Biomedical subjects

A Schmidt

Publications and source records attributed to A Schmidt.

At least 289 records · Page 16Linked to original sources

PubMed1996

A newly identified Minute locus, M(2)32D, encodes the ribosomal protein L9 in Drosophila melanogaster.

A gene encoding a ubiquitously expressed mRNA in Drosophila melanogaster was isolated and identified as the gene for ribosomal protein L9 (rpL9) by its extensive sequence homology to the corresponding gene from rat. The rpL9 gene is localized in polytene region 32D where two independent P element insertions flanking the locus are available. Remobilization of either P element generated lines with a typical Minute phenotype, e.g. thin and short bristles, prolonged development, and female semisterility in heterozygotes as well as homozygous lethality. All these characteristics can be rescued when a 3.9 kb restriction fragment containing the rpL9 gene is reintroduced by P element-mediated germline transformation. This result confirms that M(2)32D codes for ribosomal protein L9.

Amino Acid Sequence↗

PubMed1996

Protein-tyrosine phosphatase activity regulates osteoclast formation and function: inhibition by alendronate.

Alendronate (ALN), an aminobisphosphonate used in the treatment of osteoporosis, is a potent inhibitor of bone resorption. Its molecular target is still unknown. This study examines the effects of ALN on the activity of osteoclast protein-tyrosine phosphatase (PTP; protein-tyrosine-phosphate phosphohydrolase, EC 3.1.3.48), called PTPepsilon. Using osteoclast-like cells generated by coculturing mouse bone marrow cells with mouse calvaria osteoblasts, we found by molecular cloning and RNA blot hybridization that PTPepsilon is highly expressed in osteoclastic cells. A purified fusion protein of PTPepsilon expressed in bacteria was inhibited by ALN with an IC50 of 2 microM. Other PTP inhibitors--orthovanadate and phenylarsine oxide (PAO)-inhibited PTPepsilon with IC50 values of 0.3 microM and 18 microM, respectively. ALN and another bisphosphonate, etidronate, also inhibited the activities of other bacterially expressed PTPs such as PTPsigma and CD45 (also called leukocyte common antigen). The PTP inhibitors ALN, orthovanadate, and PAO suppressed in vitro formation of multinucleated osteoclasts from osteoclast precursors and in vitro bone resorption by isolated rat osteoclasts (pit formation) with estimated IC50 values of 10 microM, 3 microM, and 0.05 microM, respectively. These findings suggest that tyrosine phosphatase activity plays an important role in osteoclast formation and function and is a putative molecular target of bisphosphonate action.

Alendronate↗

PubMed1996

Accelerated immune response in transgenic mice expressing rat CD44v4-v7 on T cells.

Splice variants of the glycoprotein CD44 (CD44v) that confer metastatic behavior to noninvasively growing rat tumor cells are transiently expressed on lymphocytes during activation. A mAb directed against an epitope encoded by CD44 exon v6 blocks both metastasis formations and lymphocyte activation, implicating CD44v in normal immune function. To explore the nature of this function of CD44v, transgenic mice were generated that constitutively express rat CD44v4-v7 on thymocytes and peripheral T cells. The number of lymphocytes as well as the distribution of lymphocyte subpopulations were similar in nontransgenic and rat CD44v4-v7 transgenic mice. T cells of the transgenic mice, however, responded faster to activating stimuli, particularly during primary stimulations with T cell mitogens and T-dependent Ags in vivo and in vitro. This accelerated response depended on the expression of the transgene product, since a rat CD44v6-specific Ab reverted the response profiles of the transgenic mice to those of nontransgenic mice. Since the transgene gained in vivo and in vitro functional activity only upon antigenic stimulation, it is likely that CD44 variant isoforms are involved in the process of signal transduction during lymphocyte activation.

Animals↗

PubMed1996

Structural constraints on the ternary complex of 5-enolpyruvylshikimate-3-phosphate synthase from rotational-echo double-resonance NMR.

The 46 kDa enzyme 5-enolpyruvylshikimate-3-phosphate (EPSP) synthase catalyzes the condensation of shikimate-3-phosphate (S3P) and phosphoenolpyruvate to form EPSP. The reaction is inhibited by N-(phosphonomethyl)-glycine (Glp), which in the presence of S3P, binds to EPSP synthase to form a stable ternary complex. As part of a solid-state NMR characterization of this structure, 15N labels were introduced selectively into the lysine, arginine and histidine residues of EPSP synthase and distances to a 13C label in Glp and to the 31P in S3P and Glp were measured by rotational-echo double-resonance NMR. Three lysine and four arginine residues are in the proximity of the phosphate group of S3P and the carboxyl and phosphonate groups of Glp. A single histidine residue is in the vicinity of the binding site (closer to Glp than to S3P) but is more distant than the lysine and arginine residues.

