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J Büttner

Publications and source records attributed to J Büttner.

At least 19 recordsLinked to original sources
PubMed2000

Mutation analysis of replicative genes encoding the large subunits of DNA polymerase alpha and replication factors A and C in human sporadic colorectal cancers.

We examined cDNAs of the catalytic subunit of DNA polymerase alpha (185 kDa), the 70 kDa subunit of replication protein A (single-stranded DNA-binding protein) and the 140 kDa subunit of replication factor C for mutations. Surgical specimens from 12 patients with sporadic colon cancer and normal mucosae from the same patients were investigated. In addition, we analyzed 3 human colon cancer cell lines that exhibited defects in mismatch repair (DLD-1, HCT116, SW48) and 3 colon cancer cell lines without such a defect (HT29, SW480 and SW620). For detection of mutations, we used reverse transcription of mRNA, amplification of cDNAs by PCR, analysis of single-strand conformation polymorphism and DNA sequencing. Eleven colon cancers and 6 colon cancer cell lines were analyzed for DNA polymerase alpha. Only 2 silent point mutations were detected, in 1 colon carcinoma and in cell line HCT116. Two sequence alterations of the 70 kDa subunit of replication factor A were identified in 15 specimens (9 colon carcinomas and 6 cell lines). Colon carcinomas from 2 patients (CC5MA and CC25HN) exhibited an ACA-->GCA transition in codon 351, which caused a Thr-->Ala exchange. In carcinomas CC5MA and CC8MA, a TCC-->TCT (Ser-->Ser) transition in codon 352 was observed. The deviations in codons 351 and 352 occurred in both cancer tissues and normal mucosae, suggesting a genetic polymorphism. No mutation was found in the 140 kDa subunit of replication factor C from 16 specimens (10 tumors and 6 cell lines). Point mutations were identified in the p53 tumor-suppressor gene in 4 of the 6 colon cancer cell lines and 3 of the 8 carcinoma specimens. We did not find tumor-associated DNA sequence alterations that resulted in amino acid changes in the DNA replication genes analyzed. We infer that the scarcity of mutations found is due to stringent selection, eliminating functionally impaired replication proteins.

Colorectal Neoplasms↗
PubMed2000

Effect of cholestyramine on bile acid pattern and synthesis during administration of ursodeoxycholic acid in man.

BACKGROUND: Cholestyramine is the first-line treatment for cholestasis-induced pruritus and is prescribed along with ursodeoxycholic acid (UDCA) in patients with cholestatic liver diseases. Impairment of the intestinal absorption of endogenous hydrophobic bile acids by cholestyramine is well known. It is unclear, however, whether cholestyramine also impairs the absorption of the hydrophilic bile acid, UDCA, in man. AIMS: To study serum levels of UDCA and endogenous bile acids as well as endogenous bile acid synthesis during simultaneous or separate administration of UDCA and cholestyramine in vivo; and absorption of UDCA both in the presence and absence of its hydrophobic epimer, chenodeoxycholic acid (CDCA), by cholestyramine in vitro. PATIENTS AND METHODS: Five healthy subjects received UDCA (12.5 +/- 0.5 mg kg-1 daily) as a single dose for periods of 14 days with or without cholestyramine (4 g daily). Fasting serum levels of bile acids and of 7alpha-hydroxy-4-cholesten-3-one (alpha-HC), a measure of endogenous bile acid synthesis, were determined by gas chromatography and high pressure liquid chromatography, respectively. In vitro, bile acid solutions were incubated for 24 h in the presence or absence of cholestyramine, and bile acid concentrations were determined in the supernatant. RESULTS: Simultaneous administration of UDCA and cholestyramine in man led to a decrease of fasting serum levels of UDCA by 60% when compared to UDCA serum levels during administration of UDCA alone. In contrast, serum levels of endogenous bile acids were not affected and alpha-HC serum levels were found increased 2. 7-fold indicating stimulation of endogenous bile acid synthesis by cholestyramine. Administration of cholestyramine and UDCA at an interval of 5 h tended to diminish the effect of cholestyramine on UDCA serum levels. In vitro, conjugated and unconjugated UDCA were effectively bound by cholestyramine both in the presence and absence of hydrophobic bile acids. CONCLUSIONS: The results strongly support the recommendation to administer UDCA and cholestyramine at different times of day.

