Biomedical subjects

G Scholz

Publications and source records attributed to G Scholz.

At least 37 records · Page 2Linked to original sources

PubMed1999

[Development of prediction models for three in vitro embryotoxicity tests which are evaluated in an ECVAM validation study]

In 1997 ZEBET started the co-ordination of a study funded by the European Centre for the Validation of Alternative Methods (ECVAM) with the aim of pre-validation and validation of three in vitro embryotoxicity tests. These tests employ the cultivation of post-implantation whole rat embryos (WEC test), cultures of primary limb bud cells of rat embryos (micromass test, MM-Test), and cultures of a pluripotent mouse embryonic stem cell line (embryonic stem cell test, EST). In the current Validation Study each of the tests is evaluated in four laboratories under blind conditions. In an initial phase of the validation study six out of 20 test chemicals comprising different embryotoxic potential (non, weak and strong embryotoxic) were tested. The results were used to improve the prediction models (PM) for the WEC test and the MM-Test in order to identify the embryotoxic potential of test chemicals. In addition, the existing PM for the EST was evaluated using the results from testing of the initial six chemicals. The PM for the EST was developed using the results of a previous pre-validation study (Scholz et al.,1999), in which 94% of the learning sample were classified correctly. Applying this PM to the results of the initial phase of the current validation study, 80% of the experiments were classified correctly according to the embryotoxic potential of the tested chemicals in vivo. Applying the PM for the MM-Test and the WEC test that were developed during the current validation study in both tests, 79% correct classifications were achieved. Since the PM of the WEC-Test took into account only parameters of growth and development, but not cytotoxicity data, a second PM (PM2) for the WEC was developed that was improved by incorporating cytotoxicity data of the differentiated mouse fibroblast cell line 3T3 derived from the EST. This approach, which has previously never been used as an adjunct to the WEC test, resulted in an increase of correct classifications to 96%.

Journal Article↗

PubMed1998

[Frequency specificity of acoustic evoked potentials of early and intermediate latency with high-pass noise masking].

In contrast to auditory brainstem potentials (ABR), a more apically defined generation of middle latency evoked potentials (MLR) can be assumed due to the longer integration time present. To date, the effect of stimulus frequency spectra on MLR has not been completely known. By using masking with high-pass filtered white noise in 12 normal-hearing adults, we measured the dependence of amplitude and latency of ABR and MLR on filter frequency. No differences were found between the masking effects for ABR and MLR evoked by tone bursts of 0.5 and 2 kHz. The high-pass masking of click stimuli showed a greater influence on the amplitude and latency of the ABR than on MLR. These different qualities have to be considered clinically when evaluating the methods and results of hearing testing with acoustic evoked potentials. Our findings show that MLR evoked by tone bursts should be used for the frequency selective diagnosis of hearing thresholds, especially in low-frequency ranges.

Acoustic Stimulation↗

PubMed1998

Low-frequency masking for detection of endolymphatic hydrops in patients with glaucoma.

The coincidence of various eye and ear abnormalities has been described in the literature. Some authors discuss the possible existence of endolymphatic hydrops in patients with glaucoma. Whereas the current diagnostic tests for glaucoma are well-defined and evident, those for endolymphatic hydrops are not so reliable. This has made it difficult to accurately study the coincidence of endolymphatic hydrops and glaucoma. For better detection of endolymphatic hydrops, we performed low-frequency masking tests in 23 patients with primary open-angle glaucoma without signs of Ménìere's disease. The phase dependent sensitivity of the organ of Corti to a short test stimulus can be measured by applying a low-frequency masker tone to determine the modulation depth. Whereas the modulation depth in the normal hearing population is around 20-35 dB, the modulation depth in patients with Ménière's disease may be significantly decreased (5-10 dB), depending on the stage of disease. A decreased modulation depth was found in at least one ear in 19 of our 23 patients with glaucoma. Correlations between homeostatic mechanisms and their histological characteristics, e.g. melanocytes and their hormonal and enzymatic regulation, will be discussed. If the common pathogenesis of eye and ear lesions is better understood, it may be possible to develop new and more effective strategies for prevention and therapy.

Acoustic Stimulation↗

PubMed1997

Efficient transcription of an immunoglobulin kappa promoter requires specific sequence elements overlapping with and downstream of the transcriptional start site.

