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J Stahl

Publications and source records attributed to J Stahl.

At least 37 records · Page 2Linked to original sources
PubMed2000

Preparation of response force and movement direction: onset effects on the lateralized readiness potential.

Two experiments assessed the preparatory effects of advance information about response force and movement direction on the lateralized readiness potential (LRP). In a choice reaction time (RT) task, an imperative stimulus required an isometric flexion or an extension of the left or right index finger. Prior information about response force or about movement direction reduced RT and shortened the interval from the onset of the imperative stimulus up to the onset of the LRP. Advance information, however, about direction but not about force decreased the interval from LRP onset to the onset of the overt response. The identical pattern of results was obtained in a second experiment, in which each participant performed both precue conditions. The findings of both experiments support the notion that response force is specified before movement direction. These results are consistent with the view accordingly different mechanisms are involved in the specification of muscle force and movement direction.

Adult↗
PubMed2000

CAT administration of language placement examinations.

This article describes the development of a computerized adaptive test for Cegep de Jonquiere, a community college located in Quebec, Canada. Computerized language proficiency testing allows the simultaneous presentation of sound stimuli as the question is being presented to the test-taker. With a properly calibrated bank of items, the language proficiency test can be offered in an adaptive framework. By adapting the test to the test-taker's level of ability, an assessment can be made with significantly fewer items. We also describe our initial attempt to detect instances in which "cheating low" is occurring. In the "cheating low" situation, test-takers deliberately answer questions incorrectly, questions that they are fully capable of answering correctly had they been taking the test honestly.

Aptitude Tests↗
PubMed1999

Photoaffinity labeling and mass spectrometry identify ribosomal protein S3 as a potential target for hybrid polar cytodifferentiation agents.

The ability of a novel class of hybrid polar compounds (HPCs) to induce differentiation and consequent cessation of proliferation of transformed cells has led to their development as potential chemotherapeutic agents in the treatment of cancer. Suberoylanilide hydroxamic acid (SAHA) is a prototype of a family of hydroxamic acid based compounds (SAHA-like HPCs) that can, at micromolar concentrations, induce a variety of transformed cell lines to differentiate. The mechanism of action of the HPCs is not entirely understood. Searching for a cellular target of the SAHA-like HPCs, we synthesized a photoaffinity labeling reagent structurally based on SAHA, and probed for SAHA-binding proteins in murine erythroleukemia (MEL) cells. Photoaffinity labeling in cell free extracts identified a 32-kDa protein (p32) that was specifically labeled by the photoaffinity reagent. Cell fractionation assays localized p32 to the P100 fraction. p32 was partially purified and identified by mass spectrometry as the 40 S ribosomal protein S3. Expression of epitope-tagged S3 in bacterial lysates followed by photoaffinity labeling confirmed its specific labeling. Identification of a cytodifferentiation agent target may shed light on the mechanism by which the SAHA-like HPCs exert their antitumor effects.

Affinity Labels↗
PubMed1999

Preoperative treatment of rectal cancer with radiation, chemotherapy and hyperthermia: analysis of treatment efficacy and heat-shock response.

Preoperative treatment of locally advanced rectal cancer with radiation, chemotherapy and hyperthermia is analyzed with regard to heat-shock response. In 23 patients with locally advanced rectal cancer (uT3/uT4), hyperthermia was administered in combination with radiotherapy and chemotherapy. In parallel, the effect of the treatment on levels of the heat-shock proteins HSP27 and inducible HSP70 in tumors and surrounding tissues was investigated by Western blotting. The patients' sera were also examined for autoantibodies against HSPs. HSP27 and inducible HSP70 were detected in most rectal tumors and surrounding tissues before and after treatment. HSP27 and inducible HSP70 levels had changed in 10 tumors after treatment. However, prior to treatment, there existed an unexpected diversity in HSP levels in the tumors and surrounding tissue. Hyperthermia doses in cumulative minutes for which 90% of the tumor is above the reference temperature (cum min T90 > or = 15 min) led to increased survival and response compared to that of a control group of patients treated without or with low-dose hyperthermia (cum min T90 < 15 min). However, there was no correlation to different expression of the HSPs. Hyperthermia as used in this setting does not lead to any sustained expression of HSPs in either the tumor or the surrounding tissue.

