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Biomedical subjects

E Beck

Publications and source records attributed to E Beck.

At least 55 records · Page 3Linked to original sources
PubMed1998

cDNA sequence and heterologous expression of monomeric spinach pullulanase: multiple isomeric forms arise from the same polypeptide.

The spinach pullulanase gene was cloned and sequenced using peptide sequences of the purified enzyme as a starting point and employing PCR techniques and cDNA library screening. Its open reading frame codes for a protein of 964 amino acids which represents a precursor of the pullulanase. The N-terminal transit peptide consists of 65 amino acids, and the mature protein, comprising 899 amino acids, has a calculated molecular mass of 99kDa. Pullulanase is a member of the alpha-amylase family. In addition to a characteristic catalytic (beta/alpha)8-barrel domain, it contains a domain, F, that is specific for branching and debranching enzymes. Pullulanase cDNA was expressed in Escherichia coli, and the purified protein was compared with the enzyme from spinach leaves. Identity of the two proteins was confirmed in terms of catalytic properties, N-terminal amino acid sequences and molecular masses. The pullulanase produced by E. coli showed the same microheterogeneity as the spinach leaf enzyme: it could be resolved into two substrate-induced forms by electrophoresis in amylopectin-containing polyacrylamide gels, and, in the absence of substrate, into several free forms (charge isomers) by isoelectric focusing or chromatofocusing. Rechromatofocusing of single free forms resulted in the originally observed pattern of molecular forms. However, heterogeneity of the protein disappeared on isoelectric focusing under completely denaturing conditions when only one protein band was observed. Post-translational modifications such as glycosylation and phosphorylation could be excluded as potential explanations for the protein heterogeneity. Therefore the microheterogeneity of spinach leaf pullulanase results from neither genetic variation nor post-translational modifications, but is a property of the single unmodified gene product. The different interconvertible forms of the pullulanase represent protein populations of different tertiary structure of the same polypeptide.

Amino Acid Sequence↗
PubMed1998

Multiple virulence determinants of foot-and-mouth disease virus in cell culture.

Hypervirulent variants of foot-and-mouth disease virus (FMDV) of serotype C arise upon serial cytolytic or persistent infections in cell culture. A specific mutation in the internal ribosome entry site of persistent FMDV was previously associated with enhanced translation initiation activity that could contribute to the hypervirulent phenotype for BHK-21 cells. Here we report that several hypervirulent FMDV variants arising upon serial cytolytic passage show an invariant internal ribosome entry site but have a number of mutations affecting structural and nonstructural viral proteins. The construction of chimeric type O-type C infectious transcripts has allowed the mapping of a major determinant of hypervirulence to the viral capsid. Tissue culture-adapted FMDV displayed enhanced affinity for heparin, but binding to cell surface heparan sulfate moieties was not required for expression of the hypervirulent phenotype in Chinese hamster ovary (CHO) cells. Virulence was identical or even higher for glycosaminoglycan-deficient CHO cells than for wild-type CHO cells. FMDV variants with decreased affinity for heparin were selected from a high-binding parental population and analyzed. Substitutions associated with decreased heparin binding were located at positions 173 of capsid protein VP3 and 144 of capsid protein VP1. These substitutions had a moderate effect on virulence for BHK-21 cells but completely abrogated infection of CHO cells. The comparative results with several FMDV isolates show that (i) increased affinity for heparin and alterations in cell tropism may be mediated by a number of independent sites on the viral capsid and (ii) the same capsid modifications may have different effects on different cell types.

Amino Acid Substitution↗
PubMed1998

CA 125 elevations in patients with malignant lymphomas.

Circulating CA 125 serum levels were measured in 60 patients with several hematological malignancies. Using 35 U/ml as cutoff level, elevated CA 125 concentrations were found in 3 of 18 patients with acute leukemia, 1 of 5 patients with chronic myelocytic leukemia, 2 of 9 patients with Hodgkin's lymphoma and in 14 of 28 patients with non-Hodgkin's lymphoma. None of the healthy control group had CA 125 serum levels above 35 U/ml. In patients with malignant lymphoma, elevated CA 125 serum concentrations were associated with abdominal involvement (p < 0.01). 15 of 19 patients with abdominal tumor masses had CA 125 concentrations above 35 U/ml, but only 1 of 18 patients with supradiaphragmatic involvement. Serial determinations of CA 125 were performed in 3 patients with malignant lymphoma during chemotherapy. Disease regression was associated with decreasing CA 125 serum levels. Thus, CA 125 may be a useful indicator of abdominal involvement in patients with malignant lymphoma. Moreover, serial CA 125 measurement may be of value in monitoring response to chemotherapy in these patients.