Adult↗
PubMed2000

Impacts of laboratory methodology on medical thinking in the 19th century.

During the 19th century the first laboratories at hospitals and clinics in Central Europe were established. These 'clinical laboratories' were devoted to chemical analyses in practical medicine and clinical research. A characteristic feature of these laboratories was the use of measuring instruments, e.g. volumetric apparatus, polarimeters, spectroscopes, colorimeters, photometers. Using these techniques new phenomena were introduced into clinical medicine which could serve as signs in the diagnosis of diseases. Many of the new diagnostic signs were quantitative data as results of measurements. Their main advantage was the greater differentiation in the description of phenomena compared with the qualitative data used before. Another important characteristic of the new diagnostic signs was the discovery of causal relations to physiological and pathological processes in the organism. The physicians and chemists in the clinical laboratory were eager not only to collect empirical data but also to find causal relationships by research work similar to that carried out in physics and chemistry. Once causal chains were been identified more general relationships became clear. One example is the concept of metabolism which comprised a dynamic view on chemical processes in the body and the variation of "Stoff" (material) in time, including a quantitative input-output analysis of the body in health and disease. In the second half of the 19th century, scientifically based diagnostic signs began to replace the traditional symptoms and signs used since antiquity.

Clinical Laboratory Techniques↗
PubMed1999

Detection of mutations in the DNA polymerase delta gene of human sporadic colorectal cancers and colon cancer cell lines.

To test the hypothesis whether DNA polymerases acquire mutator properties during tumor development (mutator hypothesis), we examined DNA polymerase delta mRNA in 6 colon cancer cell lines (DLD-1, HCT116, SW48, HT29, SW480 and SW620) and 7 sporadic human colorectal cancers. For analysis we used amplification of cDNA by polymerase chain reaction, single-strand conformation polymorphism and sequencing techniques. In 5 of the cell lines, 9 mutations leading to changes of the amino acid sequence of DNA polymerase delta were detected. Most mutations were found in the cell lines DLD-1, HCT116 and SW48 for which defects in mismatch repair genes had been identified previously. In the majority of cases, wild type and mutated sequences were present. In 2 cell lines (HCT116 and SW48), a single-nucleotide deletion occurred at the same position. This resulted in a premature termination codon by which the DNA interaction domain of the enzyme was eliminated. Furthermore, sequence deviations were found in the tumor tissues of 4 colon cancer patients. Wild-type and altered sequences were present simultaneously. The deviations included missense mutations (2 cases) and silent mutations (2 cases). The missense mutations and one of the silent mutations were found in normal mucosa as well. In addition, the mutation clustered region of a tumor suppressor gene, often found to be defective in colon cancer, the adenomatous polyposis coli (APC) gene, was investigated in surgical specimens and cell lines. One carcinoma and 2 cell lines exhibited amino acid changes in both the DNA polymerase delta gene and in the mutation clustered region of the APC gene. Since most of the mutations detected in the DNA polymerase delta mRNA are likely to alter the structure of the protein, the enzyme is expected to be functionally impaired. In particular, copying fidelity might be decreased, thus contributing to the high mutation rate observed in colorectal cancer.

Aged↗
PubMed1999

Lipopeptides as dimerization inhibitors of HIV-1 protease.

In AIDS therapy, attempts have been made to inhibit the virus-encoded enzymes, e.g. HIV-1 protease, using active site-directed inhibitors. This approach is questionable, however, due to virus mutations and the high toxicity of the drugs. An alternative method to inhibit the dimeric HIV protease is the targeting of the interface region of the protease subunits in order to prevent subunit dimerization and enzyme activity. This approach should be less prone to inactivation by mutation. A list of improved 'dimerization inhibitors' of HIV-1 protease is presented. The main structural features are a short 'interface' peptide segment, including non-natural amino acids, and an aliphatic N-terminal blocking group. The high inhibitory power of some of the lipopeptides [e.g. palmitoyl-Tyr-Glu-Leu-OH, palmitoyl-Tyr-Glu-(L-thyronine)-OH, palmitoyl-Tyr-Glu-(L-biphenyl-alanine)-OH] with low nanomolar Ki values in the enzyme test suggests that mimetics with good bio-availability can be derived for AIDS therapy.

