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Biomedical subjects

M Weitz

Publications and source records attributed to M Weitz.

At least 19 recordsLinked to original sources
PubMed1998

A prospective evaluation of magnetic resonance cholangiopancreatography in patients with suspected bile duct obstruction.

BACKGROUND: The value of magnetic resonance cholangiopancreatography (MRCP) is under debate. AIMS: To assess the diagnostic accuracy of MRCP and endoscopic retrograde cholangiopancreatography (ERCP) and to determine whether MRCP may help to prevent unnecessary interventional procedures. METHODS: Eighty six patients with suspected common bile duct obstruction who presented between January and December 1996 were enrolled. Twenty six were excluded due to anatomical reasons or because MRCP or ERCP could not be performed successfully. Results of MRCP were interpreted by two radiologists and a gastroenterologist unaware of clinical diagnosis. Final diagnosis was determined by ERCP and histopathological findings or a follow up of at least 12 months. RESULTS: MRCP images of diagnostic quality were obtained in all 60 patients. Thirteen patients had a clear bile duct. Sensitivity and specificity for the detection of any abnormality (n=47) were 89% and 92%, and for the detection of malignancy (n=27) 81% and 100%, respectively. These results were equivalent to the respective figures of ERCP (91% and 92% for any abnormality, and 93% and 94% for malignant diseases). CONCLUSIONS: MRCP is as sensitive as ERCP in the evaluation of bilary tract diseases. As the specificity of this non-invasive technique is close to 100%, MRCP may prevent inappropriate invasive explorations of the common bile duct and pancreatic duct.

Adolescent↗
PubMed1997

Value of magnetic-resonance cholangio-pancreatography (MRCP) after unsuccessful endoscopic-retrograde cholangio-pancreatography (ERCP).

BACKGROUND AND STUDY AIMS: The present study tries to evaluate the success rate of MRCP when two attempts by experts to perform ERCP in a center failed. PATIENTS AND METHODS: From March 1996 to December 1996 thirteen patients fulfilled the inclusion criteria. The MR cholangiopancreatograms were acquired using commercially available software in a clinical MR scanner (Magnetom Expert 1 T-Scanner, Siemens, Erlangen, Germany). MRCP utilized heavily T2-weighted turbo-spin echo sequences with fat supression (HASTE). Maximum intensity projection (MIP) of the pancreatic duct and biliary tree was then carried out. Additionally, T1-weighted sequences were obtained using the breath-hold technique. RESULTS: The MRCP images were of diagnostic quality in all 13 patients. In five cases the diagnoses detected by MRCP were followed by an interventional procedure. One patient showed a pancreatic pseudocyst, that was percutaneously drained using ultrasound guidance. In three cases we found benign bile duct obstruction, all of which were successfully treated by percutaneous transhepatic drainage. In one patient choledocholithiasis was diagnosed, the stone was successfully managed by percutaneous transhepatic extraction. CONCLUSION: MRCP is the method of choice in cases where ERCP is incomplete or where duct cannulation is not possible. A further advantage of MR imaging is the fact that it may provide complementary information about the whole region of interest, thus detecting the cause of duct pathology in many cases.

Aged↗
PubMed1997

Identification of active-site residues in protease 3C of hepatitis A virus by site-directed mutagenesis.

Picornavirus 3C proteases (3Cpro) are cysteine proteases related by amino acid sequence to trypsin-like serine proteases. Comparisons of 3Cpro of hepatitis A virus (HAV) to those of other picornaviruses have resulted in prediction of active-site residues: histidine at position 44 (H44), aspartic acid (D98), and cysteine (C172). To test whether these residues are key members of a putative catalytic triad, oligonucleotide-directed mutagenesis was targeted to 3Cpro in the context of natural polypeptide precursor P3. Autocatalytic processing of the polyprotein containing wild-type or variant 3Cpro was tested by in vivo expression of vaccinia virus-HAV chimeras in an animal cell-T7 hybrid system and by in vitro translation of corresponding RNAs. Comparison with proteins present in HAV-infected cells showed that both expression systems mimicked authentic polyprotein processing. Individual substitutions of H44 by tyrosine and of C172 by glycine or serine resulted in complete loss of the virus-specific proteolytic cascade. In contrast, a P3 polyprotein in which D98 was substituted by asparagine underwent only slightly delayed processing, while an additional substitution of valine (V47) by glycine within putative protein 3A caused a more pronounced loss of processing. Therefore, apparently H44 and C172 are active-site constituents whereas D98 is not. The results, furthermore, suggest that substitution of amino acid residues distant from polyprotein cleavage sites may reduce proteolytic activity, presumably by altering substrate conformation.

