Biomedical subjects

U Fischer

Publications and source records attributed to U Fischer.

At least 37 records · Page 2Linked to original sources

PubMed2002

Clinical relevance of urokinase-type plasminogen activator and its inhibitor type 1 (PAI-1) in squamous cell carcinoma of the uterine cervix.

OBJECTIVE: The expression of uPA and PAI-1 as parameters of tumour-associated proteolysis has been implicated in the process of tumour cell invasion and the metastatic process. However, there is limited information on the impact of these parameters in cervical carcinoma. METHODS: Quantitative levels for uPA (n = 114) and PAI-1 (n = 103) were researched in operatively treated, surgically staged squamous cell cancer of the uterine cervix, using an ELISA-technique. Results were assessed regarding their impact in predicting pelvic lymph nodes metastases, tumour recurrence rate and recurrence free survival (RFS) using uni- and multivariate analysis. RESULTS: Median levels of both parameters were significantly higher in tumour tissue than in normal cervical tissue (p < 0.001). Detection of uPA gave no useful prognostic information. PAI-1 concentration showed a positive correlation with advanced tumour stage (p = 0.008), but no significant correlation with nodal status (pN0: 2.6 vs. pN1: 4.0 ng/mg protein; p = 0.092). Using a cut-off level of 2.4 ng/mg protein, patients with elevated PAI-1 levels demonstrated reduced RFS (45.9 versus 52.9 months; p = 0.1). Multivariate analysis, including nodal status, tumour stage, lymphovascular space involvement and grading failed to demonstrate any prognostic impact of uPA and PAI-1. CONCLUSIONS: The results indicate, that PAI-1 expression is of some prognostic impact in cervical cancer, indicating an association of elevated PAI levels with local tumour progression and reduced recurrence-free survival.

Adult↗

PubMed2001

Methylation of Sm proteins by a complex containing PRMT5 and the putative U snRNP assembly factor pICln.

Seven Sm proteins, termed B/B', D1, D2, D3, E, F, and G, assemble in an ordered manner onto U snRNAs to form the Sm core of the spliceosomal snRNPs U1, U2, U4/U6, and U5. The survival of motor neuron (SMN) protein binds to Sm proteins and mediates in the context of a macromolecular (SMN-) complex the assembly of the Sm core. Binding of SMN to Sm proteins is enhanced by modification of specific arginine residues in the Sm proteins D1 and D3 to symmetrical dimethylarginines (sDMAs), suggesting that assembly might be regulated at the posttranslational level. Here we provide evidence that the previously described pICln-complex, consisting of Sm proteins, the methyltransferase PRMT5, pICln, and two novel factors, catalyzes the sDMA modification of Sm proteins. In vitro studies further revealed that the pICln complex inhibits the spontaneous assembly of Sm proteins onto a U snRNA. This effect is mediated by pICln via its binding to the Sm fold of Sm proteins, thereby preventing specific interactions between Sm proteins required for the formation of the Sm core. Our data suggest that the pICln complex regulates an early step in the assembly of U snRNPs, possibly the transfer of Sm proteins to the SMN-complex.

Animals↗

PubMed2001

Preoperative local MRI-staging of patients with a suspected pancreatic mass.

The aim of this study was to define the value of MRI of the pancreas for preoperative local staging of patients with a suspected pancreatic mass. Ninety-four patients (41 women, 53 men; age range 32-87 years) with a suspected pancreatic tumor underwent preoperative staging with MRI on a 1.5-T system. The MRI protocol included breath-hold MR cholangiopancreatography in turbo spin-echo technique, biphasic contrast-enhanced 3D MR angiography, and MRI of the upper abdomen with breath-hold T2-weighted half-Fourier acquired single-shot turbo spin-echo and T1-weighted fast-low-angle-shot (pre- and postcontrast) sequences. Data were collected prospectively and analyzed by two radiologists in agreement modality. Evaluation criteria were vascular involvement, resectability, and a characterization benign vs malignant. Results were compared to histopathology in 78 patients. Sixteen patients were followed-up. In 74 of 94 patients a solid tumor or an inflammation of the pancreas ( n=62) or the papilla ( n=12) was detected. In this group, MRI had a sensitivity of 98%, a specificity of 92%, and an accuracy of 96% in the characterization of malignant tumors. Regarding only the solid tumors, the positive predictive value of MRI was 87% with respect to resectability. Other pathologic findings included adenoma or inflammation of the duodenum ( n=5), carcinoma or benign stenosis of the choledochus duct ( n=7) and carcinoma of the gall bladder ( n=2). In 6 patients MRI did not depict any pathologic findings, and follow-up confirmed this interpretation. Magnetic resonance imaging allows a local preoperative staging in patients with suspected pancreatic tumor. Limitations, however, concern to the diagnostics of peritoneal and/or liver metastases.

