Quantitative analysis of MDR-1 gene expression in acute myeloid leukemia by reverse transcription polymerase chain reaction.
Explore the source record for details and available documents.
ATP Binding Cassette Transporter, Subfamily B, Mem↗
Biomedical subjects
A Schulz
Publications and source records attributed to A Schulz.
Modulation of pain perception in man by a vasopressin analogue.
The aim of the present experiment was to test whether vasopressin modulates pain perception in man. Twenty-four male volunteers participated in four sessions, each 2 weeks apart. After an adaptation session the subjects were treated intranasally with either 30 or 60 micrograms desmopressin (DDAVP) or placebo according to a cross-over double-blind design. Pain induction involved mechanical, thermal, and ischemic stimulation DDAVP had no unitary effects on pain perception in the different pain tests. The 30 micrograms dose induced sensitization to thermal stimuli. Neither treatment influenced ischemic pain perception. The mechanical pain threshold of the index finger was increased by the 60 micrograms dose only. After treatment with either dosage of DDAVP the subjects generally tolerated the pressure on their index finger for a longer time than after placebo treatment.
Administration, Intranasal↗
The similarities of bar and pat gene products make them equally applicable for plant engineers.
The bar and pat genes, isolated from different Streptomyces species, both encode a phosphinothricin acetyltransferase (PAT) and are widely applied in plant genetic engineering. The genes were expressed in Escherichia coli and the corresponding proteins were purified and used for functional and structural comparison. Both proteins are homodimers regardless of whether they are expressed in microorganisms or in plants. They have comparable molecular weights and show immuno-cross-reactivity to their respective polyclonal antisera. The enzymes have a similar substrate affinity towards L-phosphinothricin and do not acetylate any of the other L-amino acids tested. In model digestion experiments using simulated human gastric fluids, their enzymatic activity is decreased within seconds, accompanied by a rapid and complete breakdown of both proteins. These data demonstrate the structural and functional equivalence of the PAT proteins, which is also reflected in the comparable performance of transgenic plants carrying the bar or pat gene.
Acetyltransferases↗
EcoRV-T94V: a mutant restriction endonuclease with an altered substrate specificity towards modified oligodeoxynucleotides.
Synthetic oligodeoxynucleotides with single methyl phosphonate (mp) substitutions were used for an analysis of the contribution of phosphate contacts to the recognition of the cleavage site by the restriction endonuclease EcoRV. Only in the last position within the recognition sequence, is the methyl phosphonate substitution tolerated by the enzyme. The wild-type enzyme cleaves the Sp diastereomer of the oligodeoxynucleotide GACGATATmpCGTC and the unmodified sequence with equal rates, whereas the Rp diastereomer is cleaved much more slowly. Inspection of the crystal structure of an EcoRV-DNA complex revealed that the non-bridging oxygen atoms of the phosphodiester bond between the T and C bases are in hydrogen bonding distance of the hydroxyl group of the amino acid Thr94. We therefore tried to engineer a variant of EcoRV that would prefer a methyl phosphonate linkage over a normal phosphodiester bond and produced mutants with amino acid exchanges at position 94. One of them, Thr94Val, shows a dramatically reduced activity towards the unmodified DNA and does not accept the Rp diastereomer, but cleaves the Sp diastereomer with the same rate as wild-type EcoRV. Its selectivity, i.e. the ratio of cleavage rates determined for the unmodified and modified substrates, differs by three orders of magnitude from that of the wild-type enzyme.
DNA↗
Stereospecific production of the herbicide phosphinothricin (glufosinate): purification of aspartate transaminase from Bacillus stearothermophilus, cloning of the corresponding gene, aspC, and application in a coupled transaminase process.
