Biomedical subjects

S Huber

Publications and source records attributed to S Huber.

At least 19 recordsLinked to original sources

PubMed1998

Human myelin basic protein specific T cell lines display differential cytotoxicity against astrocytes, but are consistently cytotoxic against monocytes.

In experimental allergic encephalomyelitis (EAE) myelin basic protein (MBP) specific T cells differ in their encephalitogenic potential. To investigate the functional diversity of human MBP specific T cell lines, we analysed their cytotoxic activity against human astrocytes and monocytes. Five out of 14 MBP specific T cell lines killed astrocytes in the presence of MBP. Nevertheless, all lines lysed blood derived monocytes. T cell lines that lysed astrocytes efficiently in the presence of MBP, recognized peptide aa 80-99/86-105 in context with HLA-DRB5 * 0101, peptide aa 50-69/61-83 in context with HLA-DRB1 * 1501 and peptides aa 139-153, and aa 148-162 in context with HLA-DRB1 * 0101. There was no correlation of MBP-mediated lysis of astrocytes with TCR-Vbeta usage, HLA-restriction and the production of tumor-necrosis-factor-alpha (TNF-alpha), lymphotoxin (LT) and interferon-gamma (IFN-gamma). Different lysis of astrocytes, however, revealed a functional heterogeneity of MBP specific T cells, which was not observed by using monocytes as targets.

Astrocytes↗

PubMed1998

CFTR mRNA and its truncated splice variant (TRN-CFTR) are differentially expressed during collecting duct ontogeny.

The collecting duct epithelium originates from the embryonic ureter by branching morphogenesis. Ontogeny-dependent changes of CFTR mRNA expression were assessed by quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) in primary monolayer cultures of rat ureteric buds (UB) and cortical collecting ducts, microdissected at different embryonic and postnatal developmental stages. The amount of wild-type CFTR-specific PCR product in UB declined to 20% of the initial value between embryonic gestational day E15 and postnatal day P1. After birth the CFTR product increased transiently between P1 and P7 by a factor of 10 and decreased towards day P14. PCR products specific for TRN-CFTR, a truncated splice variant, however, were low in early embryonic cells, increased markedly between day E17 and P2, and reached a plateau postnatally. Therefore, mRNA encoding TRN-CFTR does not appear to have a specific embryonic-morphogenetic function. By contrast, such function is suggested for wild-type CFTR mRNA as its abundance was high in early embryonic nephrogenesis, as well as during a postnatal period shortly before branching morphogenesis is completed.

Alternative Splicing↗

PubMed1998

[A multicenter study of diagnostic criteria in ultrasound mammography. Statistical pitfalls and ways out of the data jungle].

This paper demonstrates the statistical processing of data from a multicentric study. Seventy-seven patients with a suspicious, solid breast mass were included in a two-center study, using clinical breast examination, mammography, and ultrasound. The assessment was formalized using a uniform evaluation sheet. The prospective results of the examinations were compared with the histologic report. The degree of interdependence between an examiner's diagnosis and diagnostic criteria used was assessed using the kappa statistics. We found that despite standardized assessment, the criteria used for differential diagnosis were at least partially different. The criteria valid for diagnosis at both centers were whether a mass fitted well into the surrounding tissue or led to destruction or architectural distortion, whether it was fixed to the surrounding tissues, as assessed by sonopalpation maneuvers, and whether it borders were sharp or unsharp.

Artifacts↗

PubMed1998

[Dynamic assessment of contrast medium enhancement in Doppler ultrasound imaging. Current status].

Findings in previous investigations have indicated that contrast-enhanced colour Doppler can be used to improve the demonstration of characteristic features of neovascularization in malignancies. However, the quantification of the increase in colour Doppler signals after Levovist in the cited study relied only on descriptive criteria defined by the investigator, resulting in a subjective scoring system. A computer-assisted method for quantitative analysis of colour pixel density is a novel approach for objective assessment of colour Doppler signal after contrast media administration. After microbubble contrast medium injection in 47 patients, carcinomas and benign lesions behaved differently regarding degree, onset and duration of Doppler enhancement. However, the high interindividual variability and the temporal variations of the Doppler signal still limit the value of these criteria for prospective diagnosis. Rapid image-acquisition techniques are needed in order to apply the necessary mathematical processing before the contrast kinetics can be used for diagnostic purposes.

Adult↗

PubMed1998

Colour Doppler sonography in breast tumours: an update.

