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Biomedical subjects

M Uhl

Publications and source records attributed to M Uhl.

At least 19 recordsLinked to original sources
PubMed2000

Evaluation of the single cell gel electrophoresis assay with human hepatoma (Hep G2) cells.

Human Hep G2 cells have retained the activities of phase I and phase II enzymes which are involved in the metabolism of environmental genotoxins. The present study describes the results of single cell gel electrophoresis (SCGE) assays with a panel of different model compounds with these cells. With genotoxic carcinogens such as aflatoxin B(1) (AFB(1)), benzo(a)pyrene (B(a)P), nitrosodimethylamine (NDMA) and cyclophosphamide (CP), statistically significant dose dependent induction of DNA migration was measured. With the two heterocyclic amines, 2-amino-3-methyl-3H-imidazo[4, 5-f]quinoline (IQ) and 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1), and also with rodent carcinogens such as safrole, hexamethylphosphoramide (HMPA) and the pyrrolizidine alkaloid isatidine, which give negative results in other in vitro genotoxicity tests, positive results were obtained in Hep G2/SCGE assays. Nitrosomethylurea (NMU) was the only directly acting compound tested in the study and was by far (ca. 10(3)-fold) more active than the corresponding nitrosamine. The exposure concentrations required to cause significant effects varied over a broad range. The most pronounced effect was seen with AFB(1) (0.008 microM) followed by HMPA (15 microM), B(a)P (25 microM), NMU (100 microM), isatidin (500 microM), CP (900 microM), IQ (1200 microM), safrol (4000 microM), and NDMA (90x10(3) microM). Numbers in parenthesis give the lowest concentrations, which caused a significant increase of DNA migration. With two compounds, namely, the non-carcinogen pyrene and the synthetic hormone tamoxifen (TF), negative results were obtained under all test conditions. These findings are in agreement with the results of recent investigations which indicated that human hepatocytes are unable to convert TF to DNA reactive metabolites, whereas it is activated by rat liver cells and causes DNA adducts in these cells. Comparisons of the present results with data from earlier experiments indicate that the Hep G2/SCGE assay enables the detection of genotoxins including compounds which give misleading results in other in vitro genotoxicity tests and appears to be an alternative to tests with primary liver cells from laboratory animals.

Biotransformation↗
PubMed2000

A public domain image-analysis program for the single-cell gel-electrophoresis (comet) assay.

The single-cell gel electrophoresis (or comet) assay has gained widespread acceptance as a cheap and simple genotoxicity test, but it requires a computer-assisted image-analysis system. As commercial programs are expensive and inflexible, we decided to develop an image-analysis system based on public domain programs and make it publicly available for the scientific community. Our system is based on the scientific image-processing program NIH Image, and was written in its Pascal-like macro language. User interaction was kept as simple as possible, to enable the measurement of a large number of cells with a few keystrokes. Therefore, the time for image analysis is very low, even on slow computers. The comet macro can be obtained from http://mailbox.univie.ac.at/christoph.helma++ +/comet/, NIH Image is available at http://rsb.info.nih.gov/nih-image/. Both programs are free of charge.

Comet Assay↗
PubMed2000

Tandem mass spectrometry: a helpful tool in hair analysis for the forensic expert.

The Bavarian State Bureau of Investigation in Munich has the exclusive responsibility for investigation of criminal acts. One considerable expertise is that of hair analysis. According to the legal system in Germany, there is a special interest when some clients' hair tested positive for illicit drugs. An accused with a lot of drugs in his hair will be treated as a supposed addict and will be guaranteed extenuating circumstances. The instrumentation used for hair analysis is a powerful analytical tool: a Varian 3400 gas chromatograph linked to a Finnigan Tandem-MS (TSQ 700). The methanol extraction method is used for the detection of illegal drugs and metabolites: amphetamine, methamphetamine, MDA, MDMA (ecstasy), MDE, MBDB, methadone, THC, EDDP (metabolite of methadone), cocaine, benzoylecgonine, cocaethylene, opiates (dihydrocodeine, codeine, heroin, 6-monoacetylmorphine, morphine, acetylcodeine). For the detection of 9-carboxy-THC by negative chemical ionization the hair sample is hydrolyzed under alkaline conditions. Solid-phase extraction is used for clean-up. The LOQ for the determination of 11-nor-delta-9-tetrahydrocannabinol-9-carboxylic-acid is 0.16 pg/mg hair. An unsurpassed combination for rendering an expert opinion based on hair analysis may be: a forensic expert using diligence and experience, coupled with the performance of a sophisticated analytical instrument.