Adult↗

PubMed2001

Direct interaction of the spinal muscular atrophy disease protein SMN with the small nucleolar RNA-associated protein fibrillarin.

Disruption of the survival motor neuron (SMN) gene leads to selective loss of spinal motor neurons, resulting in the fatal human neurodegenerative disorder spinal muscular atrophy (SMA). SMN has been shown to function in spliceosomal small nuclear ribonucleoprotein (snRNP) biogenesis and pre-mRNA splicing. We have demonstrated that SMN also interacts with fibrillarin, a highly conserved nucleolar protein that is associated with all Box C/D small nucleolar RNAs and functions in processing and modification of rRNA. Fibrillarin and SMN co-immunoprecipitate from HeLa cell extracts indicating that the proteins exist as a complex in vivo. Furthermore, in vitro binding studies indicate that the interaction between SMN and fibrillarin is direct and salt-stable. We show that the glycine/arginine-rich domain of fibrillarin is necessary and sufficient for SMN binding and that the region of SMN encoded by exon 3, including the Tudor domain, mediates the binding of fibrillarin. Tudor domain missense mutations, including one found in an SMA patient, impair the interaction between SMN and fibrillarin (as well as the common snRNP protein SmB). Our results suggest a function for SMN in small nucleolar RNP biogenesis (akin to its known role as an snRNP assembly factor) and reveal a potential link between small nucleolar RNP biogenesis and SMA.

Amino Acid Sequence↗

PubMed2001

Contrast-enhanced MR angiography of abdominal vessels: is there still a role for angiography?

The purpose of this review article is to describe recent advantages in contrast-enhanced (CE) three-dimensional (3D) magnetic resonance angiography (MRA) in comparison with other vascular imaging techniques, and to discuss their current clinical applications for the imaging of abdominal vessels. Principles and technical considerations are presented and clinical applications are reviewed for different vascular diseases. In ruptured aortic aneurysms and acute dissections CT is the method of first choice. Contrast-enhanced 3D MRA can be well used for therapeutic planning and follow-up in patients with stable disease. A comprehensive MR examination including CE 3D MRA, MR urography and MR nephrogram has the potential to replace the conventional studies for the evaluation of renal vascular disease. It is an accurate method for imaging the origins of coeliac and superior mesenteric arteries, although the image resolution is too low for reliable assessment of the inferior mesenteric artery. Contrast-enhanced 3D MRA has emerged as the method of choice for studying the portal venous system in liver transplant recipients, in patients with portal hypertension and in cases with abdominal tumours for preoperative evaluation. Additional non-invasive flow measurements are useful in monitoring portal hypertension. The abdominal veins can be well imaged using unenhanced MR techniques. Imaging may be facilitated with intravascular contrast media. Contrast-enhanced 3D MRA can replace intra-arterial DSA for diagnosis, therapy planning and follow-up in patients with abdominal vascular disease. Catheter-based arteriography will still be used for interventional procedures such as percutaneous transluminal angioplasty, stent placement and embolisation.

Abdomen↗

PubMed2001

Expression, cellular distribution and protein binding of the glioma amplified sequence (GAS41), a highly conserved putative transcription factor.