3-Phosphoshikimate 1-Carboxyvinyltransferase↗

PubMed1996

Tyrosinase is a new marker for cell populations in the mouse neural tube.

Tyrosinase, the key enzyme in melanin synthesis, is expressed in pigment cells derived from both neural crest and neuroectoderm. The present study was performed to detect tyrosinase promoter activity and tyrosinase gene expression during murine brain development. Mouse tyrosinase 5' region (6.1 Kb) was used to direct lacZ expression in transgenic mice. During embryogenesis, the transgene reproduced tyrosinase expression in pigment cells but was also observed in embryonic neuroectoderm and migrating neural crest cells. Both tyrosinase and lacZ were detected in cell populations often organized in columnar arrangements and found throughout the entire neural tube, in the cranial region as well as in the spinal chord. In the developing brain, the highest density of positive cells was localized to ventricular and subventricular zones and to evaginations of the neural tube such as optic vesicle, pineal gland, and olfactory bulbs. These results demonstrate that tyrosinase promoter activity and tyrosinase expression are not restricted to differentiated pigment cells. We suggest that tyrosinase is a new marker for cell populations in the neural tube, and that expression is correlated to regions undergoing rapid cell proliferation.

Animals↗

PubMed1996

Profile characteristics of the MPI-DLV clusters of pain patients.

This article describes an investigation into whether there are differences among the four pain patient clusters from the Multidimensional Pain Inventory-Dutch Language Version (MPI-DLV) in terms of prescribed analgesic medication, level of intelligence, and other personality characteristics (MMPI). Dysfunctional patients were distinguished by a higher level of analgesic medication intake. Patients classified as Average were marked by a marginally significant higher level of intelligence. The personality profile of Interpersonally Distressed patients indicated an elevated neurotic triad and a tendency to a passive-aggressive personality structure. Patients classified as Dysfunctional and Average were marked by the so-called conversion-V. There were no elevated scales in the Adaptive Coper profile. Clinical implications are discussed.

Adaptation, Psychological↗

PubMed1996

Human protein tyrosine phosphatase-sigma: alternative splicing and inhibition by bisphosphonates.

Two forms of the transmembrane human protein tyrosine phosphatase (PTP sigma), generated by alternative splicing, were identified by cDNA cloning and Northern hybridization with selective cDNA probes. The larger form of PTP sigma is expressed in various human tissues, human osteosarcoma, and rat tibia. The hPTP sigma cDNA codes for a protein of 1911 amino acid residues and is composed of a cytoplasmic region with two PTP domains and an extracellular region that can be organized into three tandem repeats of immunoglobulin-like domains and eight tandem repeats of fibronectin type III-like domains. In the brain, the major transcript of PTP sigma is an alternatively spliced mRNA, in which the coding region for the fibronectin type III-like domains number four to seven are spliced out, thus coding for a protein of 1502 amino acid residues similar to the rat PTP sigma and rat PTP-NE3. Using in situ hybridization, we assigned hPTP sigma to chromosome 6, arm 6q and band 6q15. The bacterial-expressed hPTP sigma exhibits PTPase activity that was inhibited by orthovanadate (IC50 = 0.02 microM) and by two bisphosphonates used for the treatment of bone diseases, alendronate (ALN) (IC50 = 0.5 microM) and etidronate (IC50 = 0.2 microM). In quiescent calvaria osteoblasts, micromolar concentrations of vanadate, ALN and etidronate stimulate cellular proliferation. These findings show tissue-specific alternative splicing of PTP sigma and suggest that PTPs are putative targets of bisphosphonate action.

Alternative Splicing↗

PubMed1996

Identification of fatty acid methyl ester as naturally occurring transcriptional regulators of the members of the peroxisome proliferator-activated receptor family.