3C Viral Proteases↗
PubMed1996

[Infectious diseases: current diagnostic methods].

A microbiological diagnosis is either based on the direct detection of the pathogen or on the reaction of the host (indirect method). Many pathogens, in particular small bacteria and viruses, are undetectable by culture or difficult to cultivate. Molecular methods for the direct detection of such pathogens have been widely introduced in many microbiological laboratories within the past few years. The characteristics of these methods as well as new culture methods and new serological tests are discussed.

Bacteria↗
PubMed1996

Identification of hepatitis A virus non-structural protein 2B and its release by the major virus protease 3C.

The RNA genome of hepatitis A virus (HAV) encodes a giant polyprotein that is putatively cleaved proteolytically into four structural and seven non-structural proteins. So far, most of the proposed non-structural proteins and their respective cleavage sites have not been identified. A vaccinia virus recombinant (vRGORF) containing the complete HAV ORF under the control of the bacteriophage T7 promoter was used to express HAV in recombinant animal cells (BT7-H) that constitutively expressed T7 DNA-dependent RNA polymerase. A HAV-specific 27.5 kDa expression product was identified as peptide 2B. The 27.5 kDa 2B antigen was also found in HAV-infected MRC-5 cells. The N-terminal amino acid residues of the new peptide 2B are Ala-Lys-Ile-Ser-Leu-Phe and polyprotein cleavage between 2A and 2B occurred at amino acids 836-837 (Gln-Ala). Furthermore, heterologous expression in the same system of regions P1-P2 and of the protease 3C (3Cpro) gene, showed that P1-P2 polyprotein is not cleaved autocatalytically but by 3Cpro. Hence, 3Cpro is effective in cleaving the polyprotein 2A-2B junction.

3C Viral Proteases↗
PubMed1996

RNA-protein interactions at the 3' end of the hepatitis A virus RNA.

The regulative cis-acting terminal RNA structures and the proteins involved in the amplification of the hepatitis A virus (HAV) genome are unknown. By UV cross-linking/label transfer experiments, we have analyzed sequences of the 3'-nontranslated region (3'NTR) and preceding domains of the viral genome for their ability to interact with host proteins. A series of cDNA constructs were used to create genomic- and antigenomic-sense transcripts. The results show that the 3'-NTR-poly(A) interacted with host cell proteins with molecular masses of 38, 45, 57, 84, and 110 kDa only weakly, compared with RNA structures also consisting of 3D-coding regions. Protein p38 was most efficiently labeled after interaction with secondary-structure elements located at the 3' end of the HAV RNA, p38 also interacted with a 5'-terminal RNA probe. Optimal RNA binding was found to be dependent on the salt concentration. The specificity of the RNA-protein interaction was proven by competition assays. These data might indicate that a higher-order structure formed at the junction of the 3Dpol-coding sequence and the 3'-NTR of the HAV genome (putative RNA pseudoknot) significantly improves binding of host proteins and thus suggests that this structure might be essential for the formation of the replication complex initiating minus-strand RNA synthesis.

Animals↗
PubMed1993

Human parvovirus B19 infections: routine diagnosis by a new nested polymerase chain reaction assay.

A nested primer PCR assay was developed to detect human parvovirus B19 in various clinical specimens in a routine diagnostic laboratory. Under optimized conditions the highly specific assay had a sensitivity of less than 10 genome units. For practical reasons, however, this sensitivity was adjusted to 10-100 virus genomes in diagnostic applications. Using clinical specimens from 200 patients with suspected B19 infection, nested PCR was shown to have important diagnostic advantages over the detection of B19 specific antibodies. The data suggest that on the basis of serological data as obtained with currently available test systems a considerable proportion of B19 infections would be misdiagnosed. Examples for the usefulness of the PCR assay in routine diagnosis are given.

Adolescent↗