Amino Acid Sequence↗
PubMed1998

Biological variation and quantification of health: the emergence of the concept of normality.

Historical research on the concept of normality, its roots and its development show that this concept has its sources in very different areas of scientific and medical thinking. Of great importance were: (i) a new theory of disease arising early in the 19th century supposing a continuous change from the healthy to the diseased state; (ii) the examination of variation within and between species of plants and animals; (iii) the clinical theory of constitution developed to describe the wholeness of the individual determined both by genetic factors and the influence of the environment; (iv) the development of mathematical and statistical tools starting with the adaptation of Bernoulli's "law of the great numbers" and Gauss' and Legendre's "law of errors" to biological measurements by the Belgian astronomer Quetelet. At the end of the First World War the concept of normality was first discussed. An idealistic "value norm" was set against a statistical "frequency norm". Between 1920 and 1930 the principles of our present concept were accepted and the mathematical tools developed. It took several decades to introduce this concept into practical medicine finally being recommended by the IFCC.

Clinical Laboratory Techniques↗
PubMed1998

Determination of pH in human erythrocytes. Sources of systematic error.

The results of a simultaneous determination of the pH value in erythrocytes with the potentiometric measuring method and the 14C-labelled 5,5-dimethyl-2,4-oxazolidinedione (DMO) method showed a mean method difference of 0.026 pH units. The cause of this discrepancy was assumed to be matrix-inherent liquid-junction potentials in the potentiometric measurement. Taking these into account in the calculation leads to consistent values for the methods investigated. The DMO method proved to be free of systematic errors. Another indication of this is that its mean ratio of extracellular to intracellular H+ ion concentration (H+e/H+i) substantially agreed with the distribution ratios of other freely diffusible ions and their pH dependence.

Erythrocytes↗
PubMed1997

Diagnostic validity as a theoretical concept and as a measurable quantity.

The analytical result of a laboratory examination is a scientific fact and has no medical meaning as such. It must be interpreted to become a medical finding. To explain the very complex cognitive procedure of the interpretation a three-level model is used. In an environment of cost containment in health care systems the quality of medical laboratory findings is very important. Analytical results are monitored by quality control procedures. For measuring the performance of medical findings the concept of the 'validity' of a laboratory test is used. Validity means the 'degree of achieving the objective'. Accordingly, a valid laboratory finding is one which correctly answers the question which the physician at the sick-bed directs to the laboratory. Quantitative measures for the validity of interpretation can be developed by an analysis of the underlying classification processes. Characteristic indices describing the validity quantitatively in terms of conditional probabilities can be derived from decision tables. Examples of 'validity indices' are diagnostic (or prognostic) 'sensitivity' and 'specificity'. These indices are powerful tools for developing strategies for the clinical use of laboratory examinations in diagnosis, prognosis and therapy management. Moreover, validity indices are appropriate output quantities for the estimation of effectiveness and efficiency of a diagnostic or prognostic examination.

Clinical Laboratory Techniques↗
PubMed1997

Screening of inhibitors of HIV-1 protease using an Escherichia coli cell assay.

To evaluate the available peptidic and pseudopeptidic inhibitors of HIV protease for their possible in vivo activity, a screening test using Escherichia coli was established. E. coli cells carrying the plasmid pET9c-PR containing the gene for HIV-1 protease under the control of a T7-promotor are grown in the absence and in the presence of inhibitors. The action of the toxic protease produced by the cells is counteracted by the inhibitors. Provided sufficient membrane permeability of the inhibitors exists, this results in accelerated cell growth. From the peptides known to be active in an in-vitro enzyme test, most compounds inhibit HIV protease only to a limited degree in this test. However, two short peptides (Ac-Ser-Tyr-Glu-Leu and Lys-Ile-Ser-Tyr-Asp-Tyr) protect cell growth to an considerabe extent, thus indicating that they reach the E. coli cytosol and there block HIV protease. Two pseudopeptides known to be very potent in the enzyme test (SDZ PRI 053 and CIBA 61755) also inhibit HIV-1 protease strongly in this cell growth test.

