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Biomedical subjects

M Walther

Publications and source records attributed to M Walther.

At least 37 records · Page 2Linked to original sources
PubMed2001

Exogenous antigen containing perivascular phagocytes induce a non-encephalitogenic extravasation of primed lymphocytes.

Recent evidence suggests that T-lymphocyte extravasation and CNS-parenchymal infiltration during autoimmune disease might be regulated by antigen-presenting (ED2(+)) cerebral/spinal perivascular phagocytes (CPP/SPP). Since the massive erythrocytic and leukocytic infiltrates in the CNS of rats with experimental allergic encephalomyelitis do not allow a precise differentiation between CPP/SPP and the invading cells in the Virchow-Robin space, we developed a new immune-response model whereby the extravasation of T-lymphocytes was not followed by other blood cells. Adult Lewis rats were sensitized to horseradish peroxidase (HRP). Subsequent intracerebroventricular (i.c.v.) injections of HRP and/or Fluoro-Emerald (FE) served to: (1) challenge the primed T-lymphocytes and (2) label the CPP/SPP for additional immunocytochemical analysis. We found that 24 h and 3 days after single, double, or triple antigen boosting T-lymphocytes (R73(+), W3/25(+), OX50(+)) entered the Virchow-Robin space but did not break through the astrocytic glia limitans. Instead they adhered to HRP-containing activated CPP/SPP (mabs OX-6(+), SILK6(+), CD40(+), CD80(+), CD86(+)). This selective contact was mediated neither by cell adhesion molecules (P-selectin, ICAM-1, VCAM-1), nor promoted by chemokine receptors (CCR1, CCR5) or chemokines (monocyte chemoattractant protein (MCP)-1, MIP-1alpha, MIP-1beta, RANTES). This non-inflammatory, but antigen-dependent lymphocyte extravasation provides optimal conditions to further study the CNS immune response.

Animals↗
PubMed2001

Bis(1,3,4-thiadiazolo)-1,3,5-triazinium halides. 2. Intramolecular ring transformation and synthesis of novel highly substituted guanidines.

Bis(1,3,4-thiadiazolo)-1,3,5-triazinium halides 6 can be easily attacked by nucleophiles at either the C(3a) or the C(4a) position of the central six-membered (cationic) ring. Nucleophilic attack leads to at least two reaction channels, one of which has been previously detected (pathway a) and leads to novel aminals 19. In this paper we report on a second channel (pathway b). Attack of primary or secondary amines 8 at C(3a) or C(4a) in 6 (and their analogues 7) leads to the weakly stabilized intermediates 14. A cascade of several proton shifts, ring openings, rearrangements, and ring closure processes is initiated which finally leads via 17 and 18 to novel highly substituted guanidines 9, 10, 12, and 13. Pathway b seems to be the result of well-balanced negative-hyperconjugative effects in 14 and/or 17 which control the highly selective opening of a relatively stable central 1,3,5-triazinium ring to yield the crucial intermediate 18. Some representatives of the guanidines have been characterized by X-ray analyses. Since some of the guanidines contain one or two chirality centers, an effort was made to investigate the stereochemistry of these compounds.

Journal Article↗
PubMed2001

Structural basis for lipoxygenase specificity. Conversion of the human leukocyte 5-lipoxygenase to a 15-lipoxygenating enzyme species by site-directed mutagenesis.