The aim of our study was to reassess the diagnostic performance of image-based, computer-assisted colour Doppler quantification under routine conditions. We used a computer-assisted protocol that quantitatively characterises a colour Doppler image by the relative amount of colour pixels (colour pixel density, CPD) and the colour hues (numerically expressed by the mean colour value, MCV) in 44 patients with breast carcinoma and 49 patients with benign breast lesions. Studies were carried out over two periods by two examiners, subsequently in charge of breast ultrasound. During the first period, the sensitivity of the MCV was 92 %, the specificity 75 %; the sensitivity of the CPD was 80 %, the specificity 81 %. During the second period, the sensitivity of the MCV was 58 %, the specificity 77 %; the sensitivity of the CPD was 68 %, the specificity 71 %. Despite measures to create uniform examination conditions, the diagnostic performance of this method may decline under routine conditions.

Adult↗

PubMed1998

Analysis of mouse glomerular podocyte mRNA by single-cell reverse transcription-polymerase chain reaction.

Selective investigation of glomerular podocytes is not possible using conventional methods in vivo. Analysis of glomerular epithelium-derived cells in culture yields dubious results because of the rapid dedifferentiation of podocytes. We developed a modification of the polymerase chain reaction (PCR) method previously used to analyze cultured neurons. Podocytes harvested from freshly dissected glomeruli are ideal target cells for this modified, single cell reverse transcription-PCR method to reproducibly identify specific mRNA species from resident intact podocytes in vivo.

Actins↗

PubMed1998

Expression of glucose transporters in human peritoneal mesothelial cells.

Glucose containing solutions, the basis of peritoneal dialysis fluids, affect the proliferation and regeneration of peritoneal mesothelial cells (MsC). The aim of this study was to examine mechanisms of glucose transport into MsC, that is, the expression of facilitative glucose transporters (GLUT) and the Na(+)-dependent glucose transporter (SGLT1) in human primary MsC and a transfected MsC line. Since expression of both transporters is differentiation dependent, we investigated the effects of cell differentiation induced by culturing MsC on membranes or by addition of hexamethylene bisacetamide (HMBA; 6 mM), which enhances SGLT1 expression in LLC-PK1 cells. Levels of mRNA for GLUT1 through GLUT4 and SGLT1 were evaluated by reverse transcriptase-polymerase chain reaction (RT-PCR). The presence of the corresponding proteins was examined by Western blotting and localized by immunofluorescence. Active, Na(+)-dependent glucose transport was assessed by alpha-methyl-D-[14C]glucopyranoside (AMG) with and without the SGLT1-specific inhibitor phlorizin and by patch clamp experiments in NaCl or choline-chloride, For Na(+) dependent glucose uptake choline chloride instead of NaCl served as negative control. Facilitative transport was assessed using 2-fluoro-2-deoxy-[14C]-D-glucose (FDG) with and without the inhibitors cytochalasin B or phloretin. Primary and transfected MsC express GLUT1 and GLUT3 mRNA while no transcripts were found for GLUT2 and GLUT4. No SGLT1 transcript was detectable in subconfluent cells. Semiquantitative RT-PCR analysis documented that the addition of the differentiation inducer HMBA to confluent cultures or growth of MsC on membranes for seven days produced a down-regulation of mRNA for GLUT1, no change for GLUT3, and a substantial increase for SGLT1 mRNA. Under these conditions MsC express SGLT1 protein and possess a Na(+)-dependent glucose uptake as assessed by AMG. Phlorizin (1 mM) inhibits AMG uptake by 30 to 40%. In patch clamp experiments the addition of extracellular glucose depolarized the membrane potential only in the presence of sodium. These results indicate that differentiated MsC express GLUT1, GLUT3, and SGLT1. Further characterization of these transport mechanisms and their regulation may help to understand the cellular effects of glucose on MsC in peritoneal dialysis.

Base Sequence↗

PubMed1998

Computer assisted retraining of attentional impairments in patients with multiple sclerosis.

OBJECTIVE: To evaluate the efficacy of a computer based retraining of specific impairments of four different attentional domains in patients with multiple sclerosis. METHODS: Twenty two outpatients with multiple sclerosis received consecutively a specific training comprising 12 sessions in each of the two most impaired attention functions. The baseline of attentional deficits, the performance after each training period, and the course of performance in the next nine weeks was assessed by a computerised attention test battery. Additionally, the impact of the training on daily functioning was evaluated with a self rating inventory. RESULTS: Subgroups of patients with multiple sclerosis showing different patterns of attentional impairment could be separated. Significant improvements of performance could almost exclusively be achieved by the specific training programmes. The increase of performance remained stable for at least nine weeks. For quality of life patients reported less attention related problems in everyday situations. CONCLUSIONS: In patients with multiple sclerosis it seems worthwhile to assess attentional functions in detail and to train specific attention impairments selectively.

