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Biomedical subjects

D Meyer

Publications and source records attributed to D Meyer.

At least 271 records · Page 15Linked to original sources
PubMed1994

A novel case of compound heterozygosity with "Normandy"/type I von Willebrand disease (vWD). Direct demonstration of the segregation of one allele with a defective expression at the mRNA level causing type I vWD.

We report the case of a family with type I von Willebrand disease (vWD), characterized by a quantitative defect in von Willebrand factor (vWF), associated with a defective binding of vWF to factor VIII (FVIII) also called the "Normandy" variant of vWD. PCR products from genomic DNA of the family members were analysed in the region coding for the binding domain of vWF to FVIII. It showed that the proposita and one of her sons were heterozygous for the Arg91Gln missense mutation, abolishing an MspI restriction enzyme site located in exon 20. The transcription of the normal and mutated alleles was tested by the amplification of cDNA after reverse transcription of platelet mRNA in this region. A total lack of expression of the normal allele was observed in the proposita, who appeared as a compound heterozygous with one allele mutated at Arg91 and a "silent" expression of the other one. The segregation of the "silent" allele was studied in the family with the exonic BstEII RFLP both at the DNA and mRNA levels. The proposita has transmitted her "silent" allele to her daughter and to another son. As this son was informative for this RFLP, the absence of expression of the allele could be demonstrated at the mRNA level, providing evidence that this defect was responsible for his type I vWD.

Alleles↗
PubMed1994

New direct assay of free protein S antigen using two distinct monoclonal antibodies specific for the free form.

Monoclonal antibodies (mAbs) specific for free protein S and devoid of reactivity with protein S-C4b-BP complexes, have been produced. A one-step sandwich-type enzyme-linked immunoassay (ELISA) has been developed with two mAbs reacting with distinct epitopes of free protein S. F(ab')2 fragments from mAb 15C4 were coated on microplates and mAb 34G2 conjugated with horseradish peroxidase (HRP) was added immediately before diluted plasma. The presence of calcium in the sample diluent prevented dissociation of complexes during the assay. This assay was specific as demonstrated by good recovery of purified protein S added to plasma and the lack of influence of C4B-BP-protein S complexes. Thus, addition of increasing amounts of purified C4B-BP to human citrated plasma induced a dose-dependent decrease of free protein S. The assay was sensitive, allowing measurement of 5-500 ng/ml of free protein S, with a detection threshold of 2 ng/ml. It was also reproducible with inter-assay and intra-assay variation coefficients of 2.5-5.1% and 3.1-5.0%, respectively. Thus, this new ELISA of free protein S antigen in plasma has the advantages of being fast, accurate and reproducible. It appears to be extremely useful for routine studies as no preliminary treatment of plasma is required.

Antibodies, Monoclonal↗
PubMed1994

Clinical applications of a direct assay of free protein S antigen using monoclonal antibodies. A study of 59 cases.

A new one-step ELISA using two monoclonal antibodies specific for distinct epitopes of the free form of protein S (ELISA-m) has been developed for the direct measurement of free protein S in untreated plasma. This assay has been compared with the classic method using polyclonal antibodies to protein S (ELISA-p). The latter method has the drawback of requiring PEG precipitation of plasma which is time-consuming, difficult to perform with accuracy and therefore poorly reproducible in most laboratories. Results of both ELISAs were compared with those of a functional assay. In 30 normal subjects, there was an excellent correlation between ELISA-m and ELISA-p (r = 0.95) as well as between ELISA-m and the functional assay (r = 0.96). In twelve patients with a congenital deficiency, the levels of free protein S antigen were similarly decreased with ELISA-m and ELISA-p and in good agreement with those of protein S activity. In 20 patients with miscellaneous inflammatory diseases, the levels of free proteins S were normal with good correlation between both ELISAs and PS activity, despite high levels of C4bBP-protein S complexes. As expected, in 15 dicoumarol-treated patients, there was a significant and parallel decrease of free protein S antigen with both ELISAs, with even lower levels of protein S activity. In 14 patients with liver cirrhosis, the mean values for free protein S antigen were normal using both assays, but with wide extreme values, whereas protein S activity was significantly lower.(ABSTRACT TRUNCATED AT 250 WORDS)

Antibodies, Monoclonal↗
PubMed1994

The role of platelet von Willebrand factor in platelet adhesion and thrombus formation: a study of 34 patients with various subtypes of type I von Willebrand disease.