Acute Disease↗
PubMed1998

Hospital compliance on a budget.

Community and rural hospitals are among the last to jump aboard the compliance program bandwagon. For many, it boils down to a belief that a plan either is not necessary or not affordable. Ellen Beck profiles the special issues that smaller hospitals face amid government investigations into Medicare/Medicaid billing fraud.

Aged↗
PubMed1998

Government officials redefine use of False Claims Act to combat fraudulent hospital bills.

The Dept. of Justice and the Dept. of Health and Human Services Office of Inspector General have issued new guidelines for using the False Claims Act to prosecute hospitals that fraudulently bill Medicare. Ellen Beck explains the changes and why the American Hospital Association calls the more lenient use of the FCA a "positive first step" toward acknowledging that the act is being used improperly.

American Hospital Association↗
PubMed1997

Randomised trial of roxithromycin in non-Q-wave coronary syndromes: ROXIS Pilot Study. ROXIS Study Group.

BACKGROUND: There is serological evidence for an association between Chlamydia pneumoniae and coronary heart disease. We investigated the hypothesis that an antichlamydial macrolide antibiotic, roxithromycin, can prevent or reduce recurrent major ischaemic events in patients with unstable angina. METHODS: The effect of roxithromycin was assessed in a double-blind, randomised, prospective, multicentre, parallel-group, placebo-controlled pilot study of 202 patients with unstable angina or non-Q-wave myocardial infarction. Patients were randomly assigned either roxithromycin 150 mg orally twice a day (n = 102) or placebo orally twice a day (n = 100). The treatment was for 30 days. Patients were followed up for 6 months. We report the primary clinical endpoints (cardiac ischaemic death, myocardial infarction, and severe recurrent ischaemia), assessed at day 31, in 202 patients on an intention-to-treat basis. FINDINGS: A statistically significant reduction in the primary composite triple endpoint rates was observed in the roxithromycin group: p = 0.032. The rate of severe recurrent ischaemia, myocardial infarction, and ischaemic death was 5.4%, 2.2%, and 2.2% in the placebo group and 1.1%, 0%, and 0%, in the roxithromycin group, respectively. No major drug-related adverse effects were observed. INTERPRETATION: Antichlamydial antibiotics may be useful in therapeutic intervention in addition to standard medication in patients with coronary-artery disease. Large-scale trials are needed to confirm these preliminary observations.

Adult↗
PubMed1997

Sensitivity of transformed fibroblasts for intercellular induction of apoptosis is determined by their transformed phenotype.

Intercellular induction of apoptosis defines a potential control mechanism of oncogenesis. It is based on induction of apoptosis in transformed fibroblasts by neighboring nontransformed fibroblasts. Transforming growth factor type beta (TGF-beta) represents the initial triggering molecule to induce nontransformed cells to release apoptosis-inducing factors. To test whether sensitivity for intercellular induction of apoptosis is directly dependent on the transformed phenotype, v-src-transformed rat fibroblasts and emerging revertants were tested for their sensitivity. All transformed cell clones were sensitive, whereas all revertant clones had lost their sensitivity in parallel with the loss of the transformed phenotype. In addition, revertants had regained the potential to induce apoptosis in transformed cells. Sensitivity to intercellular induction of apoptosis is therefore directly dependent on the transformed phenotype, whereas the ability to induce apoptosis is a specific feature of nontransformed fibroblasts.

Animals↗
PubMed1997

[The conservative and surgical therapy of traumatic humeral shaft fractures].

Sixty-three patients with humeral shaft fractures were evaluated clinically and radiographically 18 months after injury; 27 patients were treated surgically (group A) and 36 patients conservatively (group B). Analysis of the results according to a score by Kwasny revealed 6.2 points in group A and 2.2 points in group B (P < 0.0001; F = 46.9). The results of these two comparable groups suggest that conservative treatment of humeral shaft fractures is superior regarding mobility of the shoulder and elbow, strength, the incidence of neurological complications, pain, subjective rating and cosmesis. There were no differences on roentgenograms between the two groups (P = 0.48).

Adolescent↗
PubMed1997

Functional involvement of polypyrimidine tract-binding protein in translation initiation complexes with the internal ribosome entry site of foot-and-mouth disease virus.