The expression of immunoglobulin (Ig) genes depends on tissue-specific elements in the promoter and enhancer regions of light chain and heavy chain genes. In contrast to the complex modular character of Ig enhancers, the promoters appear to be simple, depending primarily on a conserved TATA box and octamer elements. We have analyzed the role of proximal sequences for Igkappa promoter function. Igkappa promoter transcription critically depends on initiator-like sequences and on a downstream element located at +24 to +39 relative to the start site. Replacement of these sequences resulted in strong reduction of promoter activity. In vitro, these elements were found to be more effective in extracts of lymphoid than of non-lymphoid origin. Deletion of the downstream and initiation site regions had a comparable effect on promoter activity to obliteration of the TATA box or octamer element. The downstream sequence was bound by two nuclear proteins, identical to the previously identified Ig-specific C5 and C6 complexes. Whereas C5 is found in HeLa cells and in lymphoid cells, C6 is lymphoid specific. Thus, further specific sequences in addition to the previously characterized elements, the octamer and the TATA box, are required for efficient kappa promoter expression in B lymphocytes.

Animals↗

PubMed1996

Genetic analysis of two tomato mutants affected in the regulation of iron metabolism.

Iron is one of the most important micronutrients for plants. Like other organisms, plants have developed active mechanisms for the acquisition of sufficient iron from the soil. Nevertheless, very little is known about the genetic mechanisms that control the active uptake. In tomato, two spontaneously derived mutants are available, which are defective in key steps that control this process. The recessive mutation chloronerva (chln) affects a gene which controls the synthesis of the non-protein amino acid nicotianamine (NA), a key component in the iron physiology of plants. The root system of the recessive mutant fer is unable to induce any of the characteristic responses to iron deficiency and iron uptake is thus completely blocked. We present a characterization of the double mutant, showing that the fer gene is epistatic over the chln gene and thus very likely to be one of the major genetic elements controlling iron physiology in tomato. In order to gain access to these two genes at the molecular level, both mutants were precisely mapped onto the high density RFLP map of tomato. The chln gene is located on chromosome 1 and the fer gene is on chromosome 6 of tomato. Using this high-resolution map, a chromosome walk has been started to isolate the fer gene by map-based cloning. The isolation of the fer gene will provide new insights into the molecular mechanisms of iron uptake control in plants.

Algal Proteins↗

PubMed1996

[Evaluation of contact and care centers, a low-threshold daytime source in the city of Zurich].

Contact centres for drug addicts in the city of Zurich exist since 1988 to provide decentralized low threshold meeting places and contact centres for persons who are poor and neglected. The main objective of these survival aids is to counteract further impoverishment of the addicted in order to actively contribute to their subsistence. Since its foundation the project as a whole has been submitted to numerous structural changes which had to be taken into consideration by the evaluation. Subject of the examination was the description of the target group and the staff, the use of the services and their effect on the users as well as the environment. Only quantitative methods were used. The target group of the centers consisted of drug consumers who at the best are partially socially integrated, but usually completely desintegrated. The evaluation revealed that the different contact centers services were extensively used and met the focal needs of their users. However, the high fluctuation of personnel within the project teams turned out to be a permanent problem for the project management and the operators. As main reasons for the fluctuation the authors discuss the great dependency on political decisions and the problematical demanding clientele. Due to these structural turbulences some staff related offers are only accepted to a limited degree, and the contact centers tend to become simple supply services. Therefore the authors endorse the redefinition of long term objectives of the institution under consideration of the necessary framework.

Adult↗

PubMed1996

[Evaluation of Haus Breitenstein, a low-threshold living facility in the city of Zurich].

Early in 1991 in attended home with 13 rooms was established in the Breitenstein House in Zurich as a low threshold survival aid for most severe drug addicts. After only 17 months, before the end of the two-year pilot phase, the facility was closed for economic reasons. The objective of the project laid emphasis on the furtherance of each resident's initiative and responsibility. Using quantitative as well as qualitative instruments the accompanying evaluation came to the conclusion that the attended low threshold accommodation is a realistic completion and an alternative to already existing aids. The institution enabled the drug addicts to reestablish their damaged autonomy and take a constructive step towards the shaping of their lives. In individual cases a substantial structural change could be initiated. The authors concluded that the Breitenstein House was a successful attempt to develop comprehensive strategies in drug policy.

Adult↗

PubMed1996

Transcriptional down regulation of the nov proto-oncogene in fibroblasts transformed by p60v-src.