Activities of Daily Living↗

PubMed1998

Effects of a microbubble contrast agent on breast tumors: computer-assisted quantitative assessment with color Doppler US--early experience.

PURPOSE: To evaluate objectively the effects of a microbubble contrast agent on the color Doppler ultrasound (US) examination of breast lesions. MATERIALS AND METHODS: Forty-seven patients aged 23-71 years underwent color Doppler US before and after intravenous injection of a microbubble contrast agent. A 3-minute computer-assisted assessment of the color pixel density (CPD) was used to evaluate objectively the increase in the number of color Doppler US signals, the transit time of the microbubble bolus, and the potential additional differential diagnostic information. RESULTS: Peak CPD at contrast agent-enhanced color Doppler US was 14.3% +/- 8.1 (mean +/- 1 standard deviation) for carcinomas and 9.3% +/- 4.9 for benign lesions (P = .04). The time to peak enhancement was shorter in carcinomas (38 seconds +/- 20) than in benign tumors (71 seconds +/- 48, P = .02). Final CPD was close or equal to baseline values. With the median of 13% for peak CPD as a threshold, the sensitivity for this parameter was 55%, the specificity was 79%, and the accuracy was 62% (P = .04). For a median time to peak of 50 seconds, the sensitivity was 84%, the specificity was 57%, and the accuracy was 76%. CONCLUSION: After microbubble contrast agent injection, carcinomas and benign lesions behave differently in degree, onset, and duration of Doppler US enhancement. High interindividual variability and temporal variations in the Doppler US signal still limit the value of these criteria for prospective diagnosis.

Adult↗

PubMed1998

Single cell RT-PCR analysis of ClC-2 mRNA expression in ureteric bud tip.

Embryonic epithelia at the tip of the ureteric bud (UB) face the interspace between epithelial and mesenchymal cells and are fundamentally involved in reciprocal signaling during early nephrogenesis. To characterize their membrane conductive proteins, patch-clamp and single cell RT-PCR techniques were applied to embryonic rat UBs [embryonic day 17 (day E17)] microdissected from the outer cortex. Cells at the UB tip had a high whole cell conductance (14 +/- 2 nS/10 pF, n = 8). The main fractional conductance resembled that of Ca-activated Cl channels in nonepithelial cells, with its time-dependent activation at depolarizing and inactivation at hyperpolarizing voltages. A second Cl-selective current fraction, by contrast, activated slowly during strong hyperpolarization, suggestive of a ClC-2-mediated conductance. To determine the origin of this current, cytoplasm was harvested into the patch pipette, RNA was reverse transcribed, and cDNA encoding the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) housekeeper gene or the ClC-2 Cl channel was amplified by polymerase chain reaction (PCR). GAPDH and ClC-2 PCR products were identified in 23 and 8 (out of a total of 57) single cell cDNA samples, respectively. ClC-2 PCR products with two different lengths were obtained, which might be due to two alternatively spliced ClC-2 mRNA isoforms. This first and combined approach by patch-clamp and single cell RT-PCR techniques to embryonic epithelia indicates that 1) cells at the UB tip express a phenotype remarkably different from that of postembryonic collecting duct principal cells and that 2) ClC-2 is likely to have a key role in early nephrogenesis.

Animals↗

PubMed1998

Activated protein C resistance assay performance: improvement by sample dilution with factor V-deficient plasma.

