Biomedical subjects

M Mahler

Publications and source records attributed to M Mahler.

At least 19 recordsLinked to original sources

PubMed2005

Improved serological differentiation between systemic lupus erythematosus and mixed connective tissue disease by use of an SmD3 peptide-based immunoassay.

Autoantibodies to the Sm antigens are specifically found in 5 to 30% of patients with systemic lupus erythematosus (SLE) depending on the detection system and the patient group. Several immunoassays designed for research and diagnostic laboratory use have been developed. The autoantigens employed in these tests include purified native proteins, recombinant polypeptides, and synthetic peptides. In the present study, we compared the clinical accuracy of anti-Sm autoantibody assays from commercial suppliers including different conventional enzyme-linked immunosorbent assay (ELISA) systems based on purified Sm antigens, an addressable laser bead assay and a newly developed anti-Sm peptide assay. Although the clinical sensitivity of all assays under investigation was comparable, relatively poor correlations and significant differences in specificity were found with a patient cohort of 150 patients. The sensitivity and specificity were 10 and 94%, respectively, for the anti-Sm ELISA from Euroimmun, 10 and 90%, respectively, for the QuantaLite Sm (INOVA), 12 and 88%, respectively, for the Sm assay in the Varelisa ReCombi ANA profile (Pharmacia Diagnostics), 10 and 94%, respectively, for the QuantaPlex Sm (INOVA), and 12 and 100%, respectively, for the new SmD3 peptide-based ELISA (Varelisa Sm Antibodies). The majority of positive test results within the control groups were found in patients with mixed connective tissue disease. Based on the results, we conclude that the detection of anti-Sm antibodies strongly depends both on the nature of the antigen and on the detection system. Finally, we conclude that the recently identified SmD peptide containing a symmetrical dimethylarginine at position 112 of D3 represents a promising tool for the detection of a highly specific subpopulation of anti-Sm antibodies.

Antibodies, Antinuclear↗

PubMed2005

Environment as a critical factor for the pathogenesis and outcome of gastrointestinal disease: experimental and human inflammatory bowel disease and helicobacter-induced gastritis.

Environmental factors play an important role in the manifestation, course, and prognosis of diseases of the gastrointestinal tract such as inflammatory bowel disease (IBD) and Helicobacter pylori-induced gastritis. These two disease complexes were chosen for a discussion of the contribution of environmental factors to the disease outcome in humans and animal models. Dissecting complex diseases like IBD and Helicobacter-induced gastritis has shown that the outcome of disease depends on the allelic constellation of a host and the microbial and physical environments. Host alleles predisposing to a disease in one genomic and/or environmental milieu may not be deleterious in other constellations; on the other hand, microbes can have different effects in different hosts and under different environmental conditions. The impact of the complex interaction between host genetics and environmental factors, particularly microflora, also underlines the importance of a defined genetic background and defined environments in animal studies and is indicative of the difficulties in analyzing complex diseases in humans.

Animals↗

PubMed2004

Technical and clinical evaluation of anti-ribosomal P protein immunoassays.

Autoantibodies to the three ribosomal phospho (-P) proteins P0, P1, P2, referred to as Rib-P, are specifically found in 10-40% of patients with systemic lupus erythematosus. The variations in the observed frequency of these autoantibodies is related to a number of factors such as the test system used to detect the antibodies. Several immunoassays that were designed for research and diagnostic laboratory use have been developed. The autoantigens employed in these tests include native proteins, recombinant polypeptides, and synthetic peptides. In this study, we compared the technical and clinical accuracy of anti-Rib-P antibody assays from different commercial suppliers including ELISA systems and a novel addressable laser bead assay (from Euroimmun, MBL, Pharmacia Diagnostics, INOVA). Although the assays from all suppliers used in this study performed well in the technical part of the study, relatively poor correlations and significant differences in the clinical accuracy were found. Based on the results, we conclude that the detection of anti-Rib-P antibodies strongly depends on both the nature of the antigen and the detection system. We recommend that anti-Rib-P assays should be standardized on an international level. The Varelisa Rib-P profile and the addressable laser bead Rib-P assays represent promising tools and platforms for the detection of anti-Rib-P antibodies in the future.

