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Biomedical subjects

M Adam

Publications and source records attributed to M Adam.

At least 55 records · Page 3Linked to original sources
PubMed1994

[The effect of aluminum on the structure and metabolism of collagen].

BACKGROUND: Aluminium is considered to be the etiopathogenetic factor in various pathological conditions. It was demonstrated already previously that metals even under conditions in vivo link with collagen structures and influence the protein metabolism. Therefore the authors investigated the effect of aluminium (Al) on collagen and its metabolism. METHODS AND RESULTS: In the described trial aluminium was used as potassium alum, oxalate alum or aluminium chloride. The effect of aluminium was investigated in rats, chick embryos or fibroblast cultures. Using electron microscopy of collagen from the tail tendons, the development of transverse striation following 15 weeks of i.m. Al administration was revealed. At the same time also an increase of temperature of contraction of this collagen by 1.5 to 3.7 degrees C occurred, depending on the Al compound used. On fibroblast cultures an inhibitory effect of Al on their proliferation was found. In chick embryos Al caused a decline of the radioactive hydroxyproline concentration and thus also a reduced collagen synthesis practically in all investigated tissues. Similarly the negative effect of Al was manifested in the incorporation of radioactive glucosamine into proteoglycans of granulation tissue of rats. In cartilaginous collagen enhanced proline hydroxylation caused by Al was observed. CONCLUSIONS: Although it is not possible, with regard to Al chemistry, to make an unequivocal statement on the nature of the compound which causes a particular reaction in biological systems, it may be said, based on the achieved results, that Al has an effect on biological systems due to its bond with collagen structures and by influencing their metabolism.

Aluminum↗
PubMed1994

Cloning, functional expression, and characterization of the human prostaglandin E2 receptor EP2 subtype.

A cDNA clone encoding the human prostaglandin (PG) E2 receptor EP2 subtype has been isolated from a human lung cDNA library. The 1.9-kilobase pair cDNA, hEP2, encodes for a 488-amino acid protein with a predicted molecular mass of 53,115 and has the seven putative transmembrane domains characteristic of G protein-coupled receptors. The specific binding of [3H]PGE2 to COS cell membranes transfected with the hEP2 cDNA was of high affinity with an equilibrium dissociation constant (Kd) of 1 nM and the rank order of potency for prostaglandins in competition for [3H]PGE2 specific binding was PGE1 = PGE2 >> iloprost > PGF2 alpha > PGD2. In competition studies using more selective prostanoid-receptor agonist and antagonists, the [3H]PGE2 specific binding was competed by MB28767, an EP3 agonist, but not by the EP1-preferring antagonists AH6809 and SC19220, or by the EP2 agonist butaprost. Electrophysiological studies of Xenopus oocytes co-injected with hEP2 and cystic fibrosis transmembrane conductance regulator (cAMP-activated Cl- channel) cDNAs detected PGE2-specific inward Cl- currents, demonstrating that the hEP2 cDNA encoded a functional receptor which produced an increase in cAMP levels. Thus, we have cloned the human EP2 receptor subtype which is functionally coupled to increase in cAMP. Northern blot analysis showed that hEP2 is expressed as a 3.8-kilobase mRNA in a number of human tissues with the highest expression levels present in the small intestine.

Amino Acid Sequence↗
PubMed1994

[Intracavernous auto-injection of prostaglandin E1 for diagnosis and treatment of erectile dysfunction].

34 impotent patients were offered auto-injection therapy and all except 2 began treatment. Prostaglandin E1 (PGE) was used regularly for a mean of 8.6 months (range 1-17). The onset of erection was 5-30 min (mean 11.4) after injection and it lasted at least 30 min (mean 106.2 min). An erection of more than 6 hrs was observed in only 2 patients. There have been no instances of cavernous fibrosis, chemical priapism or systemic reactions. The most common adverse effect was penile pain after injection. It was mild to moderate in 4 patients (11.7%), and in only 1 was it severe enough to require stopping treatment. Overall, intracavernous injection of PGE appears to be safe, effective treatment for impotence without side-effects other than pain after injection in a few.

