Observation of the shadows of the Moon and Sun using 100 TeV cosmic rays.
Explore the source record for details and available documents.
Journal Article↗
Biomedical subjects
D Müller
Publications and source records attributed to D Müller.
Characterization and direct glycoform profiling of a hybrid plasminogen activator by matrix-assisted laser desorption and electrospray mass spectrometry: correlation with high-performance liquid chromatographic and nuclear magnetic resonance analyses of the released glycans.
Mass spectrometric analysis of the recombinant hybrid plasminogen activator K2tu-PA at the native glycoprotein and corresponding protein level (high mass) is described. For gross structural characterization of the major glycotypes present, a combination of enzymatic degradation and matrix-assisted laser desorption mass spectrometric analysis of the products proved convenient. In this way, mono- and di-N-glycosylated one- and two-chain molecules, unresolved in the spectra of native material, were identified, with the one-chain type monoglycosylated at Asn247 representing the major component. Actual detection of resolved original glycoforms, or unresolved groups thereof when composed of isobaric species, and their assignments regarding antennicity and degree of sialylation were possible by electrospray mass spectrometry for the major monoglycosylated one-chain species. Desialation also allowed detection of the diglycosylated one-chain species as a minor constituent. The electrospray mass spectrometric results were correlated with structural and quantitative data available from a parallel high-performance liquid chromatographic and 1H nuclear magnetic resonance study performed at the glycan (low mass) level on liberated individual carbohydrate components. The results of the two studies showed full consistency in all respects amenable to evaluation, i.e. excellent agreement for the assignments of antennicity and degree of sialylation of the major glycan components, and good agreement for the determination of proportions in which these were present in the major monoglycosylated glycoprotein. This provided a sound basis for direct glycoform profiling by electrospray mass spectrometry. Typical applications to batch quality control and to structural characterization of non-standard material are shown.
Amino Acid Sequence↗
Desensitization of brain insulin receptor. Effect on glucose/energy and related metabolism.
The overall majority of cases of Alzheimer disease are not caused by genetic abnormalities. A pluricausal etiology is assumed, and the age factor may be of pivotal significance. Aging leads to inherent changes in basic metabolic principles, including the functionally most important cerebral glucose/energy metabolism. Experimentally induced perturbation of the neuronal control over the glucose metabolism by means of intracerebroventricular administration of streptozotocin leads to cascade-like abnormalities in glucose breakdown and energy formation and in membrane phospholipid and monoaminergic catecholamine metabolism, which closely resemble the disturbances found in sporadic Alzheimer disease. It is concluded that this model is a good tool for in vivo study of the cellular events characteristic for this human neurodegenerative disorder.
Aging↗
Role of the cytoskeleton in the regulation of Cl- channels in human embryonic skeletal muscle cells.
The effects of volume change and cytoskeleton manipulation on the Cl- channels in human embryonic skeletal muscle cells were studied. Trypsination, used for production of myoballs, changes the channel properties only a little. When the external osmolarity was reduced from 300 to 270 mosmol/l, the specific Cl- conductance, gCl, (at -80 mV) of myoballs increased from 5.1 +/- 1.9 to 30.4 +/- 12.2 microS/cm2 (SD; n = 6) within 15 min. Concomitantly, the kinetics of Cl- currents, elicited by clamping the membrane potential from a negative to positive values, changed from activation and subsequent slow inactivation to instantaneous activation with fast inactivation. G protein activation, protein kinase action or [Ca2+]i elevation seemed not to be involved in these effects. Similar changes were produced in the absence of a transmembrane osmotic gradient by 500 nM intracellular cytochalasin D (gCl = 34.3 +/- 10.3 microS/cm2; n = 6) or 12.5 microM colchicine (gCl = 15.4 +/- 1.4 microS/cm2; n = 5). When the external osmolarity was increased to 418 mosmol/l, 1 microM cytochalasin D did not affect gCl. In four of six cell-attached patches the open probability of the intermediate Cl- channel was increased after reduction of the bath osmolarity. In inside-out patches, the drugs increased the open probability of the channels. It is concluded that the Cl- channels are under control of the cytoskeleton.
Calcium↗
Natriuretic peptides elevate cyclic 3',5'-guanosine monophosphate levels in cultured rat pinealocytes: evidence for guanylate cyclase-linked membrane receptors.
