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Biomedical subjects

W Reith

Publications and source records attributed to W Reith.

At least 55 records · Page 3Linked to original sources
PubMed1997

Dynamic contrast-enhanced T2*-weighted MRI in patients with cerebrovascular disease.

Our purpose was to investigate the potential of dynamic susceptibility contrast-enhanced MRI in assessing regional haemodynamics in patients with cerebrovascular disease. T2*-weighted FLASH sequences were performed on a control group of 10 healthy subjects, 13 patients with unilateral stenosis or occlusion of the internal carotid artery and 6 patients with acute onset of neurological symptoms, the observed signal intensities being converted into concentration-time curves. A gamma-variate function was fitted to the measured concentration-time curves to eliminate effects of tracer recirculation. In each patient the two cerebral hemispheres were compared and the difference between the mean transit times and the percental change of the regional cerebral blood volume, calculated for each side. Patients with haemodynamically significant unilateral carotid obstruction can be divided into two subgroups: those with good and those with poor collateral supply. Patients with good collateral supply had a slight but not statistically significant increase in mean transit time and cerebral blood volume on the diseased side, whereas those with poor collaterals had a significant increase compared with the control group. In patients with acute onset of neurological symptoms perfusion maps clearly demonstrated the disturbed perfusion at a time when T2-weighted images were still normal. Perfusion imaging is a reliable and noninvasive method of assessing changes in cerebral perfusion in patients with unilateral carotid stenosis. This MR technique permits monitoring of haemodynamic changes during therapy and thus may become an alternative to SPECT and PET scanning. In patients with acute occlusion of a cerebral artery, perfusion imaging reveals the entire, perfusion deficit before conventional MRI and thus allows early intervention.

Acute Disease↗
PubMed1997

MHC class II deficiency: definition of a new complementation group.

MHC class II deficiency is a severe primary immunodeficiency characterized by the absence of Major Histocompatibility Complex class II gene expression. It is genetically heterogenous and can result from defects in several different transacting regulatory factors required for transcription of MHC class II genes. Cell lines from MHC class II deficiency patients have been assigned to three complementation groups (A, B, C). An in vitro generated cell line (6.1.6) was reported to be the sole representative of a fourth group (group D). The molecular defect in 6.1.6 resides in the recently cloned RFXAP gene. Direct complementation experiments and mutation analysis were performed with cell lines from several MHC class II deficiency patients in which the affected gene had not been identified. These experiments have allowed us to define a previously unrecognized MHC class II deficiency complementation group containing patients having mutations in the RFXAP gene.

Animals↗
PubMed1997

[Diffusion-weighted magnetic resonance tomography: a highly promising MR technic for the early recognition of cerebral ischemia].

PURPOSE: The aim of this study was to show the temporal and spatial evolution of ischaemia over time using diffusion-weighted magnetic resonance imaging, and to correlate the extent of ischaemia with the postmortem infarct size after 24 hours. MATERIAL AND METHODS: In 8 rats focal cerebral ischaemia was induced by intravascular occlusion of the middle cerebral artery. The evolution of the ischaemic lesion was examined over 180 min with an experimental MR scanner. RESULTS: Diffusion-weighted magnetic resonance imaging displayed a hyperintense area in the lateral part of the putamen as early as 5 min after the onset of ischaemia. The mean volume of ischaemia on diffusion mapping after 5 min was 62.5 +/- 12.9 microliters and increased to 224.4 +/- 48.5 microliters after 180 min. This correlated well with the corrected infarct volume at postmortem examination (194.0 +/- 23.1 microliters, r = 0.72, p < 0.05) using the TTC staining. CONCLUSION: Diffusion-weighted magnetic resonance imaging is a reliable tool in the early diagnosis of cerebral ischaemia. Due to the non-invasiveness this method can be used for therapy monitoring and might help to develop new therapeutic strategies.

Animals↗
PubMed1997

[Diffusion-weighted MR tomography in focal cerebral ischemia: possibilities offered by a 1.0 Tesla clinical device].

