bluuebunny/crossref_metadata_2025 huggingface dataset:
Identification of Colon Cancer Patients by Molecular Diagnosis
Authors: Müller Oliver
Published: January 2004

The important causal role of DNA alterations in cancer development implies that tumor specific DNA mutations might serve as ideal parameters in cancer diagnosis. In particular the detection of mutations in fecal DNA might complement the current methods for the early diagnosis of colorectal tumors. Over the last 10 years many studies have proven the usefulness and the feasibility of the fecal DNA analysis in the diagnosis of colon tumors. Technical difficulties have circumvented the clinical application and the broad screening by molecular diagnosis. Recent methodological proceedings together with the promising outcomes of first multi-target studies have put a diagnostic stool-based DNA test for colorectal tumors within close reach.
Molecular Detection of Circulating Cancer Cells
Authors: Molnar Bela , Sipos Ferenc , Galamb Orsolya , Tulassay Zsolt
Published: January 2004

Background: The circulating tumor cells of solid tumors, especially in colorectal cancer has been of great interest in medical research in the last and new century. Methods: A literature review of recent data for the preparation of a proposal for clinical applications. Results: Starting with the detection of circulating cancer cells in histological sections, lymph nodes, bone marrow and peripheral blood have recently become the primary samples for these investigations. Following microscopic morphology, new methods were and are being developed continuously including rare cell enrichment and detection techniques like fluorescence microscopy, flow cytometry, RT-PCR and methylated DNA PCR. The sensitivity of these assays reaches down to the concentration of 1 tumor cell/ml blood. A lot of different epithelial markers were targeted with these techniques e.g. cytokeratins, EGFR, CEA, and EMA. Clinically, circulating tumor cells were found as independent prognostic factors in lymph nodes and bone marrow. In blood, their presence appears to be an early marker for recurrence and relapse. The change in their quantitative number is supposed to reflect the chemotherapeutic sensitivity and metastatic growth activity of the tumor. Conclusions: The detection of circulating tumor cells in bone marrow and lymph nodes is of clinical significance. Their presence in peripheral blood and therapeutic applications need further clarification.
Alterations of the CARD15/NOD2 Gene and the Impact on Management and Treatment of Crohn’s Disease Patients
Authors: Török Helga-Paula , Glas Jürgen , Lohse Peter , Folwaczny Christian
Published: January 2004

The recent identification of the CARD15/NOD2 gene as a susceptibility locus for Crohn’s disease represents an important step towards the delineation of the immunopathogenesis of inflammatory bowel disease. CARD15 functions as an intracellular receptor for bacterial components and thus represents an important link between inflammatory bowel disease and innate immunity. Three major CARD15/NOD2 gene mutations have been associated with Crohn’s disease in Caucasians in several independent studies. Together, they explain about 20% of the genetic susceptibility for Crohn’s disease. Genotype-phenotype analyses demonstrated an association of these mutations with ileum-specific disease, an increased incidence of the fibrostenotic phenotype and an earlier age of disease onset. Beside these associations, no other relationship between the CARD15/NOD2 genotype and disease behavior or response to treatment has been detailed so far. Thus, the clinical impact of knowing the patient’s genotype is limited at this time. Screening for CARD15 mutations in order to identify high-risk individuals or to introduce an individualized disease management is therefore currently not recommended.
Identification of Patients with Genetic Risk Factors of Pancreatitis: Impact on Treatment and Cancer Prevention
Authors: Keim Volker
Published: January 2004

Recent data suggest that genetic alterations are relevant risk factors for chronic pancreatitis. The highest risk is associated with autosomal-dominant mutations (N29I, R122H) of the cationic trypsinogen (PRSS1). Further mutations were identified in the genes of the pancreatic trypsin inhibitor (SPINK1) and in the cystic fibrosis transmembrane conductance regulator (CFTR). A remarkable finding was that both molecules were also mutated in patients suffering from alcoholic chronic pancreatitis. According to recent estimations, genetic alterations may be regarded as more severe risk factors than chronic alcohol consumption. To identify patients with mutations, a positive family history could be of help, but mutations were also found in a significant number of those with a negative family history. On the other hand, in approximately 40% of the patients with a positive family history no mutations were found up to now. The age at onset is lower in patients with genetic risk factors; however, no clear limit can be denominated above which a screening is not appropriate. Therefore, in our department genetic screening is offered to all patients with chronic pancreatitis of unclear origin. There is no specific treatment in patients with a genetically based disease. The patients with familial pancreatitis-increased rates of pancreas cancer were described but there is no agreement concerning the prophylactic strategy. Prevention of cancer by routine pancreatectomy, though performed recently, is not justified at the moment. Clinical criteria may be more appropriate to decide the timing and the extent of the operation.
Informed Consent for Molecular-Based Diagnostic and Prognostic Studies in the Cancer Patient
Authors: Reymond Marc A. , Allal Abdelkarim S. , Steinert Ralf , Eder Frank , Halangk Walter , Lippert Hans
Published: January 2004

The consent of the subject who is donating cells, tissues, organs or body fluid for research purposes is a precondition for biomedical research on human samples. Before such consent can be obtained, comprehensive information should be provided to the subject by the investigators. This information includes the scope of research, the intended use of subject’s body parts and associated data, the risks and benefits associated with the research project, and the right to withdraw their consent at any time without prejudice. Consent is free and must not be obtained under any kind of pressure. The traditional practice of obtaining consent for unspecified future use of biological samples and data generated from clinical trials is no longer adequate for genetic research. However, the investigator has still interest to keep the definition of the field of research as broad as possible. Privacy is an issue of importance for the patient, who is the primary supplier of human samples, and for his relatives. Benefit sharing is another field of contradictory discussion. Normal volunteers are a special case among gene expression studies. An example of informed consent form is provided.
Microsatellite Alterations in Free-Circulating Serum DNA in Patients with Ulcerative Colitis
Authors: Rauh Peter , Rickes Steffen , Fleischhacker Michael
Published: January 2004