The glioma amplified sequence 41 (GAS41) was previously isolated by microdissection mediated cDNA capture from the glioblastoma multiforme cell line TX3868 and shown to be frequently amplified in human gliomas. We determined the complete cDNA sequence of the GAS41 gene, demonstrated that the GAS41 protein is evolutionarily conserved, specifically at the N-terminus, and identified the yeast transcription factor tf2f domain within the GAS41 sequence. A human multiple-tissue Northern blot revealed ubiquitous expression of GAS41 with the highest expression in human brain. After generating polyclonal antibodies we found GAS41 protein expression in the nucleus of the TX3868 cell line by Western blot analysis and immunofluorescence microscopy. The nuclear localization was confirmed for several human tumors including gliomas of different grades of malignancy. In neuroblastoma however, GAS41 was found in the nucleoli but not in the nucleoplasm. Yeast two-hybrid screening of the TX3868 cell line identified the nuclear mitotic apparatus protein (NuMA), the KIAA1009 protein, and prefoldin subunit 1 (PFDN1) as potential interacting partners of GAS41. We generated a polyclonal antibody against the KIAA1009 protein and we demonstrated that the KIAA1009 protein is a nuclear protein, which appears to be co-localized with the GAS41 protein and NuMA.

Amino Acid Sequence↗

PubMed2001

PHF3-specific antibody responses in over 60% of patients with glioblastoma multiforme.

Glioblastoma multiforme (GBM), a malignant astrocytic tumour, represents the most frequent tumour of the human brain. Nevertheless, its molecular pathology is not well understood. We utilized the immune system, which contributes to cancer protection, to help identify new GBM-related genes. By screening a human GBM cDNA library with autologous patient serum (SEREX-approach), we isolated a gene termed PHF3 (PHD finger protein 3). The gene product of PHF3 is immunogenic in GBM as tested in an allogenic patient serum screening demonstrating antibodies in 24 of 39 (61.53%) sera, whereas none of the 14 healthy persons had antibodies against PHF3. While previous SEREX studies revealed allogenic antibody responses up to 40%, our results for PHF3 represent the highest reported rate for a specific antibody response. We show that GBM patients with an antibody response against PHF3 show significant better survival than patients without PHF3-specific antibodies. Because the amino acid sequence of PHF3 contains a PHD finger (also termed LAP motif), a TFIIS homology, a proline rich region and nuclear localization signals, it supposedly functions as a transcription factor. A polyclonal antibody generated against PHF3 shows nuclear expression in most investigated formalin-fixed, paraffin embedded tissues. In GBM, PHF3 expression is concentrated in cells surrounding necroses.

Adult↗

PubMed2001

SMNrp is an essential pre-mRNA splicing factor required for the formation of the mature spliceosome.

SMNrp, also termed SPF30, has recently been identified in spliceosomes assembled in vitro. We have functionally characterized this protein and show that it is an essential splicing factor. We show that SMNrp is a 17S U2 snRNP-associated protein that appears in the pre-spliceosome (complex A) and the mature spliceosome (complex B) during splicing. Immunodepletion of SMNrp from nuclear extract inhibits the first step of pre-mRNA splicing by preventing the formation of complex B. Re-addition of recombinant SMNrp to immunodepleted extract reconstitutes both spliceosome formation and splicing. Mutations in two domains of SMNrp, although similarly deleterious for splicing, differed in their consequences on U2 snRNP binding, suggesting that SMNrp may also engage in interactions with splicing factors other than the U2 snRNP. In agreement with this, we present evidence for an additional interaction between SMNrp and the [U4/U6.U5] tri-snRNP. A candidate that may mediate this interaction, namely the U4/U6-90 kDa protein, has been identified. We suggest that SMNrp, as a U2 snRNP-associated protein, facilitates the recruitment of the [U4/U6.U5] tri-snRNP to the pre-spliceosome.

Animals↗

PubMed2001

Co-regulation of survival of motor neuron (SMN) protein and its interactor SIP1 during development and in spinal muscular atrophy.

