bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cocaethylene Affects Human Microvascular Endothelial Cell p38 Mitogen-Activated Protein Kinase Activation and Nuclear Factor-κB DNA-Binding Activity
Authors: Tacker Danyel Hermes , Herzog Norbert K , Okorodudu Anthony O 
Published: August 2006

AbstractBackground: Cocaethylene (CE) is known to increase the permeability of human microvascular endothelial cell monolayers. The molecular mechanism underlying this increase may involve calcium-modulated signaling pathways such as the p38 mitogen-activated protein kinase (p38 MAPK) and the nuclear factor-κB (NF-κB) family of transcription factors. The hypothesis of this study was that CE-mediated endothelial permeability change may be mediated by the p38 MAPK and consequently NF-κB dimers.Methods: We used sandwich ELISA to detect phosphorylated p38 MAPK in the cell line human microvascular endothelial cell 1 (HMEC-1) after treatment with 1 mmol/L CE. We used electrophoretic mobility shift assay to detect changes in NF-κB dimers present in HMEC-1 and their DNA-binding activity after treatment with CE. Lipopolysaccharide (LPS) from Salmonella typhosa was used as a positive control for all experiments.Results: Treatment with CE and LPS had similar effects on HMEC-1 p38 MAPK phosphorylation and NF-κB DNA-binding activity. Both treatments increased the phosphorylation of p38 MAPK, consistent with activation of proinflammatory cell signaling. Treatment of HMEC-1 with CE decreased DNA binding of both the RelA/p50 and p50/p50 dimers of the NF-κB transcription factor family, whereas treatment with LPS decreased and then increased the DNA binding of these dimers.Conclusion: In addition to increasing HMEC-1 monolayer permeability, CE also alters transcription factor and kinase activity related to inflammation. Thus, CE causes endothelial activation that can elicit a prolonged and organized cellular response, rather than being directly toxic to endothelial cells.
Multilectin Affinity Chromatography for Characterization of Multiple Glycoprotein Biomarker Candidates in Serum from Breast Cancer Patients
Authors: Yang Ziping , Harris Lyndsay E , Palmer-Toy Darryl E , Hancock William S 
Published: August 2006

Abstract
Background: Glycoproteins are often associated with cancer and are important in serum studies, for which glycosylation is a common posttranslational modification.
Methods: We used multilectin affinity chromatography (M-LAC) to isolate glycoproteins from the sera of breast cancer patients and controls. The proteins were identified by HPLC–tandem mass spectrometry (MS/MS) analysis of the corresponding tryptic digests. We used the FuncAssociate Gene Ontology program for association analysis of the identified proteins. Biomarker candidates in these groups were comparatively quantitated by use of peak area measurements, with inclusion of an internal standard. We analyzed data for concordance within the ontology association groups for vector of change with the development of breast cancer.
Results: Detection of the known low-concentration biomarker HER-2 (8–24 μg/L) enabled us to establish a dynamic range of 106, relative to the amount of albumin, for the depletion step. We then used ELISA to confirm this range. Proteins associated with lipid transport and metabolism, cell growth and maintenance, ion homeostasis, and protease inhibition were found to be differentially regulated in serum from women with breast cancer compared with serum from women without breast cancer.
Conclusions: M-LAC for isolation of the serum glycoproteome, coupled with liquid chromatography–MS/MS and the use of gene ontology associations, can be used to characterize large panels of candidate markers, which can then be evaluated in a particular patient population.
The Relationships of Plasma Adiponectin with a Favorable Lipid Profile, Decreased Inflammation, and Less Ectopic Fat Accumulation Depend on Adiposity
Authors: Kantartzis Konstantinos , Rittig Killian , Balletshofer Bernd , Machann Jürgen , Schick Fritz , Porubska Katarina , Fritsche Andreas , Häring Hans-Ulrich , Stefan Norbert 
Published: August 2006