Background/Aims: The aim of this study was to find out whether there is free-circulating DNA in the serum of patients with ulcerative colitis and whether it is possible to establish a panel of genetic alterations which might be useful markers for diagnostic purposes, staging, or follow up. Methods: A set of 11 microsatellite markers located on different chromosomes were analyzed in a group of 59 patients with ulcerative colitis. Radiolabeled PCR products from serum and white blood cell DNA were analyzed by autoradiography and correlated with the clinical data from these patients. Results: Seven of 59 patients showed one or more microsatellite alteration(s) in their serum DNA. Six patients had an alteration in one marker, and two patients had changes in 4 markers, respectively. There was no correlation between the frequency of microsatellite alterations and the clinical data. Conclusion: From the panel of 11 microsatellite markers used for these studies, 6 of them seem to be well suited for the detection of ulcerative colitis-associated alterations in free circulating serum DNA. In order to increase the specificity of this assay, it is necessary to increase the number of patients to be analyzed and to use a different set of markers.
Effects of Reduced Dialysate Calcium on Calcium-Phosphorus Product and Bone Metabolism in Hemodialysis Patients
Authors: Fiedler Roman , Deuber Heinz J. , Langer Thomas , Osten Bernd , Mohan Subburaman , Jehle Peter M.
Published: January 2004

Background: The safety of using reduced calcium dialysate (RDC) in hemodialysis (HD) patients is controversial due to related changes in bone metabolism. In the present study we investigated whether an 18-month treatment period with RDC may induce significant changes in calcium-phosphorus product (CaxP), bone metabolism, and components of the insulin-like growth factor (IGF) system in HD patients. Study Design: In this prospective study, 13 HD patients with biochemical signs of diminished or low-normal bone turnover and high CaxP due to high serum calcium level were treated by lowering dialysate calcium from 3.5 to 2.5 mEq/l for 18 months. By specific immunometric assays, serum levels of intact parathyroid hormone (PTH), bone alkaline phosphatase (B-ALP), pyridinoline (PYR), desoxypyridinoline (D-PYR), 25-OH-vitamin D3 (25-vit D3), 1,25-(OH)2-vitamin D3 (1,25-vit D3), free IGF-I, IGF-II, and IGF-binding protein (IGFBP)-1 to -6 were measured. Results: CaxP decreased significantly from 5.62 (baseline) to 3.95 mmol2/l2 (at 18 months), whereas PTH increased from 81 ± 57 pg/ml at baseline to 236 ± 188 at 12 months (p < 0.01), remaining in this range thereafter. Parameters of bone resorption (PYR) as well as formation (B-ALP) significantly increased during RDC, with peak levels after 12 months. Despite increasing doses of oral alfacalcidol, levels of 25-vit D3 and 1,25-vit D3 subsequently declined during RDC. In parallel with the changes in bone markers, free IGF-I levels decreased (baseline: 1.9 ± 0.9 ng/ml, after 18 months: 1.1 ± 0.7; p < 0.01). The decline of free IGF-I correlated with decreasing levels of IGFBP-3 and increasing levels of IGFBP-1/-4. Conclusion: The treatment with RDC effectively lowered CaxP and stimulated bone formation and resorption. The different changes in bone markers and IGF system components mirror the complex effects on bone metabolism.
Impaired T Cell Proliferation and Zeta Chain Phosphorylation after Stimulation with Staphylococcal Enterotoxin-B in Hemodialysis Patients
Authors: Eleftheriadis Theodoros , Papazisis Konstantinos , Kortsaris Alexandros , Vayonas Georgios , Voyatzi Sultana , Vargemezis Vasilios
Published: January 2004

Background: Patients on regular hemodialysis treatment are in an immunodeficiency state. Several studies have shown defective T cell proliferation after stimulation with various agents. Staphylococcal enterotoxin B (SEB) is a MHC-dependent superantigen that triggers proliferation of a large proportion of T cells. T cell activation after stimulation with SEB parallels normal T cell signal transduction. An important and early event in this transduction pathway is the phosphorylation of the ζ chain. In this study, T cell proliferation and ζ chain phosphorylation after stimulation with SEB were evaluated. Methods: Peripheral blood mononuclear cells (PBMCs) from 24 patients and 14 healthy individuals were isolated and cultured with or without stimulation with SEB (1 ng/ml). Cell proliferation was estimated by immunoenzymatic measurement of bromodeoxyuridine uptake. PBMCs from 8 patients and 6 healthy individuals were isolated and pulsed for 2 min with or without SEB (10 µg/ml). ζ chain phosphorylation was estimated by immunoprecipitation and immunoblotting with antiphosphotyrosine antibody. Results: Lymphocyte proliferation index after SEB stimulation was lower in hemodialyzed patients. Stimulation of T cells with SEB also resulted in a lower ζ chain phosphorylation in hemodialyzed patients. Conclusions: Lymphocyte proliferation after MHC-dependent stimulation is impaired in hemodialyzed patients. This proliferation defect is due to impaired ζ chain phosphorylation.
Quality of Life in Male Hemodialysis Patients
Authors: Türk Süleyman , Guney Ibrahim , Altintepe Lutfullah , Tonbul Zeki , Yildiz Alaattin , Yeksan Mehdi
Published: January 2004