PURPOSE: To develop and to implement a fast, highly diffusion-weighted MR-sequence on a conventional 1 T MR scanner and to examine this sequence as to its capability of characterising tissue damage due to focal cerebral ischaemia. METHODS: Within a CE-FAST-sequence diffusion gradients were inserted in all gradient directions. By phantom measurements the maximal diffusion weighting was determined for different T1 and T2 values and flip angles. The optimised sequence was used for monitoring the diffusion of focal cerebral ischaemia in 14 rats following endovascular occlusion of the middle cerebral artery. RESULTS: The ischaemic lesion could be easily visualised as a bright area in the images with high diffusion weighting. The mean relative decrease of the diffusion coefficient in the ischaemic brain parenchyma was 29%. The diffusion coefficient, decreased significantly with increasing occlusion time. CONCLUSION: Our study suggests that the installed sequence is capable of producing highly diffusion weighted images without requiring a special gradient system. The sequence is potentially useful for monitoring cerebral ischaemia on the basis of calculating the change of the diffusion coefficient.

Animals↗
PubMed1997

[Studies on contrast medium dosage in perfusion-weighted MR imaging].

PURPOSE: In this study we investigated, whether increasing the dosage of a paramagnetic contrast agent results in a stronger signal decrease in T2*-weighted perfusion sequences and therefore more meaningful parameter maps. MATERIAL AND METHODS: In a prospective study bolus injection of gadolinium-DTPA was performed at dosages of 0.1, 0.2, and 0.3 mmol/kg body weight (BW) in 10 patients each. Before, during and after bolus injection 40 T2*-weighted images of a reference brain slice were acquired within 65.6 seconds on a 1.0 T clinical scanner and perfusion parameters were calculated. RESULTS: Due to the limited signal decrease during bolus passage and the resulting low signal-difference-to-noise ratio (delta S/N) no reliable differentiation of gray and white matter was possible at a contrast agent dosage of 0.1 mmol/kg BW. Only at higher dosages, both, signal decrease and delta S/N were strong enough to allow differentiation of gray and white matter and to yield reliable parameter maps. CONCLUSION: For meaningful MR perfusion imaging at 1.0 T and with the given sequence a contrast agent dosage of at least 0.2 mmol/kg BW is necessary, if a 0.5-molar contrast agent is used.

Adult↗
PubMed1997

Bolus injection of MR contrast agents: hemodynamic effects evaluated by intracerebral laser Doppler flowmetry in rats.

PURPOSE: To determine the effects on arterial blood pressure and cerebral blood flow of intravenous bolus injection of three MR contrast agents: gadopentetate dimeglumine, polylysine-Gd-DTPA, and superparamagnetic iron particles (SPIO). METHODS: A single-fiber laser Doppler flowmetry probe was placed intracerebrally in 56 anesthetized rats. Cerebral blood flow and mean arterial blood pressure were measured before (baseline), during, and up to 30 minutes after intravenous bolus administration of the three contrast agents: 0.1 mmol/kg and 0.3 mmol/kg gadopentetate dimeglumine (n = 18 per group), 0.3 mmol/kg polylysine-Gd-DTPA (n = 10), and 0.03 mmol/kg SPIO (n = 10). RESULTS: Neither the higher nor lower dose of gadopentetate dimeglumine had any statistically significant effect on cerebral blood flow, and there was no change in blood pressure during administration of either dose of gadopentetate dimeglumine. Administration of polylysine-Gd-DTPA caused a transient drop in blood pressure in two animals, marked in one (decrease to 21% of baseline values) and mild in the other (84% of baseline). After administration of SPIO, a significant decrease in blood pressure occurred in one animal (41% of baseline). Despite this decrease in mean arterial blood pressure, there were no statistically significant changes in cerebral blood flow after administration of polylysine-Gd-DTPA or SPIO. CONCLUSION: Our results suggest that bolus injection of these contrast agents at clinically relevant doses causes no significant alteration in cerebral blood flow. We conclude that gadopentetate dimeglumine is well suited for cerebral MR perfusion imaging without inherent influence on cerebral blood flow and that the same is probably true for polylysine-Gd-DTPA and SPIO.

