Biomedical subjects

K Richter

Publications and source records attributed to K Richter.

At least 55 records · Page 3Linked to original sources

PubMed2001

The alternatively folded state of the antibody C(H)3 domain.

The C(H)3 domain of antibodies is characterized by two antiparallel beta-sheets forming a disulfide-linked sandwich-like structure. At acidic pH values and low ionic strength, C(H)3 becomes completely unfolded. The addition of salt transforms the acid-unfolded protein into an alternatively folded state exhibiting a characteristic secondary structure. The transition from native to alternatively folded C(H)3 is a fast reaction. Interestingly, this reaction involves the formation of a defined oligomer consisting of 12-14 subunits. Association is completely reversible and the native dimer is quantitatively reformed at neutral pH. This alternatively folded protein is remarkably stable against thermal and chemical denaturation and the unfolding transitions are highly cooperative. With a t(m) of 80 degrees C, the stability of the alternatively folded state is comparable to that of the native state of C(H)3. The defined oligomeric structure of C(H)3 at pH 2 seems to be a prerequisite for the cooperative unfolding transitions.

Acids↗

PubMed2001

Software based digital signal processing and spectrum deconvolution in X-ray spectroscopy.

Approaches for software based digital signal processing and numerical deconvolution of measured signals which overcome limitations of state-of-the-art systems are described. The basic technical equipment for digital signal processing consists of an energy resolving detector with a preamplifier followed by a fast sampling analogue-to-digital converter (ADC). The main idea is the numerical decomposition of the measured signal into contributions caused by single photon absorption using standard pulses. The latter can be obtained by measurements under definite conditions. The maximum pulse rate is then limited only by the ratio of sampling time to the time between two pulses which should be attributed to single events. Thus pulse overlaps do not require pulse rejection. At sampling rates of 10(8) samples per second theoretically a comparable photon rate can be detected at throughputs of 100%. Beyond that it is outlined that in a comparable manner a numerical deconvolution of measured energy spectra (statistic distribution functions of single events) into combinations of standard spectra, which can likewise be determined by measurement, offers outstanding possibilities, too. On the one hand the energy resolution attainable for individual events for a given detector can be improved drastically by the statistical treatment of spectra. On the other hand an energy resolving work principle becomes possible for certain detectors, which do not permit this conventionally due to their poor signal to noise ratio.

Journal Article↗

PubMed2001

Indoor and outdoor BTX levels in German cities.

On the basis of the ongoing study INGA (INdoor exposure and Genetics in Asthma), Germany's most detailed and standardized epidemiological study on indoor exposure to both allergens in house dust and volatile compounds in the air of the home environment has been performed. The purpose of this paper is to describe the spatial and seasonal variability of indoor and outdoor BTX (Benzene, toluene, ethyl benzene, ortho-xylene, meta- and para-xylene) concentrations for the study period from June 1995 to November 1996. Within this framework, air concentrations of volatile organic compounds (BTX) were measured in 204 households in Erfurt (Eastern Germany) and 201 households in Hamburg (Western Germany). BTX sampling was conducted over one week using OVM 3500 passive diffusion sampling devices in the indoor (living room and bedroom) and outdoor environment (outside the window of the living room). Indoor and outdoor median BTX concentrations in Erfurt were slightly, but significantly higher than those in Hamburg. This gap was most pronounced in the levels of indoor toluene (37.3 microg/m3 for Erfurt and 20.5 microg/m3 for Hamburg, P < 0.0001). In both cities, winter indoor and outdoor concentrations for the five compounds exceeded the summer values. Outdoor concentrations of ethyl benzene and ortho-xylene were very low (50% < L.D.). In general, the indoor BTX air concentrations were significantly higher than the outdoor concentra- tions, the lowest I/O ratios were found in the case of benzene. Living room and bedroom values for the five compounds were highly correlated (Spearman coefficient 0.5-0.9). Despite the better insulation of the homes in West Germany, no indication for the expected higher indoor concentrations of BTX in the West could be found. The strong and yet undiscovered indoor source for toluene in East Germany might lead to a further increase in the indoor air load in those homes in the East, which undergo renovations which will lead to improved insulation.