In order to investigate the respective role of plasma and platelet von Willebrand factor (vWF) in mediating platelet adhesion and thrombus formation, we performed ex vivo perfusion studies with native blood from patients with various subtypes of type I von Willebrand disease (vWD). We studied 34 patients with type I vWD (19 'platelet normal', five 'platelet low', two 'platelet discordant', eight 'Vicenza'). Parallel studies were carried out on nine patients with severe vWD (type III). At high shear rate (2600 s-1) we found that the defect in platelet-vessel wall interactions in patients having a normal platelet vWF content ('platelet normal' and 'Vicenza') involved thrombus formation, whereas platelet adhesion was normal. At this high shear rate, platelet adhesion and thrombus volume were significantly decreased in patients with subtypes 'platelet low' and 'platelet discordant', i.e. when platelet vWF is either low or dysfunctional. These results indicate that platelet vWF may substitute for plasma vWF to promote platelet adhesion, emphasizing the important role of platelet vWF. They also confirm the role of vWF in thrombus formation at high shear rate because an abnormal thrombus volume was observed in all patients, even when platelet adhesion was normal.

Blood Platelets↗
PubMed1994

Fine epitope mapping of monoclonal antibodies to the NH2-terminal part of von Willebrand factor (vWF) by using recombinant and synthetic peptides: interest for the localization of the factor VIII binding domain.

Two different approaches were used in order to define the epitope of three monoclonal antibodies (MoAbs) against the NH2-terminal part of the mature subunit of von Willebrand factor (vWF) which contains its factor VIII (FVIII) binding site. First, a vWF cDNA fragment library using the bacteriophage lambda gt11 expression vector was screened with radiolabelled MoAbs. The epitope of each MoAb was defined, following sequence analysis, by the overlapping DNA sequence of immunoreactive clones. MoAb 32B12, a potent inhibitor of FVIII/vWF interaction, binds within the Glu35-Ile81 sequence of vWF subunit. MoAb 14A12, a non-inhibitory antibody, recognizes a sequence within Thr141-Val220. MoAb 31H3, a partial inhibitory antibody, gives no positive clone. In the second method, a panel of 24 synthetic pentadecapeptides corresponding to the first NH2-terminal 105 amino acid residues was used to block the binding of inhibitor MoAbs to immobilized vWF in an ELISA system. The localization of MoAb 32B12 epitope was confirmed and restricted to the Met51-Ala60 sequence. The MoAb 31H3 binding to vWF is inhibited by two synthetic peptides with the overlapping sequence Cys66-Gly76. All these data confirm that the FVIII binding site of vWF is not limited to the binding area (Thr78-Thr96) of the previously described MoAbs inhibiting FVIII/vWF interaction but is composed of several key sequences.

Antibodies, Monoclonal↗
PubMed1994

Persistent hyperplastic primary vitreous of the eye: imaging findings with pathologic correlation.

Persistent hyperplastic primary vitreous is a rare benign developmental disorder of the globe in which the embryonic hyaloid artery fails to regress normally, resulting in abnormal lenticular development and secondary changes of the retina and globe. This entity often presents a diagnostic challenge as it is the most common intraocular abnormality to be confused with retinoblastoma. Sonography, CT, and MR imaging have been used alone and in combination to diagnose this condition. This essay illustrates the imaging characteristics of persistent hyperplastic primary vitreous in two children with unilateral disease and in one child with bilateral disease as representative cases (Figs. 1-3). Available pathologic correlates also are presented.

Diagnosis, Differential↗
PubMed1994

What women say about their birth experiences: a cross-cultural study.

