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Biomedical subjects

D Leroy

Publications and source records attributed to D Leroy.

At least 73 records · Page 4Linked to original sources
PubMed1996

Characterisation of human cdc2 lysine 33 mutations expressed in the fission yeast Schizosaccharomyces pombe.

The mammalian p34cdc2 protein kinase, a universal cell cycle regulator, complements cdc2/CDC28 temperature-sensitive mutations in yeasts. We report the biochemical characterisation of two substitutions of human cdc2 at lysine 33, a residue involved in nucleotide binding, that differently alter the fission yeast cell cycle. K33A-hscdc2 and K33R-hscdc2 mutants are both catalytically inactive, but overexpression of K33R-cdc2 is lethal while K33A-cdc2 is not. We show that human K33R-cdc2 acts as a dominant negative allele that associates yeast cdc13/cyclinB and therefore renders endogeneous Schizosaccharomyces pombe cdc2 unactivatable. These results are discussed on the light of the molecular modeling of the mutants in the cdc2 model structure.

Betaine↗
PubMed1996

Precision-cut dog renal cortical slices in dynamic organ culture for the study of cisplatin nephrotoxicity.

The dog is the non-rodent species the most often used in preclinical drug safety evaluation. In this study, we established a new system of precision-cut dog renal cortical slices, evaluated their biochemical, functional, and morphological integrity, and determined the effects of cisplatin (cis-diamminedichloroplatinum (II), CDDP), a very potent nephrotoxic antineoplastic agent used to treat a variety of solid tumors, on the viability and histopathology of slices. Precision-cut renal cortical slices were made perpendicular to the cortical-papillary axis. Slices were incubated in DMEM/Ham's F12 culture medium containing 1 g/L glucose, 2 mmol/L glutamine, and 2 mmol/L heptanoic acid at 37 degrees C in an atmosphere of 5% CO2-70% O2-25% N2 in dynamic organ culture. Our results showed that slices maintained ATP and GSH content, protein synthesis, Na(+)-dependent uptake of glucose inhibited by phlorizin, PAH (p-aminohippuric acid) uptake inhibited by probenecid, and TEA (tetraethylammonium) uptake inhibited by mepiperphenidol for at least 6 h of culture, and morphological integrity up to 24 h. After 6 h of exposure, CDDP induced vacuolation and cell necrosis in the epithelial tubular cells of slices with a concentration-related increase in extension but not in severity. The development of the lesions started in the proximal tubules and extended to the distal tubules. The location and the extension of the lesions confirmed the observations in dog kidneys after in vivo treatment with CDDP by the intravenous route. The concentration-related decrease in slice viability after 6 h exposure to CDDP was in keeping with the extension of the histopathological lesions in the renal parenchyma. The slice viability was unaffected up to 0.63 mmol/L CDDP. At 1.25 and 2.5 mmol/L CDDP, slice viability fell by 35% and 75%, respectively. These results suggest that precision-cut dog renal cortical slices in culture may be suitable for addressing the specific nephrotoxicity issues encountered in this species.

Animals↗
PubMed1996

Cryotherapy of PUVA lentigines.

PUVA lentigines do not subside after discontinuation of photochemotherapy. Cryotherapy with liquid nitrogen was done on a small area in a patient who presented with numerous PUVA lentigines. Five years after this treatment, the PUVA lentigines had not returned. Cosmetically disturbing PUVA lentigines can be treated with cryotherapy.

Cryotherapy↗
PubMed1996

Heterologous expression of the human cyclin-dependent kinase inhibitor p21Cip1 in the fission yeast, Schizosaccharomyces pombe reveals a role for PCNA in the chk1+ cell cycle checkpoint pathway.