We have sought to identify genes whose expression is altered as a consequence of transformation by p60v-src. Using the mRNA differential display method, we have identified the nov proto-oncogene as one gene that is down regulated in chicken embryo fibroblasts (CEFs) transformed by p60v-src. nov transcripts were also found to be present at only very low levels in proliferating CEFs in comparison with quiescent CEFs. Serum stimulation of quiescent CEFs also resulted in a decline in the steady-state level of nov transcripts. Taken together, these findings suggest that the nov gene is expressed only in quiescent fibroblasts and that its down regulation may contribute to cellular transformation by the v-src oncogene. Down regulation of the nov gene appears to occur at both the transcriptional and posttranscriptional levels. Results obtained from experiments with a protein kinase inhibitor suggest that protein kinase C may be a key downstream effector in mediating the down regulation of nov transcripts in response to activation of p60src or serum stimulation. In addition, we found that transcription of an unknown gene is required for the decline in the steady-state level of nov transcripts in response to serum stimulation.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗

PubMed1995

Activation of YRP kinase by v-Src and protein kinase C-mediated signal transduction pathways.

We have previously reported that a serine(threonine) protein kinase that phosphorylates histone H1 in vitro is activated by tyrosine phosphorylation in v-Src-transformed rat 3Y1 fibroblasts. We now refer to this kinase as YRP kinase, for tyrosine-regulated protein kinase. Since YRP kinase may play a role in mediating the growth-stimulatory and morphology-altering effects of v-Src, we have further examined the signal transduction involved in the activation of YRP kinase. Although YRP kinase is constitutively activated in fibroblasts transformed by v-Src, activation of protein kinase C was also found to lead to activation of YRP kinase. Activation of YRP kinase by protein kinase C was found to be potentiated by vanadate treatment or overexpression of c-Src. The activation of YRP kinase by v-Src, however, does not appear to be mediated by protein kinase C, suggesting that YRP kinase can be activated by two separate signal transduction pathways. Transformation of fibroblasts by v-Ras or v-Mil did not result in activation of YRP kinase, indicating that the MAP kinase pathway does not mediate the activation of YRP kinase by v-Src or protein kinase C.

Animals↗

PubMed1995

Masking and pitch shift of tone bursts and clicks by low-frequency tones.

From experiments in animals and investigations in humans it is known that the normally phase-dependent masking of a short stimulus by a low-frequency continuous tone does not occur in the case of endolymphatic hydrops. The recording of the masked threshold of short tone stimuli in a loud tone of 30 Hz is to be evaluated for the clinical diagnostics of Ménière's disease. To this purpose, the main parameters of the measurements (type, frequency, duration of the stimulus, and intensity of the masker) and their effect of phase-dependent masking and pitch-shift are investigated. Stimuli above 2 kHz are masked less than those of lower frequencies. Wide-band stimuli are less useful, since only the low-frequency component of their spectrum is masked. The tone stimuli should be short (1 - 2 ms) in order to make the measurement of the phase dependence more accurate. With increasing masker level the masking at phase 0 degree corresponds to the increase in level, at phase 270 degrees the amount is twice as much. The pitch shift which is perceived in low-tone masking depends on the phase of the stimulus, and on the levels of the stimulus and the masking tone. The use of brain stem recordings in the investigation of phase-dependent low tone masking is problematic since well-synchronizing stimuli with high frequency spectral components are masked poorly.

Acoustic Stimulation↗

PubMed1995

[Diagnosis of endolymphatic hydrops with low tone masked otoacoustic emissions].

BACKGROUND: A new method of diagnosis of endolymphatic hydrops by recording low-tone masked evoked otoacoustic emissions (TEOAE) is presented. METHODS: A short acoustic stimulus and a masker tone of 30 Hz are applied in an adjustable phase relation simultaneously to the same ear. In the normal hearing ear the masker shows little influence on the TEOAE at 0 degrees, whereas the suppression at phase 270 degrees (maximal rarefaction at the eardrum) is nearly complete. However, in cases of endolymphatic hydrops this masking effect is reduced or absent, indicating impaired mobility of the basilar membrane. RESULTS: The masked TEOAE were recorded of patients with normal hearing, Menière's disease, and sudden hearing loss without vertigo. In Menière cases with supposed endolymphatic hydrops, the amplitude modulation of the emissions was found to be much less than in the other groups. CONCLUSIONS: Where TEOAE can be recorded, low-tone masking is a quick, objective, and noninvasive method for the diagnosis of endolymphatic hydrops.

Acoustic Stimulation↗

PubMed1994

Identification of novel ubiquitous and cell type-specific factors that specifically recognize immunoglobulin heavy chain and kappa light chain promoters.