Background: Health-related quality of life (QOL) is affected in hemodialysis patients (HD). A number of factors such as age, anemia, and comorbidity had been implicated in decreased QOL. Erectile dysfunction (ED) is a frequent and potentially treatable complication in HD patients. In this cross-sectional study, we aimed to evaluate the possible relation between the QOL and ED in HD patients. Patients and Methods: Among the 511 chronic HD patients dialyzed in 11 outpatient HD centers, 148 male patients (mean age: 46 ± 9 years) were included. The mean time on dialysis was 41 ± 35 months (range: 3–203 months). Biochemical parameters such as BUN, creatinine, hemoglobin, serum albumin and Kt/V were measured. The QOL of the patients were measured with the short form of Medical Outcomes Study (SF-36), physical component scores (PCS) and mental component scores (MCS) were calculated. The ED was evaluated by the International Index of Erectile Function (IIEF). Results: One hundred and four of the 148 patients (70%) had ED. Hemoglobin levels were correlated with PCS (r = 0.197, p = 0.02) and MCS (r = 0.20, p = 0.019). Patients with ED had lower scores in nearly all the components related to PCS and MCS as compared to patients without ED. IIEF score was correlated with PCS (r = 0.369, p < 0.001) and MCS (r = 0.308, p < 0.001). In linear regression analysis, IIEF score and hemoglobin levels were the independent variables that predicted both PCM and MCS. Conclusion: ED, a frequent complication in HD patients, was related to QOL together with anemia. Successful treatment of ED and anemia may lead to improvement in QOL in HD patients.
Autoimmune Renal Injury in C3- and C4-Deficient Mice: A Histological and Functional Study
Authors: Kouki P. , Marsh J.E. , Sacks S.H. , Sheerin N.S.
Published: January 2004

Background: Complement deficiency predisposes to autoimmune renal disease. Since complement deficient mice are increasingly used to study the immunopathogenesis of renal disease we have determined whether mice deficient in C3 or C4 are susceptible to spontaneous immune-mediated renal injury. Methods: C3-deficient, C4-deficient and complement-sufficient, wild-type mice were maintained in standard conditions for 1 year at which stage renal function, renal histology, circulating antibody and autoantibody levels were assessed. Results: No significant decline in renal function was demonstrated in the complement-deficient mice. However, there was histological evidence of glomerular injury in both the C3- and C4-deficient mice, but of insufficient severity to alter function. Serum IgG2a concentration was significantly lower in C3- and C4-deficient mice. In contrast C3-deficient mice had higher concentrations of serum IgG2b. There was a tendency for mice from all groups, including the complement-sufficient mice, to develop autoantibodies. C4-deficient mice had higher titres of anti-dsDNA IgG but otherwise deficient mice had similar autoantibody titres to controls. Conclusion: We conclude that C4-deficient mice demonstrate a small increase in autoantibody production at 1 year of age compared to C3-deficient and wild-type mice. Furthermore, although complement-deficient mice exhibit glomerular changes, they are of minor functional significance, and are unlikely to affect the study of experimentally induced renal disease in these mice.
Stimulation of NHE3 in OKP Cells by an Autocrine Mechanism
Authors: Amemiya Morimasa , Mori Honami , Imamura Shigeki , Toyoda Akira , Funayama Izumi , Asano Yasushi , Kusano Eiji , Tabei Kaoru
Published: January 2004

Background/Aims: Chronic hypokalemia increases NHE3 activity in OKP cells. The aim of the present study was to determine whether an autocrine mechanism is involved in this activation. Methods: After incubation of OKP cells in normal-K+ and low-K+ media for 24 h, the potassium concentration in the low-K+ media was adjusted to a normal level. These conditioned media were then used as the normal-K+ and low-K+ supernatants. Other OKP cells were incubated in these normal-K+ and low-K+ supernatants and the mechanism of Na+/H+ antiporter activation was examined. Results: The EIPA-resistant Na+/H+ antiporter activity of OKP cells increased after 4 h incubation in the low-K+ supernatant, and the amount of NHE3 protein increased at 24 h. Since both BQ788 and saralasin blocked this antiporter activation, the supernatant concentration of endothelin I (ET-I) and angiotensin II (Ang-II) were measured. The ET-I concentration was reduced, but the Ang-II concentration remained unchanged. There was a significant association between a reduction in the ET-I concentration and an increase in Na+/H+ antiporter activity, but only when Ang-II was present in the supernatant. Conclusion: An autocrine mechanism is involved in the activation of NHE3 in OKP cells. Both ET-I and Ang-II play a role in this activation.
Regulation of Plasma Hemopexin Activity by Stimulated Endothelial or Mesangial Cells
Authors: Kapojos Jola J. , Poelstra Klaas , Borghuis Theo , Banas Bernhard , Bakker Winston W.
Published: January 2004