Air Pollutants↗

PubMed2001

Yersinia outer protein P of Yersinia enterocolitica simultaneously blocks the nuclear factor-kappa B pathway and exploits lipopolysaccharide signaling to trigger apoptosis in macrophages.

Exposure of macrophages to bacteria or LPS mediates activation of signaling pathways that induce expression of self defense-related genes. Pathogenic Yersinia species impair activation of transcription factor NF-kappaB and trigger apoptosis in macrophages. In this study, we dissected the mechanism of apoptosis induction by Yersinia. Selectively, Yersinia enterocolitica strains producing the effector protein Yersinia outer protein P (YopP) hampered NF-kappaB activation and subsequently conferred apoptosis to J774A.1 macrophages. Thereby, YopP bound and inhibited the macrophage NF-kappaB-activating kinase IKKbeta. YopP- and Yersinia-, but not Salmonella-induced apoptosis was specifically prevented by transient overexpression of NF-kappaB p65, giving evidence that YopP mediates cell death by disrupting the NF-kappaB signaling pathway. Transfection of J774A.1 macrophages with YopP induced a moderate, but significant degree of apoptosis (40-50% of transfected cells). This effect was strongly enhanced by additional initiation of LPS signaling (80-90%), indicating a synergism between LPS-induced signal transduction and inhibition of NF-kappaB by YopP. This reflects a strategy of a bacterial pathogen that takes advantage of LPS, serving as cofactor, to impair the macrophage.

Adhesins, Bacterial↗

PubMed2001

Determination of scopolamine in human serum and microdialysis samples by liquid chromatography-tandem mass spectrometry.

A liquid chromatographic-tandem mass spectrometric (LC-MS-MS) method with a rapid and simple sample preparation was developed for the determination of scopolamine in biological fluids. Scopolamine and the internal standard atropine in serum samples were extracted and cleaned up by using an automated solid phase extraction method. Microdialysis samples were directly injected into the LC-MS system. The mass spectrometer was operated in the multi reaction monitoring mode. A good linear response over the range of 20 pg/ml to 5 ng/ml was demonstrated. The accuracy for added scopolamine ranged from 95.0 to 104.0%. The lower limit of quantification was 20 pg/ml. This method is suitable for pharmacokinetic studies.

Calibration↗

PubMed2001

Hsp90: chaperoning signal transduction.

Hsp90 is an ATP dependent molecular chaperone involved in the folding and activation of an unknown number of substrate proteins. These substrate proteins include protein kinases and transcription factors. Consistent with this task, Hsp90 is an essential protein in all eucaryotes. The interaction of Hsp90 with its substrate proteins involves the transient formation of multiprotein complexes with a set of highly conserved partner proteins. The specific function of each component in the processing of substrates is still unknown. Large ATP-dependent conformational changes of Hsp90 occur during the hydrolysis reaction and these changes are thought to drive the chaperone cycle. Natural inhibitors of the ATPase activity, like geldanamycin and radicicol, block the processing of Hsp90 substrate proteins. As many of these substrates are critical elements in signal transduction, Hsp90 seems to introduce an additional level of regulation.

Adenosine Triphosphatases↗

PubMed2001

Effect of inhaled ciclesonide on airway responsiveness to inhaled AMP, the composition of induced sputum and exhaled nitric oxide in patients with mild asthma.

To assess the efficacy of ciclesonide, a novel corticosteroid pro-drug, we compared its effect on lung function, airway responsiveness to inhaled AMP, the composition of induced sputum, and the level of exhaled nitric oxide (NO) with the effect of budesonide in patients with asthma. Fifteen non-smoking steroid-naive patients (mean FEV(1), 94%pred) inhaled either 400 microg ciclesonide or 400 microg budesonide as a single morning dose for two weeks each separated by a > or =3 week wash-out period. The study was performed in a double-observer, randomized, cross-over design. FEV(1)increased significantly during treatment with budesonide (3.38 vs. 3.64 l P=0,003), but not after ciclesonide (3.60 vs. 3.69 l). PC(20)FEV(1)of AMP increased (P<0,001, each) after both budesonide (4.59 vs. 32.48 mg/ml, 2.8 doubling doses) and ciclesonide (3.92 vs. 20.00 mg/ml, 2.4 doubling doses). The percentage of sputum eosinophils was significantly reduced after ciclesonide (7.9 vs. 3.4% P=0.01), but not budesonide (6.0 vs. 4.3%). After both budesonide and ciclesonide, a significant (P<0.001) reduction in the level of exhaled NO occurred. In none of the parameters studied, the changes differed significantly between treatment with budesonide or ciclesonide. These data suggest that ciclesonide is equi-effective to budesonide with regard to its potency to reduce the airway responsiveness to inhaled AMP as well as airway inflammation in patients with mild asthma.