Animals↗
PubMed1996

Locations of human and mouse genes encoding the RFX1 and RFX2 transcription factor proteins.

RFX transcription factors constitute a highly conserved family of site-specific DNA binding proteins involved in the expression of a variety of cellular and viral genes, including major histocompatibility complex class II genes and genes in human hepatitis B virus. Five members of the RFX gene family have been isolated from human and mouse, and all share a highly characteristic DNA binding domain that is distinct from other known DNA binding motifs. The human RFX1 and RFX2 genes have been assigned by in situ hybridization to chromosome 19p13.1 and 19p13.3, respectively. In this paper, we present data that localize RFX1 and RFX2 precisely within the detailed physical map of human chromosome 19 and genetic data that assign Rfx1 and Rfx2 to homologous regions of mouse chromosomes 8 and 17, respectively. These data define the established relationships between these homologous mouse and human regions in further detail and provide new tools for linking cloned genes to phenotypes in both species.

Animals↗
PubMed1996

RFX proteins, a novel family of DNA binding proteins conserved in the eukaryotic kingdom.

Until recently, the RFX family of DNA binding proteins consisted exclusively of four mammalian members (RFX1-RFX4) characterized by a novel highly conserved DNA binding domain. Strong conservation of this DNA binding domain precluded a precise definition of the motif required for DNA binding. In addition, the biological systems in which these RFX proteins are implicated remained obscure. The recent identification of four new RFX genes has now shed light on the evolutionary conservation of the RFX family, contributed greatly to a detailed characterization of the RFX DNA binding motif, and provided clear evidence for the function of some of the RFX proteins. RFX proteins have been conserved throughout evolution in a wide variety of species, including Saccharomyces cerevisiae, Schizosaccharomyces pombe, Caenorhabditis elegans, mouse and man. The characteristic RFX DNA binding motif has been recruited into otherwise very divergent regulatory factors functioning in a diverse spectrum of unrelated systems, including regulation of the mitotic cell cycle in fission yeast, the control of the immune response in mammals, and infection by human hepatitis B virus.

Amino Acid Sequence↗
PubMed1996

[High dosage administration of paramagnetic contrast media in diagnosis of focal brain lesions].

In analogy with high-dose contrast-enhanced CT, there have been a few studies during recent years that have dealt with high-dose paramagnetic contrast dyes in MRI. One reason for these studies was the development of new and low-osmolar contrast agents in the MR field. Depending on the clinical problem, a high-dose contrast study in MRI is rarely indicated: (1) in metastatic disease, MR imaging with high-dose contrast material is indicated when the standard dose study is negative or only shows a solitary cerebral lesion or a number of lesions just suitable for radiosurgery; (2) in patients with malignant glioma the high-dose study allows better definition of the tumor margins. If a radical surgical approach is planned, the diagnostic potential should be fully used; if only a biopsy or subtotal debulking is planned, a standard dose study is enough. (3) in patients with MS, a high-dose study is only recommended within therapeutic trials in which the number of active plaques is a primary variable.

Brain↗
PubMed1996

Cloning and functional characterization of mammalian homologues of the COPII component Sec23.

We screened a human cDNA library with a probe derived from a partial SEC23 mouse homologue and isolated two different cDNA clones (hSec23A and hSec23B) encoding proteins of a predicted molecular mass of 85 kDa. hSec23Ap and hSec23Bp were 85% identical and shared 48% identity with the yeast Sec23p. Affinity-purified anti-hSec23A recognized a protein of approximately 85 kDa on immunoblots of human, mouse, and rat cell extracts but did not recognize yeast Sec23p. Cytosolic hSec23Ap migrated with an apparent molecular weight of 350 kDa on a gel filtration column, suggesting that it is part of a protein complex. By immunoelectron microscopy, hSec23Ap was found essentially in the ribosome-free transitional face of the endoplasmic reticulum (ER) and associated vesicles. hSec23Ap is a functional homologue of the yeast Sec23p as the hSec23A isoform complemented the temperature sensitivity of the Saccharomyces cerevisiae sec23-1 mutation at a restrictive temperature of 34 degrees C. RNase protection assays indicated that both hSec23 isoforms are coexpressed in various human tissues, although at a variable ratio. Our data demonstrate that hSec23Ap is the functional human counterpart of the yeast COPII component Sec23p and suggest that it plays a similar role in mammalian protein export from the ER. The exact function of hSec23Bp remains to be determined.