Cyclic GMP formation in the rat pinealocyte has generally been thought to involve guanylate cyclases (GC) which are activated via GTP-regulatory proteins following beta 1-adrenergic receptor stimulation. Recent studies have also pointed to a cytosolic GC in these cells whose activity can be elevated by nitric oxide donors. Little attention has been paid to the possibility that pinealocytes might express membrane-bound GC in the form of natriuretic peptide receptors. The present report demonstrates functional membrane GC in rat pinealocytes by (1) cross-linking analyses with radiolabelled atrial natriuretic peptide (ANP); (2) reverse transcriptase polymerase chain reaction (RT-PCR) and DNA blot hybridization with probes for both the GC-A and GC-B forms of the natriuretic receptor; and (3) monolayer cell cultures of pinealocytes, which accumulate cGMP in response to ANP and its related peptides. As the role for cGMP in the rat pineal gland does not appear to be directly coupled to the synthesis of melatonin, the natriuretic peptides may have other regulatory functions in this neuroendocrine tissue.
Affinity Labels↗
[Mechanical strength and chemical stability of biodegradable block-polymerized and injection molded poly-L-lactide in vitro].
Biodegradable poly-L-lactic acid rods made of block-polymerized material (BP; molecular weight 550,000) and injection-moulded rods with high (SGI; molecular weight 121,000) and low (SGA, molecular weight 118,000) molecular orientation were compared 2, 4 and 6 weeks after incubation in enzyme solutions with high hydrolytic activity (esterase, alpha chymotrypsin and peptidase) and in buffer solution (TRIS buffer). The molecular weight, modulus of elasticity, bending strength (three-point bending test), and cyclic bending load to failure applied to the rods in a newly developed testing machine (1 Hz, maximum 100,000 cycles) were compared. The molecular weight of BP material decreased to 36% after 2 weeks, in contrast to the injection-moulded materials, in which it decreased only to 66% even after 6 weeks. The bending strength of all specimen decreased significantly faster in alpha chymotrypsin than in the other media (MANOVA, P < 0.001). SGI had a significantly higher bending strength than SGA, and SGA a higher strength than BP. There was no difference after incubation in the other two enzymatic solutions. BP lost 80% of its initial bending strength (140 N/mm2) after 6 weeks, and SGI and SGA (120 N/mm2) only 20%. Under permanent cyclic loading BP initially resisted 100,000 cycles with an applied cyclic load of 12.5 N/mm2, decreasing after 6 weeks to only 9,500 cycles, in contrast to SGI and SGA, which resisted to 46,000 cycles. There was, however, no relevant difference in the mechanical characteristics of the two injection-moulded rods. These results confirm that BP is degraded significantly faster than SGA and SGI.(ABSTRACT TRUNCATED AT 250 WORDS)
Biodegradation, Environmental↗
[Inadequate therapeutic effects of implantable cardioverter/defibrillators: etiology, therapy and prevention].
With the increasing number of implantable cardioverter-defibrillator (ICD) implantations in patients with sustained ventricular tachyarrhythmias, there is a growing interest in typical complications associated with this therapy. We analyzed the reasons and the incidence of inadequate therapy deliveries in 100 patients with epicardial (n = 27) or transvenous (n = 73) ICDs during a follow-up period of 10 +/- 8 months. A total of 21 of 100 patients received inadequate therapies. The most common reason was sinus tachycardia in eleven patients. Additional unnecessary shocks were avoided by reprogramming and application of beta-blockers. Lead failures caused the erroneous detection and defibrillation of ventricular fibrillation without any preceding clinical symptoms in four patients with an epicardial and in one patient with a transvenous ICD. All patients underwent successful surgical revision of their system. Atrial fibrillation with rapid ventricular response triggered inadequate shocks in four patients. Following digoxin administration no patient had additional inadequate shocks. In one patient non-sustained tachycardias caused unnecessary defibrillations. These results demonstrate that inadequate defibrillations are a common complication in patients after ICD-placement. The performance of x-ray, stress testing, and Holter monitoring on a regular basis may facilitate early diagnosis of possible reasons for unnecessary therapy deliveries. The improvement of detection and memory functions in future ICD-generations appears to be mandatory as well.
Adolescent↗
Critical currents and microstructure of melt-textured stoichiometric YBa2Cu3O7- delta crystallites.
Explore the source record for details and available documents.
Journal Article↗
Two different mRNAs from rat liver code for the transport of bumetanide and taurocholate in Xenopus laevis oocytes.