Abstract
Background: The metabolic effects of adiponectin, including insulin sensitivity, seem to become stronger with increasing adiposity. Adiposity may also affect the relationship of adiponectin concentrations with serum lipid profile; markers of inflammation, atherosclerosis, and endothelial function; and ectopic fat accumulation.
Methods: We measured plasma adiponectin concentrations, serum lipids, and serum markers of inflammation, atherosclerosis, and endothelial function in 242 Caucasians without type 2 diabetes. We also measured visceral adipose tissue with magnetic resonance tomography and liver and intramyocellular fat with 1H magnetic resonance spectroscopy.
Results: We divided the study participants into 2 groups: lean [mean (SE) total body fat, 26% (0.6%); n = 119] and obese [36% (0.6%); n = 123]. In the obese group, plasma adiponectin concentrations showed a strong positive association with concentrations of HDL cholesterol (P &lt;0.0001) and negative associations with LDL cholesterol, triglycerides, high-sensitivity C-reactive protein, interleukin 6, apolipoprotein B100, soluble E-selectin, soluble vascular cellular adhesion molecule 1, plasminogen activator inhibitor 1, leukocyte count, and liver and intramyocellular fat (all P &lt;0.03). In the lean group, adiponectin showed a less strong association with HDL cholesterol (P = 0.005) and liver fat (P = 0.03) and no significant associations with the other variables (all P &gt;0.10). High visceral adipose tissue was a strong predictor of low adiponectin concentrations, particularly in the obese group, and attenuated many of the significant relationships.
Conclusions: High adiponectin plasma concentrations are associated with favorable lipid profiles, decreased subclinical inflammation, decreased markers of atherosclerosis and endothelial function, and low ectopic fat accumulation, particularly in obese persons. Adiponectin may also have a concentration-related effect on the relationship between visceral adipose tissue and these metabolic characteristics, especially in obese persons.
Genotype-Specific Signal Generation Based on Digestion of 3-Way DNA Junctions: Application to KRAS Variation Detection
Authors: Amicarelli Giulia , Adlerstein Daniel , Shehi Erlet , Wang Fengfei , Makrigiorgos G Mike 
Published: August 2006

AbstractBackground: Genotyping methods that reveal single-nucleotide differences are useful for a wide range of applications. We used digestion of 3-way DNA junctions in a novel technology, OneCutEventAmplificatioN (OCEAN) that allows sequence-specific signal generation and amplification. We combined OCEAN with peptide-nucleic-acid (PNA)-based variant enrichment to detect and simultaneously genotype v-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog (KRAS) codon 12 sequence variants in human tissue specimens.Materials and Methods: We analyzed KRAS codon 12 sequence variants in 106 lung cancer surgical specimens. We conducted a PNA-PCR reaction that suppresses wild-type KRAS amplification and genotyped the product with a set of OCEAN reactions carried out in fluorescence microplate format. The isothermal OCEAN assay enabled a 3-way DNA junction to form between the specific target nucleic acid, a fluorescently labeled “amplifier”, and an “anchor”. The amplifier-anchor contact contains the recognition site for a restriction enzyme. Digestion produces a cleaved amplifier and generation of a fluorescent signal. The cleaved amplifier dissociates from the 3-way DNA junction, allowing a new amplifier to bind and propagate the reaction.Results: The system detected and genotyped KRAS sequence variants down to ∼0.3% variant-to-wild-type alleles. PNA-PCR/OCEAN had a concordance rate with PNA-PCR/sequencing of 93% to 98%, depending on the exact implementation. Concordance rate with restriction endonuclease-mediated selective-PCR/sequencing was 89%.Conclusion: OCEAN is a practical and low-cost novel technology for sequence-specific signal generation. Reliable analysis of KRAS sequence alterations in human specimens circumvents the requirement for sequencing. Application is expected in genotyping KRAS codon 12 sequence variants in surgical specimens or in bodily fluids, as well as single-base variations and sequence alterations in other genes.
Pro12Ala Sequence Variant of the PPARG Gene Is Associated with Postprandial Hypertriglyceridemia in Non-E3/E3 Patients with the Metabolic Syndrome
Authors: Cardona Fernando , Morcillo Sonsoles , Gonzalo-Marín Montserrat , Garrido-Sanchez Lourdes , Macias-Gonzalez Manuel , Tinahones Francisco J 
Published: August 2006

