bluuebunny/crossref_metadata_2025_split huggingface dataset:
Colonic IgA producing cells and macrophages are reduced in recurrent and non-recurrentClostridiumdifficileassociated diarrhoea
Authors: Johal S S , Lambert C P , Hammond J , James P D , Borriello S P , Mahida Y R 
Published: August 2004

Background:InClostridium difficileassociated diarrhoea (CDAD), histological changes in the colonic mucosa range from minimal inflammation to pseudomembranous colitis (PMC). The disease also recurs in a considerable proportion of patients.Aim:To investigate mucosal immune system cells in colonic biopsies of patients with CDAD.Methods:Colonic biopsies were obtained from 12 control patients with diarrhoea, six patients with CDAD and minimal inflammation, and 10 patients with CDAD with pseudomembranous colitis (samples obtained from areas with and without inflammatory exudate). Immunohistochemical studies were performed using antibodies to T cells (CD3), macrophages (CD68), B/plasma cells (CD79α), and to IgA, IgM, and IgG. Labelled cells in lamina propria were quantified.Results:In contrast to T cells, there were significant reductions in B/plasma cell and macrophage counts in all biopsies from patients with CDAD compared with controls (p<0.001). Studies using anti-immunoglobulin antibodies showed significant reductions in IgA producing cells in CDAD biopsies (p<0.05), with the greatest reduction in samples from patients with PMC. In contrast, there was a significant increase (p<0.05) in IgG producing cells in CDAD biopsies. Only patients with PMC relapsed. In these patients, B/plasma cell and IgA producing cell counts (in biopsies with and without inflammatory exudates) were significantly lower (p<0.01) in mucosal samples from those who subsequently relapsed (five) than those who did not.Conclusions:A selective reduction in mucosal IgA producing cells and macrophages is associated with colonic disease inC difficileinfected patients. Severe reduction in colonic IgA producing cells may predispose to recurrence of CDAD.
Low-Density Lipoprotein Particle Size Loci in Familial Combined Hyperlipidemia
Authors: Badzioch Michael D. , Igo Robert P. , Gagnon France , Brunzell John D. , Krauss Ronald M. , Motulsky Arno G. , Wijsman Ellen M. , Jarvik Gail P. 
Published: August 2004

Objective—Low-density lipoprotein (LDL) size is associated with vascular disease and with familial combined hyperlipidemia (FCHL).Methods and Results—We used logarithm of odds (lod) score and Bayesian Markov chain Monte Carlo (MCMC) linkage analysis methods to perform a 10-cM genome scan of LDL size, measured as peak particle diameter (PPD) and adjusted for age, sex, body mass index, and triglycerides in 4 large families with FCHL (n=185). We identified significant evidence of linkage to a chromosome 9p locus (multipoint lodmax=3.70; MCMC intensity ratio [IR]=21) in a single family, and across all 4 families to chromosomes 16q23 (lodmax=3.00; IR=43) near cholesteryl ester transfer protein (CETP) and to 11q22 (lodmax=3.71; IR=120). Chromosome 14q24-31, a region with previous suggestive LDL PPD linkage evidence, yielded an IR of 71 but an lodmax=1.79 in the combined families.Conclusions—These results of significant evidence of linkage to 3 regions (9p, 16q, and 11q) and confirmatory support of previous reported linkage to 14q in large FCHL pedigrees demonstrate that LDL size is a trait influenced by multiple loci and illustrate the complementary use of lod score and MCMC methods in analysis of a complex trait.
Insulin-Induced Cell Cycle Progression Is Impaired in Chinese Hamster Ovary Cells Overexpressing Insulin Receptor Substrate-3
Authors: Kaburagi Yasushi , Yamashita Ryo , Ito Yuzuru , Okochi Hitoshi , Yamamoto-Honda Ritsuko , Yasuda Kazuki , Sekihara Hisahiko , Sasazuki Takehiko , Kadowaki Takashi , Yazaki Yoshio 
Published: August 2004