We have isolated and characterized an aspartate transaminase (glutamate:oxalacetate transaminase, EC 2.6.1.1) from the thermophilic microorganism Bacillus stearothermophilus. The purified enzyme has a molecular mass of 40.5 kDa by sodium dodecyl sulfate gel analysis, a temperature optimum of 95 degrees C, and a pH optimum of 8.0. The corresponding gene, aspC, was cloned and overexpressed in Escherichia coli. The recombinant glutamate:oxalacetate transaminase protein was used in immobilized form together with 4-aminobutyrate:2-ketoglutarate transaminase (EC 2.6.1.19) from E. coli for the production of L-phosphinothricin [L-homoalanin-4-yl-(methyl)phosphinic acid], the active ingredient of the herbicide Basta (AgrEvo GmbH), from its nonchiral 2-keto acid precursor 2-oxo-4-[(hydroxy)(methyl)phosphinoyl]butyric acid (PPO). In this new coupled process conversion rates of ca. 85% were obtained with substrate solutions containing 10% PPO by using only slight excesses of the amino donors glutamate and aspartate. The contamination of the reaction broth with amino acid by-products was < 3%.
4-Aminobutyrate Transaminase↗
hrcA, the first gene of the Bacillus subtilis dnaK operon encodes a negative regulator of class I heat shock genes.
Whereas in Escherichia coli only one heat shock regulon is transiently induced by mild heat stress, for Bacillus subtilis three classes of heat shock genes regulated by different mechanisms have been described. Regulation of class I heat shock genes (dnaK and groE operons) involves an inverted repeat (CIRCE element) which most probably serves as an operator for a repressor. Here, we report on the analyses of an hrcA null mutant (delta hrcA), in which hrcA, the first gene of the dnaK operon, was deleted from the B. subtilis chromosome. This strain was perfectly viable at low and high temperatures. Transcriptional analysis of the deletion mutant revealed a high level of constitutive expression of both the dnaK and groE operons even at a low temperature. A further increase in the amount of groE transcript was observed after temperature upshift, suggesting a second induction mechanism for this operon. Overproduction of HrcA protein from a second copy of hrcA derived from a plasmid (phrcA+) in B. subtilis wild-type and delta hrcA strains prevented heat shock induction of the dnaK and groE operons at the level of transcription almost completely and strongly reduced the amounts of mRNA at a low temperature as well. Whereas the wild-type strain needed 4 h to resume growth after temperature upshift, the delta hrcA strain stopped growth only for about 1 h. Overproduction of HrcA protein prior to a heat shock almost completely prevented growth at a high temperature. These data clearly demonstrate that the hrcA product serves as a negative regulator of class I heat shock genes.
Amino Acid Sequence↗
Primary plasmacytoma of the testis. Case report and review of the literature.
A primary plasmacytoma of the testis is an extremely rare tumor. To date less than 10 cases have been published. Typically a firm mass of the testis or also of the epididymis is found. Histologically, infiltration can be seen with atypical plasma cells compressing the seminiferous tubuli. Immunohistochemical examination shows a PAS-positive reaction. Monoclonality can be confirmed by identification of the IgA lambda chain. Therapy of choice is the radical orchiectomy. The prognosis of primary plasmacytoma of the testis seems to be good. Nevertheless, a follow-up of some years is necessary to detect possible intramedullar manifestations.
Humans↗
Scalp necrosis in temporal (giant cell) arteritis:implications for the dermatologic surgeon.
Temporal arteritis, a variant of giant cell arteritis, is a systemic granulomatous vasculitis of large and medium-sized arteries. Usually the clinical features are dominated by ophthalmological and neurological complications. In rare instances, ischaemic necrosis, especially of the scalp, may lead patients to the dermatologist. We report a 76-year-old woman presenting with a unilateral scalp necrosis, accompanied by a dramatic ipsilateral impairment of vision. Immediately after duplex-sonography of the extracranial vessels and after initiation of corticosteroid therapy, the diagnosis of temporal arteritis was confirmed by temporal artery biopsy. One month later, because of insufficient secondary healing of the ulcer, the defect was covered by a mesh graft. The taking of the graft was delayed due to immunosuppressive therapy, but was complete. The patient unfortunately died as a result of complications related to surgical removal of an aspergilloma in the sphenoid cavity secondary to immunosuppressive therapy. We discuss the technique of artery biopsy and the possibility of surgical management of scalp necrosis in temporal arteritis.