In a search for novel factors that interact with the octamer element, immunoglobulin kappa light chain (V kappa) and heavy chain VH) gene promoters were compared for binding to nuclear proteins from lymphoid cells. Both promoters showed a similar pattern of bound factors, which was entirely different from that seen with the human immunodeficiency virus, type I promoter. Besides OTF-1 (Oct-1) and OTF-2 (Oct-2), at least three additional complexes were observed. Two of these were functionally analyzed: C1, a slowly migrating complex, which was much more abundant in B cells than in non-lymphoid cells, and C4, which appeared to have ubiquitous distribution. As determined by several methods, the protein(s) forming the C1 complex bound on the V kappa gene to a site that partially covers the octamer element, whereas, surprisingly, in the VH gene a region spanning the TATA box up to the transcriptional initiation site was recognized. The ubiquitous C4 complex was analyzed for the light chain promoter. The protein specifically bound to a site immediately 5' to the octamer. By in vitro transcription analysis, we found that C4 augments the octamer-dependent transcription of the V kappa gene. The activation required binding of OTF-1/OTF-2, and C4 could not stimulate transcription by itself, implying a synergistic interaction. Due to the overlapping binding sites, the effect of C1 by itself could not be clearly separated from the C4 effect. However, the specific interaction of C1 with crucial control elements of light chain and heavy chain promoters strongly suggests a functional role in immunoglobulin gene transcription control.

B-Lymphocytes↗

PubMed1993

Activation of a histone H1 kinase by tyrosine phosphorylation in v-src-transformed fibroblasts.

Using anti-phosphotyrosine immunoaffinity chromatography, we have searched for serine/threonine kinases that are directly regulated by tyrosine phosphorylation in v-src-transformed rat 3Y1 fibroblasts. Tyrosine phosphoprotein preparations from v-src-transformed cells contain a kinase activity that phosphorylates histone H1 in vitro on serine residues and this activity is present at a 20-fold greater level than that in parental cell preparations. This activity elutes from a MonoQ FPLC column as a single peak and gel filtration chromatography suggests that the kinase has a molecular mass of approximately 55 kDa. Tyrosine phosphatase treatment inactivates the histone H1 kinase and this result indicates that the specific activity of the kinase is regulated by tyrosine phosphorylation. Experiments with cells transformed with a temperature-sensitive mutant of the v-src oncogene demonstrate that the tyrosine phosphorylation of the histone H1 kinase is an early event in v-src transformation. The kinase is distinct from known cdc2 family members that contain the PSTAIR motif, because the kinase can be separated almost completely from these proteins by immunoprecipitation with an antibody against p34cdc2. The profile of antibody reactivity and sensitivity to modulators of protein kinases suggests that this activity is distinct from known second messenger-regulated kinases and from previously characterized MAP kinases.

Amino Acid Sequence↗

PubMed1992

Affinity labelling of the active site of brain phosphatidylinositol 4-kinase with 5'-fluorosulphonylbenzoyl-adenosine.

5'-p-Fluorosulphonylbenzoyl-adenosine (FSO2BzAdo), an affinity labelling analogue of ATP, was used to label the active site of sheep brain phosphatidylinositol 4-kinase (PtdIns 4-kinase). The incubation of PtdIns 4-kinase with concentrations of FSO2BzAdo as low as 50 microM resulted in considerate inactivation of the enzyme. (e.g. 55% less after 60 min with 50 microM FSO2BzAdo). The kinetics of inactivation of PtdIns 4-kinase by FSO2BzAdo suggest a two-step mechanism, in which a rapid reversible binding of FSO2BzAdo to the enzyme is followed by a covalent sulphonation step. The first-order rate constant (k2) for the inactivation of PtdIns 4-kinase was calculated to be 0.063 min-1, and the steady-state constant of inactivation (Ki) to be 200 microM. Preincubation of the enzyme with either ATP plus Mg2+, or PtdIns alone, prior to addition of FSO2BzAdo reduced the degree of inactivation of the enzyme; suggesting that FSO2BzAdo binds within the active site PtdIns 4-kinase. Moreover, since ATP plus Mg2+ provided the greatest protection against inactivation, it is concluded that the main site of labelling of PtdIns 4-kinase by FSO2BzAdo is within the ATP-binding site of the enzyme. Results obtained from chemical modification experiments, which employed pyridoxal 5'-phosphate and tetranitromethane, are consistent with a catalytically-essential lysine being present within the ATP-binding site of PtdIns 4-kinase. Therefore, it is hypothesised that the inactivation of PtdIns 4-kinase by FSO2BzAdo may be due to the labelling of this lysine residue.