Mammalian lipoxygenases constitute a heterogeneous family of lipid-peroxidizing enzymes, and the various isoforms are categorized with respect to their positional specificity of arachidonic acid oxygenation into 5-, 8-, 12-, and 15-lipoxygenases. Structural modeling suggested that the substrate binding pocket of the human 5-lipoxygenase is 20% bigger than that of the reticulocyte-type 15-lipoxygenase; thus, reduction of the active-site volume was suggested to convert a 5-lipoxygenase to a 15-lipoxygenating enzyme species. To test this "space-based" hypothesis of the positional specificity, the volume of the 5-lipoxygenase substrate binding pocket was reduced by introducing space-filling amino acids at critical positions, which have previously been identified as sequence determinants for the positional specificity of other lipoxygenase isoforms. We found that single point mutants of the recombinant human 5-lipoxygenase exhibited a similar specificity as the wild-type enzyme but double, triple, and quadruple mutations led to a gradual alteration of the positional specificity from 5S- via 8S- toward 15S-lipoxygenation. The quadruple mutant F359W/A424I/N425M/A603I exhibited a major 15S-lipoxygenase activity (85-95%), with (8S,5Z,9E,11Z,14Z)-8-hydroperoxyeicosa-5,9 ,11, 14-tetraenoic acid being a minor side product. These data indicate the principle possibility of interconverting 5- and 15-lipoxygenases by site-directed mutagenesis and appear to support the space-based hypothesis of positional specificity.

Amino Acid Sequence↗
PubMed2001

Correlation of power Doppler sonography with vascularity of the synovial tissue of the knee joint in patients with osteoarthritis and rheumatoid arthritis.

OBJECTIVE: To examine the significance of power Doppler sonography (PDS) in the diagnosis of synovial hypertrophy of the knee joint by verifying and comparing the PDS findings with histopathologic findings of synovial membrane vascularity. METHODS: The knee joints of 23 patients who were undergoing arthroplasty of the knee joint because of osteoarthritis or rheumatoid arthritis were examined with ultrasound before arthroplasty. The vascularity of the synovial membrane was classified semiquantitatively using PDS. A sample of synovial tissue was obtained during the arthroplasty, and the vascularity of the synovial tissue was evaluated by immunohistochemistry (factor VIII) and was graded qualitatively by a pathologist who was unaware of the PDS findings. The visual qualitative grading by the examiner was controlled by analyzing PDS images and histologic samples using a digital image evaluation system. RESULTS: The correlation between the qualitative PDS results and the qualitative grading of the vascularity by the pathologist was 0.89 by Spearman's rho (P < 0.01). The Pearson correlation coefficient between the digital analysis of the PDS images and the digital analysis of the tissue sections was 0.81 (P < 0.01). Digital image analysis and qualitative grading by the examiner had a correlation of 0.89 by Spearman's p (P < 0.01) for the PDS images. The correlation between the qualitative estimation of vascularity by the pathologist and the digital image analysis was 0.88 by Spearman's rho (P < 0.01). CONCLUSION: In the present study, PDS proved to be a reliable diagnostic method for qualitative grading of the vascularity of the synovial tissue. In clinical practice, PDS allows further differentiation of the hypertrophic synovium.

Adult↗
PubMed2001

Molecular cytogenetic characterization of early and late renal cell carcinomas in von Hippel-Lindau disease.

Deletions of 3p25, gains of chromosomes 7 and 10, and isochromosome 17q are known cytogenetic aberrations in sporadic renal cell carcinoma (RCC). In addition, a majority of RCCs have loss of heterozygosity (LOH) of the Von Hippel-Lindau (VHL) gene located at chromosome band 3p25. Patients who inherit a germline mutation of the VHL gene can develop multifocal RCCs and other solid tumors, including malignancies of the pancreas, adrenal medulla, and brain. VHL tumors follow the two-hit model of tumorigenesis, as LOH of VHL, a classic tumor suppressor gene, is the critical event in the development of the neoplastic phenotype. In an attempt to define the cytogenetic aberrations from early tumors to late RCC further, we applied spectral karyotyping (SKY) to 23 renal tumors harvested from 6 unrelated VHL patients undergoing surgery. Cysts and low-grade solid lesions were near-diploid and contained 1-2 reciprocal translocations, dicentric chromosomes, and/or isochromosomes. A variety of sole numerical aberrations included gains of chromosomes 1, 2, 4, 7, 10, 13, 21, and the X chromosome, although no tumors had sole numerical losses. Three patients shared a breakpoint at 2p21-22, and three others shared a dicentric chromosome 9 or an isochromosome 9q. In contrast to the near-diploidy of the low-grade lesions, a high-grade lesion and its nodal metastasis were markedly aneuploid, revealed loss of VHL by fluorescence in situ hybridization (FISH), and contained recurrent unbalanced translocations and losses of chromosome arms 2q, 3p, 4q, 9p, 14q, and 19p as demonstrated by comparative genomic hybridization (CGH). By combining SKY, CGH, and FISH of multiple tumors from the same VHL kidney, we have begun to identify chromosomal aberrations in the earliest stages of VHL-related renal cell tumors. Our current findings illustrate the cytogenetic heterogeneity of different VHL lesions from the same kidney, which supports the multiclonal origins of hereditary RCCs. Published 2001 Wiley-Liss, Inc.