Animals↗

PubMed2003

Characterization of the human autoimmune response to the major C-terminal epitope of the ribosomal P proteins.

Autoantibodies to the ribosomal phospho (-P) proteins P0, P1, and P2, collectively referred to as Rib-P, are specifically found in 10-40% of patients with systemic lupus erythematosus (SLE). These antibodies are believed to be correlated with lupus nephritis, hepatitis, and central nervous system involvement. The major immunoreactive epitope of these ribosomal antigens has been localized to the carboxy terminus, which is a highly conserved domain of all three proteins and contains two phosphorylated serine residues. The phosphorylated amino acids of the P proteins are known not to be critical epitope determinants. Furthermore, epitope-mapping studies have shown that the major epitope is located within the last 11 C-terminal amino acids. Using peptide arrays we identified more precisely this shared epitope as the six C-terminal amino acids GFGLFD and elucidated the molecular recognition events of anti-Rib-P antibodies at the amino acid level. We identified Phe(111) and Phe(114) of Rib-P2 as the key residues for the interaction, with further contributions of Gly-112 and Asp-115. This amino acid stretch is also present in proteins of several pathogenic micro-organisms such as Trypanosoma cruzi, Brugia malayi, Pseudomonas aeruginosa, Candida albicans, several Leishmania species, and Bartonella henselae. Using newly developed ELISA systems with a C-terminal peptide (C22) and the recombinant proteins (P0, P1, and P2) as antigens we found a high specificity of anti-Rib-P antibodies for SLE and demonstrated positive correlations with anti-U1-C, anti-Sm-B/B' and anti-D and anti-dsDNA antibodies. The sensitivity and specificity in the peptide (C22) based assay varied between 12.8%/100% and 23.4%/96.7% for SLE, depending on the assigned cutoff. In contrast to other studies, we found no significant correlation of anti-Rib-P reactivity with central nervous system manifestations or renal involvement in SLE patients. We conclude that the epitope motif GFGLFD in the C-termini of the ribosomal P proteins is the key determinant of anti-Rib-P antibodies, and that the C22 peptide and the recombinant proteins can be used equally well for the detection of anti-Rib-P antibodies. The role of the major Rib-P epitope in the development of anti-ribosomal P antibodies and in the pathogenesis of SLE remains a subject of further investigation.

Amino Acid Sequence↗

PubMed2003

Recombinant anti-P protein autoantibodies isolated from a human autoimmune library: reactivity, specificity and epitope recognition.

The ribosomal P proteins are specific and important autoantigens in patients affected by systemic lupus erythematosus. In this study, we describe for the first time the selection and characterization of recombinant human monoclonal anti-P protein (auto)-antibody fragments from an autoimmune patient-derived phage display antibody library. The selected recombinant anti-P antibodies specifically recognize the P proteins in immunofluorescence assays on HEp-2 cells and in immunoblotting assays, and they immunoprecipitate the P proteins under native conditions. Using both anti-P-positive patient sera and the selected recombinant anti-P antibodies, the immunodominant epitope was determined and shown to be located at the C-terminal end of the P proteins (amino acids 111-115). Inhibition of in vitro protein translation demonstrated that interaction of the monoclonal patient-derived anti-P antibodies with their native epitope functionally inhibits the activity of the P proteins on the ribosome, confirming the notion that patient autoantibodies are often directed to the functional centre of their autoantigenic target.

Antibody Specificity↗

PubMed2003

Advances in B-cell epitope analysis of autoantigens in connective tissue diseases.

The characterization of autoantibody specificities in rheumatic diseases is important in both diagnostic and basic research areas. Identification of the epitopes recognized by autoantibodies and their clinical and biological significance is not a trivial task. Epitopes may range in complexity from simple linear sequences of amino acids to complex quaternary structures. In addition to this structural complexity the frequency with which an autoantigen and its epitopes are recognized in a patient population may be useful in diagnosis, defining disease subgroups, and may offer information on disease prognosis. In this review recent advances in the epitope mapping of autoantigens in connective tissue diseases are discussed, with particular emphasis placed on the methodologies used to identify epitopes and the classification of the structural features of epitopes. To illustrate the identification of epitope structure, clinically relevant autoantigens, including CENP-A, PM/Scl-100, fibrillarin, filaggrin, Ro-52, and dsDNA, are discussed as examples of each type of epitope.