OBJECTIVE: To evaluate a modification of a commercially available reagent kit (COATEST APC Resistance Kit) for functional activated protein C (APC) resistance testing, and to determine the ability of the modified assay to demonstrate APC resistance in patients receiving warfarin. DESIGN: Functional APC resistance testing was performed using both the first-generation COATEST APC Resistance Kit and a modified, or second-generation, version of the COATEST assay that uses predilution of the patient sample with factor V-deficient plasma. Factor V genotyping for APC resistance (FV R506Q) was performed using a well-characterized polymerase chain reaction-restriction fragment length polymorphism method. SETTING: University medical center. PATIENTS: Seventy-three individuals referred for hypercoagulability testing who were not receiving warfarin therapy and 29 patients with a history of venous thrombosis who were receiving warfarin therapy. MAIN OUTCOME MEASURE: Sensitivity and specificity as determined by comparing functional APC resistance to the FV R506Q genotype. RESULTS: In 73 patients referred for hypercoagulability testing, but not receiving warfarin therapy, a sensitivity of 0.86 and a specificity of 0.75 were obtained with the first-generation COATEST assay. In contrast, a sensitivity and specificity of 1.0 were obtained when the second-generation COATEST assay was employed. In 29 patients receiving warfarin, the first-generation assay exhibited a sensitivity and specificity of 0.88 and 0.95, respectively, whereas the sensitivity and specificity for the second-generation assay was 1.0. CONCLUSIONS: Predilution of patient plasma with factor V-deficient plasma results in improved sensitivity and specificity of the COATEST APC Resistance Kit, thus offering a simple modification to enhance APC resistance determination in the routine clinical laboratory setting.

Blood Coagulation Disorders↗

PubMed1997

Reciprocal expression of myelin-associated glycoprotein splice variants in the adult human peripheral and central nervous systems.

The L- and S-MAG isoforms differ only at their C-terminus and are believed to be functionally distinct. To obtain information on the relative expression of these alternatively spliced isoforms in humans, we cloned an S-MAG cDNA fragment. The deduced amino-acid sequence of the human S-MAG C-terminus shows fairly conservative substitutions of 4 out of the 10 residues compared to the rodent peptide. Using reverse transcription and a competitive polymerase chain reaction, we show that, in contrast to rodents, the L-MAG splice variant predominates in adult human brain while, like in rodents, S-MAG transcripts are most abundant in peripheral nerve. The results obtained by Western blot analysis and immunohistochemistry are in good agreement with the findings at the mRNA level. Animal experiments may thus be more representative for the role of MAG in human nerve than in brain.

Adult↗

PubMed1997

[Diagnosis and differential diagnosis in multiple sclerosis].

The diagnosis of multiple sclerosis (MS) relies on the occurrence of characteristic symptoms, on the patient's history and on the correct interpretation of nowadays very sensitive but not very specific auxillary examinations. This paper reviews the diagnostic criteria and typical signs and symptoms of the disease. The significance of cerebrospinal fluid (CSF) analysis, magnetic resonance imaging (MRI) and evoked potentials is discussed, with emphasis on their predictive value concerning the development of MS after a first episode with symptoms suggestive of MS. A wide range of other diseases mimicking MS, like infectious, autoimmune, granulomatous, metabolic and hereditary diseases is reviewed. A checklist for important points in patient history and further investigations is proposed.

Adult↗

PubMed1997

Epithelial nephrogenesis.

The metanephric, or definitive, kidney forms as a result of inductive processes between tissues of two distinct embryologic origins, the metanephric mesenchymal blastema and the ureteric bud. After inductive signalling between these primordial tissues, mesenchymal cells aggregate next to the branching ureteric bud tip, convert to epithelial cells and differentiate into the diverse cell populations of the nephron. The ureteric bud enters branching morphogenesis and gives rise to the collecting duct system. Nephrogenesis has become a target system by which to study developmental processes including embryonic inductive interactions, mesenchyme-to-epithelium conversion, cell lineage pathways, epithelial cell polarization, branching morphogenesis and spatio-temporal expression of transcription factors. This review summarizes data on cellular and extracellular events during epithelial metanephrogenesis.

Animals↗

PubMed1997

Reduced glucose metabolism in the frontal cortex and basal ganglia of multiple sclerosis patients with fatigue: a 18F-fluorodeoxyglucose positron emission tomography study.