Cannabinoids↗
PubMed2000

Appearance of autosomal recessive polycystic kidney disease in magnetic resonance imaging and RARE-MR-urography.

PURPOSE: To describe the appearance of autosomal recessive polycystic kidney disease (ARPKD) on MRI and RARE-MR urography. MATERIALS AND METHODS: Seven boys and one girl (aged 3 months to 14 years, median 2.5 years) were evaluated. Images were obtained with 0.23-T and 1.5-T MR systems using T1-weighted (T1-W) spin-echo, T2-weighted (T2-W) turbo-spin-echo and RARE-MR-urography sequences. Signal intensities, morphological appearance of the affected kidneys and, specifically, the picture of the urinary tract on RARE-MR-urography were evaluated. RESULTS: All children showed kidney enlargement, reniform but humpy kidney shape, homogeneously grainy renal parenchyma, normal renal pelvis and normal calyces. Signal intensity was hyperintense in T2-W images in all cases. In six cases (n = 7), T1-W images were hypointense. On RARE-MR urography a hyperintense, linear radial pattern was seen in the cortex and medulla which represents the characteristic microcystic dilatation of collecting ducts in ARPKD. Three boys and the girl presented with a few circumscribed small subcapsular cysts. CONCLUSIONS: In order to confirm the diagnosis of ARPKD, RARE-MR urography seems to be a non-invasive imaging tool that shows directly the microcystic dilated water-filled collecting ducts.

Adolescent↗
PubMed2000

Ultrasound demonstration of mammographically detected microcalcifications.

PURPOSE: To evaluate the capabilities of breast ultrasound (US) for identifying microcalcifications in benign breast changes, in situ carcinomas, and small nonpalpable invasive carcinomas. MATERIAL AND METHODS: Forty-six consecutive patients with 49 clustered microcalcifications detected by mammography were included in this prospective study. Patients with palpable breast lesions were excluded. Breast US was performed with knowledge of mammographic findings for presence and visibility of microcalcifications, and for parenchymal structure abnormalities. Mammographic and US findings were compared with histology. RESULTS: Nine ductal in situ carcinomas, 2 lobular in situ carcinomas, 11 invasive carcinomas and 27 benign lesions were confirmed by histology. For all lesions, US achieved a sensitivity of 75% in the detection of microcalcifications. The detection rate for microcalcification in invasive and in situ carcinomas was 100%. In 11 cases, no microcalcifications were visible on US; they all proved to be benign on histology. CONCLUSION: Microcalcifications in malignant lesions are reliably recognized by US. They are, however, difficult to detect in fibrocystic breast changes.

Adult↗
PubMed1999

Single-cell gel electrophoresis assays with human-derived hepatoma (Hep G2) cells.

The purpose of the present study was the development of a protocol for detecting chemically-induced DNA damage, using the alkaline single-cell gel electrophoresis (SCGE) assay with human-derived, metabolically competent hepatoma (Hep G2) cells. Previous studies indicated that Hep G2 cells have retained the activities of certain phase I and phase II enzymes and reflect the metabolism of genotoxins in mammals better than other in vitro models which require addition of exogenous activation mixtures. The optimal trypsin concentration for the removal of the cells from the plates were found to be 0.1%. Dimethylsulfoxide, at concentrations up to 2%, was an appropriate solvent for water-insoluble compounds. To determine the optimal exposure periods for mutagen treatment, the time kinetics of comet formation was investigated with genotoxic chemicals representing various classes of promutagens namely benzo[a]pyrene (B[a]P), 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), and N-nitrosodimethylamine (NDMA) and with N-nitrosomethylurea (NMU). All compounds caused a statistically significant induction in DNA damage. With the promutagens, comet formation increased gradually as a function of the exposure duration, and reached maximum values between 20-24 h. With NMU, comet induction maximized already after a short exposure (1 h) and remained at a constant level for up to 24 h. Based on these results, the Hep G2/SCGE assay appears to be a suitable approach for investigating DNA damaging potential of chemicals. Further experiments with IQ and B[a]P showed that the assays are highly reproducible. Comparisons of the present results with those from earlier experiments in which other endpoints (induction of sister chromatid exchanges, micronuclei and chromosomal aberrations) were measured in Hep G2 cells, indicated that the sensitivity of the SCGE assays is more or less identical. Since the SCGE assay is less time consuming than other genotoxicity assays we anticipate that it might be a suitable approach to investigate DNA damaging effects of chemicals in the human-derived, metabolically competent cell line.