Amino Acid Sequence↗

PubMed2001

A population of autoantibodies against a centromere-associated protein A major epitope motif cross-reacts with related cryptic epitopes on other nuclear autoantigens and on the Epstein-Barr nuclear antigen 1.

Autoimmune diseases arise from a host's immune response against self-antigens. The triggering events ultimately resulting in such a break of tolerance are largely unknown. It is also not known why certain molecular structures become autoantigenic. The hypothesis has long been proposed that autoimmune diseases arise from molecular mimicry followed by an epitope spreading mechanism. Recently we have shown that the anti-centromere-associated protein A (CENP-A) immune response is directed against an autoantigenic motif, G/A-P-R/S-R-R, that occurs three times in the N-terminal amino acids of CENP-A. In the present study we used mutational analyses with immobilized oligopeptide arrays to identify the amino acids in this motif that are responsible for antibody binding. In particular, we found that surprisingly mimotopes of this motif are present in a vast number of autoantigens and in the Epstein-Barr nuclear antigen 1. With affinity-purified antibodies we show that the antibodies against this motif are polyclonal and cross-react with several autoantigens. However, in these autoantigens this motif often represents a cryptic epitope explaining the obvious conflict between our results and the known high specificity of autoantibodies. The presence of such an ubiquitous structure on autoantigens suggests a novel peptide-driven mechanism for the evolution of autoantibodies.

Amino Acid Motifs↗

PubMed2001

Dy-EOB-DTPA: tolerance and pharmacokinetics in healthy volunteers and preliminary liver imaging in patients.

RATIONALE AND OBJECTIVES: To investigate the tolerance and pharmacokinetics of the new liver-specific x-ray contrast agent Dy-EOB-DTPA [(4S)-4-(4-ethoxybenzyl)-3,6,9-tris(carboxylatomethyl)-3,6,9-triazaundecanedioic acid, dysprosium (Dy) complex, disodium salt] in healthy volunteers and to obtain preliminary imaging data by abdominal spiral computed tomography (CT) in tumor patients with liver metastases. METHODS: A total of 40 healthy male volunteers received 10-minute intravenous infusions of 0.05, 0.1, 0.25, 0.375, or 0.5 mmol/kg Dy-EOB-DTPA (n = 6 per dose group) or placebo (n = 10). Blood, urine, and feces were sampled for Dy measurements by inductively coupled plasma atomic emission spectrometry (ICP-AES) and for the detection of possible metabolites by high-performance liquid chromatography analysis with ICP-AES detection. Safety parameters were determined before, during, and after the study. Two patients with suspected liver metastases first received 120 mL of iopromide (300 mg iodine/mL; approximately 0.6 mmol/kg) and, 24 or 72 hours later, Dy-EOB-DTPA at a dose of 0.25 mmol/kg. Computed tomography images were obtained 50 seconds after iopromide administration and before and 90 minutes after Dy-EOB-DTPA administration. RESULTS: Dysprosium-EOB-DTPA was well tolerated. At the higher doses (0.375 and 0.5 mmol/kg), there was a slight increase in side effect intensity. In general, nausea, headache, and paresthesia mainly were reported as mild to moderate adverse events. Laboratory parameters did not exceed the normal range. Electrocardiographic, vital sign, or hemodynamic parameters were not affected by contrast agent administration. The terminal half-life of elimination of Dy-EOB-DTPA was approximately 1.5 hours, total clearance was 2 to 3 mL x min(-1) x kg(-1), and the renal clearance was approximately 1.5 mL x min(-1) x kg(-1). There was a significant dose dependence for the following parameters: maximal concentration in blood, terminal half-life, mean residence time, total clearance, urinary excretion, and fecal excretion. The volume of distribution in the steady state and renal clearance were not dependent on dose. In the blood and urine, no metabolites of Dy-EOB-DTPA could be detected. In the tumor patients, CT scanning after Dy-EOB-DTPA injection increased the number of detected metastases from 27 (plain scan) to 40 (iopromide) and then to 41 (Dy-EOB-DTPA) in patient No. 1 and from 1 (plain scan and iopromide) to 3 (Dy-EOB-DTPA) in patient No. 2. CONCLUSIONS: Dysprosium-EOB-DTPA was shown to be a well-tolerated liver-specific contrast agent. Its pharmacokinetic profile is characterized by a terminal half-life of approximately 1.5 hours. There are indications of saturation of liver uptake at the highest dose level of 0.5 mmol/kg. In comparison with plain scans and scans performed after iodinated contrast agent administration, Dy-EOB-DTPA seems to increase the number of detectable liver lesions.