Abstract
Background: Postprandial hypertriglyceridemia, a component of the metabolic syndrome, has varied etiology and involves many genes related to triglyceride metabolism. Variations in these genes may affect postprandial hypertriglyceridemia in the context of the metabolic syndrome.
Methods: We orally administered 60 g of fat overload to 74 patients with the metabolic syndrome. We then measured baseline concentrations of cholesterol, triglycerides, HDL cholesterol, apolipoprotein AI, apolipoprotein B, uric acid, and uric acid excretion; we also performed homeostasis model assessments of insulin resistance and insulin sensitivity. At 3 h, we measured triglycerides, cholesterol, apolipoprotein AI, and apolipoprotein B. Patients were considered to have postprandial hypertriglyceridemia if the difference in plasma triglycerides between baseline and 3 h after the test was 1.71 mmol/L or more. We also measured anthropometrical variables and classified the patients according to their peroxisome proliferative activated receptor, gamma (PPARG) gene and apolipoprotein E (APOE) genotype.
Results: Postprandial hypertriglyceridemia occurred in 64.7% of patients with the Ala12 allele vs 19.9% of the Pro12Pro patients, (P = 0.00032; odds ratio, 7.6), and in 87.5% of the patients with both the Ala12 allele and the non-E3/E3 APOE genotype (odds ratio, 23.8). Logistic regression analysis showed that PPARG and APOE sequence variants were associated with the presence of postprandial hypertriglyceridemia.
Conclusion: The Pro12Ala PPARG sequence variant together with a non-E3/E3 APOE genotype is associated with a high risk for postprandial hypertriglyceridemia in patients with the metabolic syndrome, indicating a close association between these genes and the regulation of lipoproteinase clearance.
The C-Reactive Protein +1444C/T Alteration Modulates the Inflammation and Coagulation Response in Human Endotoxemia
Authors: Marsik Claudia , Sunder-Plassmann Raute , Jilma Bernd , Kovar Florian M , Mannhalter Christine , Wagner Oswald , Rumpold Helmut , Endler Georg 
Published: August 2006

AbstractBackground: C-reactive protein (CRP) plays a major role in the immune system and is an independent risk marker of cardiovascular disease. However, CRP’s role in atherogenesis as innocent bystander, causative, or even protective agent, remains unresolved. The +1444C/T alteration in the CRP gene has been reported to determine basal CRP concentrations. We hypothesized that this alteration may also be associated with the degree of inflammatory response and coagulation activation in a well-standardized model of systemic inflammation.Methods: We administered 2 ng/kg endotoxin [Escherichia coli bacterial lipopolysaccharide (LPS)] intravenously to stimulate inflammation in 91 healthy young Caucasian male paid volunteers (age range, 19–40 years). Participants were confined to bed rest and fasted for 8.5 h after LPS infusion. We collected blood samples before LPS infusion and at 0, 2, 6, and 24 h after LPS infusion to measure inflammation markers [interleukin 6 (IL6), tumor necrosis factor-α (TNFα)], temperature, and coagulation markers (prothrombin fragment F1+2, D-dimer). We analyzed the CRP 3′ untranslated variant with a mutagenic separated PCR assay.Results: Basal concentrations of high-sensitivity CRP were ∼40% lower in +1444CC alteration carriers than in T homozygous (TT) allele carriers (P = 0.04). In contrast, basal IL6 concentrations were 2-fold higher in wild-type C homozygous (CC) than in TT individuals (P = 0.01). In response to the LPS challenge, CC individuals had 4-fold higher peak TNFα concentrations (P &lt;0.01), &gt;2.5-fold higher peak IL6 concentrations (P &lt;0.01), and increased temperature (P &lt;0.01). Twenty-four hours after LPS challenge, prothrombin fragment F1+2 concentrations were 75% higher and D-dimer concentrations 50% higher in CC than in TT individuals (P &lt;0.05).Conclusions: Genetic factors regulating CRP concentrations also modulate the individual response to endotoxin-stimulated inflammation.
Postanalytical External Quality Assessment of Warfarin Monitoring in Primary Healthcare
Authors: Kristoffersen Ann-Helen , Thue Geir , Sandberg Sverre 
Published: August 2006