The pathogenesis of glomerular alterations and proteinuria in corticosteroid-responsive nephrotic syndrome (CRNS) is unknown. As an isoform of plasma hemopexin (Hx) with protease activity may be implicated in this disease, we have studied the inhibition of Hx by ADP and reactivation to its active form by endothelial or mesangial cells in vitro. We hypothesized that these cells might potentially be able to convert the inactivated form of Hx (Hxi) to active Hx (Hxa) in vitro, mediated by cellular ecto-ADPase. Since ecto-ADPase (CD39) is upregulated after stimulation of these cells with lipopolysaccharide (LPS) or certain cytokines, we postulated that this conversion might occur specifically after inflammatory stimulation of these cells. Human endothelial or mesangial cell cultures were incubated overnight with or without LPS (10.0 ng/ml) or TNFα (10.0 ng/ml), washed and subsequently incubated with Hxi (1.5 mg/ml) in serum-free conditions (Hxi was prepared by treatment of Hxa with ADP or ADP-β-S). After 60 min, supernatants were tested for their capacity to alter glomerular extracellular matrix molecules (i.e. ecto-apyrase) in vitro using standard methods, and compared with Hxi that had not been incubated with cells. Supernatants containing Hxa (1.5 mg/ml) served as positive control. The results show significant activity in supernatants with Hxi (prepared using native ADP). However, Hxi inactivated by ADP-β-S (which is non-hydrolyzable) could not be reactivated after contact with LPS-stimulated or unstimulated cells in vitro. As ecto-ADPase of these cells is upregulated by LPS, we believe that reactivation of Hxi to Hxa is mediated by cellular ecto-ADPase. Although the relevance of this inflammation-mediated activation mechanism of Hx in patients with CRNS requires further experimentation, our preliminary observations suggesting that this mechanism is corticosteroid dependent may support a potential role of Hxa in CRNS.
Low Sodium Excretion in SIADH Patients with Low Diuresis
Authors: Musch Wim , Hedeshi Ali , Decaux Guy
Published: January 2004

It is well known that during low diuresis or low effective circulating volume, salt excretion is low. The aim of this study was to find out whether salt excretion, expressed as either urinary sodium concentration (UNa) or fractional sodium excretion (FENa), and the combined use of FENa and fractional urea excretion (FEurea) still differentiate between hyponatremic SIADH and hyponatremic salt depletion (SD) patients when diuresis is low. The relationships between UNa, FENa and diuresis, indirectly estimated by the urinary to plasma creatinine ratio (U/P creat), were studied in 42 hyponatremic SIADH patients, 21 hyponatremic SD patients and 66 normonatremic controls (CO) of similar age and sex ratio. There was no significant relationship between UNa and U/P creat either in SIADH or in SD or CO patients. FENa and U/P creat were inversely correlated, both in CO (r = –0.72; p < 0.001) and in SIADH (r = –0.68; p < 0.001). SIADH and SD patients can be fairly well differentiated from one another using FENa and U/P creat. Even with high U/P creat values, SIADH patients, despite a sharp decrease in their FENa values, presented still higher FENa values than SD patients did (mean FENa = 0.3 ± 0.2% in SIADH and 0.1 ± 0.04% in SD; p < 0.05). However, FENa values of SIADH patients with low diuresis (mean FENa = 0.3 ± 0.2% for a mean U/P creat = 191 ± 40) are indistinguishable from those of SD patients with normal urine volumes (mean FENa = 0.2 ± 0.2% for a mean U/P creat = 92 ± 30). The combined use of FENa and FEurea remains a reliable way to discriminate SD patients and SIADH patients, as far as the differential limit value for FENa is narrowed to a value of 0.15%, for hyponatremic patients with U/P creat >140. Conclusion: In SIADH, FENa values are lower than 0.5%, as soon as U/P creat exceeds a value of 180. In SD patients with U/P creat values exceeding 140, FENa is lower than 0.15% and FEurea lower than 45%.
Increased Renal Fibrosis and Expression of Renal Phosphatidylinositol 4-Kinase-β and Phospholipase Cγ1 Proteins in Piglets Exposed to Ochratoxin-A
Authors: Aukema H.M. , House J.D. , Bankovic-Calic N. , Ogborn M.R.
Published: January 2004

Endemic nephropathy has been linked to exposure of ochratoxin-A (OA) in grains and animal products. The underlying events surrounding this form of renal injury are not well known, partly due to the lack of a suitable animal model of the disease. Therefore, in this study, a pig model of OA-induced renal injury was established and used to examine whether elements of the phosphoinositide signalling pathway are altered in this disease. Weanling piglets were fed diets containing 0, 2, and 4 ppm OA for 6 weeks. Serum creatinine and urea and renal fibrosis were monitored biweekly using serial blood samples and renal biopsies. At termination, the protein levels of renal phosphatidylinositol 4-kinase-β (PtdIns4Kβ) and phospholipase Cγ1 (PLCγ1) were determined using immunoblotting and scanning densitometry. Serum creatinine was elevated by 2 weeks and renal fibrosis was elevated by 4 weeks at both levels of inclusion of OA. At the end of the experimental period, kidney size and water content were elevated, as were the protein levels of renal PtdIns4Kβ and PLCγ1 in OA-exposed animals. Therefore, serial biopsies can be used to track changes in renal pathology in the OA-exposed piglet. We conclude that this is a useful model for OA-induced renal injury in which the underlying molecular events associated with this form of renal injury can be studied.
A Study of VEGF and Its Receptors in Two Rat Models of Proteinuria
Authors: Kanellis John , Levidiotis Vicki , Khong Tiffany , Cox Alison J. , Stacker Steven A. , Gilbert Richard E. , Cooper Mark E. , Power David A.
Published: January 2004