Adult↗
PubMed1994

Characterization of protein tyrosine phosphatase SH-PTP2. Study of phosphopeptide substrates and possible regulatory role of SH2 domains.

The src homology 2 (SH2) domain containing protein-tyrosine-phosphatase SH-PTP2, was over-expressed in Escherichia coli for a kinetic study employing a set of synthetic 13- to 14-mer phosphopeptide substrates. The full-length SH-PTP2 protein, as well as a truncated form, lacking the two amino terminus SH2 domains (SH-PTP2(delta SH2)), exhibited Michaelis-Menten kinetics, and demonstrated striking substrate preferences on phosphopeptides having sequences based on sites of intracellular protein tyrosine phosphorylation. For example, while a KM of 59 microM and kcat/KM of 1.1 x 10(5) were obtained using SH-PTP2(delta SH2) and PDGFRY1021, a phosphorylation site within the platelet-derived growth factor receptor, other peptides revealed no detectable phosphate release. PDGFRY1009, modeled after a sequence identified as an in vivo binding site for SH-PTP2, was also a good substrate for this enzyme. The truncated form, lacking the SH2 domains demonstrated higher catalytic efficiency than the full-length enzyme. Interestingly, soluble SH2 domains were found to inhibit the catalytic activity of SH-PTP2 in a concentration-dependent manner. There was also evidence of a non-phosphotyrosine-mediated association between the two domains. These observations suggested that the SH2 domains have a direct role in regulating the catalytic activity of SH-PTP2.

Amino Acid Sequence↗
PubMed1994

Engineering and overexpression of periplasmic forms of the penicillin-binding protein 3 of Escherichia coli.

Replacement of the 36 and 56 N-terminal amino acid residues of the 588-amino-acid-residue membrane-bound penicillin-binding protein 3 (PBP3) of Escherichia coli by the OmpA signal peptide allows export of F37-V577 PBP3 and G57-V577 PBP3 respectively into the periplasm. The modified ftsI genes were placed under the control of the fused lpp promoter and lac promoter/operator; expression of the truncated PBP3s was optimized by varying the copy number of the recombinant plasmids and the amount of LacI repressor, and export was facilitated by increasing the SecB content of the producing strain. The periplasmic PBP3s (yield 8 mg/l of culture) were purified to 70% protein homogeneity. They require the presence of 0.25 M NaCl to remain soluble. Like the membrane-bound PBP3, they undergo processing by elimination of the C-terminal decapeptide I578-S588, they bind penicillin in a 1:1 molar ratio and they catalyse hydrolysis and aminolysis of acyclic thioesters that are analogues of penicillin. The membrane-anchor-free PBP3s have ragged N-termini. The G57-V577 PBP3, however, is less prone to proteolytic degradation than the F37-V577 PBP3.

Amino Acid Sequence↗
PubMed1994

Cloning and expression of three isoforms of the human EP3 prostanoid receptor.

Functional cDNA clones coding for three isoforms of the human prostaglandin E receptor EP3 subtype have been isolated from kidney and uterus cDNA libraries. The three isoforms, designated hEP3-I, hEP3-II and hEP3-III, have open reading frames corresponding to 390, 388 and 365 amino acids, respectively. They differ only in the length and amino acid composition of their carboxy-terminal regions, beginning at position 360. The human EP3 receptor has seven predicted transmembrane spanning domains and therefore belongs to the G-protein-coupled receptor family. The rank order of potency for prostaglandins and related analogs in competition for [3H]PGE2 specific binding to membranes prepared from transfected COS cells was comparable for all three isoforms, and as predicted for the EP3 receptor, with PGE2 = PGE1 >> PGF2 alpha = iloprost > PGD2 >> U46619. In addition, the EP3-selective agonist MB28767 was a potent competing ligand with an IC50 value of 0.3 nM, whereas the EP1-selective antagonist AH6909 gave IC50 values of 2-7 microM and the EP2-selective agonist butaprost was inactive. In summary, we have cloned three isoforms of the human EP3 receptor having comparable ligand binding properties.