The nuclear hormone receptors NUC-1 (PPAR delta) and PPAR alpha are members of the peroxisome proliferator-activated receptor (PPAR) family. The members of this receptor family are activated by agents that stimulate peroxisome proliferation, free fatty acids, prostaglandin 12 metabolites, and agents considered for the therapy of insulin-independent diabetes mellitus. To identify putative physiological agents that activate NUC-1, we tested the ability of acetone extracts of various rat tissues to activate the transcription of an MMTV-luciferase reporter gene, via a GR/NUC-1 hybrid receptor. GR/NUC-1 contains the ligand binding region of the NUC-1 receptor and the DNA binding domain of the glucocorticoid receptor. Using this assay, we found stimulatory activity in the pancreas, which upon purification and characterization was identified as methyl-palmitate, known to be enriched in pancreatic lipids. In addition, we determined that ethyl esters of palmitic and oleic acids are also potent activators of this receptor. Thus, fatty acid ester formation may control the cellular concentrations of fatty acids, and acyl-ester formation may play a role in the control of metabolic pathways and the activation of the PPAR.

Animals↗

PubMed1996

Equipment configuration and procedures: preferences for interventional microtherapy.

Magnetic resonance imaging (MRI), computed tomography, and electron beam tomography scanners are built for radiologic diagnosis. With increasing frequency they are being used in the field of interventional Microtherapy to permit transparent visualization of the therapeutic field. Each of these scanners can be combined with endoscopy, fluoroscopy/digital subtraction angiography, and ultrasound units for hybrid imaging techniques as well as with therapeutic systems like lasers or radiofrequency. MRI affords 3D localization without x-ray exposure. Open access and keyhole imaging allow nearly real time guidance of instruments. Minimally invasive techniques using endoscopes and hybrid tomographic guidance result in improved tip tracking of microinstruments and reduced complications. This safer access into the body will lead to interdisciplinary cooperation with the potential for large cost reductions. This report summarizes our experience regarding which of the hybrid imaging suites is best suited for procedures including among others drug instillations, prosthesis (stent) implantation, or microoperations (endoscopic diskectomy/sequestrectomy), and physiological measurements simultaneously.

Diagnostic Imaging↗

PubMed1996

[Current status of treatment of Achilles tendon ruptures. Results of a nationwide survey in Germany].

In a standard questionnaire distributed nationwide, we questioned staff from 1307 clinics (surgical, trauma-surgical, orthopedic), of which 787 clinics (60.2%) answered by letter. In 698 clinics (88.7%), operation is considered the standard therapy. Seventy-five clinics (9.5%) use both conservative and operative therapy (average postoperative rate of complication 3.5%). Only 14 hospitals (1.8%) treat strictly conservatively. The average rate of reruptures is 1.6% in the operated group (77.7% postoperative plaster cast fixation) regardless of postoperative treatment; the conservative group (96.1% functional treatment) showed 2.7% reruptures. Thus, operation is the standard therapy for fresh ruptures of the Achilles tendon in Germany. However, there seems to be a trend towards conservative functional therapy.

Achilles Tendon↗

PubMed1996

Human recombinant insulin-like growth factor I and -II stimulate the expression of basic fibroblast growth factor but suppress the division of bovine coronary smooth muscle cells.

Insulin-like growth factor I and II (IGF-I and -II)--two 7.65- and 7.47-kDA polypeptides belonging to the somatomedine family--are regular constituents of human blood plasma. Both factors exert mitogenic activity on a variety of cell types including arterial smooth muscle cells. In the present study, the effect of IGF-I and -II on cultured bovine coronary smooth muscle cells (cSMC) was assessed. Human recombinant IGF-I and IGF-II added to cSMC cultured in a medium containing 10% fetal bovine serum (FBS) decreased the cell number and [3H]thymidine incorporation in a dose dependent fashion up to 40% and 43% compared to control cells (100%). At the same time, the expression of basic fibroblast growth factor (bFGF) increased from 60 pg/5 x 10(4) cells (control) to 75 (IGF-I) and 113 pg/5 x 10(4) cells (IGF-II). In parallel with enhanced bFGF expression, the bFGF receptor content per cell and the [35S]sulfate incorporation into extracellular and cell-associated proteoglycans also increased under the influence of IGF-I and -II. In contrast, with low serum concentration (0.1% FBS) the addition of IGF-I and -II to bovine cSMC cultures resulted in a slight increase in cell number, protein content and [3H]thymidine incorporation as described in previous studies. These results suggest that the mitogenic activity of IGF-I and -II towards coronary smooth muscle cells depends on culture conditions. In the presence of 10% fetal bovine serum that mimics in vivo conditions, IGF-I and -II did not necessarily act as mitogenic factors but inhibited the proliferation of cSMC in vitro possibly by modulating antagonizing the action of other growth factors. Irrespective of the inhibition of cell division, the cellular bFGF, the bFGF receptor and the bFGF activity-related proteoheparan sulfate were overexpressed under the influence of IGF.