Spinal muscular atrophy (SMA) is a neuromuscular disease characterized by the degeneration of motor neurons in the spinal cord. The disease is caused by mutations of the survival of motor neuron 1 gene (SMN1), resulting in a reduced production of functional SMN protein. A major question unanswered thus far is why reduced amounts of ubiquitously expressed SMN protein specifically cause the degeneration of motor neurons without affecting other somatic cell types. In a first attempt to address this issue we have investigated the Smn interacting protein 1 (Sip1), with an emphasis on its developmental expression and subcellular distribution in spinal motor neurons in relation to Smn. By confocal immunofluorescence studies we provide evidence that a significant amount of Smn does not co-localize with Sip1 in neurites of motor neurons, indicating that Smn may exert motor neuron-specific functions that are not dependent on Sip1. Sip1 is highly expressed in the spinal cord during early development and expression decreases in parallel with Smn during postnatal development. Strikingly, reduced production of Smn as observed in cell lines derived from SMA patients or in a mouse model for SMA coincides with a simultaneous reduction of Sip1. The finding that expression of Sip1 and Smn is tightly co-regulated, together with the unique localization of Smn in neurites, may help in understanding the motor neuron-specific defects observed in SMA patients.

Amino Acid Sequence↗

PubMed2001

Evidence that fragile X mental retardation protein is a negative regulator of translation.

Fragile X syndrome is a common form of inherited mental retardation. Most fragile X patients exhibit mutations in the fragile X mental retardation gene 1 (FMR1) that lead to transcriptional silencing and hence to the absence of the fragile X mental retardation protein (FMRP). Since FMRP is an RNA-binding protein which associates with polyribosomes, it had been proposed to function as a regulator of gene expression at the post-transcriptional level. In the present study, we show that FMRP strongly inhibits translation of various mRNAs at nanomolar concentrations in both rabbit reticulocyte lysate and microinjected Xenopus laevis oocytes. This effect is specific for FMRP, since other proteins with similar RNA-binding domains, including the autosomal homologues of FMRP, FXR1 and FXR2, failed to suppress translation in the same concentration range. Strikingly, a disease-causing Ile-->Asn substitution at amino acid position 304 (I304N) renders FMRP incapable of interfering with translation in both test systems. Initial studies addressing the underlying mechanism of inhibition suggest that FMRP inhibits the assembly of 80S ribosomes on the target mRNAs. The failure of FMRP I304N to suppress translation is not due to its reduced affinity for mRNA or its interacting proteins FXR1 and FXR2. Instead, the I304N point mutation severely impairs homo-oligomerization of FMRP. Our data support the notion that inhibition of translation may be a function of FMRP in vivo. We further suggest that the failure of FMRP to oligomerize, caused by the I304N mutation, may contribute to the pathophysiological events leading to fragile X syndrome.

Animals↗

PubMed2001

KUB3 amplification and overexpression in human gliomas.

Gene amplification is known to occur frequently in human glioma. Recently we reported cloning of a novel gene termed glioma-amplified sequence 16 (GAS16) by microdissection-mediated cDNA capture. In this article, we demonstrate that GAS16 results from an alternative splicing process of the Ku70 binding protein 3 (KUB3) that is essential for DNA double-strand break repair. The alternative splice product was found in glioblastoma and in normal fetal brain. We determined the amplification frequency of KUB3 in glioma with different grading. We analyzed a total of 102 glioma primary tumors and found KUB3 to be amplified in 12/82 (14%) glioblastomas, 4/13 anaplastic astrocytomas (30%), and 2/4 astrocytomas, but in none of three pilocytic astrocytomas. Northern blot analysis of glioblastoma shows a strong correlation between KUB3 amplification and overexpression. Amplification of KUB3 appears to be independent of other genetic changes frequently associated with the development of gliomas, including EGFR amplification, LOH of TP53, and LOH of chromosome 10. The KUB3 amplification and overexpression may interfere with the function of KUB3 in the DNA-PK complex involved in the maintenance of genome stability and reduction of mutation frequency.