Background: The high level of expression of vascular endothelial growth factor (VEGF) in normal podocyte foot processes suggests that VEGF has an important role in maintaining normal glomerular function. While altered VEGF expression occurs in many glomerular diseases, a direct role for VEGF in the pathogenesis of proteinuria has not been demonstrated. Methods: Expression of VEGF and its receptors (VEGFR-1 and VEGFR-2) was examined in passive Heymann nephritis (PHN) and puromycin aminonucleoside nephrosis (PAN), by immunohistochemistry, in situ hybridization, Northern and Western blotting. Inhibition of VEGF in the PAN model was performed by administration of a blocking antibody. Results: In both models, glomeruli showed upregulation of VEGF and VEGF receptors compared to control animals. VEGF mRNA was increased most significantly (5-fold) at day 5 after induction of PHN, prior to the onset of proteinuria, with persistent upregulation (3-fold) at day 21. Increased VEGF mRNA was also seen in PAN, but it was less marked. In situ hybridization and immunohistochemistry localized VEGF predominantly to podocytes. Increased expression of VEGFR-1 and VEGFR-2 protein was seen in glomerular endothelial cells of PHN and PAN rats by immunohistochemistry, as was VEGFR-2 mRNA by in situ hybridization. Upregulation of VEGFR-1 by endothelial cells was more striking in the PAN model than PHN. Administration of a blocking antibody to rats with PAN did not affect proteinuria, creatinine clearance or sodium excretion. Conclusion: The expression of VEGF and its receptors is significantly increased in the PHN and PAN rat models of proteinuria suggesting a role for VEGF in the disease process. VEGF may have an important role in promoting glomerular repair in a variety of glomerular diseases.
Activation of the Inflammatory Reaction within Minutes after Birth in Ventilated Preterm Lambs with Neonatal Respiratory Distress Syndrome
Authors: Jaarsma Anneke S. , Braaksma Margriethe A. , Geven Wil B. , van Oeveren Wim , Bambang Oetomo Sidarto
Published: January 2004

To study the activation of the inflammatory reaction within minutes after birth, we measured parameters of inflammation before and immediately after birth. To assess whether respiratory distress syndrome (RDS) or birth itself initiates activation, we compared preterm ventilated lambs with term nonventilated lambs. Preterm lambs were delivered by cesarean section at 132 days gestational age (term 145 days) and were ventilated by conventional ventilation (n = 9). Before clamping the cord, 5, 10 and 15 min after birth, blood was sampled from umbilical catheters. Term lambs (n = 9) were born spontaneously after 140–145 days gestational age. Immediately after birth, a venous umbilical catheter was inserted. Blood was sampled before the first breath and 5, 10, 15 and 20 min after birth while the lamb was breathing spontaneously. Blood was analyzed for AP50 (complement activation), number of polymorphonuclear leukocytes (PMNs) and β-glucuronidase (released from activated PMNs). In preterm lambs, we found a decreased number of PMNs and increased levels of β-glucuronidase already at 5 min after birth. In the term lambs, we found only a short-term mild decrease in PMNs and short-term increase in β-glucuronidase. We conclude that systemic activation of the inflammatory reaction can be found in ventilated preterm lambs with RDS within 5 min after birth. This very early activation is mild, transient and less pronounced in term-born spontaneously breathing lambs compared with preterm, ventilated lambs with RDS.
Statistical Evaluation of Local Alignment Features Predicting Allergenicity Using Supervised Classification Algorithms
Authors: Soeria-Atmadja D. , Zorzet A. , Gustafsson M.G. , Hammerling U.
Published: January 2004

Background: Recently, two promising alignment-based features predicting food allergenicity using the k nearest neighbor (kNN) classifier were reported. These features are the alignment score and alignment length of the best local alignment obtained in a database of known allergen sequences. Methods: In the work reported here a much more comprehensive statistical evaluation of the potential of these features was performed, this time for the prediction of allergenicity in general. The evaluation consisted of the following four key components. (1) A new high quality database consisting of 318 carefully selected, non-redundant allergens and 1,007 sequences carefully selected to be non-allergens. (2) Three different supervised algorithms: the kNN classifier, the Bayesian linear Gaussian classifier, and the Bayesian quadratic Gaussian classifier. (3) A large set of local alignment procedures defined using the FASTA3 alignment program by means of a wide range of different parameter settings. (4) Novel performance curves, alternative to conventional receiver-operating characteristic curves, to display not only average behaviors but also statistical variations due to small data sets. Results: The linear Gaussian classifier proved most useful among the tested supervised machine learning algorithms, closely followed by the quadratic Gaussian equivalent and kNN. The overall best classification results were obtained with a novel feature vector consisting of the combined alignment scores derived from local alignment procedures using different substitution matrices. Conclusions: The models reported here should be useful as a part of an integrated assessment scheme for potential protein allergenicity and for future comparisons with alternative bioinformatic approaches.
Ginsenoside Rh1 Possesses Antiallergic and Anti-Inflammatory Activities
Authors: Park Eun-Kyung , Choo Min-Kyung , Han Myung Joo , Kim Dong-Hyun
Published: January 2004

Background: Ginseng (the root of Panax ginseng C.A. Meyer, Araliaceae) has been reported to possess various biological activities, including anti-inflammatory and antitumor actions. In this study, we investigated the antiallergic activity of ginsenosides isolated from ginseng. Method: We isolated ginsenosides by silica gel column chromatograghy and examined their in vitro and in vivo antiallergic effect on rat peritoneal mast cells and on IgE-induced passive cutaneous anaphylaxis (PCA) in mice. The in vitroanti-inflammatory activity of ginsenoside Rh1 (Rh1) in RAW264.7 cells was investigated. Results: Rh1 potently inhibited histamine release from rat peritoneal mast cells and the IgE-mediated PCA reaction in mice. The inhibitory activity of Rh1 (87% inhibition at 25 mg/kg) on the PCA reaction was found to be more potent than that of disodium cromoglycate (31% inhibition at 25 mg/kg); Rh1 was also found to have a membrane-stabilizing action as revealed by differential scanning calorimetry. It also inhibited inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) protein expression in RAW 264.7 cells, and the activation of the transcription factor, NF-ĸB, in nuclear fractions. Conclusion: The antiallergic action of Rh1 may originate from its cell membrane-stabilizing and anti-inflammatory activities, and can improve the inflammation caused by allergies.
Parietaria Profilin Shows Only Limited Cross-Reactivity with Birch and Grass Profilins
Authors: Asero Riccardo , Mistrello Gianni , Roncarolo Daniela , Amato Stefano
Published: January 2004