The synthesis of picornavirus polyproteins is initiated cap independently far downstream from the 5' end of the viral RNA at the internal ribosome entry site (IRES). The cellular polypyrimidine tract-binding protein (PTB) binds to the IRES of foot-and-mouth disease virus (FMDV). In this study, we demonstrate that PTB is a component of 48S and 80S ribosomal initiation complexes formed with FMDV IRES RNA. The incorporation of PTB into these initiation complexes is dependent on the entry of the IRES RNA, since PTB and IRES RNA can be enriched in parallel either in 48S or 80S ribosomal complexes by stage-specific inhibitors of translation initiation. The formation of the ribosomal initiation complexes with the IRES occurs slowly, is temperature dependent, and correlates with the incorporation of PTB into these complexes. In a first step, PTB binds to the IRES, and then the small ribosomal subunit encounters this PTB-IRES complex. Mutations in the major PTB-binding site interfere simultaneously with the formation of initiation complexes, translation efficiency, and PTB cross-linking. PTB stimulates translation directed by the FMDV IRES in a rabbit reticulocyte lysate depleted of internal PTB, and the efficiency of translation can be restored to the original level by the addition of PTB. These results indicate that PTB plays an important role in the formation of initiation complexes with FMDV IRES RNA and in stimulation of internal translation initiation with this picornavirus.

Animals↗
PubMed1997

Point mutations within the betaG-betaH loop of foot-and-mouth disease virus O1K affect virus attachment to target cells.

The amino acid sequence Arg-Gly-Asp (RGD) is a highly conserved region located on the P1D protein of most sero- and subtypes of foot-and-mouth disease virus (FMDV)and participates in binding of FMDV to their target cells. In order to analyze the role of the RGD sequence in FMDV infection of cells in more detail, 13 mutations within or near the RGD sequence of virus type O1Kaufbeuren were designed by using a full-length cDNA plasmid. Transfection of baby hamster kidney cells (BHK-21) with in vitro-transcribed cRNAs containing mutations bordering the RGD sequence led to the production of infectious virus in most cases. In contrast, almost all of the mutants containing changes within the RGD sequence produced noninfectious viral particles indistinguishable from wild-type virus by electron microscopy. In order to demonstrate that these noninfectious progeny from the RGD mutants were defective only in their cell adsorption, the respective cRNAs were cotransfected together with a cRNA expressing the wild-type P1 protein. The resulting virus particles were able to infect BHK-21 cells. These results demonstrate the important role of the RGD sequence in FMDV binding to cells but also emphasize the influence of other amino acids in the bordering region.

Amino Acid Sequence↗
PubMed1997

[Breath condensate as a method of noninvasive assessment of inflammation mediators from the lower airways].

The detection of mediators from the lower airways still depends on invasive or provoking sampling techniques like bronchoalveolar lavage (BAL) or induced sputum, respectively. Both methods affect the specimen itself. In contrast, the breathing condensate opens the possibility to get native specimens from lower airways during breathing at rest. The breathing condensate was obtained by freezing of exhaled air. The equipment was developed in the FILT Res. Soc. Ltd.. The method is applied for a patent. Leukotriene B4 and Leukotriene C4D4E4F4 were measured in the exhalation of asthmatics, patients with different airway disorders and healthy volunteers. In an additional study the condensate was obtained before and after of a non-specific bronchial challenge test. In asthmatics a close correlation between leukotriene concentration of the condensate and the degree of asthmatic disease according to "International Consensus Report" was found, but no correlation to lung function tests. Within a bronchial challenge test applying histamine the release of leukotrienes was shown to be more sensitive to the challenge test than a lung function test. The results of the study indicate new diagnostic possibilities in lung diseases using the detection of non volatile substances in the exhaled air.

Asthma↗
PubMed1996

The chloroplast envelope is permeable for maltose but not for maltodextrins.

Permeation of [14 C]maltose into the stroma (measured as the sorbitol-impermeable space) of isolated intact spinach (Spinacia oleracea L.) chloroplasts was studied using the silicone oil centrifugation technique. Maltose uptake showed Michaelis Menten-kinetics with a K(m) of 25 mM and a Vmax of 19.5 mumol maltose. mg Chl-1. h-1 at 15 degrees C. Lack of interaction of glucose and maltose uptake suggested the presence of individual translocators for maltose and glucose in the inner chloroplast envelope. Maltose uptake was markedly inhibited by maltodextrins (maltotriose up to maltoheptaose). The corresponding [14C]maltodextrins were prepared by degradation of [14C]starch with pullulanase and alpha-amylase and purified by high performance TLC. None of these maltodextrins, when administered at a concentration of 10 mM, was transported into the sorbitol-impermeable space of the chloroplasts. The results suggest that the transport system for maltose is also accessible to maltodextrins but that only maltose can be translocated across the inner envelope of spinach chloroplasts.