AbstractTo analyze the roles of insulin receptor substrate (IRS) proteins in insulin-stimulated cell cycle progression, we examined the functions of rat IRS-1 and IRS-3 in Chinese hamster ovary cells overexpressing the human insulin receptor. In this type of cell overexpressing IRS-1 or IRS-3, we showed that: 1) overexpression of IRS-3, but not IRS-1, suppressed the G1/S transition induced by insulin; 2) IRS-3 was more preferentially localized to the nucleus than IRS-1; 3) phosphorylation of glycogen synthase kinase 3 and MAPK/ERK was unaffected by IRS-3 overexpression, whereas that of protein kinase B was enhanced by either IRS; 4) overexpressed IRS-3 suppressed cyclin D1 expression in response to insulin; 5) among the signaling molecules regulating cyclin D1 expression, activation of the small G protein Ral was unchanged, whereas insulin-induced gene expression of c-myc, a critical component for growth control and cell cycle progression, was suppressed by overexpressed IRS-3; and 6) insulin-induced expression of p21, a cyclin-dependent kinase inhibitor, was decreased by overexpressed IRS-3. These findings imply that: 1) IRS-3 may play a unique role in mitogenesis by inhibiting insulin-stimulated cell cycle progression via a decrease in cyclin D1 and p21 expressions as well as suppression of c-myc mRNA induction in a manner independent of the activation of MAPK, protein kinase B, glycogen synthase kinase 3 and Ral; and 2) the interaction of IRS-3 with nuclear proteins may be involved in this process.
“Hijacking” the Thyrotropin Receptor: A Chimeric Receptor-Lysosome Associated Membrane Protein Enhances Deoxyribonucleic Acid Vaccination and Induces Graves’ Hyperthyroidism
Authors: Pichurin Pavel N. , Chazenbalk Gregorio D. , Aliesky Holly , Pichurina Oxana , Rapoport Basil , McLachlan Sandra M. 
Published: August 2004

AbstractNaked DNA vaccination with the TSH receptor (TSHR) does not, in most studies, induce TSHR antibodies and never induces hyperthyroidism in BALB/c mice. Proteins expressed endogenously by vaccination are preferentially presented by major histocompatibility complex class I, but optimal T cell help for antibody production requires lysosomal processing and major histocompatibility complex class II presentation. To divert protein expression to lysosomes, we constructed a plasmid with the TSHR ectodomain spliced between the signal peptide and transmembrane-intracellular region of lysosome-associated membrane protein (LAMP)-1, a lysosome-associated membrane protein. BALB/c mice pretreated with cardiotoxin were primed intramuscularly using this LAMP-TSHR chimera and boosted twice with DNA encoding wild-type TSHR, TSHR A-subunit, or LAMP-TSHR. With each protocol, spleen cells responded to TSHR antigen by secreting interferon-γ, and 60% or more mice had TSHR antibodies detectable by ELISA. TSH binding inhibitory activity was present in seven, four, and two of 10 mice boosted with TSHR A-subunit, LAMP-TSHR, or wild-type TSHR, respectively. Importantly, six of 30 mice had elevated T4 levels and goiter (5 of 6 with detectable thyroid-stimulating antibodies). Injecting LAMP-TSHR intradermally without cardiotoxin pretreatment induced TSHR antibodies detectable by ELISA but not by TSH binding inhibitory activity, and none became hyperthyroid. These findings are consistent with a role for cardiotoxin-recruited macrophages in which (unlike in fibroblasts) LAMP-TSHR can be expressed intracellularly and on the cell surface. In conclusion, hijacking the TSHR to lysosomes enhances T cell responses and TSHR antibody generation and induces Graves’-like hyperthyroidism in BALB/c mice by intramuscular naked DNA vaccination.
Endocannabinoid Signaling Negatively Modulates Stress-Induced Activation of the Hypothalamic-Pituitary-Adrenal Axis
Authors: Patel Sachin , Roelke Craig T. , Rademacher David J. , Cullinan William E. , Hillard Cecilia J. 
Published: August 2004

Abstract
Activation of the hypothalamic-pituitary-adrenal (HPA) axis is critical for the adaptation and survival of animals upon exposure to stressful stimuli, and data suggest that endocannabinoid (eCB) signaling modulates neuroendocrine function. We have explored the role of eCB signaling in the modulation of stress-induced HPA axis activation. Administration of the CB1 receptor antagonist/inverse agonist SR141716 (0.01, 0.1, 1, and 5 mg/kg, ip) to male mice produced a small, dose-dependent increase in the serum corticosterone (CORT) concentration. Despite this effect, the highest dose of SR141716 did not significantly increase neuronal activity within the paraventricular nucleus of the hypothalamus, as measured by the induction of Fos protein. Similarly, exposure of mice to 30 min of restraint increased serum CORT concentrations, but did not produce a consistent, statistically significant increase in Fos expression within the PVN. However, pretreatment of mice with SR141716 before restraint stress robustly potentiated restraint-induced CORT release and Fos expression within the PVN. Pretreatment of mice with either the CB1 receptor agonist CP55940, the eCB transport inhibitor AM404, or the fatty acid amide hydrolase inhibitor URB597 significantly decreased or eliminated restraint-induced CORT release. Upon exposure to acute restraint, hypothalamic 2-arachidonylglycerol content was reduced compared with the control value; however, after 5 d of restraint exposure (which resulted in an attenuated CORT response), the hypothalamic 2-arachidonylglycerol content was increased compared with the control value. These data indicate that eCB signaling negatively modulates HPA axis function in a context-dependent manner and suggest that pharmacological augmentation of eCB signaling could serve as a novel approach to the treatment of anxiety-related disorders.
Valproic Acid Inhibits Leptin Secretion and Reduces Leptin Messenger Ribonucleic Acid Levels in Adipocytes
Authors: Lagace Diane C. , McLeod Roger S. , Nachtigal Mark W. 
Published: August 2004