Background: In patients showing multiple reactivity to seasonal airborne allergens, a skin prick test (SPT) with Parietaria pollen scores frequently negative, suggesting that profilin from Parietaria might not share IgE-binding epitopes with profilin from botanically unrelated airborne allergens. This study investigated the immunologic cross-reactivity between profilins from Parietaria, grass and birch pollen. Methods: 36 patients hypersensitive to birch profilin, Bet v 2, underwent SPT with Parietaria, and IgE to both whole Parietaria pollen and Phleum profilin were sought in their sera by ELISA. In ELISA inhibition studies, IgE reactivity to Phleum profilin was measured before and after absorption of sera from Parietaria reactors with both whole Parietaria pollen extract, and Par j 1/Par j 2 fraction. Further, their IgE reactivity to whole Parietaria pollen was measured before and after absorption with the Par j 1/Par j 2 fraction. Results: All sera showed IgE reactivity to Phleum profilin. Only 14/36 (39%) patients had a positive SPT to Parietaria and 17/36 (47%) showed IgE to Parietaria. Absorption of sera from Parietaria reactors with whole Parietaria extract caused a marked reduction in IgE reactivity to grass profilin; in contrast, absorption of sera with the Par j 1/Par j 2 fraction did not exert any inhibitory effect on IgE reactivity to grass profilin. Absorption of sera with the Par j 1/Par j 2 fraction markedly reduced IgE reactivity to whole Parietaria extract in 8/9 cases. Conclusion: Less than 50% of patients sensitized to birch and grass profilin cross-react to Parietaria profilin. In most cases, cross-reactors are hypersensitive to major, specific Parietaria allergens as well. This findings may be of practical relevance when the prescription of specific immunotherapy is considered.
Five-Year Follow-Up of a 13-Year-Old Boy with a Pituitary Adenoma Causing Gigantism – Effect of Octreotide Therapy
Authors: Schoof Ellen , Dörr Helmuth G. , Kiess Wieland , Lüdecke Dieter K. , Freitag Eduard , Zindel Volker , Rascher Wolfgang , Dötsch Jörg
Published: January 2004

Background/Aim: In children, there is little experience with octreotide therapy for pituitary tumors, especially growth hormone (GH) producing adenomas. We report on a 13-year-old boy with gigantism due to a GH-producing pituitary adenoma caused by a Gsα mutation on the basis of McCune-Albright syndrome. Methods: At the age of 6.5 years a GH- and prolactin-producing pituitary adenoma was diagnosed. The adenoma was surgically removed. Immediately thereafter, the small adenoma residuum was treated with octreotide (2 × 100 µg/day s.c.). Results: During therapy with octreotide, the growth rate dropped to normal values; however, rose again after 2 years of treatment. The insulin-like growth factor I (IGF-I) levels remained above the 95th percentile, the GH level mostly >2 µg/l. After 5 years of octreotide therapy, GH (6.9 µg/l), IGF-I (620 µg/l), IGF-binding protein 3 (5.4 mg/l), and prolactin (17.0 ng/ml) levels were still elevated. The growth velocity was +2.4 SDS (standard deviation score), the pubertal status was mature, and the bone age was 14.3 years (prospective final height 208 cm). A magnetic resonance imaging scan showed an unchanged residual 4-mm rim of adenoma at the pituitary site. Side effects from octreotide therapy were not reported by the patient or his family. The therapy was changed to the long-acting release octreotide analog octreotide-LAR. After 1 year of treatment with octreotide-LAR, the GH level was 1.0 µg/l, and the prospective final height dropped by 10 cm. Conclusions: This case demonstrates that combined surgical and medical treatment can influence the prognosis of childhood gigantism; however, the prognosis of this rare condition remains uncertain.
Pituitary Metastasis of Follicular Thyroid Carcinoma
Authors: Chrisoulidou Alexandra , Pazaitou-Panayiotou Kalliopi , Flaris Nikolaos , Drimonitis Apostolos , Giavroglou Iosifina , Ginikopoulou Eudoxia , Vainas Iraklis
Published: January 2004

Objective: The case of a 60-year-old male patient with follicular thyroid cancer who developed a pituitary mass proved to be a metastasis from thyroid cancer. Methods: Assessment with whole-body scan, ultrasound, computed tomography and thyroglobulin measurements. Results: Despite surgery and repeated doses of radioiodine, the patient developed diplopia and ptosis of the right eyelid, along with increasing thyroglobulin levels. A pituitary mass was discovered, with no signs of pituitary deficiency. The mass was removed and found to consist of neoplastic cells immunohistochemically positive to thyroglobulin. Conclusions: Distant metastases may develop in cases of follicular thyroid carcinoma, even after repeated doses of 131I. Metastatic follicular thyroid carcinoma to the pituitary is a rare entity.
Effects of Simvastatin on Plasma Lipoproteins and Response to Arterial Injury in Wild-Type and Apolipoprotein-E-Deficient Mice
Authors: Choudhury Robin P. , Carrelli Angela L. , Stern Joshua D. , Chereshnev Igor , Soccio Raymond , Elmalem Valerie I. , Fallon John T. , Fisher Edward A. , Reis Ernane D.
Published: January 2004