Adenosine Monophosphate↗

PubMed2001

Molecular and physiological characterisation of a 14-3-3 protein from lily pollen grains regulating the activity of the plasma membrane H+ ATPase during pollen grain germination and tube growth.

A 14-3-3 protein has been cloned and sequenced from a cDNA library constructed from mRNAs of mature pollen grains of Lilium longiflorum Thunb. Monoclonal antibodies (MUP 5 or MUP 15) highly specific against 14-3-3 proteins recognised a 30-kDa protein in the cytoplasmic fraction of many various lily tissues (leaves, bulbs, stems, anther filaments, pollen grains, stigmas) and in other plants (Arabidopsis seedlings, barley recombinant 14-3-3). In addition, 14-3-3 proteins were detected in a microsomal fraction isolated from pollen grains and tubes, and the amount of membrane-bound 14-3-3 proteins as well as the amount of the plasma membrane (PM) H+ ATPase increased during germination of pollen grains and tube growth. No change was observed in the cytoplasmic fraction. A further increase in the amount of 14-3-3 proteins in the microsomal fraction was observed when pollen grains were incubated in germination medium containing 1 microM fusicoccin (FC) whereas the number of 14-3-3s in the cytoplasmic fraction decreased. Fusicoccin also protected membrane-bound 14-3-3 proteins from dissociation after washing with the chaotropic salt KI. Furthermore, FC stimulated the PM H+ ATPase activity, the germination frequency and the growth rate of pollen tubes, thus indicating that a modulation of the PM H+ ATPase activity by interaction with 14-3-3 proteins may regulate germination and tube growth of lily pollen.

14-3-3 Proteins↗

PubMed2001

Daily changes in neuroendocrine control of moulting hormone secretion in the prothoracic gland of the cockroach Periplaneta americana (L.).

Time of day related changes in ecdysteroid secretion by the prothoracic gland of last instar nymphs were studied using in vitro coincubations of prothoracic glands and brains under a 12-h light:12-h dark cycle. The experiments reveal that the cells of the prothoracic gland of the cockroach nymphs do not have an endogeneous circadian oscillator determining rhythmicity of ecdysteroid secretion. PTTH release in the scotophase is responsible for the peak of ecdysteroid production during the photophase.

Journal Article↗

PubMed2001

Allogeneic recombinant soluble MHC class I molecules modify urinary odor cues in rats.

The highly polymorphic genes of the major histocompatibility complex (MHC) determine, in part, the odor cues and behavior of an individual. In animal models, MHC-associated odors that regulate distinct behavior have been identified mainly in urine. However, the underlying mechanism is still not clear. Here, we show that injected recombinant soluble (rs) MHC class I molecules (DA, rsRT1.A(a) and Lewis, rsRT1.A(l)) temporarily alter urine odor of Lewis test rats (RT1.A(l)). This change in urinary signals was observed in behavioral assays using the habituation/dishabituation test and in odor signals analysed by gas chromatography (GC) and mass spectrometry (MS). Gas chromatographic analysis revealed that these altered odor signals are caused by quantitative changes of at least two nitrogen-containing urinary compounds. The results suggest that urinary olfactory cues are directly or indirectly influenced by MHC class I gene products.

Animals↗

PubMed2001

Nitric oxide up-regulates ferritin mRNA level in snail neurons.