AbstractBackground: An increasing number of patients are treated with warfarin worldwide, and many are monitored in general practice, often with office instruments. Bleeding or thromboembolic episodes may be consequences of inadequate treatment. We have therefore examined some important aspects of general practitioners’ (GPs’) knowledge of warfarin treatment.Methods: A questionnaire including 2 case histories with familiar indications for warfarin treatment (mechanical heart valve prosthesis and pulmonary embolism) was circulated to 3781 GPs in Norway as a postanalytical quality assessment.Results: A total of 1547 GPs (41%) responded. There were substantial variations among GPs concerning the frequency of international normalized ratio (INR) monitoring, stated therapeutic ranges for arterial (but not venous) indications for anticoagulation therapy, and handling of a moderately high INR result of 5.9. Most GPs estimated an unrealistically high risk of serious bleeding in the latter situation (median, 15%; 10th and 90th percentiles, 4% and 50%, respectively). The critical difference necessary to change the warfarin dose was highly dependent on perceived therapeutic intervals, and about half of the GPs suggested a critical difference of 0.8 INR, which is attainable with office instruments. Sex and age of the GPs, practice size, and availability of an INR instrument in the office laboratory did not influence the results to any substantial degree, as variations within subgroups were similar.Conclusions: Gross variations in practice were found, especially for aspects of warfarin treatment that lacked uniform guidelines. Evidence-based and practicable recommendations for treatment and monitoring of these patients are still needed.
Unfavorable Prognostic Value of Human Kallikrein 7 Quantified by ELISA in Ovarian Cancer Cytosols
Authors: Shan Shannon J C , Scorilas Andreas , Katsaros Dionyssios , Rigault de la Longrais Irene , Massobrio Marco , Diamandis Eleftherios P 
Published: August 2006

AbstractBackground: Human tissue kallikrein 7 (gene, KLK7; protein, hK7) is a member of the kallikrein family of secreted serine proteases. Reports indicate that in ovarian cancer, KLK7 is significantly up-regulated at the mRNA level. The aim of this study was to determine whether hK7, measured quantitatively by ELISA in ovarian cancer cytosols, is a prognostic biomarker for ovarian cancer.Methods: We used a newly developed ELISA with 2 monoclonal antibodies to quantify hK7 production in 260 ovarian tumor cytosols and correlated these data with various clinicopathologic variables and patient outcomes [progression-free survival (PFS) and overall survival (OS)] over a median follow-up period of 52 months.Results: Median (range) hK7 concentration in ovarian tumor cytosols was 2.84 (0–32.8) ng/mg of total protein. Compared with healthy and benign ovarian tissues and nonovarian tumors that metastasized to the ovary, malignant ovarian tumor cytosols highly overproduced hK7 (P &lt;0.001). We used the median value as the cutoff value to categorize tumors as hK7-positive and hK7-negative. Women with hK7-positive tumors most frequently had advanced-stage disease, higher tumor grade (G3), suboptimal debulking, and serous or undifferentiated histotype (P &lt;0.001). Univariate analysis showed that hK7 positivity was associated with significantly shorter PFS (P = 0.01) but not OS. Kaplan–Meier survival curves confirmed an increased risk of relapse in women with hK7-positive tumors (P = 0.009). In multivariate analysis, hK7 was not significantly associated with either PFS or OS.Conclusions: hK7 is associated with other unfavorable characteristics of ovarian cancer, but it is not an independent prognosticator for ovarian cancer.
Paired‐End diTagging for Transcriptome and Genome Analysis
Authors: Ng Patrick , Wei Chia‐Lin , Ruan Yijun 
Published: August 2006

AbstractThe Paired‐End diTagging (PET) procedure enables one to obtain sequence information from both termini of any contiguous DNA fragment. This is achieved by a series of enzymatic manipulations that introduce MmeI sites directly flanking each DNA insert during the construction of a plasmid library. Subsequent MmeI digestion and self‐ligation results in the production of covalently‐linked paired‐end ditags (PETs) that can be extracted and then concatenated for efficient sequencing. By mapping the PET sequences to assembled genomes, the original DNA fragments from which the PETs were derived can be precisely localized. This unit details two applications of PET technology. In GIS‐PET, ditagging of mRNA converted to full‐length cDNA enables whole‐transcriptome analysis, including novel gene identification, gene prediction validation, and gene expression studies. In ChIP‐PET, ditagging of chromatin immunoprecipitation–enriched genomic DNA fragments enables the global mapping of transcription factor binding sites.
RNA Immunoprecipitation for Determining RNA‐Protein Associations In Vivo
Authors: Gilbert Chris , Svejstrup Jesper Q. 
Published: August 2006