The reactions of 221 women to their place of delivery (state or private hospital), type of delivery (vaginal or Caesarean) and associated obstetrical interventions were explored by means of a structured interview conducted at 3 months postpartum. Women were randomly selected from the birth records of three cultural groups residing in the municiple area of Johannesburg, South Africa: White (n = 72), Indian (n = 79) and Mixed cultural origin (n = 78). The type of birth experienced by women differed in the cultural groups examined. While most women experienced positive emotional reactions to birth many also expressed a variety of negative reactions. The differences in care experienced by women in the present study reflect the impact of the two systems of care, private and state, that are available to women in South Africa.

Adult↗
PubMed1994

[Pouches of the upper gastrointestinal tract].

The own findings about the functional behavior of the gastric substitute together with the results from the literature demand a jejunal J-pouch after gastrectomy, if it is not a palliative resection. It is an open question, whether the good experience with the Roux-en-Y-pouch-reconstruction--only 15 min. more time of operation, low rate of complications--allows a wider indication for this procedure.

Anastomosis, Roux-en-Y↗
PubMed1994

[The functionally inactive endocrine pancreas tumor].

Endocrine tumors of the pancreas are extremely rare (1-3%). We present two cases with functional inactive tumors--one benign and one malignant--, which were immunohistologically secreting glucagon without having clinical symptoms. Differential diagnosis, operative strategy and therapy of endocrine tumors of the pancreas are discussed.

Adult↗
PubMed1994

Optimization of the transfection of human endothelial cells by electroporation.

A method for efficient transfection of human umbilical vein endothelial cells (HUVEC) is presented here. The procedure involves cell synchronization followed by electroporation and allows the detection of the activity of low-strength promoters. Sodium butyrate, added to the culture medium after electroporation, strongly potentiates the transcriptional activity of viral promoters. The method can be successfully applied to analyze the transcriptional activity of different promoters linked to reporter genes or to express foreign genes controlled by viral promoters.

Butyrates↗
PubMed1994

MEDUSA: a fuzzy expert system for medical diagnosis of acute abdominal pain.

Even today, the diagnosis of acute abdominal pain represents a serious clinical problem. The medical knowledge in this field is characterized by uncertainty, imprecision and vagueness. This situation lends itself especially to be solved by the application of fuzzy logic. A fuzzy logic-based expert system for diagnostic decision support is presented (MEDUSA). The representation and application of uncertain and imprecise knowledge is realized by fuzzy sets and fuzzy relations. The hybrid concept of the system enables the integration of rule-based, heuristic and case-based reasoning on the basis of imprecise information. The central idea of the integration is to use case-based reasoning for the management of special cases, and rule-based reasoning for the representation of normal cases. The heuristic principle is ideally suited for making uncertain, hypothetical inferences on the basis of fuzzy data and fuzzy relations.

Abdomen, Acute↗
PubMed1994

Epitope mapping of inhibitory monoclonal antibodies to human von Willebrand factor by using recombinant cDNA libraries.

Two recombinant expression libraries containing small (300-600 base pairs) cDNA fragments of von Willebrand Factor (vWF) were screened in order to map the epitope of monoclonal antibodies (MAbs) to vWF. Among eleven MAbs tested, seven were effectively mapped. The epitopes of MAbs 418 and 522, which inhibit the binding of vWF to Factor VIII (FVIII), were localized between Leu 2 and Arg 53 and between Glu 35 and Ile 81 of the vWF subunit respectively, within the N-terminal trypsin fragment called SpIII-T4 [amino acids (aa) 1-272] which contains a binding domain for FVIII. The epitope of MAb 710, which inhibits the binding of vWF to glycoprotein Ib (GPIb), was identified between Ser 593 and Ser 678 on the tryptic 52/48 kDa fragment (aa 449-728) which contains binding domains for GPIb, collagen, heparin, sulfatides and subendothelium extracellular matrices. The epitope of MAb 723, which does not interfere with any known function of vWF, was localized between Ser 523 and Gly 588. The epitopes of MAb 505 and MAb 400, which inhibit the binding of vWF to collagen, were identified between Leu 927 and Arg 1114 within the SPI fragment (aa 911-1365) corresponding to the central part of the vWF subunit. The epitope of MAb 9, which inhibits the binding of vWF to GPIIb/IIIa, was identified in the C-terminal part of the vWF subunit between Gln 1704 and Asp 1746, the latter being the third aa of the RGD sequence common to adhesive proteins and serving as a recognition site for integrin receptors.