We cloned and sequenced the ferric ion-binding protein, ferritin, from the nervous system of the pulmonate snail, Helix pomatia. Helix H-ferritin cDNA contains a 519-bp open reading frame (ORF) and predicts an iron-responsive element (IRE) at the 5'-untranslated region (5'-UTR) of the ferritin mRNA. The deduced amino acid sequence revealed 86% similarity with Lymnaea stagnalis ferritin and about 70% similarity with vertebrate H-ferritin. While secreted ferritin isoforms contain a signalling sequence at their N-terminal end, Helix ferritin does not contain this sorting signal indicating that it is restricted to the cytoplasm. The amino acid ligands at positions Glu25, Tyr30, Glu59, Glu60, His63, Glu105 and Gln139 indicate an active ferroxidase site in Helix ferritin. In situ hybridization visualized ferritin mRNA in neuronal cell bodies but not in the neuropil. In contrast, ferritin-immunoreactive protein was localized in cell bodies and neurites. We further demonstrate that the NO donors S-nitroso-N-acetylpenicillamine (SNAP), or hydroxylamine (HA), increase the intracellular ferritin mRNA level by about 55%. In conclusion, our findings show that Helix neurons express an intracellular H-ferritin isoform and suggest that iron and NO metabolism are coupled.

Amino Acid Sequence↗

PubMed2001

[Treatment of bronchial asthma using a new adjustable combination treatment plan: Asthma Control Plan (ATACO)].

The current guideline of the German Respiratory League (Deutsche Atemwegsliga) recommends the synergistic combination therapy with long acting beta 2-agonists and inhalative corticosteroids only for patients suffering from moderate to severe persistent asthma (step 3 and 4 of the asthma severity scale). Now convenient fixed combinations of these substances are available, which could enhance patient's compliance. A large, randomised, parallel-group study in 8000 mild to moderate asthmatics was designed to compare a flexible asthma control plan with the conventional fixed-dose management with respect to quality of life, symptom control and treatment costs. The fixed combination of 6 micrograms Formoterol and 200 micrograms Budesonide per puff in a new dry powder device was applied either due to a novel flexible asthma control plan "ATACO" (group A) or as a standardised conventional dosing regimen (group B) inhaling two puffs b.i.d. In group A (ATACO) patients reduce the run-in dose after four weeks from two puffs b.i.d. to one puff b.i.d. with the option of doubling the dose immediately, if (pre-defined) asthma deterioration occurs. One week later the dose can be either doubled again or reduced due to the actual asthma symptoms of the patient. After run-in, group B patients continue to take two inhalations b.i.d. In this group, asthma exacerbations will be managed as usual by the physician. In contrast, the ATACO group flexible management plan allows the self-medication: an immediate increase in the dose of the fixed combination will lead to both a fast relief of bronchospasm and an automatically higher dosed corticosteroid treatment for the underlying asthmatic inflammation. Conversely, if later asthma symptoms improve, less reliever and controller medication will be needed and used. The immediate treatment of new onset bronchospasm and asthmatic inflammation by the patient himself could maintain at least the same grade of asthma control, as the conventional group B treatment, improve asthma-related quality of life and decrease treatment costs. If the concept works, fixed combinations of long-acting beta 2 agonists and inhalative corticosteroids could have an impact on future asthma guidelines.

Administration, Inhalation↗

PubMed2001

Arginine-143 of Yersinia enterocolitica YopP crucially determines isotype-related NF-kappaB suppression and apoptosis induction in macrophages.

Pathogenic Yersinia spp. counteract host defense mechanisms by modulating the cellular signal relay in response to infection. Subversion of the antiapoptotic NF-kappaB signaling pathway by the Yersinia enterocolitica virulence protein YopP crucially determines the induction of apoptosis in Yersinia-infected macrophages. Here, we analyzed a panel of pathogenic, phylogenetically distinct Y. enterocolitica serotypes for their abilities to trigger macrophage apoptosis. Y. enterocolitica from the highly pathogenic serogroup O8 was substantially more effective in apoptosis induction than Yersinia from the serogroups O3 and O9. Complementation of yopP-knockout mutants revealed that this effect was specifically conferred by the serogroup O8 YopP. The amino acid sequences of YopPO8 and YopPO9 share 94% identity, and both YopP isotypes were found to interact with the NF-kappaB-activating kinase IKKbeta in macrophages. However, selectively, YopPO8 mediated efficient inhibition of IKKbeta activities, which led to substantial suppression of NF-kappaB activation. To localize the YopPO8-related effector domain, we interchanged stretches of amino acids and single amino acid residues between YopPO8 and YopPO9. Functional characterization of the resulting mutants revealed a major role of the arginine-143 residue in determining the inhibitory impact of YopP on IKKbeta activity and survival of macrophages.