Cell Membrane Permeability↗
PubMed1996

The formation and stability of imidazolidinone adducts from acetaldehyde and model peptides. A kinetic study with implications for protein modification in alcohol abuse.

The kinetics of the reaction of acetaldehyde (AcH) with the alpha-amino group of several di- and tripeptides to form 2-methylimidazolidin-4-one adducts were determined at pH 7, 4, 37 degrees C, using reverse phase HPLC to separate peptides from adducts. The imidazolidin-4-one structure of the adducts was confirmed by 13C NMR spectroscopy. The reaction of val-gly-gly with AcH was shown to follow second-order kinetics over a wide range of concentrations of both reactants, with k2 = 0.734 +/- 0.032 M(-1) min(-1). Under conditions similar to those in the liver of an alcoholic during chronic ethanol oxidation ([Ach]o = 50-910 microm; [free peptide alpha-amino groups]o = 1.5 mM), the reaction proceeded until effectively all of the AcH had been consumed. The side chain of the N-terminal amino acid was shown not to have a marked effect on the rate of imidazolidinone formation. The decomposition of the imidazolidinone adduct of val-gly-gly and AcH was observed at 60-100 degrees C. Extrapolation of an Arrhenius plot to 37 degrees C provided an estimate of K(obs) of 0.002 h-1 (t1/2 approximately 14 days). Based on these kinetic studies, it is concluded that imidazolidinone adducts of AcH with proteins may be present in the liver and, possibly, in the blood of alcoholics.

Acetaldehyde↗
PubMed1996

Cathepsin B of Schistosoma mansoni. Purification and activation of the recombinant proenzyme secreted by Saccharomyces cerevisiae.

Procathepsin B from the parasitic trematode Schistosoma mansoni was expressed as a glycosylation-minus mutant in yeast cells and purified by means of a histidine affinity tag which was added to the carboxyl terminus of the recombinant protein. The purified zymogen underwent autoprocessing but required an assisting protease for activation. Pepsin-activated schistosomal cathepsin B was further characterized with the cathepsin B-specific substrates N-benzyloxycarbonyl (Z)-Arg-Arg-p-nitroanilide, Z-Arg-Arg-7-amido-4-methyl-coumarin, and Z-Phe-Arg-7-amido-4-methylcoumarin. A proteolytic activity comparable to mammalian cathepsin B was observed. In addition, we analyzed the degradation of human hemoglobin by schistosomal cathepsin B, which has been suggested to be the physiological target of the protease.

Amino Acid Sequence↗
PubMed1996

Community service needs of people with HIV infection in London.

The objectives of this study were to describe the expressed needs for community services of HIV-infected individuals by disease stage, gender and transmission category and the barriers which prevent the receipt of such services. Structured interviewer-administered questionnaires concerning a 6-month retrospective period were used to obtain information on need for community services and problems which prevented the receipt of services. The study sample included 70 homosexual men with asymptomatic HIV disease, 42 homosexual men with symptomatic non-AIDS, 53 homosexual men with AIDS, 23 heterosexual men, 29 heterosexual women, 9 male and 9 female injecting drug users. The main outcome measures were the extent to which needs for community services were met and person/service combinations for which problems or barriers prevented the receipt of community services. On average, subjects expressed a need for 10 categories of community services over the 6-month period: homosexuals expressed a mean of 10, heterosexuals 10, injecting drug users 11, subjects with asymptomatic HIV infection 9, subjects with symptomatic non-AIDS 11, subjects with AIDS 13, men 9 and women 14. A total of 58% of community service needs were always met, 6% were rarely not met, 16% were sometimes not met, 6% were often not met and 14% were not met at all. The extent to which subjects felt that their needs were met was similar for the different study groups, but the needs of women were met somewhat less frequently than those of men. Similarly, people with AIDS felt that their needs were met slightly less often. Reported levels of unmet need were high for a wide range of services. The most common reason subjects gave for not having received a community service for which they expressed a need was ignorance of where or how to obtain the service. This was mentioned in one-third of all such cases. Anxieties over the competence with which a service would be rendered was mentioned in 13% of cases and long waiting times in 11%. The frequencies of unmet need for many community services were high and often seemed to arise either from a lack of awareness on the part of subjects on how and where services could be obtained or from doubts about the relevance of services offered. Both of these barriers should be surmountable through the provision of better information to patients, extending user involvement in service development and the better co-ordination of service delivery through care management approaches.

Adult↗