Escherichia coli↗
PubMed1997

Clinical chemistry in laboratory medicine in Europe--past, present and future challenges.

European experts in clinical laboratory sciences have different backgrounds, and development of expertise in this field started more than 100 years ago. Specific national activities have created the heterogeneity that now exists amongst the academic professional membership of more than 40 scientific societies in Europe. The recent political changes have in addition contributed to the rapidly changing profile of Clinical Chemistry and related fields. Based on a questionnaire answered by 31 national representatives, the past, present and future aspects of the European Clinical Laboratory are reviewed. Of the more than 30,000 members of national societies, the majority studied medicine (40.1%), chemistry (27.2%) and pharmacy (21.1%) with large national differences in relative percentages. Post-graduate education is provided by two thirds of the national societies. In most European countries the same experts cover not only clinical chemistry but also haematology, haemostaseology, immunology and transfusiology. National quality assurance programmes are said to be established in 25 countries, but mandatory in only 11 of them. Of the future challenges, the implementation of request strategies were named most often, with interpretative reports and preanalytical aspects estimated as similarly important. It was thought that information technology and new scientific developments would make the greatest impact in the coming years, with economic pressure being the major limiting factor. Despite these limitations an increase in the number of tests is anticipated by most representatives, supporting the assumption of an increasing role of the clinical laboratory in future clinical medicine.

Chemistry, Clinical↗
PubMed1996

The inhibition of human immunodeficiency virus proteases by 'interface peptides'.

The active human immunodeficiency virus type 1 (HIV-1) protease has a homodimeric structure, the subunits are connected by an 'interface' beta-sheet formed by the NH2- and COOH-terminal amino acid segments. Short peptides derived from these segments are able to inhibit the protease activity in the range of micromolar IC50 values. We have further improved the inhibitory power of such peptides by computer modelling. The best inhibitor, the palmitoyl-blocked peptide Pam-Thr-Val-Ser-Tyr-Glu-Leu, has an IC50 value of less than 1 microM. Some of the peptides also showed very good inhibition of the HIV-2 protease. The C-terminal segment of the HIV-1 matrix protein, Acetyl-Gln-Val-Ser-Gln-Asn-Tyr, also inhibits HIV-1 protease. Kinetic studies confirmed the 'dissociative' mechanism of inhibition by the peptides. Depending on the peptide structure and ionic strength, both dimerization inhibition and competitive inhibition were observed, as well as synergistic effects between competitive inhibitors and interface peptides.

Antiviral Agents↗
PubMed1996

[Fulminant air embolism in arthroscopy of the shoulder joint using CO2].

We report on a case of an unexpected gas embolism during an arthroscopy of a newly injured shoulder joint. We used carbon dioxide as an arthroscopic medium which has not been considered hazardous up to now. The sudden decrease of the endtidal CO2 concentration in connection with a decrease of peripheral oxygen saturation as well as the clinical symptoms led to an immediate diagnosis of gas embolism. In this case external cardiac massage and aspiration of gas from the right ventricle proved an effective treatment.

Arthroscopy↗
PubMed1994

Clinical chemistry as scientific discipline: historical perspectives.

The fundamental ideas which underlie clinical chemistry as an independent scientific field were formed over the course of centuries. Exactly 200 years ago the first modern concepts for this discipline were formulated in close connection with the restructuring of medical education during the French Revolution on the one hand, and the emergence of a new idea of a 'clinic' on the other hand. However, not until 1840 was clinical chemistry institutionalized as academic subject and simultaneously integrated into medical teaching. After about 1860, clinical chemistry was practiced by the clinicians themselves in close relationship with clinical activities, yet again with emphasis on teaching. In this period, clinics and hospitals established 'clinical laboratories'. With the start of the 20th century, after biochemistry had developed into an independent scientific field, clinical chemistry continued to evolve in close relationship with that latter discipline. This was particularly true in the United States, where an 'American School of Clinical Biochemistry' emerged which was to greatly influence the field.

Chemistry, Clinical↗