AbstractTreatment of epilepsy or bipolar disorder with valproic acid (VPA) induces weight gain and increased serum levels for the satiety hormone, leptin, through an unidentified mechanism. In this study we tested the effects of VPA, a short-chain branched fatty acid (C8:0), on leptin biology and fatty acid metabolism in 3T3-L1 adipocytes. VPA significantly reduced leptin secretion in a dose-dependent manner. Because fatty acid accumulation has been hypothesized to block leptin secretion, we tested the effect of VPA on fatty acid metabolism. Using 14C-radiolabeled VPA, we found that the 14C was mainly incorporated into triacylglycerol. VPA did not alter lipogenesis from acetate, nor did it change the amount of intracellular free fatty acids available for triacylglycerol synthesis. Decreased leptin secretion was accompanied by a reduction in leptin mRNA, even though VPA treatment did not alter the protein levels for known transcription factors affecting leptin transcription including: CCAAT/enhancer binding protein-α, peroxisome proliferator-activated receptor-γ, or steroid regulatory element binding protein 1a. VPA altered levels of leptin mRNA independent of de novo protein synthesis without affecting leptin mRNA degradation. This report demonstrates that VPA decreases leptin secretion and mRNA levels in adipocytes in vitro, suggesting that VPA therapy may be associated with altered leptin homeostasis contributing to weight gain in vivo.
Characterization of the Human Corticotropin-Releasing Factor2(a) Receptor Promoter: Regulation by Glucocorticoids and the Cyclic Adenosine 5′-Monophosphate Pathway
Authors: Nanda Steven A. , Roseboom Patrick H. , Nash George A. , Speers James M. , Kalin Ned H. 
Published: August 2004

Abstract
Corticotropin-releasing factor (CRF) is a neurotransmitter and hormone believed to integrate responses to stress. Evidence suggests central CRF systems are overactive in some individuals suffering from depression and anxiety disorders. CRF receptor antagonism blocks stress-induced endocrine, autonomic, and behavioral effects in animal models, and studies have implicated the CRF2 receptor in anxiety-related behaviors. Greater understanding of the regulation of CRF2 expression may facilitate understanding mechanisms underlying anxiety. The present studies are the first to characterize the transcriptional regulation of the human CRF2(a), the predominant CRF2 isoform in brain. Four kilobase pairs of sequence immediately upstream of the first exon of CRF2(a) represented our full-length promoter region. Sequentially smaller fragments of the CRF2(a) promoter region were generated by PCR and cloned upstream of a luciferase reporter gene. Expression was monitored from these constructs within Chinese hamster ovary-K1 cells and within rat aortic A7R5 cells that express CRF2. Glucocorticoid treatment decreased expression and elevating intracellular cAMP increased expression from the human CRF2(a) promoter. The regions of the CRF2(a) promoter that regulate the inducible expression were determined, and the functional cAMP response element and glucocorticoid response element cis-regulatory elements within these regions were identified using a combination of site-directed mutagenesis and EMSAs. Given the possibility of species-specific differences in gene expression, interpretation of gene expression studies from rat and mouse model systems is difficult. Examination of expression from the human CRF2(a) promoter will provide insight into these model systems and may translate more readily to the development of therapeutics to treat human psychiatric illness.
Screening tests for abnormalities of the protein C anticoagulant pathway / Suchteste für Anomalien im antikoagulatorischen Protein C-System
Authors: Schambeck Ch. M. 
Published: August 2004