Aged↗
Use of granulocyte colony-stimulating factor (filgrastim) in the treatment of non-cytotoxic drug-induced agranulocytosis.
Five patients with non-cytotoxic drug-induced agranulocytosis were treated with recombinant human granulocyte-colony-stimulating factor (rh-G-CSF). The drugs involved were dipyrone, captopril, clozapine and carbimazole. Bone marrow examination revealed a depleted granulopoiesis with normal erythro- and megakaryocytopoiesis. After discontinuation of the suspected drug, rh-G-CSF was administered daily at 5 microg/kg subcutaneously. The neutrophil counts were recovered between day 6 and 12 and patients were discharged from hospital shortly afterwards. Compared to data from the literature, the neutrophil recovery appeared to be faster than expected without the use of haematopoietic growth factors. In conclusion, rh-G-CSF at a standard dose of 5 microg/kg seems to be an effective treatment for drug-induced agranulocytosis.
Administration, Cutaneous↗
Grading of precancerous laryngeal lesions by multiparameter image analysis at separate epithelial layers.
In order to detect early precancerous changes which do not involve the whole thickness of the epithelium, we used a novel image analysing program based on an IBAS system (Kontron, Germany) to determine nuclear DNA content (NC) as well as average nuclear area (NA) and variation of nuclear area (VA), in the entire epithelium and in three sublayers, parabasal, intermediate, and superficial. DNA aneuploidy was found in only half of the cases classified as 'high-grade' (HG) lesions, comprising carcinoma in situ (CIS) and severe epithelial dysplasias (EDIII), and was chiefly demonstrable in the parabasal third of the epithelium. The other lesions were DNA euploid. HG lesions showed highly significant increases of NA and VA at the lower levels of the epithelium when compared with 'low-grade' (LG) lesions comprising moderate and mild epithelial dysplasias (EDII and EDI). Our data show that the combination of multiparameter image analysis with conventional morphology assists in the objective grading of precancerous lesions and permits the reliable detection of high-grade lesions.
Adult↗
"True bone" in vitro?
Explore the source record for details and available documents.
Bone Development↗
Genetic characterization of a new hantavirus detected in Microtus arvalis from Slovakia.
A new hantavirus, called Malacky, has been identified in lung tissue specimens of a vole, Microtus arvalis, by the reverse transcriptase polymerase chain reaction (RT-PCR). The voles were trapped in a geographical area in Slovakia where hemorrhagic fever with renal syndrome (HFRS) is endemic in the human population. Sequence analysis of a major part of the S segment showed this virus to represent a new subtype within Tula, a new hantavirus genetic group defined very recently.
Amino Acid Sequence↗
[Channel selection for EEG-monitoring in anesthesia].