Fission yeast cells expressing the human gene encoding the cyclin-dependent kinase inhibitor protein p21Cip1 were severely compromised for cell cycle progress. The degree of cell cycle inhibition was related to the level of p21Cip1 expression. Inhibited cells had a 2C DNA content and were judged by cytology and pulsed field gel electrophoresis to be in the G2 phase of the cell cycle. p21Cip1 accumulated in the nucleus and was associated with p34cdc2 and PCNA. Thus, p21Cip1 interacts with the same targets in fission yeast as in mammalian cells. Elimination of p34cdc2 binding by mutation within the cyclin-dependent kinase binding domain of p21Cip1 exaggerated the cell cycle delay phenotype. By contrast, elimination of PCNA binding by mutation within the PCNA-binding domain completely abolished the cell cycle inhibitory effects. Yeast cells expressing wild-type p21Cip1 and the mutant form that is unable to bind p34cdc2 showed enhanced sensitivity to UV. Cell cycle inhibition by p21Cip1 was largely abolished by deletion of the chk1+ gene that monitors radiation damage and was considerably enhanced in cells deleted for the rad3+ gene that monitors both DNA damage and the completion of DNA synthesis. Overexpression of PCNA also resulted in cell cycle arrest in G2 and this phenotype was also abolished by deletion of chk1+ and enhanced in cells deleted for rad3+. These results formally establish a link between PCNA and the products of the rad3+ and chk1+ checkpoint genes.

Amino Acid Sequence↗
PubMed1996

Delayed hypersensitivity at the injection sites of a low-molecular-weight heparin.

We report a case of delayed hypersensitivity to a low-molecular-weight heparin. The patient presented with eczema at the site of subcutaneous injections and allergy tests confirmed the clinical diagnosis. There were positive tests with 3 low-molecular-weight heparins and unfractionated subcutaneous calcium heparin but negative with intravenous sodium heparin. Although delayed hypersensitivity to low-molecular-weight heparins is rare, the possibility of cross-allergy to unfractionated heparin could be a therapeutic problem for some patients.

Eczema↗
PubMed1996

Eczematous-like drug eruption induced by synergistins.

The authors report 4 cases of eczematous-like drug eruption after oral ingestion of synergistins, pristinamycin (3 cases) and virginiamycin (1 case). The lesions occurred after contact sensitization with topical virginiamycin. The clinical symptoms appeared a few hours after ingestion: a generalized maculopapular eruption, sometimes with general symptoms of anaphylactic reaction. Eczema appeared again on initial areas of contact dermatitis. There is a common allergenic group between these 2 antibiotics, which is a macrocyclic lactone. Physiopathology of this drug eruption is not clear: allergic reaction of the delayed type or anaphylactic reaction. Patients allergic to virginiamycin should be strongly cautioned against oral pristinamycin.

Adult↗
PubMed1996

Occupational contact dermatitis from propacetamol.

We report 4 cases of contact sensitization to propacetamol. They presented with lesions on the hands, forearms, crease of the elbows, and neck. They were all sensitized to multiple allergens and 2 of them were atopic. Patch tests to Pro-Dafalgan and propacetamol were positive; sodium citrate and paracetamol were negative. Our cases were similar to those published for the first time by Barbaud in 1995. The only allergen was propacetamol; patch tests with diethyglycine and paracetamol were negative. Propacetamol chlorhydrate is composed of a complex paracetamol-diethylglycine, which probably acts like a hapten capable of inducing cutaneous allergy. It is an occupational allergy affecting nurses who work in surgery departments or post-anesthesia recovery rooms, where high doses of analgesics are widely used. The patients were not allergic to oral paracetamol. Despite the usual precautions, the mixture of propacetamol chlorhydrate and solvent leaks onto the nurses' hands, suggesting that health care workers handling propacetamol chlorhydrate should wear gloves.

Acetaminophen↗
PubMed1996

Flutamide photosensitivity.

Flutamide is a non-steroid antiandrogen used in advanced prostate cancer. This drug induces liver toxicity. Other side effects have been reported, especially four cases of photosensitivity. Our patient presented a sun-exposed eruption that disappeared when flutamide was discontinued and relapsed when it was rechallenged. Photopatch tests were negative. UVA MED was much lower during the treatment and the rechallenge of flutamide. The action spectrum of the drug seems to be in the UVA range. According to the literature, this photosensitivity is probably due to a photo-allergic mechanism.

Aged↗
PubMed1995

Direct identification of a polyamine binding domain on the regulatory subunit of the protein kinase casein kinase 2 by photoaffinity labeling.