AbstractSimilar to chromatin immunoprecipitation (ChIP), RNA immunoprecipitation (RIP) can be used to detect the association of individual proteins with specific nucleic acid regions, in this case on RNA. Live cells are treated with formaldehyde to generate protein‐RNA cross‐links between molecules that are in close proximity in vivo. RNA sequences that cross‐link with a given protein are isolated by immunoprecipitation of the protein, and reversal of the formaldehyde cross‐linking permits recovery and quantitative analysis of the immunoprecipitated RNA by reverse transcription PCR. The basics of RIP are very similar to those of ChIP, but with some important caveats. This unit describes the RIP procedure for Saccharomyces cerevisiae. Although the corresponding steps for metazoan cells have not yet been worked out, it is likely that the yeast procedure can easily be adapted for use in other organisms.
Comparison of the Kid's Block Food Frequency Questionnaire to the 24‐hour recall in urban Native American youth
Authors: Smith Chery , Fila Stefanie 
Published: August 2006

AbstractThis study evaluated the appropriateness of the Kid's Block Food Frequency Questionnaire (Kid's Block FFQ) for use in a group of urban Native American youth by comparing it to a self‐reported 24‐hr recall. Subjects were 61 urban Native American youth, aged 9–13 years. Researchers assessed dietary intake using the two methods in random order on the same day. Analyses of diets revealed no significant difference between estimated total energy, protein, fat, and saturated fat between the two methods. Significant differences in estimations for the two tools occurred for carbohydrates and some micronutrients. Results suggest that the Kid's Block FFQ may be less appropriate than 24‐hr recall in measuring the dietary intake of Native American youth. A high prevalence of obesity in this population necessitates more accurate dietary assessment tools for this population. Am. J. Hum. Biol. 18:706–709, 2006. © 2006 Wiley‐Liss, Inc.
The impact of occupational injury reduction on the U.S. economy
Authors: Zaloshnja Eduard , Miller Ted R. , Waehrer Geetha 
Published: August 2006

AbstractBackgroundPreventing occupational injuries reduces labor and fringe benefit costs to employers. The related savings filter through the economy, impacting its performance. This study is a first attempt to measure the impact of occupational injury reduction on national economic output, gross domestic product, national income, and employment by using an input–output model of the U.S. economy.MethodsOccupational injury costs by industry for 1993 were used as a baseline for an input–output model, and the impact of the 38% injury rate reduction between 1993 and 2002 was measured. All computations are in year 2000 dollars.ResultsDeclining occupational injury between 1993 and 2002 increased employment by an estimated 550,000 jobs. The increase in gross domestic product (GDP) was $25.5 billion or 9% of the average annual GDP increase from 1993 to 2002.ConclusionsThese estimates represent the benefits of injury rate reduction but ignore associated prevention costs. Am. J. Ind. Med. 49:719–727, 2006. © 2006 Wiley‐Liss, Inc.
Costs differences across demographic groups and types of occupational injuries and illnesses
Authors: Leigh J. Paul , Waehrer Geetha , Miller Ted R. , McCurdy Stephen A. 
Published: August 2006

AbstractBackgroundLittle is known about cost differences for demographic groups or across occupational injuries and illnesses.MethodsIn this incidence study of nationwide data for 1993, an analysis was conducted on fatal and non‐fatal injury and illness data recorded in government data sets. Costs data were from workers' compensation records, estimates of lost wages, and jury awards.ResultsThe youngest (age ≤ 17) and oldest (age ≥ 65) workers had exceptionally high fatality costs. Whereas men's costs for non‐fatal incidents were nearly double those for women, men's costs for fatal injuries were 10 times the costs for women. The highest ranking occupation for combined fatal and non‐fatal costs—farming, forestry, and fishing—had costs‐per‐worker ($5,163) over 18 times the lowest ranking occupation—executives and managers ($279). The occupation of handlers, cleaners, and laborers, ranked highest for non‐fatal costs. Gunshot wounds generated especially high fatal costs. Compared to whites, African‐Americans had a lower percentage of costs due to carpal tunnel syndrome, circulatory, and digestive diseases.ConclusionsCosts comparisons can be drawn across age, race, gender, and occupational groups as well as categories of injuries and illnesses. Am. J. Ind. Med. 2006. © 2006 Wiley‐Liss, Inc.