Benzo(a)pyrene↗
PubMed1999

Splenic peliosis: a rare complication following liver transplantation.

In this article, we report a rare case of isolated splenic peliosis in an individual who had recently undergone liver transplantation. The disorder had remained clinically and radiologically undiagnosed until he suffered a traumatic rupture of the affected organ. The relevant literature on this topic is briefly reviewed.

Biopsy↗
PubMed1999

[Kinematic versus static MRI study of the cervical spine in patients with rheumatoid arthritis].

PURPOSE: The objective of this study was to compare the diagnostic value of cinematic magnetic resonance imaging with static MRI examinations in patients with rheumatoid arthritis and concomitant attack of the cervical spine. METHODS: Dynamic functional MRI examinations of the cervical spine were performed one five subjects without complaints and 20 patients with rheumatoid arthritis. For the functional studies, a positioning frame was used that allowed infinitely variable forward and backward inclinations of the head. RESULTS: Functional magnetic resonance imaging made possible a sufficiently good differentiation of the extension of pannus tissue cranial, ventral, and dorsal of the dens with possible displacing and impinging effects on the spinal cord during flexing and stretching movements. In addition, it is suitable for demonstration of the degree of instability in the atlanto-occipital and atlanto-axial planes. In contrast to conventional X-rays, CT, and static MRI, basilary impression as well as compressions and angulations of the cervical bone marrow are better visualized by cinematic magnetic resonance tomography. CONCLUSIONS: Functional magnetic resonance tomography is an important diagnostic method for the induction of the cervical spine in patients with rheumatoid arthritis. In particular, fusion and instabilities as well as compressions of the bone marrow often can only be detected with the help of functional MRI.

Cervical Vertebrae↗
PubMed1999

Indirect MR arthrography of the unexercised glenohumeral joint in patients with rotator cuff tears.

RATIONALE AND OBJECTIVES: To evaluate the diagnostic utility of indirect MR arthrography of the unexercised glenohumeral joint in patients with rotator cuff tears confirmed by arthroscopy or arthrotomy as the gold standard. METHODS: Twenty-six patients underwent conventional MR imaging and indirect MR arthrography of the stationary glenohumeral joint using a wrap-around surface coil. Unenhanced T1-weighted spin echo/T2-weighted fast spin echo sequences and T1-weighted gradient echo sequences, adding spectral fat suppression after intravenous administration of contrast medium, were performed in the oblique coronal and oblique sagittal planes. Images were analyzed by three experienced radiologists in consensus. Levels of diagnostic confidence were evaluated using a four-point scale of diagnostic certainty. RESULTS: Performing indirect MR arthrography of the unexercised shoulder leads to a diagnostically efficient enhancement of joint fluid (120% at 4 minutes and 145% at 8 minutes after intravenous injection of gadodiamide). In terms of soft tissue delineation, characterization of rotator cuff tears was significantly improved by using enhanced fat-suppressed T1-weighted gradient echo sequences compared with conventional MR imaging. CONCLUSIONS: Indirect MR arthrography without glenohumeral joint exercise in the diagnosis of rotator cuff tears is feasible and represents a more convenient and less time-consuming alternative to indirect MR arthrography after joint exercise.

Adult↗
PubMed1999

Cystic medial degeneration of the aorta is associated with p53 accumulation, Bax upregulation, apoptotic cell death, and cell proliferation.