Adult↗

PubMed2000

Fine-specificity of the anti-CENP-A B-cell autoimmune response.

The major targets recognized by anti-centromere autoantibodies are the three centromere-associated proteins (CENPs) A, B, and C, with apparent molecular masses of 19, 80, and 140 kDa, respectively. Previously a major epitope region on the 19-kDa CENP-A antigen was identified by synthesis of a soluble synthetic 15-mer peptide (amino acids 3-17) to be used in enzyme-linked immunosorbent assay and western blot competition assays. However, no systematic experimental scanning for epitope regions on the CENP-A autoantigen has yet been performed. In this study we scanned the complete CENP-A amino acid sequence for epitopes using 19 previously characterized autoimmune-sera. Overlapping peptides 15 amino acids in length and offset by three amino acids were synthesized on activated membranes, covering the whole CENP-A autoantigen. Probing of the membranes with various anti-centromere sera showed that all epitopes are clustered in the N-terminal 45 amino acids. For fine-mapping of this autoreactive region the N-terminus of CENP-A (amino acids 1-45) was scanned again by probing overlapping 15-mer, 12-mer, 10-mer, 8-mer, 7-mer, 6-mer, and 5-mer peptides, all offset by one amino acid, with anti-centromere sera. In this way we localized two epitope core regions within the N-terminal 45 amino acids, one covering amino acids 2-17, recognized by 17 sera, and the other covering amino acids 22-38, recognized by 18 sera. One serum did not react with CENP-A at all. Several sera seem to recognize overlapping individual epitopes within these two epitope core regions. All sera, however, recognize a sequence motif G/A-P-R/S-R-R.

Adult↗

PubMed2000

Identification of an alpha-helical epitope region on the PM/Scl-100 autoantigen with structural homology to a region on the heterochromatin p25beta autoantigen using immobilized overlapping synthetic peptides.

The polymyositis-scleroderma overlap syndrome (PM/Scl) autoantigen is a nucleolar multiprotein particle, presumably participating in the maturation of 5.8S rRNAs. The major target antigens of this particle are two polypeptides with apparent molecular masses of 100 and 75 kDa. In this study we identified the major linear epitopes along the PM/Scl-100 protein sequence by probing overlapping oligopeptides with anti-PM/Scl autoantisera. A major epitope region was identified between amino acids 231 and 245 of the PM/Scl-100 polypeptide. Mutational analysis of the corresponding peptide LDVPPALADFIHQQR by glycine-walk followed by immunodetection of the resulting peptides indicated that amino acids 234, 237, 240, and 241 of the PM/Scl-100 autoantigen are essential for binding of the corresponding antibodies. These results allow us to propose a local alpha-helical secondary structure for the PM/Scl-100 major epitope region. A homology search with the peptide LDVPPALADFIHQQR against the Swiss-Model three-dimensional database reveals some topological homology of the PM/Scl-100 major epitope region with the heterochromatin modifier protein p25beta, a known autoantigen recognized by antibodies from a subset of scleroderma patients.

Amino Acid Sequence↗

PubMed2000

[Comparison of bolus and infusion of the ultrasound contrast media levovist for color doppler ultrasound of renal arteries].