Amino Acid Sequence↗
PubMed1994

Enhancement of the efficacy of a replication-defective adenovirus-vectored vaccine by the addition of oil adjuvants.

We previously constructed a recombinant adenovirus with a defective E1A gene, which expresses high levels of the pseudorabies virus gp50 in non-transcomplementing cells. The virus is unable to replicate in mice. It elicited the production of anti-gp50 antibodies only when high concentrations (10(8) TCID50 per dose) of the virus were used and it gave mice little protection. The combination of the recombinant adenovirus at several concentrations (10(8), 10(7.4), 10(6.4) TCID50 per dose) with certain oil adjuvants in different galenic forms (water-in-oil, oil-in-water, water-in-oil-in-water) led to an increase in specific antibody responses and protection for the host when challenged with a virulent pseudorabies virus under very severe conditions, i.e. where 100% of unvaccinated mice died. A water-in-oil-in-water formulation induced a very high level of anti-gp50 antibodies even with a low concentration of adenovirus. These results could be correlated to the induction of cytokines, such as IL6, which is observed with this galenic form. The oil-adjuvanted emulsions induced IL2, suggesting that they were able to activate T-helper cells. Different oil formulations elicited the different IgG subclasses (IgG1, IgG2a, IgG2b, IgG3). These results can be extended to other live replication-defective vaccines expressing different proteins.

Adenoviruses, Human↗
PubMed1994

Vaccination of pigs with replication-defective adenovirus vectored vaccines: the example of pseudorabies.

The efficacy of a recombinant human adenovirus type 5 expressing gD, one of the immunogenic glycoprotein of pseudorabies virus, was tested in pigs. Due to the deletion of the E1a gene, the recombinant virus is unable to replicate in non transcomplementing cells but is capable of eliciting an immune response against gp50 after inoculation into animals. The virus was formulated in a water/oil/water emulsion, a strategy previously shown to enhance the immune response against the virus-induced gp50. Pigs of 18-25 kg were vaccinated twice and the recombinant virus was not isolated from nasal and rectal swabs taken after each injection of the vaccine. High levels of neutralizing antibodies were induced by the vaccination. Protection against a severe challenge was effective, as measured by growth performance (dG = 1.73), and reduction of the time of excretion of the challenge strain (mean time: 4.4 days for the vaccinated and 7.9 days for the control pigs). These results show that non replicating adenoviruses are able to induce a strong protective immune response against foreign genes in pigs, which may be of general interest for the design of pig vaccines.

Adenoviruses, Human↗
PubMed1994

The effect of different collagens and of proteoglycan on the retraction of collagen lattice.

The effect of various collagens and proteoglycan on the formation and retraction of collagen lattices was tested. The most rapid aggregation of collagen molecules was observed by the use of the least cross-linked collagen fractions (ie pepsin-digested calf skin collagen type I). Lattices formed with more cross-linked collagens (acid soluble collagen-ASC, type III) contracted slowly and less intensively. Unpurified pepsinized cartilage extract containing collagen types II, IX and XI, some glycoproteins and proteoglycans formed lattices rather well. On the contrary, purified collagen type II as well as polymeric collagen (solubilized by denaturing conditions) did not form lattices at all. The lattice formation and retraction was intensified by addition of proteoglycan into the culture medium. The authors suggest that the kinetics of the lattice formation and retraction depends on the amount of collagen cross-links and the concentration of proteoglycan in the culture medium.

Collagen↗
PubMed1994

Anaplastic Wilms' tumour, a subtype displaying poor prognosis, harbours p53 gene mutations.

The genetics of Wilms' tumour (WT), a paediatric malignancy of the kidney, is complex. Inactivation of the tumour suppressor gene, WT1, is associated with tumour aetiology in approximately 10-15% of WTs. Chromosome 17p changes have been noted in cytogenetic studies of WTs, prompting us to screen 140 WTs for p53 mutations. When histopathology reports were available, p53 mutations were present in eight of eleven anaplastic WTs, a tumour subtype associated with poor prognosis. Amplification of MDM2, a gene whose product binds and sequesters p53, was excluded. Our results indicate that p53 alterations provide a molecular marker for anaplastic WTs.