1-Phosphatidylinositol 4-Kinase↗

PubMed1991

Purification and chemical modification of a phosphatidylinositol kinase from sheep brain.

A membrane-bound phosphatidylinositol (PtdIns) kinase has been purified approximately 9500-fold to apparent homogeneity from sheep brains. The purification procedure involves: solubilisation of the membrane fraction with Triton X-100, ammonium sulphate fractionation and a number of ion-exchange and gel-filtration chromatography steps. The purified enzyme exhibited a final specific activity of 1149 nmol.min-1.mg-1. The molecular mass of the enzyme was estimated to be 55 kDa by SDS/PAGE and 150 +/- 10 kDa by HPLC gel filtration in the presence of Triton X-100. Kinetic measurements have shown that the apparent Km value of PtdIns kinase for the utilisation of PtdIns is 22 microM and for ATP 67 microM. Mg2+ was the most effective divalent cation activator of PtdIns kinase, with maximal enzymatic activity reached at a concentration of 10 mM Mg2+. In addition to adenosine and ADP, the 2'(3')-O-(2,4,6-trinitrophenyl) derivative of ATP was found to be a strong competitive inhibitor of the enzyme, with a Ki of 32 microM. Enzymatic activity was found to be stimulated by Triton X-100 but inhibited by deoxycholate.

1-Phosphatidylinositol 4-Kinase↗

PubMed1990

Binding of a photoaffinity analogue of pyridoxal to pyridoxal kinase.

The binding of pyridoxal analogues to the structural domains of pyridoxal kinase was studied by fluorescence spectroscopy and chromatographic techniques. Two fragments of 24 and 16 kDa, arising from limited proteolysis of the native enzyme, were separated by ion-exchange chromatography and used for binding studies with pyridoxal oxime. Fluorometric titrations yielded dissociation constants of 6 and 12.4 MicroM for pyridoxal oxime bound to the native enzyme and 24-kDa fragment, respectively. 4-(4-Azido-2-nitrophenyl)-pyridoxamine, a new photolabeling reagent, binds irreversibly to the kinase with concomitant loss of catalytic activity. The modified kinase (2.1 mol label/mol dimer) yields two fragments upon limited proteolysis with chymotrypsin. The two fragments were separated by reverse-phase HPLC and SDS/polyacrylamide gel electrophoresis. Radiolabeled ligand was detected only in the 24-kDa fragment. It is postulated that the pyridoxal binding site is located in the 24-kDa structural domain.

Affinity Labels↗

PubMed1990

[The effect of verapamil on adrenocortical hormone secretion].

Variations in circadian cortisol rhythm, morning ACTH concentration and the 24-hour cortisol excretion were examined in 15 healthy volunteers and 9 untreated patients with cardiovascular diseases. These variations were measured under basal conditions and during low dose (ACTHL-test) and also a high dose (ACTHH-test) ACTH stimulation. Additionally, in the patients with cardiovascular diseases the effect of verapamil-administration was investigated. Under basal conditions no significant differences were to be seen between normal subjects and the patients, either before or after therapy. The two functional tests resulted in almost identical increases of cortisol and ACTH levels, both in healthy subjects and the patients before verapamil. It was found that after verapamil application during the ACTHL-test, the levels of plasma cortisol decreased in comparison with the values noted before verapamil (p less than 0.05). However, in the ACTHH-test a significant increase of the plasma cortisol (p less than 0.05) was evident. Our results lead to the conclusion that after verapamil therapy, under slight loading, the plasma cortisol concentration can be expected to decrease slightly, and in extremely high stress situations it will increase.

Adrenocorticotropic Hormone↗

PubMed1990

[Does the calcium antagonist verapamil modify calcium metabolism?].

In 11 healthy volunteers the effects of oral verapamil administration (4 X 80 mg/d) on plasma or serum concentrations of ionized, pH-corrected ionized and total calcium, on intact and midregional parathyroid hormone and on 24 h urinary excretion of calcium were investigated at rest. In addition the concentrations of ionized, pH-corrected ionized and total serum calcium were analyzed under bicycle exercise. Verapamil resulted in an increase of total serum calcium (p less than 0.02) and a fall in midregional parathyroid hormone concentrations (p less than 0.02) at rest. There was also found a statistically significant elevation of both ionized (p less than 0.05) and pH-corrected ionized (p less than 0.05) plasma calcium, and a slight increase in the total calcium under exercise (p less than 0.1). Before verapamil a significant negative correlation between total calcium and midregional parathyroid hormone was evident (r = -0.618; p less than 0.05), which however could not be clearly discerned after verapamil.

Adult↗