Antibodies↗
PubMed1998

The 80S rat liver ribosome at 25 A resolution by electron cryomicroscopy and angular reconstitution.

BACKGROUND: The ribosome is central to protein synthesis in all living organisms. Single-particle electron cryomicroscopy has recently led to the determination of three-dimensional structures of bacterial ribosomes to approximately 20 A, which have since revolutionised our understanding of ribosomal function. The structure we present here of the 80S rat liver ribosome leads the way to similar progress for mammalian ribosomes. RESULTS: Among the new details revealed by our 25 A structure of the 80S rat liver ribosome are channels within the subunits, a large 'flat ribosomal surface' (FRS) on the outer surface of the large subunit and structural extensions of the mammalian compared to the bacterial ribosome. The main large subunit channel in both the bacterial and the mammalian species starts at the peptidyl transferase centre, below the central protuberance, and ends in the FRS, at the lower back of the large subunit. Structurally, the channels of both species can be directly superimposed. CONCLUSIONS: The mammalian structural extensions--none of which trespass the FRS--can be interpreted in terms of rRNA inserts and additional protein content over that of bacterial ribosomes. The main large subunit channel, which ends at the FRS, is the best candidate for the exit channel for proteins targeted for the endoplasmic reticulum.

Animals↗
PubMed1998

Colocalization of single ribosomes with intermediate filaments in puromycin-treated and serum-starved mouse embryo fibroblasts.

Previous experiments have revealed a relatively weak electrostatic binding capacity of in vitro reconstituted intermediate filaments (IFs) as well as of natural IFs of whole cell mount preparations for purified ribosomal particles of mammalian origin. In order to demonstrate that such associations also occur in vivo, intact cells were subjected to double immunofluorescence microscopy using antibodies directed against vimentin and ribosomal protein S17. Since in proliferating cells the majority of the ribosomal particles are assembled into polyribosomes and these are to a great extent associated with microfilaments, in vitro cultured mouse embryo skin fibroblasts (MSF cells) were treated with puromycin to allow the formation of single ribosomes. Employing confocal laser scanning microscopy, the ribosomes were detected in colocalization with vimentin IFs. Disassembly of polyribosomes was also achieved by serum starvation of cultured cells. In this case, MSF cells of a low passage attained an extended and flattened appearance with the vimentin IFs being directly associated with the cell nuclei, radiating into the peripheral areas of the cells or showing a stress fiber-like distribution. In both cases, considerable quantities of ribosomal material were seen in close neighborhood to vimentin IFs. Frequently, these ribosome-IF associations were coaligned with microtubules and they also surrounded myosin I-decorated stress fibers. Double labeling with the vital, RNA-specific fluorochrome SYTO 14 produced a fluorescence pattern largely superimposable on that of ribosomal protein S17. Treatment of the starved cells with either demecolcine or cytochalasin D had an only moderately disturbing effect on vimentin IF distribution and the ribosomes stayed in contact with the vimentin IFs. On the basis of these results, it is conceivable that IFs play a role in the storage of ribonucleoprotein particles in general and non-translating ribosomes in particular in the cytoplasm of animal cells. In addition, the often seen coalignment of IFs with microtubules and microfilaments might serve facilitated and directional transport of ribonucleoprotein particles from the nucleus to peripheral areas of the cell.

Actin Cytoskeleton↗
PubMed1997

Abundance and location of the small heat shock proteins HSP25 and alphaB-crystallin in rat and human heart.