PURPOSE: To investigate if the duration of Doppler enhancement of the ultrasound contrast agent Levovist can be prolonged by continuous infusion compared to bolus injections in patients. METHODS: 12 patients with suspected renal artery stenosis were included. Each patient received 1 or 2 bolus injections of 4 g each and a "fast" infusion of 3 ml/min (average dose: 7 g) of Levovist. In 8 patients an additional "slow" infusion of 1 ml/min (average dose 4.2 g) was given. The duration of "strong" Doppler signal enhancement and the visualization of vessels were compared. RESULTS: The duration of strong enhancement was substantially prolonged by the slow infusion (bolus 6:21 min, fast infusion: 8:57 min, slow infusion: 15:12 min, p < 0.001). The dose effectiveness (duration of strong enhancement per 1 g Levovist) was markedly improved from 0:57 min (bolus) to 3:36 min (slow infusion). Visualisation of the renal arteries was more complete with the slow infusion. CONCLUSION: Levovist infusions at 1 ml/min are superior to bolus injections in the assessment of renal arteries. They prolong the enhancement, make more efficient use of a given dose of contrast material and thus allow additional clinically relevant information to be obtained at reduced cost.

Adult↗

PubMed1999

Screening for dementia: comparison of three commonly used instruments.

The sensitivity and specificity of three cognitive screening measures - the Mini-Mental State Exam (MMSE), Mattis Dementia Rating Scale (MDRS), and Neurobehavioral Cognitive Status Examination (NCSE) - were compared in a cohort of subjects with dementia as well as normal elderly individuals. Twenty-two patients met criteria for probable Alzheimer' s disease (AD), 19 for vascular dementia (VaD), and 12 were normal control subjects. The use of standard cutpoints resulted in poor to good classification accuracy for the three measures, but measurable improvement in sensitivity was obtained by adjusting the cutpoints for each measure. Discriminatory power was maximized with an MMSE cutpoint of < or = 26, an MDRS cutpoint of < or = 134, and requiring one or more NCSE subtests to be in the impaired range. Use of age and education adjusted norms resulted in good to excellent classification accuracy for the MMSE and MDRS. The NCSE subtest score pattern failed to differentiate between AD and VaD.

Aged↗

PubMed1998

Prolongation and optimization of Doppler enhancement with a microbubble US contrast agent by using continuous infusion: preliminary experience.

PURPOSE: To investigate whether continuous infusion of an echo-enhancing contrast agent for up to 15 minutes can provide uniform and prolonged enhancement. MATERIALS AND METHODS: Six volunteers each received one bolus and three infusions of a microbubble contrast agent over 6-15 minutes at (a) a standard rate (mean, 2.08 mL/min), (b) a fast rate at twice the standard rate, and (c) a slow rate at half the standard rate. Spectral Doppler intensitometry of the femoral artery was performed for all infusions. Spectral Doppler ultrasound (US) scans of the femoral artery and color Doppler US scans of the carotid artery were subjectively assessed. RESULTS: All infusions provided an equilibrium plateau of constant prolonged enhancement starting after 1-2 minutes and lasting until the end of the infusion. Enhancement at the plateau was +13 dB (slow rate), +17.1 dB (standard rate), and +18.3 dB (fast rate) compared with baseline. Saturation artifacts with infusions were markedly fewer than those with bolus injections. Dose effectiveness (duration of enhancement that measured at least 7.5 dB per gram of contrast agent) was markedly improved with the infusions, from 0.8 min/g for the bolus to 2.6 min/g for the slow infusion. CONCLUSION: Continuous infusion of the microbubble contrast agent provided prolonged and uniform enhancement of Doppler signals and improved image quality by minimizing saturation artifacts.

Adult↗

PubMed1997

Differential expression of fibronectin, tenascin-C and NCAMs in cultured hippocampal astrocytes activated by kainate, bacterial lipopolysaccharide or basic fibroblast growth factor.

