeal granuloma biopsies. Epithelial changes were classified according to the Ljubljana classification into normal epithelium; simple, abnormal, or atypical hyperplasia; and carinoma in situ. Atrophic epithelium, not evaluated separately in the Ljubljana classification, was additionally assessed. Simple hyperplasia was found in 98 cases (65.8 per cent), abnormal hyperplasia in seven (4.7 per cent), atrophic epithelium in 24 (16.1 per cent), and normal squamous epithelium in 20 (13.4 per cent). Atypical hyperplasia and carcinoma in situ were not observed.The results of our study clearly showed that the proliferation of the covering epithelium mostly in the form of simple hyperplasia, is entirely reactive and therefore reversible. No epithelial hyperplastic lesions were found that were previously described to be associated with an increased risk of malignant alteration, namely atypical hyperplasia and carcinoma in situ. However, since an initial growth of an invasive malignant neoplasm might macroscopically imitate the appearance of laryngeal granuloma, a histological examination in all aetiological forms of laryngeal granulomas is required. By clearly discerning the benign nature of epithelial changes in laryngeal granulomas, the recently re-evaluated and further formulated Ljubljana classification may also influence the clinical handling of patients.
The arcuate eminence is not a protrusion of the superior semi-circular canal but a trace of sulcus on the temporal lobe
Authors: Tsunoda Atsunobu , Kimura Yurika , Sumi Takuro , Komatsuzaki Atsushi , Sato Tatsuo 
Published: October 2002

Thirteen cadavers (26 temporal bones) were examined to show the arcuate eminence, especially its correspondence to the superior semicircular canal and inferior surface of the temporal lobe. Arc-like eminences on the petrous bone were observed in 92 per cent of specimens, however, they did not exactly correspond to the superior semi-circular canal. Some eminences corresponded to sulci of the temporal lobe of which most were traces of the occipitotemporal sulcus.On the other hand, a dull, smooth and even domed eminence existed in nine temporal bones independently of the arc-like eminence. Those eminences corresponded to each superior semi-circular canal in only three out of nine specimens.The arcuate eminence was listed as an important landmark in the middle cranial fossa approach. However, in order to drill out the internal auditory canal safely, surgeons should rely on other landmarks or apply other methods from our data.
Effect of packing on nasal mucosa of sheep
Authors: Shaw Chi-kee Leslie , Dymock Robert B. , Cowin Allison , Wormald Peter-John 
Published: October 2002

The effects of packing with ribbon gauze and neuropatties on the nasal mucosa was assessed using sheep as an animal model. Fourteen sheep either underwent ribbon gauze or neuropattie nasal packing. Trauma to nasal mucosa caused by ribbon gauze and neuropatties was compared to mucosa on the lateral aspect of the middle turbinate which was not in contact with any packing. This tissue was used as a control. Ribbon gauze packing resulted in significant loss of 68 per cent of the ciliated surface of the mucosa when compared with the control group with a 15 per cent loss of ciliated surface (p < 0.005). Neuropattie packing also resulted in significant loss of 50 per cent of the ciliated surface of the mucosa when compared with the control group (p < 0.005). There was no significant difference in loss of ciliated mucosa in the specimens packed with ribbon gauze or neuropatties (p = 0.25).Nasal packing results in a significant mucosal injury with loss of cilia. This may influence the mucociliary clearance of the nose in the post-operative healing phase. Pre-operative nasal packing should be used circumspectly and if possible avoided.
Evaluation of functional outcomes (speech, swallowing and voice) in patients attending speech pathology after head and neck cancer treatment(s): development of a multi-centre database
Authors: Perry Alison R. , Shaw Margaret A. 
Published: October 2002

Since April 1997, in Melbourne, Australia, speech pathologists have collaborated to establish a prospective database of functional outcomes of speech, swallowing and voice for patients undergoing head and neck cancer treatments.Staff at eight acute care hospitals, all of which offer speech pathology for head and neck cancer services in Victoria, are contributing data, collated centrally, in an agreed pro forma.Early results are given (after 12 months’ data collection). The implications for clinically-based research, and the future potential for benchmarking outcomes – by expansion of the rehabilitation database beyond the current participating sites – is discussed.This paper outlines the rationale of establishing the database is multicentred, and explores some of the complexities involved, including the challenges inherent in long-term accurate data collection in the head and neck cancer patient population. This work represents the development of an appropriate, usable tool for data collection on functional outcomes.