The aim of this study was to elucidate whether bumetanide, which is a competitive inhibitor of carrier mediated bile acid uptake in liver cells, is transported by bile acid carriers. The expression of hepatocellular transport proteins for bile acid uptake and the uptake of the loop diuretic bumetanide was therefore studied in Xenopus laevis oocytes by injection of rat liver poly(A)(+)-RNA. Three hours after injection, a 70% increase in [3H]taurocholate uptake versus noninjected oocytes was accompanied by an increase in only 24% in the uptake of [3H]bumetanide. Size fractionation of the poly(A)(+)-RNA yielded 33 mRNA fractions of which fraction 21 accounted for an 800% increase of taurocholate transport with only a slight increase in bumetanide uptake. Bumetanide transport was coded by mRNA-fraction 18, which stimulated uptake by 160-200% with a concomitant small increase in taurocholate uptake. Uptake of cholate was induced by both mRNA fractions with almost 2.5 fold greater expression by the bumetanide fraction. Oocyte transport of taurocholate (expressed by fraction 21) and bumetanide transport (expressed by fraction 18) were characterized in terms of Na+ dependency, inhibition by 4,4'-diisothiocyano-1,2-diphenylethane-2,2'-disulfonic acid (DIDS) and mutual competition. The results indicate that the bumetanide transporter mRNA is clearly different from the mRNA for the taurocholate transport protein. The mRNA fraction 18 was used for the construction of a cDNA library.
Animals↗
Cysteine phosphorylation of the glucose transporter of Escherichia coli.
The glucose transporter (IIBCGlc/IIAGlc complex) of the bacterial phosphotransferase system couples vectorial translocation to phosphorylation of the transported sugar. The IIAGlc subunit transfers the phosphoryl group from the phosphoryl carrier protein P-HPr to the IIBCGlc subunit. IIBCGlc translocates and phosphorylates glucose. The site of IIBCGlc phosphorylation is cysteine 421 as shown by mass spectrometric and biochemical analyses of phosphorylated peptides. Site-directed mutagenesis of Cys421 (C421S) afforded a stable but completely inactive protein (Nuoffer, C., Zanolari, B., and Erni, B. (1988) J. Biol. Chem. 263, 6647-6655). Cys421 is located in the C-terminal cytoplasmic domain of the IIBCGlc subunit in a sequence context (LDACITRL) which is well conserved in other transporters of the bacterial phosphotransferase system. Phosphocysteine has been shown previously to be the catalytic intermediate of the mannitol transporter (Pas, H. H., Meyer, G. H., Kruizinga, W. H., Tamminga, K. S., van Weeghel, R. P., and Robillard, G. T.
Amino Acid Sequence↗
The rat insulin-degrading enzyme. Molecular cloning and characterization of tissue-specific transcripts.
The primary structure of the rat insulin-degrading enzyme (IDE) was determined by cDNA analysis. Rat IDE, as well as the previously characterized homologs from human and Drosophila, contain the carboxyl-terminal consensus sequence A/S-K-L, for peroxisome targeting. A stretch of 43 bp surrounding an alternatively used polyadenylation site is highly conserved between rat and human, suggesting that it may contain important regulatory information. Northern blot analysis revealed two IDE transcripts of 3.7 and 5.5 kb in various tissues. Testis was found to be exceptional in having three different RNAs (3.7, 4.1 and 6.1 kb) at a relatively high abundance. The expression of the IDE gene in testis is correlated with sexual maturation.
Amino Acid Sequence↗
Dephosphorylation of phosphorylated atrial natriuretic peptide by protein phosphatase 2A.
A phosphatase which exhibits strong activity toward phosphorylated atrial natriuretic peptide (ANP) was identified in the soluble fraction of rat brain homogenate. This ANP phosphatase has a neutral pH optimum, does not require divalent cations for activity, is inhibited by low concentrations of okadaic acid (50% inhibition at 1 nM) and preferentially dephosphorylates the alpha subunit of phosphorylase kinase. These properties are characteristic of serine/threonine protein phosphatase type 2A (PP2A). The apparent molecular mass of the ANP phosphatase (160 kDa), as estimated by gel filtration, is similar to that of the native heterotrimeric form of PP2A. In addition, phosphorylated ANP is an excellent substrate for the purified catalytic subunit of PP2A (Km = 42 microM, Vmax = 10.3 mumol x min-1 x mg-1). In contrast, protein phosphatase 2B (PP2B) has only very low ANP phosphatase activity (Km = 2.5 microM, Vmax = 0.008 mumol x min-1 x mg-1), and the catalytic subunit of protein phosphatase type 1 (PP1) as well as purified protein phosphatase type 2C (PP2C) are essentially inactive on ANP. These findings are consistent with the observation that PP2A-like activity accounts for virtually all ANP dephosphorylation in brain homogenate. While the phosphorylation of ANP in vitro by cAMP-dependent protein kinase is well documented, this is a first report on a phosphatase that efficiently can reverse this modification.