BACKGROUND: In the heart, there are high constitutive levels of the two related small heat shock proteins, HSP25 and alphaB-crystallin. To gain insight into their functional role, we have analyzed abundance and location of both proteins in rat and human hearts at different stages of development and in diseased state. METHODS AND RESULTS: Immunoblotting analysis of rat ventricular tissue at fetal, neonatal, and adult stages reveals the level of HSP25 to decline strongly during development, whereas the level of alphaB-crystallin remains nearly constant. In parallel, the portion of phosphorylated isoforms of HSP25 decreases as shown by two-dimensional polyacrylamide gel electrophoresis. HSP25 is detected in cardiomyocytes and endothelial and vascular smooth muscle cells, whereas alphaB-crystallin is detected in cardiomyocytes only by immunofluorescence and immunoelectron microscopy. Both proteins colocalize in the I-band and M-line region of myofibrils in cardiomyocytes. In diseased and transplanted adult human hearts, HSP25 and alphaB-crystallin levels are considerably elevated compared with fetal hearts. In failing adult human hearts, phosphorylated isoforms of HSP25 predominate, and cardiomyocytes with a partial dislocation of HSP25 and alphaB-crystallin are observed. CONCLUSIONS: Differential accumulation and location of HSP25 and alphaB-crystallin in heart tissue during development imply distinct functions of both proteins, which seem to be involved in organization of cytoskeletal structures. As judged by level, phosphorylation state, and location of both small heat shock proteins, diseased adult human hearts share features with fetal hearts.

Aging↗
PubMed1997

The signal transducer gp130--bacterial expression, refolding and properties of the carboxy-terminal domain of the cytokine-binding module.

Gp130 is the signal transducing receptor subunit of the so-called interleukin-6-type cytokines. This transmembrane protein is a member of the cytokine-receptor superfamily predicted to consist of six fibronectin-type-III-like domains in its extracellular part. The second and the third domain constitute the so-called cytokine-binding module. Domain 2 is characterized by a set of four conserved Cys residues, domain 3 by a conserved WSXWS motif. As a first approach to a more detailed characterization of the cytokine-binding domains of human gp130, we have expressed in Escherichia coli two forms of domain 3 differing in length. Both proteins were purified and refolded in a single step applying size-exclusion chromatography. According to the rotational correlation times deduced from fluorescence anisotropy decay, they do not form aggregates. CD and fluorescence spectroscopy were used to study thermal unfolding and denaturation by guanidinium hydrochloride. It was shown that N- and C-terminal extension by residues of the adjacent hinge regions substantially increase the thermal stability of the domain, which is conceivable from a molecular model. These results are the basis for further structural investigation by NMR spectroscopy.

Binding Sites↗
PubMed1997

Protein environment of mRNA at the decoding site of 80S ribosomes from human placenta as revealed from affinity labeling with mRNA analogs--derivatives of oligoribonucleotides.

Affinity labeling of 80S ribosomes from human placenta has been studied using various mRNA analogs, namely, 2',3'-O-[4-(N-2-chloroethyl)-N-methylamino]benzylidene derivatives of oligoribonucleotides (Up)(n-1)U[32P]pC (n = 3, 6 or 12) and AUGU3[32P]pC as well as ([4-(N-2-chloroethyl)-N-methylamino]benzylmethyl-[5'-32P]-phospham ide derivatives of pAUGUn (n = 3 or 6). Labeling of 80S ribosomes with the derivatives of oligouridylates was carried out in complexes obtained nonenzymatically in the presence of saturating amounts of Phe-tRNA(Phe). Complexes with derivatives bearing AUG codon were obtained using a fractionated lysate from rabbit reticulocytes which contained protein translation factors and was deprived from endogeneous ribosomes and mRNAs. In all cases, 40S subunits were labeled preferentially. Within the subunits, both 18S rRNA and proteins were found to be modified. Sites of cross-linking in 18S rRNA have been identified earlier. Here, it is shown that the main targets of cross-linking among the ribosomal proteins were S3 and S3a (with minor modification of S26) for the 3'-derivatives of (Up)5UpC and (Up)11UpC. For the same derivative of (Up)2UpC, the reverse modification pattern was observed. 5'-derivatives of pAUGUn were cross-linked to proteins S3 and S3a in comparable extent; 3'-derivative of AUGU3pC modified protein S3a preferentially.

Affinity Labels↗
PubMed1997

Identification and characterization of heat shock protein 27 protein species in human myocardial two-dimensional electrophoresis patterns.