The conventional multichannel electroencephalogram is quite inconvenient for long-term monitoring in the operating theatre or intensive care unit. Recording of the EEG would be easier if a small number of channels was sufficient. Aiming at reduction of channels, leads from different regions of the scalp were analysed visually and with regard to their spectral content. METHODS. Electrode placements corresponded to the International 10/20 System (Fig. 1). EEG recordings were made with a conventional device (ES 12,000), a personal computer, and a spectral analyser. TWO-CHANNEL RECORDINGS. Retrospective analysis was performed on data from 392 patients (age 14-90 years) whose anaesthesia was induced with various anaesthetics/narcotics, for instance thiopental, ketamine, etomidate, halothane, and enflurane. The EEG was recorded using C3-P3 and Cz-A1. For each patient the changes of spectral parameters during the course of the induction were plotted and visually analysed. For statistical analyses a 30-s epoch of each patient was randomly selected from the first few minutes after the beginning of induction. TEN-CHANNEL RECORDINGS. In ten gynaecological patients (age 26-55 years) EEG recordings were performed during induction of anaesthesia with thiopental in combination with fentanyl, N2O and O2. The set of channels consisted of Fz'-Cb1, F3'-Cb1, Cz-Cb1, C3-Cb1, P3-Cb1, Oz-Cb1, Fz'-F3', F3'-C3, C3-P3, and P3-Oz. The electrodes Fz' and F3' were positioned on the forehead near to Fz and F3, respectively. These sites were chosen because they allow easy application of electrodes. The relationship between channels was calculated with Bravais-Person's coefficient of correlation for the power and the absolute power in the frequency bands delta (0.5-3.5 Hz), theta (3.5-7.5 Hz), alpha (7.5-12.5 Hz), and beta (> 12.5 Hz). RESULTS. In visual and statistical analyses of the two- and ten-channel recordings under the influence of anaesthetics/narcotics, similar changes of EEG activity could be observed in all channels. Although differences in the absolute power of the frequency bands were present, there was high conformity in the composition of the spectral content of the different channels. Classification of the EEG into stages of anaesthesia by means of a single channel led to consistent results for all channels. Alpha activity as leading feature of the awake state predominated occipitally. In channels including the region around the ears, contamination with EKG artifacts was observed. CONCLUSIONS. EEG patterns under the influence of different anaesthetics/narcotics are adequately represented by a reduced number of channels. For the choice of an appropriate set of channels the following aspects should be considered. Contamination with artifacts should be as low as possible, electrode sites should easily be accessible, and special features of the awake state should be identifiable. Experience with routinely conducted EEG recordings in the operating theatre and the intensive care unit showed that the channels C3-P3 or C4-P4 provide a sufficient basis for automatic staging of the depth of anaesthesia, which is implemented in the EEG monitor Narkograph.
Adolescent↗
Adamantinoma of long bones. A histopathological and immunohistochemical study of 23 cases.
The clinical and histological data of twenty-three cases of adamantinomas of the long bone collected by the Working Group on Bone Tumors at the DKFZ/FRG are reported including immunohistochemical observations in twenty-one of the cases. Females and males between 5 and 67 years (mean, 25.4 years) were affected equally (11/12). All adamantinomas were positive for cytokeratins often in coexpression with vimentin, at least focally. Although exhibiting varying histological patterns, no correlation between histology and clinical course was seen. However, sex and mode of initial therapy seem to influence an unfavorable clinical outcome. All three decreased patients were males receiving marginal or delayed surgery. This underlines the low-grade malignant character of adamantinoma. To assure the histological diagnosis pathologists should employ immunohistochemistry for demonstrating the sometimes sparse epithelial cell nests when radiology is suggestive for adamantinoma. Correct diagnosis should lead to resection with wide surgical margins.
Adolescent↗
Isolation and analysis of mutants of the dnaK operon of Bacillus subtilis.
Bacillus subtilis contains at least three classes of heat-shock genes regulated by different mechanisms. We are studying class I heat-shock genes encoded by the operons dnaK and groE. These two operons are both expressed from a vegetative promoter, and their regulation involves a novel heat-shock element designated CIRCE. Here we show that induction of both operons results from enhanced synthesis of mRNA and is independent of de novo protein synthesis. To answer the question of whether dnaK is involved in the deregulation of the heat-shock response as reported for Escherichia coli, two different insertion mutations were isolated within the tetracistronic dnaK operon (orf39-grpE-dnaK-dnaJ). In one mutant a cat cassette was inserted at the beginning of orf39. Transcriptional analysis revealed that this mutation abolished expression of the whole operon. In contrast, the basal level of groE mRNA was significantly increased at 37 degrees C, followed by a prolonged delay in the shut off after temperature upshift. These data point to a crucial role for the orf39 gene in the regulation of class I heat-shock genes. In the other mutant an internal 0.8 kb Bg/II fragment of dnaK was replaced by the cat cassette. In contrast to E. coli dnaK null mutants, the two B. subtilis dnaK operon mutants could grow within a temperature range from 16-52 degrees C. At temperatures above 52 degrees C, they failed to form colonies on agar plates, started to filament, and lost motility. Furthermore, the induction profile of the groE and dnaK operons was not impaired in the dnaK::cat mutant.(ABSTRACT TRUNCATED AT 250 WORDS)
Bacillus subtilis↗
Aerosol bolus dispersion and effective airway diameters in mildly asthmatic children.