Animals↗
Two- to one-phonon E3 transition strength in 148Gd.
Explore the source record for details and available documents.
Journal Article↗
Characterization of succinimide-type dehydration products of recombinant hirudin variant 1 by electrospray tandem mass spectrometry.
Two congeners Q4 and Q5 inferred in earlier analyses to be cyclic succinimide-type dehydration products of recombinant hirudin (variant 1) were structurally fully characterized. After isotopic labeling by ring-opening with H2(18)O, the suspected anhydro-positions Asp53 and Asp33 were confirmed by tandem mass spectrometry (MS/MS) sequencing of relevant smaller peptides directly in the enzymatic hydrolysates (V8 protease) using electrospray MS/MS. The chosen strategy proved highly efficient and sensitive.
Hirudins↗
Temperature-controlled radiofrequency catheter ablation of AV conduction: first clinical experience.
A new technique for catheter ablation of atrioventricular (AV) conduction, using temperature-controlled radiofrequency energy and a bipolar asymmetrical electrode configuration, was applied to 12 patients (mean age, 48 +/- 15 years; range, 18-69 years) with medically refractory atrioventricular nodal reentrant tachycardia (AVNRT) or rapid atrial rhythms. The energy source was a 500 kHz generator with automatic power regulation to a preselected temperature of 80 degrees C. A specially designed 7 F bipolar asymmetric thermo-catheter was used for ablation in all cases. The endpoints of the procedure were: first-degree AV block in patients with AVNRT and third-degree block in patients with atrial fibrillation or flutter. Energy was applied over a range of 1-14 times per patient. After a mean follow-up of 8 +/- 4 months, third- or first-degree AV block persisted in eight patients. In comparison to constant-power radiofrequency ablation, where impedance rises are commonly observed, no impedance rise or coating of the electrode occurred during any of the 97 energy applications in this study. Variable wall contact of the electrode was identified in 20 of 97 applications by a slow temperature rise or a drop in temperature and frequent power adjustments. Thus, monitoring temperature and automatic power regulation may help to reduce the total delivered energy. Temperature control during radiofrequency energy avoids coagulum formation and consequently the associated potential hazards of constant-power application.
Adolescent↗
Lamb-shift measurement in hydrogenic phosphorus.
Explore the source record for details and available documents.
Journal Article↗
Antigenicity of three synthetic peptides from the C-terminus of HIV-1 gp 120 correlates with the fragmentation pattern obtained in FAB mass spectrometry.
The proteolytic cleavage site of HIV-1 envelope glycoprotein precursor gp160 is sensitive to mutations. In this study we observed that the antigenicity of synthetic peptides from the C-terminus of gp120 is dependent on the length of the peptide, suggesting a conformational restriction. The physical properties of the peptides evaluated by FAB mass spectrometry correlated with the serological data, but differed from predictions based on the linear sequence. From our results we conclude that the C-terminus of HIV-1 gp120 is conformationally restricted. Furthermore FAB mass spectrometry seems to possess the ability to provide information concerning the conformation of synthetic peptides.
Epitopes↗
Aging, training and exercise. A review of effects on plasma glutathione and lipid peroxides.
Changes in plasma glutathione (reduced form: GSH; oxidised form: GSSG) and lipid peroxides (LPO) levels occur with aging, training and acute physical exercise. Sources of plasma GSH, GSSG and LPO include the liver and skeletal muscle. Aging appears to be accelerated because of a decrease in the antioxidant capacity of tissues reflected in a decreased plasma GSH level. This age-dependent change could be partly compensated by physical training. Skeletal muscle appears to be able to deliver GSH into circulation with the adaptation of muscle to exercise training reflected in an increased plasma GSH level in the trained subject. Decreased plasma GSH concentration following physical exercise demonstrates increased GSH consumption in skeletal muscle resulting in a reduced export rate from muscle into plasma. The GSH system is able to effectively protect tissues against lipid peroxidation initiated by oxygen-derived free radicals produced in the intermediate metabolism during exercise. It can be assumed that the rate of this free radical production is a function of oxygen flow through organ and muscle tissue.
Aging↗