Abstract
There is an increasing demand for a rational diagnosis of
patients with thrombophilia by clinical laboratories. The
protein C pathway plays an important part in this system.
To rationalize the diagnostics, methods have been introduced
to test the anticoagulant capacity of the whole
protein C pathway. These screening assays should reliably
detect a factor V Leiden mutation, protein C and S
deficiency. These assays differ in detection methods,
interfering factors and even in their responsiveness to
protein S. The methodological approach is based on the
activation of endogenous plasma protein C by a snake
venom extract. Clincial evaluations demonstrated the
validity of these tests to screen for a factor V Leiden
mutation and protein C deficiency. Protein S deficiency
cannot be identified sufficiently. The assays, however,
seem to have additional positive effects. They could be
used to assess the risk for thrombosis in the future. Initial
studies confirm this hypothesis. Hence, a positive test
result in protein S-deficient patients correlated with a
higher risk for thrombosis.
Ein Modell zur Lieferantenbewertung im medizinischen Laboratorium / A Model for the evaluation of suppliers in medical laboratories
Authors: Kern S. , Hubmann M. , Semmelrock H.-J. , Tiran A. 
Published: August 2004

Abstract
Periodic evaluation of suppliers is an integral component
of all relevant quality management systems established
in medical laboratories. Performing such evaluation is
always laborious, but rarely entails a perceptible benefit
for laboratories and their suppliers. Therefore, we worked
out 19 criteria for the assessment of suppliers and a
model for the analysis of results in cooperation with 5
representative manufacturers for in vitro diagnostics and
11 medical laboratories. The criteria consider the special
needs of medical laboratories combined with an effective
procedure for continual quality improvement. In a pilot
scheme the model was evaluated for its suitability in
practical use. Various methods for data analysis enable
suppliers to get relevant information about individual and
collective customer satisfaction. In the case of centralised
data collection and analysis a suitable tool for
benchmarking is available.
Diagnostik von Chlamydien-Infektionen / Diagnosis of chlamydial infections
Authors: Schütt S. , Essig A. 
Published: August 2004

Abstract
Chlamydiae are obligate intracellular bacteria undergoing
a unique growth cycle which is important for the special
features of diagnosis, therapy and pathogenesis of chlamydial
infections. Mainly Chlamydia trachomatis, Chlamydia
pneumoniae and Chlamydia psittaci cause
infections in humans. The clinical relevance of chlamydia-
like species such as Simkania, Waddlia and Parachlamydia
is unclear and currently under investigation. In
principle, laboratory diagnosis of chlamydial infections
can be achieved by direct detection of the organisms
from clinical specimens and antibody detection. The
selection of suitable laboratory tests depends on the
chlamydial species and the clinical presentation of infection.
Culture tests have been proven to be highly specific,
however due to the high level of technical expertise necessary,
chlamydial cultures are generally performed in
specialized reference laboratories. Nucleic acid amplification
technologies are characterized by their high sensitivity
and have been broadly evaluated for the detection
of C. trachomatis from urogenital specimens. In contrast,
standardization of DNA amplification methods for detection
of C. pneumonia is urgently needed to evaluate the
importance of the organism in respiratory diseases, as
well as in associated extrapulmonary diseases like atherosclerosis.
Serodiagnosis of chlamydial infections
should be based on species-specific methods like the
technically demanding microimmunofluorescence test
(MIF). Commercially developed MIF assays, peptidebased
EIAs and immunoblot assays are still in a phase
of evaluation and may contribute to a more standardized
serodiagnosis of Chlamydia infections.
Anwendungsgebiete der Massenspektrometrie in der Klinischen Chemie und Laboratoriumsmedizin / Application of mass spectrometry in clinical chemistry and laboratory medicine
Authors: Fiedler G.M. , Ceglarek U. , Lembcke J. , Baumann S. , Leichtle A. , Thiery J. 
Published: August 2004

Abstract
The use of mass spectrometry (MS) as an analytical tool
becomes increasingly important in clinical laboratory
diagnostics. In the last decade there has been an exponential
growth in clinical laboratory, pharmacology, and
toxicology applications. At present, the spectrum of MS
applications ranges from the measurement of single analytes
to qualitative and quantitative multiparametric
analyses.
Combining tandem mass spectrometry with atmospheric
pressure ionization techniques offers high analytic
sensitivity and specificity, robustness, and the option for
high throughput analyses at low running costs. Today,
this analytical platform is already used in routine clinical
laboratory to screen for inherited metabolic disorders in
newborns, for therapeutic drug monitoring, and for hormone
profiling. In addition, SELDI-TOF and MALDI-TOF
techniques are used for clinical proteomic pattern diagnostics.
This short review will give an overview of the
current status of mass spectrometry techniques, established
applications in clinical laboratory diagnostics, and
the perspectives for future developments in laboratory
medicine.
DNA methylation analysis: a promising diagnostic tool / DNA-Methylierungs-Analyse: ein vielversprechendes diagnostisches Werkzeug
Authors: Schuster M. 
Published: August 2004