The contribution of aerosol techniques, the estimation of aerosol bolus dispersion and effective airway dimensions, to the clinical diagnosis of paediatric asthma was studied. In 47 children, aged 11 +/- 2 yrs, with mild asthma (forced expiratory volume in one second (FEV1) 83 +/- 9% of forced vital capacity (FVC)) effective airway diameters were derived from the recovery of inhaled 1 micron sebacate droplets. Intrapulmonary dispersion of inhaled boluses of 0.4 micron droplets was studied, by characterizing the concentration distributions of droplets in the exhaled air by their standard deviation and skewness. Effective airway diameters increased in asthmatic subjects with increasing body size, and did not differ from those obtained in 16 healthy children of similar age and height. Standard deviation and skewness of particle boluses exhaled from shallow lung depths were higher in the asthmatic children than the healthy children (e.g. standard deviation 91 +/- 17 ml vs 79 +/- 15 ml, skewness 0.38 +/- 0.16 vs 0.23 +/- 0.16, respectively, for boluses in 140 ml lung depth). The sensitivity and specificity of bolus dispersion to detect alterations in lung function was comparable to that of FEV1/FVC, the most sensitive conventional lung function parameter in the present study. There was no correlation between body height or lung function and bolus parameters. We conclude that aerosol measurements do not provide an obvious benefit for the clinical diagnosis of mild paediatric asthma, but bolus dispersion supplies additional information on alterations in convective gas transport in the diseased lung.
Aerosols↗
Immune electron-microscopy study of the trimeric glycoprotein (CD 103) of hairy cell leukemia using the Ber-ACT8 monoclonal antibody.
We describe a method which makes it possible to study the expression of the trimeric glycoprotein (TGP, CD 103) on the surface of hairy cells (HCs) by electron microscopy (e.m.). The monoclonal antibody (mAb) Ber-ACT8 was used to identify the TGP. By testing mucosa of the small intestine, we found that a prefixation incubation with Ber-ACT8 was necessary. After isolation with a density gradient, Ber-ACT8 incubation, and fixation, the peroxidase-anti-peroxidase (PAP) method was used to make the TGP visible. The HCs showed a clearly discernible stain on their surface, a fairly constant distribution of the TGP, and a very good ultrastructural preservation. This method makes it possible to demonstrate the TGP or other unfixable antigens by e.m.
Aged↗
Pulmonary dead space and airway dimensions in dogs at different levels of lung inflation.
The quasi-stationary front that separates inspired gas from mixed alveolar gas is largely determined by the balance between diffusive and convective forces of gas transport. To investigate parameters influencing this balance, a study was performed on eight anesthetized ventilated beagle dogs. Measurements were made of the volume of pulmonary dead space corresponding to four end-inspiratory lung volumes. Aerosol recovery techniques were used to determine airway sizes at lung depths corresponding to those respective dead space volumes as well as at fixed volumetric depths between 70 and 250 ml. Mean dead space volumes as measured by a single inhalation of He (and SF6) were 112 +/- 15 (SD) ml (127 +/- 15 ml), 120 +/- 18 ml (137 +/- 20 ml), 127 +/- 18 ml (145 +/- 20 ml), and 133 +/- 19 ml (155 +/- 21 ml) at end-inspiratory lung volumes of 64, 71, 79, and 86%, respectively, of total lung capacity. At fixed lung depths the airway diameters increased with higher levels of lung inflation. However, airway diameters "at the end of the dead space" did not change significantly. They were approximately 0.5 mm for SF6 dead space and approximately 0.75 mm for He dead space. These findings support the theoretical prediction that the position of the diffusion front during breathing is strongly dependent on airway geometry and much less dependent on parameters of the breathing maneuver.
Animals↗