Amino Acid Sequence↗
PubMed1994

Interaction of von Willebrand factor with platelets activated by thrombin or a synthetic 7-amino acid peptide derived from the cleaved thrombin receptor.

Thrombin and the 7-mer agonist peptide from its receptor (SFLLRNP) were compared for their ability to promote the binding of vWF to platelets. Identical Ca(2+)-dependence and kinetics of activation were observed. Studies of inhibition of the binding by a series of monoclonal antibodies to GPIb, GPIIb/IIIa and vWF and experiments performed using platelets from patients with Glanzmann thrombasthenia or Bernard-Soulier syndrome enabled to identify GPIIb/IIIa as the receptor of vWF. Binding isotherms of vWF in the presence of an excess of either agonist yielded a similar number of binding sites but an apparent dissociation constant slightly but consistently higher with the 7-mer peptide than with thrombin. The latter point was confirmed by studying the binding of limiting amounts of vWF to platelets as a function of the agonist concentration. The lower affinity in the presence of 7-mer peptide was not corrected by adding increasing amounts of FPR-thrombin, a derivative with irreversibly blocked active site but retaining the binding properties of the active enzyme. Conversely, the higher affinity observed with thrombin was decreased when platelets were treated with Serratia protease which selectively cleaved GPIb but did not affect the function of the thrombin receptor and GPIIb/IIa. Our data thus suggest that both the 7-mer peptide and thrombin are able to induce the assembly of functional GPIIb/IIIa.(ABSTRACT TRUNCATED AT 250 WORDS)

Bernard-Soulier Syndrome↗
PubMed1994

[Social control of medical-technical progress].

The analysis emphasises the dependence of medical-technological progress on the zero fees for standard health services. Technologies with an additional or total self-payment are discriminated against by the patients and the physicians. Besides, a tendency to apply medical maximum standards was discerned. The enforcement of novel methods of diagnosis and therapy has been made substantially easier. In contrast, there are scarcely any incentives to introduce cost-saving technologies from the patient's point of view. The fixed budgets of the Health Structure Act will exercise greater pressure on introducing cost-cutting innovations.

Cost Savings↗
PubMed1994

Binding of heparin fractions to von Willebrand factor: effect of molecular weight and affinity for antithrombin III.

To investigate the influence of the structure of heparin on its binding to vWF, we compared heparin fractions of different molecular weight (MW) or affinity for antithrombin III (ATIII). We studied the interaction of purified 125I-vWF or plasma vWF, labeled with a pool of 125I-monoclonal antibodies to vWF, with unfractionated heparin immobilized on agarose beads. Fractions were compared as competitors of these interactions and their effect was quantitated by their half-maximal inhibition (IC50). When the MW of the fractions decreased, especially below 7500, their IC50 increased, indicating that the affinity of the fractions for vWF decreased with their MW. Using heparin-derived oligosaccharides, we also demonstrated that a minimal chain length of 18 monosaccharides was required for heparin binding to vWF. In addition, different fractions with low affinity for ATIII were compared as competitors of 125I-vWF binding to heparin-agarose. Despite a very low content of ATIII binding sites, some fractions retained a low IC50. Thus, heparin interaction with vWF is independent of the presence of the ATIII binding site and is mostly dependent on the length of the heparin chain. These data suggest that unfractionated heparin is a more potent inhibitor of vWF-dependent functions than low MW heparin fractions.

Antibodies, Monoclonal↗