To investigate the pathophysiology of fatigue in MS, we assessed cerebral glucose metabolism (CMR-Glu) in 47 MS patients using PET and 18F-fluorodeoxyglucose. Applying the Fatigue Severity Scale (FSS), we first compared MS patients with severe fatigue (MS-FAT, n = 19, FSS > 4.9) and MS patients without fatigue (MS-NOF, n = 16, FSS < 3.7) on a pixel-by-pixel basis using Statistical Parametric Mapping (SPM95). Second, we compared FSS values of all 47 patients covering the whole range of this scale with CMRGlu using an analysis of covariance (SPM95). In addition, we determined global CMRGlu by region-of-interest analysis. Sixteen healthy subjects served as control subjects (CON). Global CMRGlu was significantly lower in both MS groups compared with CON (CON 43.3 +/- 6.9 mumol/100 mL/min, MS-FAT 34.7 +/- 4.4, MS-NOF 35.4 +/- 4.5) but was not related to fatigue severity. Comparing the two MS groups, SPM95 analysis revealed predominant CMRGlu reductions bilaterally in a prefrontal area involving the lateral and medial prefrontal cortex and adjacent white matter, in the premotor cortex, putamen, and in the right supplementary motor area of MS-FAT. In addition, there were CMRGlu reductions in the white matter extending from the rostral putamen toward the lateral head of the caudate nucleus. FSS values were inversely related to CMRGlu in the right prefrontal cortex. CMRGlu in the cerebellar vermis and anterior cingulate was relatively higher in MS-FAT than in MS-NOF patients. CMRGlu of both regions showed positive correlations with FSS values. Our data suggest that fatigue in MS is associated with frontal cortex and basal ganglia dysfunction that could result from demyelination of the frontal white matter.

Adult↗

PubMed1996

[Multiple sclerosis: therapy with recombinant beta-1b interferon: initial results with 30 multiple sclerosis patients in northwest Switzerland].

Recombinant interferon beta-1b has been registered with the Swiss health authorities since August 1995. Due to a special arrangement with health insurances it has been possible to prescribe this medication since spring 1995. We report on our experience with the first 30 consecutively treated multiple sclerosis patients. Indication, adverse event profile and clinical response to treatment are described. The most common side effects were local injection site reactions (63%), influenza-like symptoms (50%) and fatigue (33%). As compared to the prestudy period we observed a 49% reduction in the exacerbation rate. Compliance was excellent, possibly due to strict selection and extensive information about possible effects and side effects.

Adjuvants, Immunologic↗

PubMed1996

Differential expression of perforin in muscle-infiltrating T cells in polymyositis and dermatomyositis.

Polymyositis (PM) and dermatomyositis (DM) are the prototypical inflammatory diseases of skeletal muscle. In PM, CD8+ T cells invade and destroy muscle fibers, whereas humoral effector mechanisms prevail in DM. We studied the expression of the cytotoxic mediator perforin in inflammatory cells in PM and DM muscle by semiquantitative PCR, immunohistochemistry and confocal laser microscopy. Similar levels of perforin mRNA were expressed in PM and DM, and abundant perforin-expressing CD3+CD8+ and CD3+ CD4+ T cells were observed in both diseases. However, there was a striking difference in the intracellular localization of perforin. In DM, perforin was distributed randomly in the cytoplasm of the inflammatory T cells. In contrast, 43% of the CD8+ T cells that contacted a muscle fiber in PM showed perforin located vectorially towards the target muscle fiber. The results suggest (a) that the random distribution of perforin in the cytoplasm of muscle-infiltrating T cells observed in DM reflects nonspecific activation, and (b) that the vectorial orientation observed only in PM reflects the specific recognition via the T cell receptor of an antigen on the muscle fiber surface, pointing to a perforin- and secretion-dependent mechanism of muscle fiber injury.

Base Sequence↗

PubMed1996

Factor V R506Q gene mutation analysis by PCR-RFLP: optimization, comparison with functional testing for resistance to activated protein C, and establishment of cell line controls.

Resistance to activated protein C (APC) has been recently identified as a highly prevalent risk factor for the development of venous thrombosis. In the majority of cases, APC resistance correlates with the presence of a single point mutation in the factor V gene (FV R506Q). The mutation is present in 3% to 5% of the general population and in up to 50% of patients with a personal and family history of venous thrombosis. In the current study, the authors have optimized and implemented for clinical diagnosis a method for detection of FV R506Q using the polymerase chain reaction coupled with restriction fragment length polymorphism analysis (PCR-RFLP). Forty-one healthy adults and 139 patients referred for hypercoagulability testing were genotyped and their APC resistance ratios determined using commercially available reagents (COATEST APC Resistance Kit). Comparative analysis indicated that if functional APC resistance was defined as per manufacturer's guidelines, a significant number of individuals with a normal factor V genotype were categorized as APC resistant and conversely, a significant number of individuals heterozygous for FV R506Q were categorized as non-APC resistant. These results indicate that comparative functional and genotypic analyses in the individual clinical laboratory setting are critical for establishing normal ranges and cut-off values for functional APC resistance due to FV R506Q. To facilitate molecular evaluation of APC resistance, Epstein-Barr virus (EBV) immortalized B-lymphocyte cell lines were established from individuals heterozygous and homozygous for FV R506Q.

Adult↗