Alternative Splicing↗

PubMed2001

A monoclonal antibody recognising a surface marker on rainbow trout (Oncorhynchus mykiss) monocytes.

The characterisation of a monoclonal antibody (mab 45) reacting with phagocytic leucocytes isolated from blood and spleen of rainbow trout (Oncorhynchus mykiss, Walbaum) is described. The surface marker labelled by this mab is expressed at relative low levels on the membrane of large, nearly nongranulated trout leucocytes, and having the typical morphology of monocytes in flow cytometry (Kfoury et al., 1999, Fish Pathology, 34, 1-6). No reaction of mab 45 with granulocytes, lymphocytes or thrombocytes was detected. In spleen and head kidney, large, polymorphonuclear leucocytes were immunostained. The mab most strongly recognised an antigen of 48 kDa prepared from trout leucocytes of different organs, but not in trout plasma. In an in vitro phagocytosis assay trout monocytes were stained with mab 45 after phagocytosis of Aeromonas salmonicida labelled with the lipophilic fluorescent cell surface linker PKH26. However, previous binding of mab 45 on trout leucocytes did not inhibit the phagocytosis of A. salmonicida particles. Using mab 45, the dynamics of monocytes in blood, spleen and peritoneal cavity could be demonstrated after intraperitoneal injection of trout with inactivated A. salmonicida. The described mab serves as a useful tool to investigate the involvement of monocytes/macrophages in immune reactions of trout to a variety of pathogens.

Aeromonas↗

PubMed2001

Exogenous antigens and the stimulation of MHC class I restricted cell-mediated cytotoxicity: possible strategies for fish vaccines.

An MHC class I restricted cytotoxic T lymphocyte (CTL) activity assay has recently been established for rainbow trout. MHC class I restricted cytotoxicity probably plays a critical role in immunity to most viral diseases in mammals and may play a similar role in fish. Therefore, it is very important to investigate what types of vaccines can stimulate this immune response. Although logical candidates for vaccine components that can stimulate an MHC class I restricted response are live attenuated viruses and DNA vaccines, these materials are generally not allowed in fish for commercial vaccine use due to potential safety issues. In mammals, however, a number of interesting vaccination strategies based on exogenous antigens that stimulate MHC class I restricted cytotoxicity have been described. Several of these strategies are discussed in this review in the context of fish vaccination.

Animals↗

PubMed2001

[Value and significance of digital full-field mammography within the scope of mammography screening].

To evaluate the digital technique for screening mammography various experimental studies and clinical examinations were performed by using direct full-field digital mammography (FFDM). The findings concerning the detectability and characterization of microcalcifications and soft tissue masses as well as the radiation exposure were compared to the state-of-the-art conventional screen-film mammography (SFM). The results of these studies revealed a high performance of the digital images, which are at least equivalent to the conventional images, whereas digital spot views were significantly superior to conventional ones particularly in the detection of microcalcifications. This was especially true, when the potential of post-processing was used. In addition, the sensitivity of FFDM should be increased, if computer-aided-diagnosis (CAD) is available. Furthermore, the patient radiation dose can be significantly reduced. Additional advantages are quick and easy handling, efficient data transfer and digital archiving. Thus, FFDM will become an important tool in screening mammography.

Breast Neoplasms↗

PubMed2001

[MR-angiography in portal hypertension. State of the art].

When imaging the portal vascular system colour-coded sonography and CT angiography are used as an alternative to intraarterial DSA. Today fast CE 3D-MRA is the imaging modality of choice in the diagnostic of patients with portal hypertension. It enables a clear picture of the portalvenous vascular system and portalsytemic shunts. Statements regarding velocity and direction of blood flow can be made with additional use of TOF and PC techniques. CE-3D MRA can be used preoperatively before liver transplantation or shunt surgery. In the follow-up of interventional or surgical procedures exact statements concerning haemodynamically significant pathologies e.g. vascular thromboses, stenoses and occlusions can be made. Patient consent is symplify and the numbers of complications can be reduced as well as duration of surgery minimized applying this technique.