Alleles↗
PubMed1994

Design and validation of a new immunoassay for soluble forms of thrombomodulin and studies on plasma.

Thrombomodulin (TM), purified from human placental homogeneate by affinity chromatography on DIP-Thrombin agarose, was used to develop monoclonal antibodies (MAbs). Two of them, 3E2 and 24FM (both IgG1, K), which were not calcium-dependent, were found convenient for developing a two-site enzyme immunoassay. Testing of recombinant and truncated forms of TM26 demonstrated that the species containing the amino terminus including the lectin-like domain and the epidermal growth factor (EGF)-like domains 1-4 were fully measured. The working range was from 2 to 100 ng/ml with a detection threshold of 2 ng/ml. Intraassay and interassay reproducibilities were, respectively, below 7.4% and 8.6%, whereas recovery of purified TM was between 88 and 114% in plasma. Mean plasma concentration was 42.1 (+/- 11.3) ng/ml (males 51.8 +/- 7.9 ng/ml, females 34.8 +/- 7.8 ng/ml) and it was established on 62 normal individuals between the ages of 21 and 55 (28 males and 34 females). This new assay is a convenient tool for measuring plasma TM and establishing its diagnostic and predictive value in diseases associated to endothelial damage.

Adult↗
PubMed1994

Patient perceptions of the experience of electrophysiologic studies.

This descriptive study explored perceptions, feelings and attitudes related to electrophysiologic studies (EPS). Structured one hour taped interviews were conducted with 11 participants and data was subjected to qualitative analysis. Powerlessness, anticipation and total exhaustion were the categories identified from the data. Overall, participants generally perceived their EPS as a negative experience and illustrated this with words like invasion, horrible, uncomfortable, etc. Anxiety was predominant throughout the entire experience. Nurses are in an ideal position to impact these negative perceptions by helping patients incorporate the threatening nature of EPS into a cognitive plan, thus reducing anxiety. Strategies and interventions to help patients cope with this aversive event are offered as well as suggestions for future research.

Aged↗
PubMed1994

Transgenic mice harboring Aujeszky's disease virus gD gene.

Plasmid pMLP10 gp50.5 (gD) (Eloit et al., 1990) provided the sequence to integrate. The construct has been injected into the male pronucleus of one-cell mouse embryos (CBA/C5B1). A total of 1,567 microinjected embryos were transferred into the oviduct of pseudopregnant females (Hogan et al., 1986). PCR revealed that 3 out of 10 mice were born with an integrated construct; among them, 2 mice have integrated the construct in their gonads because 20 out of 107 (18.7%) of their offspring were also transgenic. This low percentage could be explained by germline mosaicism and/or differential mortality between transgenic and non-transgenic embryos.

Animals↗
PubMed1993

Cloning and expression of a cDNA for the human prostaglandin E receptor EP1 subtype.

A functional cDNA clone coding for the human prostaglandin E receptor EP1 subtype has been isolated from a human erythroleukemia cell cDNA library probed by low-stringency hybridization using a polymerase chain reaction fragment of the human thromboxane receptor. The human EP1 receptor is comprised of 402 amino acids with a predicted molecular mass of 41,858 and has the topography common to all G-protein-coupled receptors with seven predicted transmembrane spanning domains. Prostaglandin (PG) E2 challenge of Xenopus oocytes injected with EP1 cDNA resulted in an increase in intracellular Ca2+. In addition, the rank order of potency for prostaglandins in competition for [3H]PGE2 specific binding to membranes prepared from EP1 cDNA transfected COS cells was PGE2 > PGE1 > PGF2 alpha > PGD2. Furthermore, the EP1 receptor-selective antagonists AH 6809 and SC19220 were more potent than the EP2 receptor-selective agonist butaprost in these competition binding assays. In summary, therefore, we have cloned the human EP1 receptor subtype which is functionally coupled to an increase in intracellular Ca2+.