Phosphorylation of many protein substrates by the protein kinase casein kinase 2 (CK2) is stimulated severalfold in the presence of polyamines such as spermine. Previous experiments have shown that CK2 is a polyamine binding protein and that the regulatory beta subunit is required for this binding activity. To delineate the spermine binding site of CK2, we have applied a photoaffinity labeling method using a tritiated photoactivable analog of spermine, [3H]sperminediazonium. The photoaffinity labeled beta subunit was cleaved with cyanogen bromide, and two labeled peptides were separated by high performance liquid chromatography. The major one was the peptide T72EQAAEM78 and the minor one was a 22-amino acid peptide comprising residues Ile98 to Met119. Thr72 and His108 were identified as the labeled amino acids of the Thr72-Met78 and Ile98-Met119 peptides, respectively. In the same manner, we succeeded in determining the residue Leu220 as an alpha subunit residue covalently bound to the probe. The photoaffinity labeling method described here enabled the first elucidation, by direct microsequencing, of a polyamine binding site on CK2 for which we propose a provisional structural model. These observations suggest a possible mechanism for CK2 activation by polyamines at the molecular level.

Affinity Labels↗
PubMed1995

Effect of phenylarsine oxide on the fission yeast Schizosaccharomyces pombe cell cycle.

Phosphotyrosyl turnover is an essential regulatory mechanism for many biological processes, and the balance between tyrosine kinases and phosphatases plays a major role in the control of cell proliferation. Phenylarsine oxide (PAO), a potent inhibitor of tyrosine phosphatases (PTPase), was used to investigate the involvement of PTPase in the growth and control of the cell cycle of the fission yeast Schizosaccharomyces pombe. Cell proliferation was arrested by treatment with PAO, which was found to inhibit cdc25 PTPase in vitro but appeared not to act in vivo on this mitosis inducer. The PAO-treated cells displayed a mono- or binucleated phenotype and a DNA content that was either 2C or 4C, indicating a cell cycle arrest with a failure to complete cytokinesis. Entry into the cell division cycle from the G0 quiescent stage was also delayed by treatment with PAO. These results suggest that a number of key events in the mitotic cell cycle are regulated by as yet unidentified PTPases.

Arsenicals↗
PubMed1995

Phototoxic and photoprotective effects of topical isothipendyl.

Isothipendyl, an H1-receptor antagonist, is an antihistamine used as an antiallergic drug. Its molecular structure is close to the phenothiazines, but it has different photobiological properties. Like phenothiazines, isothipendyl is phototoxic with ultraviolet A (UVA) but it reduces the erythema response to UVB radiation. Isothipenyl seems to have the same protective properties as a sunscreen because its UVB protective properties are effective only if the substance is applied prior to exposure.

Anti-Allergic Agents↗
PubMed1995

[Has protein kinase CK2 a role in the intracellular mitogenic signalling?].

The protein kinase CK2 is an ubiquitous serine-threonine kinase found in all eukaryotic cells. Although well characterized on a biochemical ground, its role and regulation in the intact cell are not clearly understood. Its possible implication in the control of cell proliferation has been examined by several different approaches. (i) Immunocytochemical detection of CK2 revealed that whereas the signal was evenly distributed throughout cycle arrested cells in primary culture, it accumulates rapidly (30-90 min) in the nuclear compartment in cells stimulated to grow. (ii) CK2 biosynthesis is activated as an early response to growth factors in quiescent cells. The neo-synthesized kinase accumulates as the cells progress through the G1 phase. This growth factor-activated biosynthesis concerns in parallel the two kinase subunits. (iii) The kinase is activated in vitro by polyamines, which are increased in cells challenged by growth factors. Spermine binds to a specific domain of the beta subunit of CK2. (iv) In addition to phosphorylation CK2 forms a molecular complex with p53, a major negative regulator of the cell cycle. The complex was demonstrated in intact cells and reconstituted in vitro (Kd 70 nM) with purified components and shown to require the beta subunit and to result in the inhibition of p53 DNA-annealing activity. These observations suggest that CK2 and p53 may play a coordinated role in the cell response to mitogenic stimuli.

Animals↗