Objective: To test the non-lipid-lowering effects of simvastatin on the response to injury in normolipidemic and hyperlipidemic mice. Methods and Results: Wild-type (WT) mice (n = 40) and hyperlipidemic apolipoprotein-E-deficient (apoE–/–) mice (n = 40) received normal chow or chow containing simvastatin 100 mg/kg/day prior to bilateral femoral artery wire injury. Intimal hyperplasia and plasma cholesterol concentration were quantified after 4 weeks. Plasma cholesterol in WT mice treated or untreated with simvastatin was similar (100.9 ± 6.6 vs. 94.3 ± 17.5 mg/dl). Simvastatin did not affect intimal hyperplasia. In apoE–/– mice, intimal hyperplasia was increased 2.3-fold relative to WT mice (17,090 ± 4,998 vs. 39,490 ± 16,190; p < 0.001). In apoE–/– mice, simvastatin caused a paradoxical increase in plasma cholesterol (1,094 ± 60.3 vs. 658 ± 66.8 mg/dl; p < 0.001), confirmed by FPLC. This was associated with a further increase in intimal area (39,490 ± 16,190 vs. 55,420 ± 22,590 mm2; p < 0.01). Conclusions: (1) Simvastatin had no effect on plasma cholesterol or the response to arterial injury in normolipidemic WT mice; (2) hyperlipidemia was associated with markedly increased intimal hyperplasia, and (3) simvastatin treatment of apoE–/– mice caused paradoxical hyperlipidemia and increased intimal hyperplasia.
Expression of Costimulatory Molecules (4-1BBL and Fas) and Major Histocompatibility Class I Chain-Related A (MICA) in Aortic Tissue with Takayasu’s Arteritis
Authors: Seko Yoshinori , Sugishita Kazuyuki , Sato Osamu , Takagi Atsuhiko , Tada Yusuke , Matsuo Hiroshi , Yagita Hideo , Okumura Ko , Nagai Ryozo
Published: January 2004

To further investigate the immunological mechanisms involved, we analyzed the expression of costimulatory molecules in aortic tissue and their counterpart molecules on infiltrating cells of patients with Takayasu’s arteritis. We also examined the expression of major histocompatibility complex (MHC) class I chain-related (MIC) A in aortic tissue, which is known to be induced by external stress, and its counterpart NKG2D receptors on infiltrating cells. Among these costimulatory molecules, strong expression of 4-1BBL and Fas was induced in the aortic tissue, and most of the infiltrating cells expressed 4-1BB and FasL, suggesting these pathways play critical roles in T-cell-mediated vascular injury. We also found that MICA was strongly induced in the aortic tissue and that at least part of the infiltrating cells expressed NKG2D receptors. Some infiltrating cells – but not vascular smooth muscle cells – seemed to have undergone apoptosis. Our findings strongly suggest that 4-1BB/4-1BBL and Fas/FasL pathways play important roles in vascular injury in Takayasu’s arteritis. We assume that γδ T cells infiltrated aortic tissue recognizing MICA, resulting in the induction of MHC antigens and costimulatory molecules, and then αβ T-cells infiltrated recognizing some auto-antigens presented by MHC antigens, leading to chronic inflammation.
Two-Dimensional IgE-Binding Spectrum of Japanese Cedar (Cryptomeria japonica) Pollen Allergens
Authors: Fujimura Takashi , Shigeta Seiko , Kawamoto Seiji , Aki Tsunehiro , Masubuchi Masanori , Hayashi Takaharu , Yoshizato Katsutoshi , Ono Kazuhisa
Published: January 2004

Background: Japanese cedar (Cryptomeria japonica) pollen is one of the most prevalent sources of the allergens that elicit rhinitis and conjunctivitis. Only Cry j 1 and Cry j 2 have been well characterized as the major allergens of this pollen. Objectives: The aims of this study were to complete the repertoire of C. japonica pollen allergens, to investigate their variability with respect to IgE-reactive patterns and to identify the isoforms of Cry j 1 and Cry j 2 by proteome analysis. Methods: Proteins in C. japonica pollen separated on two-dimensional (2-D) polyacrylamide gel electrophoresis were immunodetected with IgE in sera of 40 subjects allergic to C. japonica pollen. Mass fingerprinting was used to elucidate the diversity of the major allergens. Results: 2-D immunolabeling with individual patients’ sera showed the distinguishable IgE-binding patterns inlaid with 4–87 spots from a total of 131 IgE-binding protein spots. At least 12 Cry j 1 (27.5–75% of IgE-binding frequency) and 3 Cry j 2 (32.5–40%) isoforms were localized. In total, 31 spots were found to be more reactive than the highest IgE-reactive isoform of Cry j 2. Conclusions: The proteomics approaches showed great interindividual variation of IgE-binding patterns to C. japonica proteins and contributed to the repertoire of numerous C. japonica allergens other than Cry j 1 and Cry j 2. Protein microsequencing demonstrated more complicated multiplicity in Cry j 1 than previously known and new isoforms in Cry j 2.
Partial Amino Acid Sequence of a Cellulase-Like Component with IgE-Binding Properties from Stachybotrys chartarum
Authors: Kärkkäinen Marja , Raunio Päivi , Rautiainen Jaakko , Auriola Seppo , Hinke Kaj , Pasanen Anna-Liisa
Published: January 2004