Adult↗
PubMed2001

Transient bone marrow oedema of the foot.

We treated ten patients who on the basis of MRI were suspected to have transient bone marrow oedema. In eight cases the talus was affected, in one the cuboid and in one the navicular bone. All patients had acute onset pain at the ankle. Four were treated with core decompression and had an immediate pain relief. Six were treated conservatively and became also pain-free but with considerable delay.

Acute Disease↗
PubMed2001

Alterations of lipoxygenase specificity by targeted substrate modification and site-directed mutagenesis.

BACKGROUND: Mammalian lipoxygenases (LOXs) are categorised with respect to their positional specificity of arachidonic acid oxygenation. However, the mechanistic basis for this classification is not well understood. To gain a deeper insight into the structural basis of LOX specificity we determined the reaction characteristics of wild-type and mutant mammalian LOX isoforms with native and synthetic fatty acids substrates. RESULTS: The rabbit 15-LOX is capable of catalysing major 12-lipoxygenation when the volume of the substrate-binding pocket is enlarged. These alterations in the positional specificity can be reversed when bulky residues are introduced at the omega end of the substrate. Simultaneous derivatisation of both ends of fatty acids forces a 15-LOX-catalysed 5-lipoxygenation and this reaction involves an inverse head-to-tail substrate orientation. In contrast, for arachidonic acid 5-lipoxygenation by the human 5-LOX the substrate fatty acid may not be inversely aligned. The positional specificity of this isoenzyme may be related to its voluminous substrate-binding pocket. Site-directed mutagenesis, which leads to a reduction of active site volume, converts the 5-LOX to a 15-lipoxygenating enzyme species. CONCLUSIONS: The positional specificity of LOXs is not an invariant enzyme property but depends on the substrate structure and the volume of the substrate-binding pocket. 15-LOX-catalysed 5-lipoxygenation involves an inverse substrate alignment but this may not be the case for 5-LOXs. Thus, both theories for the mechanistic basis of 5-lipoxygenation (straight and inverse substrate orientation) appear to be correct for different LOX isoforms.

Animals↗
PubMed2001

The genetic basis of renal epithelial tumors: advances in research and its impact on prognosis and therapy.

The genetics of renal cell carcinoma continues to elucidate the pathways of kidney tumorigenesis. The relationship between the VHL gene and clear cell carcinoma, MET and papillary carcinoma, and the families of genes that they regulate, continues to be unraveled. New hereditary kidney cancer syndromes, like familial oncocytoma and the Birt-Hogg-Dubé syndrome, have been identified and the search for the genes that cause them is under way. Researching the genetics of these disorders is essential for an understanding of sporadic kidney cancer genetics. This chapter will review the current knowledge of the hereditary kidney cancer syndromes, the genes that cause them, new advances in genetic research and techniques, and how this information impacts upon diagnostic, prognostic, and therapeutic methods of the future.

Carcinoma, Renal Cell↗
PubMed2000

Galectin-3 is upregulated in microglial cells in response to ischemic brain lesions, but not to facial nerve axotomy.