Animals↗

PubMed1996

Characterization of Escherichia coli isolated from foals.

Serotype, biotype, antibiogram, hemolysin production, fimbrial hemagglutinins, select toxin genes (STb, STaP, LT, slt1 and slt2) and the attaching effacing (eae) gene were determined for 99 foal strains of E. coli. E. coli from diarrheic and normal foals could not be distinguished by serotype, biotype, or antibiogram. Differences (P < or = 0.05) were observed in hemolysin production (11.5% vs 0%) and the expression of mannose-resistant hemagglutinins (23% vs 13%) among E. coli from diarrheic and healthy foals, respectively. Three of the E. coli strains from diarrheic foals were positive with probes for slt genes and one was positive for STb and LT genes. One strain from a healthy foal possessed the STb gene. As determined by the polymerase chain reaction, 8 strains possessed the eae gene. Seven of the 8 strains were from diarrheic foals and one eae-positive strain was from a healthy foal. The slt-positive strains did not possess eae genes and the eae-positive strains did not possess slt genes. These results indicate that enterotoxigenic strains of E. coli are not implicated in any substantial degree in sporadic foal diarrhea. However, the identification of slt-positive and eae-positive strains in foal feces indicate the presence of potentially virulent strains among foals.

Animals↗

PubMed1996

Microbial biosensor for free fatty acids using an oxygen electrode based on thick film technology.

A microbial biosensor based on thick film technology was developed. The microorganisms, Arthrobacter nicotianae, were immobilized in Ca-alginate directly on the electrode surface. For the stability of the calcium alginate gel the addition of 0.5 mM CaCl2 to the assay buffer was necessary. The respiratory activity of the microorganisms was monitored by oxygen consumption at -600 mV vs. Ag/AgCl reference electrode. The sensor was used in a batch system and was applied to the determination of free fatty acids in milk. Short-chain fatty acids (C4:0-C12:0) were the preferential substrates, with butyric acid being the main substrate. Consequently, the concentration of free short-chain fatty acids was represented as the butyric acid equivalent. The sensor showed linearity over the concentration range 9.5-165.5 microM (correlation coefficient, r = 0.99920). The response time of the sensor was approximately 3 min. No additional dialysis membrane was necessary, which led to a high sensitivity of the sensor and fast response times. Recovery rates of 98-113% were found for butyric acid in milk samples using the sensor without any additional membrane and a sample dilution of 200 by the assay. Two widespread disadvantages of microbial sensors, long response times and long times to return to the baseline signal after use, could be overcome.

Acyl-CoA Oxidase↗

PubMed1996

Differentiation processes in the amphibian brain with special emphasis on heterochronies.

Amphibians and caecilians exhibit a great variety of adult morphologies, life histories, and developmental strategies (biphasic development, direct development, viviparity, and neoteny). While early brain development and the differentiation of neural tissues in the three amphibian orders follow a basic pattern, differences exist in the onset and offset as well as the rate of growth and differentiation processes. These differences are described within a phylogenetic framework, and special emphasis is laid on the relationship between altered ontogenies and phylogenetic diversity. We concentrate on ontogenetic differentiation processes in the motor, olfactory, and visual system. We discuss the morphological consequences of secondary simplification of the brain in the context of paedomorphosis, which has happened several times independently among amphibians and consists in the abbreviation or truncation of late developmental processes. We deal with the cellular and molecular basis of brain development and the consequences for the adult nervous system in representative species of the three amphibian orders. Our analysis reveals that differences in brain morphology are largely due to heterochrony (i.e., the desynchronization of ontogenetic processes), a phenomenon that in turn is related to changes in genome sizes and life histories.

Amphibians↗

PubMed1996

Contribution of the intramolecular disulfide bridge to the folding stability of REIv, the variable domain of a human immunoglobulin kappa light chain.