Immunostaining of heat shock protein 27 (Hsp27) protein species on two-dimensional electrophoresis (2-DE) gels with enhanced sensitivity yields 59 spots reacting with anti-Hsp27 antibodies. Recombinant Hsp27 exists in 2-DE as two major protein species which comigrate in the human myocardial pattern with Hsp27 spots C754 and D899 as defined in the heart high-performance 2-DE database (http://www.mdc-berlin.de/emu/heart/). Preparative electrophoresis of human myocardial proteins and analysis of the enriched mass range 20-30 kDa by 2-DE revealed eight protein spots (C438, C582, C658, C697, C754, C595, C750) from the human myocardial database and a new spot not previously detected on silver-stained gels. These spots were identified as Hsp27 protein species by enzymatic in-gel-digestion and analysis by matrix assisted laser desorption-ionization (MALDI) peptide mass fingerprinting and, in part, MALDI-post source decay sequencing of single fragments. Possible post-translational modifications were investigated: immunostaining tests with anti-phospho-serine/-threonine/-tyrosine antibodies, although positive for other myocardial proteins, were negative for presumed Hsp27 protein species; likewise, periodate-glycostaining assays and biotinylation screening did not detect modifications in the investigated Hsp27 protein species.

Amino Acid Sequence↗
PubMed1997

Oncocytic cardiomyopathy: a rare cause of unexpected early childhood death associated with fitting.

A 15-month-old girl died unexpectedly in hospital following a five-day history of intermittent cardiac arrhythmias and convulsions preceded by several weeks of occasional vomiting. Autopsy revealed subendocardial nodules in the left ventricle, and tricuspid and mitral valves that were composed of aggregated large cells with foamy, pale pink cytoplasm characteristic of oncocytic cardiomyopathy. Fat stains were positive for neutral lipid and phospholipid and electron microscopic examination revealed numerous irregular mitochondria within affected cells. Examination of the brain revealed no structural or histologic abnormalities, anoxic damage or thromboembolic material. Oncocytic cardiomyopathy, though rare, may cause unexpected death in previously well young children with quite variable preceding clinical symptoms and signs which include fitting. Although the aetiology is unknown there is evidence that mitochondrial dysfunction may be involved.

Arrhythmias, Cardiac↗
PubMed1997

On-line performance assessment using rating scales.

The purpose of this paper is to report on the development of the on-line performance assessment instrument--the Assessment of Motor and Process Skills (AMPS). Issues that will be addressed in the paper include: (a) the establishment of the scoring rubric and its implementation in an extended Rasch model, (b) training of raters, (c) validation of the scoring rubric and procedures for monitoring the internal consistency of raters, and (d) technological implementation of the assessment instrument in a computerized program.

Activities of Daily Living↗
PubMed1996

[Development of a clinical-radiological CD-ROM database for focal lesions of the liver].

PURPOSE: The development of a CD-ROM database for continuous acquisition and archiving of interesting radiologic cases is described. As a pilot study radiologic images of focal lesions of the liver were collected. METHODS: The database runs on a PC (80486 DX2, 66 MHz, 16 MB RAM), which is connected to the clinic network, a high quality x-ray scanner and a CD-ROM writer. Radiologic images can be inserted into the database either by scanner or by image transfer via network. The database is designed using Access 2.0. Well documented cases were chosen retrospectively, manipulated and archived. RESULTS: 308 images of 12 focal lesions with a capacity of 802 MB were stored on CD-ROM. Per case a storage capacity of 40-75 MB is necessary for image raw material, 15-25 MB are necessary for the postprocessed data. Spatial resolution is 190 dpi (3000 pixel/40 cm). Clinical data, radiological diagnosis coded by modified ACR index and a compressed icon of the images are stored on PC, the image data are stored on CD-ROM. 2.5 hours per case are needed for the complete acquisition, manipulation and archiving procedure, but improved hardware configuration can reduce the time significantly. CONCLUSION: The described database allows continuous archiving of interesting radiologic cases on PC. ACR index is suited for a quick search of a specific combination of radiologic images and diagnosis from different modalities.