OBJECTIVE: To address a potential role for p53, Bcl2 associated protein X (Bax), and apoptosis in the processes associated with cell turnover during cystic medial degeneration (CMD) of the aorta. METHODS: Histochemical, immunohistochemical, biochemical, and morphometric methods were used to assess the presence and distribution of p53 immunoreactivity (p53-IR) and Bax immunoreactivity (Bax-IR), as well as the presence of apoptosis and tissue repair processes. RESULTS: Immunohistochemical staining disclosed evidence for p53-IR in all specimens in 26.1 (11.5)% of vascular smooth muscle cells (VSMCs) (controls 0.8 (1.3)%; p < 0. 001). Bax-IR was present in all specimens in 10 (5.4)% of medial cells (controls 0.3 (0.5)%; p < 0.001). Medial VSMCs (alpha-actin positive) with cytoplasmic staining for an apoptosis specific protein (c-jun/ASP) were present in 20/20 specimens (0.7 (0.6)% of VSMCs, controls 0%, p < 0.001), whereas terminal deoxynucleotidyl transferase mediated dUTP-biotin nick end labeling (TUNEL) positive VSMCs were present in 17/20 specimens (1 (1.5)% of VSMCs, controls 0%, p < 0.001). The presence of apoptosis was confirmed by electron microscopy and the demonstration of oligonucleosomal DNA fragments after agarose gel electrophoresis. As shown by double labeling and investigation of serial sections, p53-IR, Bax-IR, c-jun/ASP-IR, and positive TUNEL labeling localised to the same compartments of the aortic media, raising a possible role for p53 and Bax in the triggering of apoptosis of VSMC during CMD. MIB1/Ki-67 positive medial VSMCs (alpha-actin positive) and mesenchymal cells (vimentin positive) were present in all specimens (2.5 (2.8)% of medial cells; controls 0.3 (0.9)%, p < 0.001) mainly in the region around the vasa vasorum, indicating that cell regeneration during CMD may originate mainly from the mesenchyme surrounding the vasa vasorum. CONCLUSION: This study shows that the formal pathogenesis of CMD is characterised by p53 accumulation, Bax upregulation, cell death by apoptosis, and cell regeneration. Nevertheless, the precise stimuli of p53 activation and Bax upregulation as well as the role of p53 and apoptosis in the dissection process itself remain elusive.

Adult↗
PubMed1999

Rare-MR-urography--a new diagnostic method in autosomal recessive polycystic kidney disease.

PURPOSE: To describe the appearance of autosomal recessive polycystic kidney disease (ARPKD) by using a new diagnostic method: RARE-MR-urography. MATERIAL AND METHODS: Eight children were evaluated using MR images from 0.23 T and 1.5 T MR units, using T1-weighted spin-echo and T2-weighted turbo spin-echo sequences and RARE-MR-urography. Signal intensities, morphological appearance of the affected kidneys and, specifically, the picture of the urinary tract in RARE-MR-urography, were evaluated. RESULTS: All children showed enlargement, reniform but humpy kidney shape, homogeneous-grainy renal parenchyma, normal renal pelvis and calyces. Although ARPKD is always associated with some degree of congenital hepatic fibrosis, there was no bile duct dilatation or liver fibrosis at the time of examination. Signal intensity was hyperintense in T2-weighted images in all cases. In 5 cases, T1-weighted images were hypointense. In RARE-MR-urography, hyperintense, linear, radial patterns in cortex and medulla were seen, which represent microcystic dilatation of collecting ducts and are therefore characteristic of ARPKD. Four patients presented with a few circumscribed small subcapsular cysts. CONCLUSION: RARE-MR-urography is a noninvasive method which demonstrates the pathognomonic water-filled cystic structures throughout the kindeys in ARPKD.

Adolescent↗
PubMed1998

Use of metabolically competent human hepatoma cells for the detection of mutagens and antimutagens.