Amino Acid Sequence↗

PubMed2001

Respiratory symptoms in relation to indoor exposure to mite and cat allergens and endotoxins. Indoor Factors and Genetics in Asthma (INGA) Study Group.

The authors investigated the relationship between respiratory symptoms in adults and exposure to mite and cat allergens, the role of endotoxins in house dust, the effects of mixtures of several allergens, and interactions between allergen exposure and allergic sensitization. Within a nested case-control study, 405 subjects aged 25-50 yrs from two German cities answered a standardized questionnaire. Allergen-specific immunoglobulin-E was measured. Dust samples were taken from the subjects' homes to determine exposure to mite (Dermatophagoides pteronyssinus antigen 1 Der p 1) and (D. farinae antigen 1 Der f l) and cat (cat antigen d1 Fel d 1) allergen and endotoxin content in settled house dust. Exposure to Der f 1 and Der p 1 plus Der f 1 >10 microg x g(-1) of mattress dust, respectively, increased the risk of wheeze and breathlessness (odds ratios (OR): 4.04, 95% confidence interval (CI): 1.53-10.64, OR: 2.78, 95% CI: 1.06-7.28). Fel d 1 >8 microg x g(-1) was positively associated with cough at night (OR: 2.74, 95%, CI: 1.22-.17), noteworthy also in the nonsensitized subjects. Subjects exposed to elevated concentrations of more than one allergen had an up to seven-fold increase in the risk of respiratory symptoms, compared to nonexposed subjects. Sensitized subjects exposed to elevated concentrations of Der f 1 or Fel d 1 were found to have the highest risk of asthma attacks and respiratory symptoms. No statistically significant association was found between exposure to endotoxins and respiratory health. Indoor exposure to Dermatophagoides farinae antigen 1 and cat antigen d1 is a risk factor for respiratory symptoms in adults, and for cat antigen d 1 even in nonsensitized subjects. The risk is increased if subjects are exposed to a mixture of allergens or if they are sensitized in addition to high exposure.

Adult↗

PubMed2001

Beta(1-->3)-glucan in house dust of German homes: housing characteristics, occupant behavior, and relations with endotoxins, allergens, and molds.

beta(1-->3)-Glucans are potent proinflammatory agents that have been suggested to play a role in indoor-related respiratory health effects. The aim of this study was to assess whether beta(1-->3)-glucan concentrations in house dust are correlated with levels of endotoxins, allergens, and culturable mold spore counts in house dust. Further, the associations of beta(1-->3)-glucan with housing characteristics and occupant behavior were assessed. beta(1-->3)-Glucan was measured in settled house dust from living room floors of 395 homes of two German cities, Erfurt and Hamburg, with a specific enzyme immunoassay. Concentrations ranged from below the limit of detection to 19,013 microg/m(2) (22,588 microg/g dust). Concentrations per square meter were found to be correlated with endotoxins, mite and cat allergens, and culturable mold spores. Correlations were weaker when concentrations were expressed per gram of dust, indicating that variance in concentrations of all factors is largely determined by the amount of dust sampled. Associations between beta(1-->3)-glucan, housing characteristics, and occupant behavior were found for concentrations per square meter but not for concentrations per gram of dust. The following characteristics were associated with a significant increase in beta(1-->3)-glucan levels: carpets in the living room [means ratio (MR) = 1.9-2.1], keeping a dog inside (MR = 1.4), use of the home by four or more persons (MR = 1.4), use of the living room for > 180 hr/week (MR = 2.1), lower frequency of vacuum cleaning (MR = 1.6-3.0) and dust cleaning (MR = 1.2 and 1.4, respectively), and presence of mold spots during the past 12 months (MR = 1.4). We conclude that that the amount of dust sampled can be used as a proxy for hygiene and that beta(1-->3)-glucan concentrations per square meter are related to the amount of dust sampled.