We have recently demonstrated that the beta-galactoside-specific lectin galectin-3 is expressed by microglial cells in vitro, but not by normal resting microglia in vivo. In the present study, we have analyzed the expression of galectin-3 by microglia under traumatic conditions in vivo using two experimental rat models which substantially differ in the severity of lesion related to a breakdown of the blood-brain barrier (BBB) and the occurrence of inflammatory processes. These two features are absent after peripheral nerve lesion and present after cerebral ischemia. Here we show that, following facial nerve axotomy under conditions allowing (nerve anastomosis) or not subsequent regeneration (nerve resection), galectin-3 is not expressed by microglia in the corresponding facial nucleus 1-112 days after lesion. Galectin-3 is also absent in microglia at sites of a defective BBB in the normal brain, such as the circumventricular organs. Following experimental ischemia (i.e., permanent occlusion of the middle cerebral artery), in contrast, galectin-3 becomes strongly expressed by activated microglia as early as 48 hours after trauma, as determined by immunohistochemistry and Western blot analysis. Our findings suggest that the expression of galectin-3 by microglia in vivo correlates with the state of microglial activation.

Animals↗
PubMed2000

[5- to 8-year results of total hip endoprosthesis implantation with the Müller straight shaft prosthesis (cemented TiAlNb shaft)].

The aim of this study was to obtain mid-term results after total hip arthroplasty (THA) with cemented titanium stems. In all, 184 patients with a total of 202 THAs (cemented titanium stem) were clinically and radiologically examined after an average follow-up of 6 years (5-8). The recruitment was 86%. The Harris score was determined clinically. Radiologically, the directly postoperative radiographs were compared to the control radiographs according to the recommendations of Gruen et al. and Johnston et al. In 2 cases (1%) septical complications appeared after 2 years, which were treated in two-stage surgery. To date, revisions after loosening have been carried out in 3 cases (1.5%). This is equivalent to a revision rate of 2.5%. Three further cases showed evidence of loosening in more than 5 radiolucent lines (RLL), according to Gruen, making close-meshed controls necessary. Clinically, in all of the 6 cases of aseptic loosening, the Harris score remained above 80 points. In 36 cases, more than one RLL, compared to the postoperative radiographs, was ascertained and mainly found in zones 1, 7, 8, and 14. Substantial subsidence or varus could only be observed in one case. The clinical results in the Harris score were good or excellent in 78% and satisfactory in 20%. With an average of 75 at the time of follow-up, the age of the patients was, according to the indication that only patients above age 60 are to receive cemented-stem prostheses, clearly advanced. The body weight was significantly higher (82 kg; d = 2.4) in those 6 patients having evident RLL, than in patients without RLL. The ratio body weight to surface of the stem was especially different (1.5 kg/cm2 versus 1 kg/cm2; P < 0.005) in the two groups. This did not apply to sex or activity of the patient, size or kind of stem, Harris score, ectopic ossification, or body weight index. The biggest possible stem should be implanted. Not all cemented titanium stem prostheses are necessarily affected with a high rate of loosening at a mid-term follow-up.

Aged↗
PubMed2000

Results of posterior cruciate-retaining unconstrained total knee arthroplasty after proximal tibial osteotomy for osteoarthritis. A prospective cohort study.

The purpose of the study was to investigate the results of posterior cruciate ligament (PCL)-retaining total knee arthroplasty (TKA) after previous proximal closed wedge tibial osteotomy for degenerative arthritis according to the technique recommended by Coventry and Insall. Thirty-five patients with previous proximal tibial osteotomy were matched to 35 patients without previous osteotomy according to age, sex, and Knee Society patient category. TKA after osteotomy was technically more difficult, often requiring a more extensive exposure. Complications were similar in the two groups, but an extended hematoma was more common in patients with previous osteotomies (2 patients) than in those with primary arthroplasty (1 patient). Knee Society scores were significantly lower in those with previous osteotomy, primarily because of decreased anteroposterior stability and pain. No significant differences were found in function scores, range of motion, or alignment. However, overall results of this PCL-retaining unconstrained TKA did not match those reported after PCL-substituting TKA.