Amino Acid Sequence↗
PubMed1993

Cloning, expression and mutational analysis of SH-PTP2, human protein-tyrosine phosphatase.

A human cDNA clone encoding a nonreceptor protein-tyrosine-phosphatase (PTP) has been isolated and sequenced. The 2.1 kilobase pair cDNA encodes for a 593 amino acid protein that contains a single tyrosine phosphatase catalytic domain at the C-terminus. At the N-terminus the protein has two adjacent copies of Src homology region (SH2 domain) which show 61% and 73% identity at the amino acid level to the SH2 domains of the human PTP1C and Drosophila corkscrew protein, respectively. The overall homology between SH-PTP2 and PTP1C or to corkscrew protein is 58%. When this protein (or its catalytic domain) was expressed in E. coli as a glutathione-S-transferase fusion protein tyrosine-phosphatase activity was detected in bacterial cell extracts. Site-directed mutation made at the conserved cysteine (459) residue to serine within the highly conserved VHCXAGXXR sequence in the PTP catalytic domain resulted in complete loss of enzymatic activity demonstrating the importance of this cysteine residue in catalysis. Northern blot analysis showed that SH-PTP2 is expressed as a 6.5 kilobase mRNA in a number of fetal and adult human tissues and cell lines. The highest levels of its mRNA were detected in fetal brain and in adult heart tissue. The identification of SH-PTP2 along with PTP1C and corkscrew protein suggest that there exist a family of nonreceptor PTP containing SH2-domain which will participate in specific signal transduction pathways involving tyrosine phosphorylation-dephosphorylation.

Amino Acid Sequence↗
PubMed1993

Enantioselective activation of the peroxisome proliferator-activated receptor.

A cell-based transactivation assay was established using the mouse full-length peroxisome proliferator-activated receptor (PPAR) cDNA sequence and the positive peroxisome proliferator-responsive regulatory element (-578 to -553) of the rat acyl-CoA oxidase gene promoter. Activation of the reporter plasmid was dependent on co-transfection of the full-length PPAR cDNA, and the response was greatly stimulated, up to 100-fold, by peroxisome proliferators such as Wy-14,643 ([4-chloro-6-(2,3-xylidino)-2-pyrimidinylthio] acetic acid), nafenopin (2-methyl-2[p-(1,2,3,4-tetrahydro-1-naphthyl)phenoxy]-propionic acid), and clofibric acid (2-([p]-chlorophenoxy)-2-methylpropionic acid). Activation of the reporter plasmid promoter by the full-length PPAR cDNA also occurred at peroxisomal proliferator concentrations 40 times lower than that required for similar stimulation by a glucocorticoid-PPAR chimeric receptor. By using the stereoisomers of MK-571 ((+-)-3-(((3-(2-(7-chloro-2-quinolinyl)ethenyl)-phenyl)((3- (dimethylamino)-3-oxopropyl)-thio)methyl)-thio)propanoic acid), a potent leukotriene D4 receptor antagonist, we could show enantioselective activation of PPAR. The use of this compound in mice results in peroxisome proliferation; however, nearly all of the peroxisome proliferating activity can be attributed to the S enantiomer. Our results show a similar enantiomeric discrimination in PPAR activation of the reporter plasmid promoter, where again most of the activity can be attributed to the S enantiomer. The equivalent activities of these stereoisomers both in vivo and in the PPAR transactivation assay strongly implicate PPAR as a major component of the peroxisome proliferating mechanism in rodents.

Acyl-CoA Oxidase↗
PubMed1993

Poly(HEMA)-collagen composite as a biomaterial for hard tissue replacement.

This article briefly reviews the possibilities for hard tissue replacement with a new biomaterial. The basic differences found experimentally for polymer (HEMA) and collagen composite at the biological environment are stressed. The influence of the collagen distribution and matrix porosity of composite material on biodegradation is also discussed.

Animals↗