Air Pollution, Indoor↗

PubMed2001

Pharmacokinetics of oral talinolol following a single dose and during steady state in patients with chronic renal failure and healthy volunteers.

OBJECTIVE: The objective of this study was to investigate the effect of renal impairment on the pharmacokinetics of the selective beta1-receptor antagonist talinolol. METHODS: Pharmacokinetic data were obtained in 12 healthy volunteers, 12 patients with renal impairment and 8 patients with terminal renal insufficiency after the oral administration of 100 mg talinolol and under steady state conditions (100 mg talinolol daily). Concentrations of talinolol in plasma, urine and dialysate during hemodialysis were measured with a validated HPLC-method. RESULTS: Talinolol is absorbed quite rapidly from the gastrointestinal tract (tmax 2.5-4 h). Steady state conditions were reached within 3-4 days depending on renal function. The calculated mean elimination half-life (t(1/2z)) in healthy volunteers (11 male, 1 female) was about 12 h. After an oral dose of 100 mg, about 55% of the bioavailable talinolol is eliminated unchanged in the urine. This fraction is reduced to 25% in patients with moderate to severe renal failure. A strong correlation was found between the renal elimination of talinolol and creatinine clearance. In patients with renal failure, the delayed elimination leads to an increase in t(1/2z) and to a decrease in the apparent total body clearance. Steady state trough levels (c(min)ss) in these patients are about 2.2-fold higher than in volunteers. The hemodialysability of talinolol was low. CONCLUSION: The disposition of talinolol shows a strong dependence on the renal function. On the basis of the kinetic data for talinolol, dose reductions of 30-50% are recommended in subjects with moderate to severe renal impairment.

Adrenergic beta-Antagonists↗

PubMed2000

C-terminal regions of Hsp90 are important for trapping the nucleotide during the ATPase cycle.

Hsp90 is an abundant molecular chaperone that functions in an ATP-dependent manner in vivo. The ATP-binding site is located in the N-terminal domain of Hsp90. Here, we dissect the ATPase cycle of Hsp90 kinetically. We find that Hsp90 binds ATP with a two-step mechanism. The rate-limiting step of the ATPase cycle is the hydrolysis of ATP. Importantly, ATP becomes trapped and committed to hydrolyze during the cycle. In the isolated ATP-binding domain of Hsp90, however, the bound ATP was not committed and the turnover numbers were markedly reduced. Analysis of a series of truncation mutants of Hsp90 showed that C-terminal regions far apart in sequence from the ATP-binding domain are essential for trapping the bound ATP and for maximum hydrolysis rates. Our results suggest that ATP binding and hydrolysis drive conformational changes that involve the entire molecule and lead to repositioning of the N and C-terminal domains of Hsp90.

Adenosine Diphosphate↗

PubMed2000

Cpr6 and Cpr7, two closely related Hsp90-associated immunophilins from Saccharomyces cerevisiae, differ in their functional properties.

Hsp90 is an abundant cytosolic molecular chaperone. It controls the folding of target proteins including steroid hormone receptors and kinases in complex with several partner proteins. Prominent members of this protein family are large peptidyl prolyl cis/trans isomerases (PPIases), which catalyze the cis/trans isomerization of prolyl peptide bonds in proteins and possess chaperone activity. In Saccharomyces cerevisiae, two closely related large Hsp90-associated PPIases, Cpr6 and Cpr7, exist. We show here that these homologous proteins bind with comparable affinity to Hsp90 but exhibit significant structural and functional differences. Cpr6 is more stable than Cpr7 against thermal denaturation and displays an up to 100-fold higher PPIase activity. In contrast, the chaperone activity of Cpr6 is much lower than that of Cpr7. Based on these results we suggest that the two immunophilins perform overlapping but not identical tasks in the Hsp90 chaperone cycle.

Carrier Proteins↗