Adult↗
PubMed2000

Balance sheets of knee and functional scores 5 years after total knee arthroplasty for osteoarthritis: a source for patient information.

To improve patient information on the results of total knee arthroplasty (TKA) for osteoarthritis, 253 primary TKAs of a prospective study with a mean follow-up of 5.3 years were analyzed. The increase or decrease of the individual variables of the Knee Society knee and function score and the percentage of operated knees in which these variables increased were determined. Improvement in pain rating had the largest increase of all variables, contributing 60% to the knee score increase. Pain improved in 95% of the knees. Alignment improved in about 90% of knees and accounted for 25% of knee score increase. Improvement in level walking contributed more to increase of function score than better stair-climbing abilities. Level walking improved in 80% of knees and stair climbing in 55%. Pain is the most rewarding indication for TKA, followed by deformity and poor walking ability. Key words: total knee arthroplasty, knee score, functional score, patient information.

Aged↗
PubMed2000

[Resection of the head of the ulna in patients with rheumatoid arthritis].

PURPOSE: The purpose of the study was to investigate value and effect of distal ulna resection (Darrach procedure) on the wrist in rheumatoid arthritis. CLINICAL MATERIAL: From 1985 to 1995, resection of the distal ulna was performed in 47 patients (54 hands) with rheumatoid arthritis. 38 patients (6 men, 32 women, 43 hands) had an average clinical and radiological follow up of 4.7 years (range 3 to 13 years). RESULTS: 15 of the cases with rheumatoid arthritis were pain-free, 19 complained about pain after severe stress, seven complained about pain after mild stress and two about continuous pain. 85% of the patients had a range of motion (pro- and supination) of more than 120 degrees. 75% of the patients had a good or very good wrist function. In 32 cases, the patients were absolutely satisfied with the operative outcome, in five cases, they had expected more improvement. Six reported, that there was no change due to the operation. CONCLUSION: The results demonstrate that the main indication for the resection of the distal ulna is rheumatoid arthritis. This operation does not influence the progress of rheumatoid arthritis. Preoperative radiological changes (Larsen's classification) have the main prognostic value. In Larsen's II and higher, a partial arthrodesis is recommended.

Adult↗
PubMed2000

Chronopharmaceutical drug delivery from a pulsatile capsule device based on programmable erosion.

We report the development of a chronopharmaceutical capsule drug delivery system capable of releasing drug after pre-determined time delays. The drug formulation is sealed inside the insoluble capsule body by an erodible tablet (ET). The release time is determined by ET erosion rate and increases as the content of an insoluble excipient (dibasic calcium phosphate) and of gel-forming excipient (hydroxypropylmethylcellulose; HPMC) increases. The time-delayed release of a model drug (propranolol HCI) was investigated by dissolution testing (USP XXIII paddle method). Both composition and weight of ET influence the time of drug release. Moreover it was found that drug release was controlled by the quantity of HPMC, irrespective of lactose content within the tablet weight range 80-160 mg, when above a threshold concentration of 20% HPMC. Programmable pulsatile release has been achieved from a capsule device over a 2-12-h period, consistent with the demands of chronotherapeutic drug delivery. The time of drug release can be controlled by manipulation of tablet formulation.

Delayed-Action Preparations↗
PubMed2000

Plasma metanephrines: a novel and cost-effective test for pheochromocytoma.