Background: The aim of this study was to characterize the amino acid sequence of a selected Stachybotrys chartarum component and to investigate human IgE reactivity against components of S. chartarum and nine other fungal species. Methods: Human IgE reactivity against S. chartarum and nine other fungal extracts was investigated by the immunoblotting method. For automated amino acid sequencing analyses, the S. chartarum extract was purified by ion exchange chromatography prior to in-gel alkylation and digestion with modified trypsin. Results: Human IgE reactivity was detected against eight components in the S. chartarum extract. Over 80% of the sera from the exposed subjects and less than 50% of the control sera recognized the 33-, 48- and 50-kD S. chartarum components. The human sera detected a 48- to 50-kD component from the extracts of eight fungal species. Nineteen peptide sequences were identified from the 48-kD component of S. chartarum. An analysis of the peptide sequences revealed homology with known fungal glycoside hydrolase enzymes (cellulases). Conclusions: The data showed human IgE reactivity against several S. chartarum components, including one at 48 kD. On the other hand, the human sera recognized 48- to 50-kD components from seven other fungal species, suggesting shared antigenic components (e.g. enolase) between the fungi. Thus, to our knowledge, this is the first antigen identified from S. chartarum.
Antigen-Specific versus Total Immunoglobulin Synthesis: Total IgE and IgG1, but Not IgG2a Levels Predict Murine Antigen-Specific Responses
Authors: Lewkowich Ian P. , Rempel Julia D. , HayGlass Kent T.
Published: January 2004

Background: Induction of an effective antibody (Ab) response requires delivery of multiple signals to B cells. Cross-linking of the B cell antigen receptor (BCR), signaling through CD40 and CD80/86 and cytokine signals combine to induce class switching and expression of specific isotypes. These signals are principally derived from activated, antigen (Ag)-specific T cells. In contrast, IFNγ, the only cytokine known to induce class switch to IgG2a, can be produced systemically by activated NK or NKT cells, suggesting that Ag-nonspecific signals may also regulate IgG2a production. Methods: Given the potential differences in regulation between IgE/IgG1 versus IgG2a, we immunized mice on day 0 with ovalbumin (OVA) in the presence of strong type-1- or type-2-immunity-inducing adjuvants and boosted mice 4 weeks later. Mice were bled during the primary immune response and after boost to assess primary and recall Ab responses. Results: Regardless of strain of mice used, phenotype (type 1 versus type 2 dominated) or nature of the immune response induced (primary versus recall), strong correlations between OVA-specific and total IgE and IgG1 were demonstrated. In contrast, a consistent lack of correlation between OVA-specific and total IgG2a levels was observed in all but BALB/c mice. Conclusion: These data indicate that the increase in total levels of IgE/IgG1 isotypes is primarily a result of increased levels of OVA-specific Ab. In contrast, the lack of correlation between total and OVA-specific IgG2a suggests broader activation of IgG2a-producing B cells routinely occurs following exogenous Ag immunization.
Induction of TNF-α Autoantibody Production by AutoVac TNF106: A Novel Therapeutic Approach for the Treatment of Allergic Diseases
Authors: Zuany-Amorim Claudia , Manlius Corinne , Dalum Iben , Jensen Martin R. , Gautam Anand , Pay Graham , Mouritsen Søren , Walker Christoph
Published: January 2004

Background: Cytokines play an integral role in the coordination and persistence of allergic inflammatory processes and therefore represent prime targets for novel therapies in diseases such as asthma. Multiple attempts to generate low-molecular-weight cytokine inhibitors have failed, and the main attention has focused on the development of biological agents such as neutralizing antibodies. The present work describes a simple and effective method to induce the production of therapeutic anti-cytokine autoantibodies by active immunization against a modified endogenous cytokine. Methods: Balb/c mice were subcutaneously injected with AutoVac TNF106, a recombinant murine TNF-α molecule containing a foreign immunogenic T helper epitope, and the induction of neutralizing anti-TNF-α autoantibodies was analysed. These mice were then sensitized with ovalbumin (OVA), and the effect of neutralizing anti-TNF-α autoantibodies on the allergen-induced airway inflammation was analysed. Results: AutoVac TNF106-immunized mice developed high titres of neutralizing anti-TNF-α autoantibodies, which were maintained for at least 4 weeks after the last booster injection. Mice vaccinated with AutoVac TNF106 and further immunized against OVA showed diminished TNF-α levels in the bronchoalveolar lavage (BAL) fluid after OVA challenge. Moreover, pretreatment with AutoVac TNF106 resulted in significantly reduced numbers of eosinophils and neutrophils in BAL fluid in response to single or multiple allergen exposure. Conclusion: The induction of anti-TNF-α autoantibody production by the AutoVac TNF106 technology not only confirmed the role of TNF-α in the induction of allergic inflammation but also offers a novel approach to block the activity of cytokines in order to treat allergic inflammatory conditions.
Transitive Behavior in Hippocampal-Lesioned Pigeons
Authors: Strasser Rosemary , Ehrlinger Joyce M. , Bingman Verner P.
Published: January 2004

The hippocampus of birds and mammals is critical for the learning of map-like memory representations of environmental space. It has been suggested that the hippocampus of rats also participates in non-spatial relational learning, including the learning of non-spatial transitive relationships among odor stimuli [Bunsey and Eichenbaum, Nature 1996]. Although transitive-like learning has been demonstrated in a variety of vertebrate species, from a comparative perspective the role of the hippocampus in this form of learning has not been tested in other amniote groups. We trained control and hippocampal-lesioned homing pigeons on a series of visual, non-spatial, go/no-go conditional discriminations and then tested them on novel transitivity probe trials. The hippocampal-lesioned pigeons were as successful as control pigeons in responding appropriately to correct and incorrect transitivity pairs. The finding that the homing pigeon hippocampal formation is not necessary for solving this serial, conditional discrimination task is important for further understanding hippocampal function across species, and represents one of the few studies that have attempted to localize a brain region responsible for the phenomenon of transitive behavior learning.