Animals↗
PubMed1996

A consensus motif in the RFX DNA binding domain and binding domain mutants with altered specificity.

The RFX DNA binding domain is a novel motif that has been conserved in a growing number of dimeric DNA-binding proteins, having diverse regulatory functions, in eukaryotic organisms ranging from yeasts to humans. To characterize this novel motif, we have performed a detailed dissection of the site-specific DNA binding activity of RFX1, a prototypical member of the RFX family. First, we have performed a site selection procedure to define the consensus binding site of RFX1. Second, we have developed a new mutagenesis-selection procedure to derive a precise consensus motif, and to test the accuracy of a secondary structure prediction, for the RFX domain. Third, a modification of this procedure has allowed us to isolate altered-specificity RFX1 mutants. These results should facilitate the identification both of additional candidate genes controlled by RFX1 and of new members of the RFX family. Moreover, the altered-specificity RFX1 mutants represent valuable tools that will permit the function of RFX1 to be analyzed in vivo without interference from the ubiquitously expressed endogenous protein. Finally, the simplicity, efficiency, and versatility of the selection procedure we have developed make it of general value for the determination of consensus motifs, and for the isolation of mutants exhibiting altered functional properties, for large protein domains involved in protein-DNA as well as protein-protein interactions.

Amino Acid Sequence↗
PubMed1996

Regulation of MHC class II genes: lessons from a disease.

Precise regulation of major histocompatibility complex class II (MHC-II) gene expression plays a crucial role in the control of the immune response. A major breakthrough in the elucidation of the molecular mechanisms involved in MHC-II regulation has recently come from the study of patients that suffer from a primary immunodeficiency resulting from regulatory defects in MHC-II expression. A genetic complementation cloning approach has led to the isolation of CIITA and RFX5, two essential MHC-II gene transactivators. CIITA and RFX5 are mutated in these patients, and the wild-type genes are capable of correcting their defect in MHC-II expression. The identification of these regulatory factors has furthered our understanding of the molecular mechanisms that regulate MHC-II genes. CIITA was found to be a non-DNA binding transactivator that functions as a molecular switch controlling both constitutive and inducible MHC-II expression. The finding that RFX5 is a subunit of the nuclear RFX-complex has confirmed that a deficiency in the binding of this complex is indeed the molecular basis for MHC-II deficiency in the majority of patients. Furthermore, the study of RFX has demonstrated that MHC-II promoter activity is dependent on the binding of higher-order complexes that are formed by highly specific cooperative binding interactions between certain MHC-II promoter-binding proteins. Two of these proteins belong to families of which the other members, although capable of binding to the same DNA motifs, are probably not directly involved in the control of MHC-II expression. Finally, the facts that CIITA and RFX5 are both essential and highly specific for MHC-II genes make possible novel strategies designed to achieve immunomodulation via transcriptional intervention.

Animals↗
PubMed1996

Decompressive craniectomy in a rat model of "malignant" cerebral hemispheric stroke: experimental support for an aggressive therapeutic approach.