Pheochromocytomas are rare chromaffin cell tumors that nevertheless must be excluded in large numbers of patients who develop sustained or episodic hypertension as well as in many others with suggestive symptoms or with a familial history of pheochromocytoma. Diagnosis of pheochromocytoma depends importantly on biochemical evidence of excess catecholamine production by a tumor. Imperfect sensitivity and specificity of commonly available biochemical tests and the low incidence of the tumor among the tested population mean that considerable time and effort can be expended in confirming or ruling out pheochromocytoma in patients where the tumor is suspected. Measurements of plasma free metanephrines provide a superior test compared to other available tests for diagnosis of pheochromocytoma. In particular, the high sensitivity of plasma free metanephrines means that a normal test result reliably excludes all but the smallest of pheochromocytomas so that no other tests are necessary. Measurements of plasma free metanephrines, when systematically combined with other diagnostic procedures outlined in this review, provide a more efficient, reliable and cost-effective approach for diagnosis of pheochromocytoma than offered by previously available approaches.

Adrenal Gland Neoplasms↗
PubMed1999

Conversion of cucumber linoleate 13-lipoxygenase to a 9-lipoxygenating species by site-directed mutagenesis.

Multiple lipoxygenase sequence alignments and structural modeling of the enzyme/substrate interaction of the cucumber lipid body lipoxygenase suggested histidine 608 as the primary determinant of positional specificity. Replacement of this amino acid by a less-space-filling valine altered the positional specificity of this linoleate 13-lipoxygenase in favor of 9-lipoxygenation. These alterations may be explained by the fact that H608V mutation may demask the positively charged guanidino group of R758, which, in turn, may force an inverse head-to-tail orientation of the fatty acid substrate. The R758L+H608V double mutant exhibited a strongly reduced reaction rate and a random positional specificity. Trilinolein, which lacks free carboxylic groups, was oxygenated to the corresponding (13S)-hydro(pero)xy derivatives by both the wild-type enzyme and the linoleate 9-lipoxygenating H608V mutant. These data indicate the complete conversion of a linoleate 13-lipoxygenase to a 9-lipoxygenating species by a single point mutation. It is hypothesized that H608V exchange may alter the orientation of the substrate at the active site and/or its steric configuration in such a way that a stereospecific dioxygen insertion at C-9 may exclusively take place.

Amino Acid Substitution↗
PubMed1999

Contralateral trigeminal nerve lesion reduces polyneuronal muscle innervation after facial nerve repair in rats.

Functional recovery after facial nerve surgery is poor. Axotomized motoneurons (hyperexcitable upon intracellular current injections, but unable to discharge upon afferent stimulation) outgrow supernumerary branches which are misrouted towards improper muscles. We hypothesized that alterations in the trigeminal input to axotomized electrophysiologically silent facial motoneurons might improve specificity of reinnervation. To test this we compared, in the rat, behavioural, electrophysiological, and morphological parameters after transection and suture of the buccal facial nerve (buccal-buccal anastomosis, BBA) with those after BBA plus excision of the ipsi- or contralateral infraorbital nerve (ION). After BBA, the mystacial vibrissae dropped and remained motionless until 18-21 days post operation (days PO). After BBA plus ipsilateral ION excision, there was no recovery of vibrissae whisking at all. Following BBA plus contralateral ION excision, full restoration of whisking occurred at 7-10 days PO. Electromyography of whiskerpad muscles showed normal waveform and amplitude was also most rapidly restored after BBA plus contralateral ION excision. Neuron counts after retrograde tracing showed that the intact buccal nerve contained axons of the superior (91%) and inferior (9%) buccolabial nerves. After BBA, the superior nerve comprised 56%, the inferior 21%, and 23% of the motoneurons projected within both nerves. After BBA plus ipsilateral ION excision, misdirection worsened and values changed to 48, 39 and 13%, respectively. After BBA plus contralateral ION excision, portions improved to 69, 23 and 8%. We conclude that, by reducing the redundant axon branching, lesion of contralateral ION provides the best conditions for recovery of vibrissae rhythmical whisking after reconstructive surgery on the facial nerve.

Anastomosis, Surgical↗