Angiography↗
PubMed1996

Isolation and purification of functional bovine lung mast cells (BLMCs).

Purified pulmonary mast cells were obtained from bovine lung using a combination of enzymatic digestion of tissue, density gradient centrifugation using Percoll, and centrifugal elutriation. In the initial procedure, lung tissue was enzymatically digested with collagenase, hyaluronidase, protease and elastase in three 30 min incubations at 37 degrees C. Monodispersed cell suspensions contained between 2 and 6% mast cells. Further purification of these mast cells by Percoll gradients and elutriation consistently yielded mast cells of > 90% purity. These cells were morphologically intact, viable and functional, as determined by histamine release evoked by secretagogue challenge. Incubation of BLMCs with Pasteurella haemolytica A1 culture supernate containing leucotoxin (LCT) alone, resulted in increased histamine release compared to controls. LCT also potentiated calcium ionophore (CaI)-induced histamine release from BLMCs.

Animals↗
PubMed1996

Early pathologic and biochemical changes in Creutzfeldt-Jakob disease: study of brain biopsies.

We examined brain biopsy tissue from five patients with a neurologic syndrome consistent with Creutzfeldt-Jakob disease using Western blot analysis and immunohistochemistry for the detection of protease-resistant prion protein, in addition to histopathologic examination. Our results indicate that the formation of protease-resistant prion protein is an early event in disease pathogenesis and Western blot analysis can detect protease-resistant prion protein in the absence of structural lesions using a small amount of brain biopsy tissue.

Adult↗
PubMed1995

The human ribosomal protein S7-encoding gene: isolation, structure and localization in 2p25.

We have identified a gene encoding the human ribosomal protein (r-protein) S7. The S7 gene contains seven exons and six introns spanning about 6 kb. Organization of the gene is similar to that of Xenopus laevis S8, the only homologous intron-containing gene isolated so far. An mRNA transcribed from this gene has an open reading frame (ORF) of 582 nucleotides (nt), which encodes a protein of 194 amino acids (22.1 kDa). The transcription start point (tsp) was mapped by a primer extension assay to a C residue within a pyrimidine-rich tract. Human S7 (hS7) is identical to rat S7 (rS7) and exhibits significant similarity with the X. laevis, insect and plant homologs. We have used fluorescence in situ hybridization (FISH) to localize S7 to chromosome 2p25.

Amino Acid Sequence↗
PubMed1995

Implication of mammalian ribosomal protein S3 in the processing of DNA damage.

A human apurinic/apyrimidinic endonuclease activity, called AP endonuclease I, is missing from or altered specifically in cells cultured from Xeroderma pigmentosum group-D individuals (XP-D cells) (Kuhnlein, U., Lee, B., Penhoet, E. E., and Linn, S. (1978) Nucleic Acids Res. 5,951-960). We have now observed that another nuclease activity, UV endonuclease III, is similarly not detected in XP-D cells and is inseparable from the AP endonuclease I activity. This activity preferentially cleaves the phosphodiester backbone of heavily ultraviolet-irradiated DNA at unknown lesions as well as at one of the phosphodiester bonds within a cyclobutane pyrimidine dimer. The nuclease activities have been purified from mouse cells to yield a peptide of M(r) = 32,000, whose sequence indicates identity with ribosomal protein S3. The nuclease activities all cross-react with immunopurified antibody directed against authentic rat ribosomal protein S3, and, upon expression in Escherichia coli of a cloned rat cDNA for ribosomal protein S3, each of the activities was recovered and was indistinguishable from those of the mammalian UV endonuclease III. Moreover, the protein expressed in E. coli and its activities cross-react with the rat protein antibody. Ribosomal protein S3 contains a potential nuclear localization signal, and the protein isolated as a nuclease also has a glycosylation pattern consistent with a nuclear localization as determined by lectin binding. The unexpected role of a ribosomal protein in DNA damage processing and the unexplained inability to detect the nuclease activities in extracts from XP-D cells are discussed.

Amino Acid Sequence↗