BACKGROUND: Immunoglobulin domains contain about 100 amino acid residues folded into two beta-sheets and stabilized in a sandwich by a conserved central disulfide bridge. Whether antibodies actually require disulfide bonds for stability has long been a matter of debate. The contribution made by the central disulfide bridge to the overall folding stability of the immunoglobulin REIv, the variable domain of a human kappa light chain, was investigated by introducing stabilizing amino acid replacements followed by removal of the disulfide bridge via chemical reduction or genetic substitution of the cysteine residues. RESULTS: Nine REIv variants were constructed by methods of protein engineering that have folding stabilities elevated relative wild-type REIv by (up to) 16.0 kJ mol-1. Eight of these variants can be cooperatively refolded after unfolding and chemical reduction of the disulfide bridge-in contrast to wildtype REIv. The stabilizing effect of one of these residue replacements (T39K) was rationalized by determining the structure of the respective REIv variant at 1.7 A. The loss of folding stability caused by reduction of the intramolecular disulfide bond is on average 19 kJ mol-1. Removal of the disulfide bridge by genetic substitution of C23 for valine resulted in a stable immunoglobulin domain in the context of the stabilizing Y32H amino acid exchange; again, REIv-C23V/Y32H has 18 kJ mol-1 less folding stability than REIv-Y32H. The data are consistent with the notion that all variants studied have the same overall three-dimensional structure with the disulfide bridge opened or closed. CONCLUSIONS: A comparison of the magnitude of the stabilizing effect exerted by the disulfide bond and the length of the mainchain loop framed by it suggests lowering of the entropy of the unfolded state as the sole source of the effect. Disulfide bonds are not necessary for proper folding of immunoglobulin variable domains and can be removed, provided the loss of folding stability is at least partly compensated by stabilizing amino acid exchanges.

Disulfides↗

PubMed1996

Early death due to defective neonatal lung liquid clearance in alpha-ENaC-deficient mice.

The amiloride-sensitive epithelial sodium channel, ENaC, is a heteromultimeric protein made up of three homologous subunits (alpha, beta and gamma) (1,2). In vitro, assembly and expression of functional active sodium channels in the Xenopus oocyte is strictly dependent on alpha-ENaC--the beta and gamma subunits by themselves are unable to induce an amiloride-sensitive sodium current in this heterologous expression system (2). In vivo, ENaC constitutes the limiting step for sodium absorption in epithelial cells that line the distal renal tubule, distal colon and the duct of several exocrine glands. The adult lung expresses alpha, beta and gamma ENaC (3,4), and an amiloride-sensitive electrogenic sodium reabsorption has been documented in upper and lower airways (3-7), but it is not established whether this sodium transport is mediated by ENaC in vivo. We inactivated the mouse alpha-ENaC gene by gene targeting. Amiloride-sensitive electrogenic Na+ transport was abolished in airway epithelia from alpha-ENaC(-/-) mice. Alpha-ENaC(-/-) neonates developed respiratory distress and died within 40 h of birth from failure to clear their lungs of liquid. This study shows that ENaC plays a critical role in the adaptation of the newborn lung to air breathing.

Animals↗

PubMed1996

The Politano-Leadbetter antireflux plasty. Investigation of complications in 245 children.

From 1981 to 1988 245 children aged 2 months to 15 years underwent ureteral reimplantation in 326 renal units in our department. The VUR was primary in 180 patients (253 ureters) and 65 patients (73 ureters) showed associated pathologies. The follow-up program, including physical examination, urine sampling and isotope studies (including NMCU), was from 4 to 11 years after surgery. Intraoperative and early surgical complications were found in 29 cases, but no complications affected the late results, especially kidney function. Late complications have been observed in 15 cases, while in 10 cases reoperation because of persistent reflux (n = 4) obstruction (n = 3) and loss of kidney function (nephrectomy n = 3) was required. In 4 cases bladder stones required ESWL, and one patient developed hypertension.

Adolescent↗

PubMed1996

Influence of the mechanism of regurgitation on the quantification of mitral regurgitation by the proximal flow convergence method and the jet area method.

In 84 patients mitral regurgitation was quantified by angiography. The mechanism of regurgitation was determined by echocardiography (organic, n = 54, functional, n = 30). The radii of the proximal isovelocity surface areas in the flow convergence region for 28 and 41 cm.s-1 blood flow velocity and the area and length of the regurgitant jet were measured using colour flow Doppler imaging. The radii of the proximal isovelocity surface areas correlated more closely with the angiographic grade than the jet parameters irrespective of the mechanism of regurgitation. In more than 90% of the patients, grades I-II mitral regurgitation were correctly differentiated from grades III-IV by means of the radii of the proximal isovelocity surface areas. Using the jet parameters, the differentiation was correct in 50-90% of the patients depending on the mechanism of regurgitation. The jet area method particularly failed to identify grades III-IV organic mitral regurgitation due to a high prevalence of eccentric jets in these patients. It is concluded that the proximal flow convergence method was suitable for the quantification of mitral regurgitation irrespective of the mechanism of mitral regurgitation. On the other hand, the value of the jet area method depended largely on the regurgitation mechanism.

Adult↗