Different reports demonstrated that reactive glial cells express increased amounts of adhesion and matrix molecules. Despite a wealth of information on the expression of these molecules during development and after lesion, very little is known of how this expression is regulated. In the present report we used Western blots and immunocytochemistry to investigate the expression of neural cell adhesion molecule (NCAM), fibronectin and tenascin-C in cultured astrocytes from rat hippocampus. The effects of three different extracellular signals were analyzed: the glutamatergic receptor agonist kainic acid, the basic fibroblast growth factor (bFGF) and the bacterial lipopolysaccharide. Each treatment had a specific pattern of glial activation and differentially modified the expression of these proteins. Treatment of astrocytes with kainic acid resulted in an increase of tenascin-C, a decrease of fibronectin and a shift of NCAMs isoforms: NCAM 140 and PSA-NCAM (polysialic acid-rich NCAMs) were increased while NCAM 120 was decreased, bFGF increased fibronectin, tenascin-C and NCAM 120, while decreasing PSA-NCAM. Finally, the treatment of astrocytes with lipopolysaccharide induced a significant increase of fibronectin, tenascin-C and NCAM 120 but did not modify the expression of NCAM 140 and PSA-NCAM. These data suggest different mechanisms for modulation of cell surface interactions. They suggest that glial activation by bFGF and lipopolysaccharide are associated with an increase of the adhesive properties, while kainate action is rather associated with a decrease of the adhesiveness of astrocytes.

Animals↗

PubMed1997

Effects of tenascin-C in cultured hippocampal astrocytes: NCAM and fibronectin immunoreactivity changes.

Tenascin-C is an extracellular matrix glycoprotein with trophic and repulsive properties on neuronal cells, involved in migratory processes of immature neurons. Previous reports demonstrated that this molecule is produced and secreted by astrocytes, in vitro after activation by bFGF or in vivo after CNS lesion. In injured brain the expression of tenascin-C has been correlated with the glial reaction since it was observed in regions suffering a dramatic glial proliferation and hypertrophy. In this report we show that the treatment of cultured hippocampal astrocytes with tenascin-C results in an increased fibronectin and NCAM immunoreactivities. In addition, treated astrocytes form longer extensions than control ones. The number of cells as well as the levels of GFAP mRNA and protein immunoreactivity are not modified after tenascin-C treatment. The present changes may, therefore, be related to the modification of the adhesive properties of astrocytes to the substrate. These observations are compatible with the hypothesis that tenascin-C may contribute to the glial scarring process.

Animals↗

PubMed1997

Pharmacokinetics of the liver-specific contrast agent Gd-EOB-DTPA in relation to contrast-enhanced liver imaging in humans.

This study was performed to evaluate the effect of dose on the pharmacokinetics and efficacy of the gadolinium-based contrast medium gadoxetic acid, disodium, [gadolinium (4S)-4-(4-ethoxybenzyl)-3,6,9-tris(carboxylatomethyl)-3,6, 9-triazaundecandioic acid-disodium salt] (Gd-EOB-DTPA) as a liver-specific hepatobiliary contrast medium for computed tomography. Pharmacokinetics in serum and the pattern of elimination were investigated in 18 healthy volunteers up to 6 days after a 10-minute infusion of 0.2 mmol, 0.35 mmol, and 0.5 mmol of gadolinium per kilogram of body weight. Pharmacokinetic behavior was compared with the compute tomographic attenuation data in the liver parenchyma after the same doses in patients. Urinary and fecal excretion accounted for approximately equal portions of the administered dose. The degree of renal elimination increased with increasing doses, whereas renal clearance and half-life from urine data were not affected by dose. Dose-normalized area under the concentration-time curve was significantly increased with increasing doses indicating saturation in liver uptake for the highest dose. This finding was in agreement with the measured net increase in liver attenuation by computed tomography. Hepatic disposition revealed slight saturation phenomena for the highest dose (0.5 mmol gadolinium/kg). Nevertheless, this dose resulted in sufficient uptake by human liver, allowing for computed tomographic imaging.

Adult↗

PubMed1997

Adverse events and discomfort in studies on healthy subjects: the volunteer's perspective. A survey conducted by the German Association for Applied Human Pharmacology.