Collateral Circulation↗

PubMed2001

[Intramural hematomas of the aorta].

Intramural hematomas (IMH) are regarded as a hemorrhage into the aortic wall. In general a traumatic form can de differentiated from a spontaneous non-traumatic. There is a predisposition of IMH with arterial hypertension and mesoectodermal dysplastic syndromes. The diagnosis is established with the clinical presentation in combination with the findings of different imaging modalities. Acute and subacute discomfort associated with a tendency of collaps are considered as typical presenting complaints. A semicircular or concentric thickening of the aortic wall with the absence of blood flow or a dissection membrane are typical findings in transesophageal echocardiography, computed tomography and magnetic resonance tomography. From a differential diagnostic point of view a distinction from atherosclerotic wall changes, intraluminal thrombi and inflammatory aortic diseases is essential. The IMH is considered as an early presentation of aortic dissection, put into and treated according to the Stanford classification. In the course of natural history an IMH can disappear, rupture or progress into a classic aortic dissection.

Aortic Dissection↗

PubMed2001

Contrast-enhanced color Doppler ultrasound characteristics in hypervascular breast tumors: comparison with MRI.

The aim of this study was to evaluate the accuracy of contrast-enhanced color Doppler ultrasound (CE-US) in comparison with contrast-enhanced MR imaging (CE-MRI) in the discrimination of hypervascularized breast tumors. An additional CE-US of the breast was preoperatively performed in 40 patients with a hypervascular breast lesion detected on CE-MRI. The presence of blood flow signals and the morphological characteristics of the vessels in the breast lesions were evaluated pre- and post-contrast administration, as well as the dynamic aspects of the Doppler signal, including time interval to maximum signal enhancement and persistence of the signal enhancement. Twenty-three carcinomas and 17 fibroadenomas were explored. Considering initial signal enhancement > 100% after the administration of contrast material as a criterion suggesting malignancy, CE-MRI showed a sensitivity of 100% and a specificity of 76.5% in the detection of malignant breast tumors. Color Doppler signals were consistently demonstrated in all carcinomas and in 68.7% of fibroadenomas after the administration of Levovist, with CE-US showing a sensitivity of 95.6% and a specificity of 5.9%. Neither the mean number of vessels per tumor, nor the location of vessels, the time to maximum increase of the Doppler signal or the persistence of signal enhancement showed significant differences between benign and malignant lesions. Additional CE-US does not increase the low specificity of MRI in patients with hypervascularized breast tumors.

Adult↗

PubMed2001

International investigation of breast MRI: results of a multicentre study (11 sites) concerning diagnostic parameters for contrast-enhanced MRI based on 519 histopathologically correlated lesions.

A multicentre study was undertaken to provide fundamentals for improved standardization and optimized interpretation guidelines of dynamic contrast-enhanced MRI. Only patients scheduled for biopsy of a clinical or imaging abnormality were included. They underwent standardized dynamic MRI on Siemens 1.0 (163 valid lesions > or = 5 mm) or 1.5 T (395 valid lesions > or = 5 mm) using 3D fast low-angle shot (FLASH; 87 s) before and five times after standardized bolus of 0.2 mmol Gd-DTPA/kg. One-Tesla and 1.5 T data were analysed separately using a discriminant analysis. Only histologically correlated lesions entered the statistical evaluation. Histopathology and imaging were correlated in retrospect and in open. The best results were achieved by combining up to five wash-in or wash-out parameters. Different weighting of false-negative vs false-positive calls allowed formulation of a statistically based interpretation scheme yielding optimized rules for the highest possible sensitivity (specificity 30%), for moderate (50%) or high (64-71%) specificity. The sensitivities obtained at the above specificity levels were better at 1.0 T (98, 97, or 96%) than at 1.5 T (96, 93, 86%). Using a widely available standardized MR technique definition of statistically founded interpretation rules is possible. Choice of an optimum interpretation rule may vary with the clinical question. Prospective testing remains necessary. Differences of 1.0 and 1.5 T are not statistically significant but may be due to pulse sequences.

Adult↗