The human hepatoma line (Hep G2) has retained the activities of various phase I and phase II enzymes which play a crucial role in the activation/detoxification of genotoxic procarcinogens and reflect the metabolism of such compounds in vivo better than experimental models with metabolically incompetent cells and exogenous activation mixtures. In the last years, methodologies have been developed which enable the detection of genotoxic effects in Hep G2 cells. Appropriate endpoints are the induction of 6-TGr mutants, of micronuclei and of comets (single cell gel electrophoresis assay). It has been demonstrated that various classes of environmental carcinogens such as nitrosamines, aflatoxins, aromatic and heterocyclic amines and polycyclic aromatic hydrocarbons can be detected in genotoxicity assays with Hep G2 cells. Furthermore, it has been shown that these assays can distinguish between structurally related carcinogens and non-carcinogens, and positive results have been obtained with rodent carcinogens (such as safrole and hexamethylphosphoramide) which give false negative results in conventional in vitro assays with rat liver homogenates. Hep G2 cells have also been used in antimutagenicity studies and can identify mechanisms not detected in conventional in vitro systems such as induction of detoxifying enzymes, inactivation of endogenously formed DNA-reactive metabolites and intracellular inhibition of activating enzymes.

Animals↗
PubMed1998

MR findings in pontocerebellar hypoplasia.

We present four cases with combined hypoplasia of the cerebellum and the ventral pons-pontocerebellar hypoplasia (PCH). PCH represents an autosomal recessive neurodegenerative disorder with fetal onset. The disease is rare, with less than 20 cases having been reported. The main findings of PCH and the inclusion criteria for our cases can be summarised as progressive microcephaly from birth, pontocerebellar hypoplasia documented by MRI and marked chorea, which may change, later in childhood, to more dystonic patterns. The cerebral cortex becomes progressively atrophic. Motor and mental development are delayed, and epilepsy, mainly tonic-clonic seizures, is frequent. The MRI features in all of our cases were: (1) Hypoplastic cerebellum situated close to the tentorium. The hypoplastic cerebellum has a reduced number of folia, in contrast to the normal number of thin folia in simple cerebellar atrophy. (2) The cerebellar hemispheres are reduced to bean-like or wing-like structures. The cerebellar hemispheres appear to 'float' in the posterior fossa. (3) Markedly hypoplastic ventral pons. (4) Slight atrophy of the supratentorial gyral pattern. (5) Dilated cerebromedullary cistern and fourth ventricle. (6) Delayed myelination of the white matter. (7) No significant disorganisation of brain architecture and no severe corpus callosum defect.

Cerebellum↗
PubMed1998

Comparison of turbo inversion recovery magnitude (TIRM) with T2-weighted turbo spin-echo and T1-weighted spin-echo MR imaging in the early diagnosis of acute osteomyelitis in children.

OBJECTIVE: To compare turbo inversion recovery magnitude (TIRM) with standard T1-weighted (T1-W) and T2-weighted (T2-W) MR sequences in the very early detection of acute osteomyelitis in children. MATERIALS AND METHODS: In 15 children with osteomyelitis, 15 sets of T1-W spin-echo (SE) (TR/TE, 400-640/12-17), T2-W turbo spin-echo (TSE) (TR/TE/ETL, 3290-4465/112-120/11), and TIRM (TR/TE/TI, 4000-6120/60/160) images were acquired with a 1.0-T magnet. Contrast-to-noise (C/N) ratios and percentage of signal between lesion and normal bone marrow were analysed with a computer-assisted image analysing system in a region of interest (ROI). RESULTS: In 13 of 15 patients, the absolute signal enhancement in a ROI on the TIRM images was better than on the T1-W SE and T2-W TSE images and in 14 of 15 cases, C/N ratios were also better on the TIRM images than on the other sequences. In the other cases, the TIRM signal was diagnostically equivalent. On the TIRM images, the signal difference between normal and pathological tissue was increased to 43-281% (mean 124%). On the T2-W TSE images, this signal difference was 4-79% (mean 36%) and on the T1-W SE images 6-77% (mean 37%). Conclusion. The TIRM sequence is highly sensitive for detecting bone marrow oedema in the very early stage of acute osteomyelitis in children. MRI utilising the TIRM sequence allowed for an early diagnosis. With scan time of less than 4 minutes, this sequence is superior to T1-W SE and T2-W TSE images for detecting early osteomyelitis-associated bone marrow oedema.

Acute Disease↗