OBJECTIVE: The various good clinical practice (GCP) guidelines do not define the volunteering subject as an active party. The present survey addresses the volunteer's perception of study-related inconvenience and risk and its impact on their decision to enroll. METHODS: The survey consisted of a questionnaire to be filled out voluntarily and anonymously by healthy subjects who volunteered for enrollment in human pharmacology studies and who had participated in at least one previous study. Twenty-five categorised multiple-choice questions covered previous study experience, motives for volunteering, perception of and compliance with study directives and restrictions, past experience with adverse events, impact of the study environment on perceived well-being and the nature of adverse events likely to discourage them from enrollment. RESULTS: Seven centres contributed by providing at least 30 (range 30-100) evaluable questionnaires. The database consists of a total of 440 healthy subjects (30.5% females, 69.5% males), from 18 to over 60 years of age. Two hundred and seven subjects (47.1%) were company employees and 233 (52.9%) were external volunteers. Eighty nine percent only participated in studies at one particular centre. Some 53.3% indicated financial motives, 27.8% 'contribution to an improvement of pharmacotherapy', 12.7% 'social responsibility', while 6.2% indicated other motives, mainly the opportunity of a free medical check-up. Thirteen subjects (3%) admitted to not answering correctly to the recruitment questions; this limited reliability is suspected to be even larger when the answer might preclude enrollment. From the volunteers' perspective, the environmental study conditions clearly appeared to have a highly relevant impact on their personal well-being. Some 17.1% of the subjects reported to have suffered adverse events occasionally and 2.7% frequently; but 14% admitted not reporting adverse events promptly and about 20% indicated that, with respect to previous adverse events, they first sought advice from other volunteers rather than from the investigator. CONCLUSIONS: Adverse events and inconveniences are inherent to nontherapeutic studies in healthy subjects. From the volunteer's perspective it appears that the incidence of adverse experiences in such studies exceeds the reported frequencies from investigators considerably. This finding suggests that investigators are usually not aware or able to ascertain the true incidence of adverse events. The present survey also confirms that pertinent information on the personal history may be unreliable. Volunteers are reluctant to answer questions regarding, in particular, their smoking habits, caffeine and alcohol consumption. Regarding the matter of informed consent, a noteworthy contradiction between the volunteers' attitude and behaviour became apparent. Although the volunteers admit that even rather minor adverse events ordinarily would discourage them, they still consent to enrollment. In view of this apparent contradiction, there is no alternative to the investigator's personal responsibility to counsel and protect the subject. Surveys such as this one may contribute to the awareness that the explicitness of GCP guidelines merely define the format, but not the content quality of these fundamental ethical values, which remain the unique burden and challenge of the investigator.

Adolescent↗

PubMed1996

Tenascin-C mRNA and tenascin-C protein immunoreactivity increase in astrocytes after activation by bFGF.

Tenascin-C is an extracellular matrix glycoprotein with trophic and repulsive properties, involved in migratory processes in CNS. Previous reports demonstrated that this molecule is produced and secreted by astrocytes. Preliminary data on fibroblasts and astrocytes have suggested that bFGF may modulate tenascin-C expression. bFGF is a mitogenic growth factor, involved in cell differentiation and neovascularization. In the present study, we examined whether bFGF modulates the expression of tenascin-C in hippocampal astrocytes from newborn rats. Our results suggest that bFGF increases the production of tenascin-C by cultured hippocampal astrocytes. We found that both tenascin-C mRNA and protein immunoreactivity were increased after bFGF treatment. Our results also demonstrated that tenascin-C polypeptides were secreted into the extracellular medium. In agreement with previous studies, we suggest that secreted tenascin-C is mainly the high molecular weight form. In addition, our results suggest that a cleavage of the high molecular weight form. In addition, our results suggest that a cleavage of the high molecular weight form may occur in the extracellular medium causing production of proteolytic fragments, that may modify the biological properties of tenascin-C. The present results may be relevant to the understanding of lesion scarring and regeneration process.

Animals↗