Acute ischemia in the complete territory of the carotid artery may lead to massive cerebral edema with raised intracranial pressure and progression to coma and death due to uncal, cingulate, or tonsillar herniation. Although clinical data suggest that patients benefit from undergoing decompressive surgery for acute ischemia, little data about the effect of this procedure on experimental ischemia are available. In this article the authors present results of an experimental study on the effects of decompressive craniectomy performed at various time points after endovascular middle cerebral artery (MCA) occlusion in rats. Focal cerebral ischemia was induced in 68 rats using an endovascular occlusion technique focused on the MCA. Decompressive craniectomy was performed in 48 animals (in groups of 12 rats each) 4, 12, 24, or 36 hours after vessel occlusion. Twenty animals (control group) were not treated by decompressive craniectomy. The authors used the infarct volume and neurological performance at Day 7 as study endpoints. Although the mortality rate in the untreated group was 35%, none of the animals treated by decompressive craniectomy died (mortality 0%). Neurological behavior was significantly better in all animals treated by decompressive craniectomy, regardless of whether they were treated early or late. Neurological behavior and infarction size were significantly better in animals treated very early by decompressive craniectomy (4 hours) after endovascular MCA occlusion (p < 0.01); surgery performed at later time points did not significantly reduce infarction size. The results suggest that use of decompressive craniectomy in treating cerebral ischemia reduces mortality and significantly improves outcome. If performed early after vessel occlusion, it also significantly reduces infarction size. By performing decompressive craniectomy neurosurgeons will play a major role in the management of stroke patients.

Animals↗
PubMed1995

Purified X2 binding protein (X2BP) cooperatively binds the class II MHC X box region in the presence of purified RFX, the X box factor deficient in the bare lymphocyte syndrome.

The conserved X2 box sequence of MHC class II promoters is homologous to TRE/CRE elements, and is required for B cell expression and IFN-gamma induction of MHC class II genes. The X2 binding protein (X2BP) was initially identified as a DNA-binding activity that specifically interacts with the conserved X2 box sequence in both the MHC HLA-DRA and HLA-DRB promoters. To begin to demonstrate that X2BP is the X2 box factor responsible for class II expression in B cells, we have purified X2BP to homogeneity from B cell nuclear extracts using DNA-affinity chromatography. X-box DNA-affinity purification indicates that X2BP is most likely composed of two polypeptides of 120 kDa and 46 kDa. The 120-kDa protein was specifically cross-linked to an X-box probe by exposure to UV irradiation. The 46-kDa subunit of X2BP cross-reacted with anti-rat CREB polyclonal Abs but not to anti-human CREB Abs in Western analysis and supershift assays, indicating that it may be a novel member of the ATF/CREB family. Purified X2BP interacted with purified RFX, a factor that binds to the adjacent X1 box and is absent in some cell lines that are mutant for MHC class II transcription. This interaction increases the DNA-binding half-life of RFX from 5 to at least 60 min, suggesting that X2BP functions in class II MHC gene expression by forming a stable complex with RFX.

Base Sequence↗
PubMed1995

A novel DNA-binding regulatory factor is mutated in primary MHC class II deficiency (bare lymphocyte syndrome).

Regulation of MHC class II gene expression is an essential aspect of the control of the immune response. Primary MHC class II deficiency is a genetically heterogeneous disease of gene regulation that offers the unique opportunity of a genetic approach for the identification of the functionally relevant regulatory genes and factors. Most patients exhibit a characteristic defect in the binding of a nuclear complex, RFX, to the X box motif of MHC class II promoters. Genetic complementation of a B-lymphocyte cell line from such a patient with a cDNA expression library has allowed us to isolate RFX5, the regulatory gene responsible for the MHC class II deficiency. This gene encodes a novel DNA-binding protein that is indeed a subunit of the RFX complex. Mutations in the RFX5 gene have been characterized in two patients. Transfection of the patient's cells with the RFX5 cDNA repairs the binding defect and fully restores expression of all the endogenous MHC class II genes in vivo.

Amino Acid Sequence↗
PubMed1995

Molecular defects in the bare lymphocyte syndrome and regulation of MHC class II genes.

The complex pattern of expression of major histocompatibility complex (MHC) class II molecules plays an essential role in the control of the immune response. Our understanding of the molecular mechanisms controlling this expression has benefited greatly from the identification of the regulatory factors defective in two forms of a hereditary disease of MHC class II regulation: bare lymphocyte syndrome. This has also provided new tools for the experimental modulation of MHC class II expression.

Gene Expression Regulation↗