Abstract
Since the investigation of molecular biological mechanisms
was extended to the genomic level and after the
sequence of the human genome was established, interest
in DNA-based diagnostics has increased tremendously.
While most research is currently being devoted
to the discovery and clinical application of genetic, proteomic
and RNA expression markers, DNA methylation
is rapidly emerging as a new level of cellular information
with biomarker potential. DNA methylation is involved in
the regulation of gene activity. Aberrant DNA methylation
has been associated with a variety of human diseases.
As such, DNA methylation markers hold a great promise
as a diagnostic tool, e.g. in early cancer screening [1].
Methods are available for the genome-wide discovery of
methylation markers that differentiate between different
types of tissue samples. In contrast to expression markers,
DNA methylation markers are accessible at the DNA
level, which enables their analysis even in archived paraffin-
embedded tissue samples. Methods are available to
detect a few copies of aberrantly methylated DNA in a
background of a vast excess of normally methylated
DNA, which is the prerequisite for assaying methylation
markers also in body fluids that are readily accessible in
clinical practice.
Trace Analysis of Peroxide Explosives by High Performance Liquid Chromatography-Atmospheric Pressure Chemical Ionization-Tandem Mass Spectrometry (HPLC-APCI-MS/MS) for Forensic Applications
Authors: Xu X , van de Craats AM , Kok EM , de Bruyn PCAM 
Published: August 2004

Abstract
An HPLC-APCI-MS(/MS) method for the (trace) analysis of the most commonly encountered peroxide explosives, hexamethylenetriperoxidediamine (HMTD) and triacetonetriperoxide (TATP), has been developed. With this method, HMTD and TATP have been analyzed in the same run. (Pseudo-)molecular ions of these peroxides have been obtained as base peak under the same condition. A series of product ions was produced from these pseudo-molecular ions ([HMTD-1]+ and [TATP + NH4]+) in the MS/MS analysis. We also pioneeredin showing that TATP molecular ion [TATP-H]+ can be observed with HPLC-MS/MS. The limit of detection for HMTD and TATP was 0.26 and 3.3 ng, respectively, on column by HPLC-MS in the Full Scan mode and 0.08 and 0.8, respectively, by HPLC-APCI-MS/MS in Selected Reaction Monitoring (single ass unit) mode. The method presented has been applied successfully for the identification of peroxides in the bulk solid state (powder sample), as well as in post-blast extracts originating from a forensic case. For the post-blast extracts, the use of tandem MS has been shown clearly to be of crucial importance for the identification and detection of the peroxide explosives.
Y-Chromosome STR System, Y-PLEX™ 12, for Forensic Casework: Development and Validation
Authors: Shewale JG , Nasir H , Schneida E , Gross AM , Budowle B , Sinha SK 
Published: August 2004

Abstract
The Y-PLEX™ 12 system, developed for use in human identification, enables simultaneous amplification of eleven polymorphic short tandem repeat (STR) loci, namely DYS392, DYS390, DYS385 a/b, DYS393, DYS389I, DYS391, DYS389II, DYS19, DYS439 and DYS438, residing on the Y chromosome and Amelogenin. Amelogenin provides results for gender identification and serves as internal control for PCR. The validation studies were performed according to the DNA Advisory Board's (DAB) Quality Assurance Standards. The minimal sensitivity of the Y-PLEX™ 12 system was 0.1 ng of male DNA. The mean stutter values ranged between 3.76–15.72%. A full male profile was observed in mixture samples containing 0.5 ng of male DNA and up to 400 ng of female DNA. Amelogenin did not adversely affect the amplification of Y-STRs in mixture samples containing male and female DNA. The primers for the Y-STR loci present in Y-PLEX™ 12 are specific for human DNA and some higher primates. None of the primate samples tested provided a complete profile at all 11 Y-STR loci amplified with the Y-PLEX™ 12 system. Y-PLEX™ 12 is a sensitive, valid, reliable, and robust multiplex system for forensic analysis, and it can be used in human forensic and male lineage identification cases.