Magnitude of ornithine decarboxylase induction by epidermal mitogens: Effect of the assay technique

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/47234/1/403_2004_Article_BF00414009.pd

Intact epidermal cell assay for ornithine decarboxylase activity

A procedure measuring the ornithine decarboxylase (ODC) activity and polyamine formation of intact neonatal mouse epidermal cells in culture has been developed and tested. Basal cells prepared from neonatal mouse epidermis were plated on round 15-mm Lux coverslips, placed in Costar 24 well culture clusters and grown at 32°C in M-199 + 13% fetal bovine serum. Before assay the cells were rendered permeable to ornithine 14 C and ODC inhibitors using the buffer described by Berger et al. [3]. The slides, covered with adhering cell layers, were then placed in vials, covered with assay buffer and assayed intact for ODC activity. The ODC reaction was terminated by addition of citric acid to the buffer and the amount of 14 CO 2 released was determined by scintillation counting of a center well filled with trapping agent. The baseline ODC activity of the intact cells was 500–1,000 pmol 14 CO 2 /mg protein/45 min. The validity of this ODC assay procedure using intact neonatal mouse keratinocytes was tested by use of three specific ODC inhibitors and by measuring the formation of polyamines from uniform labeled ornithine. The results indicated that authentic ODC activity was measured and preserved in this intact neonatal mouse epidermal cell assay. This technique holds promise for future studies of epidermal cell regulation of ODC and polyamine synthesis and studies of the multiple ornithine metabolites and conjugates formed, using a highly manipulable in vitro system.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/47233/1/403_2004_Article_BF00509038.pd

Glycerol treatment as recovery procedure for cryopreserved human skin allografts positive for bacteria and fungi

Human donor skin allografts are suitable and much used temporary biological (burn) wound dressings. They prepare the excised wound bed for final autografting and form an excellent substrate for revascularisation and for the formation of granulation tissue. Two preservation methods, glycerol preservation and cryopreservation, are commonly used by tissue banks for the long-term storage of skin grafts. The burn surgeons of the Queen Astrid Military Hospital preferentially use partly viable cryopreserved skin allografts. After mandatory 14-day bacterial and mycological culture, however, approximately 15% of the cryopreserved skin allografts cannot be released from quarantine because of positive culture. To maximize the use of our scarce and precious donor skin, we developed a glycerolisation-based recovery method for these culture positive cryopreserved allografts. The inactivation and preservation method, described in this paper, allowed for an efficient inactivation of the colonising bacteria and fungi, with the exception of spore-formers, and did not influence the structural and functional aspects of the skin allografts

Potential release of aluminum and other metals by food-grade aluminum foil used for skin allograft cryo preservation

Since 1991, the skin bank of the Queen Astrid Military Hospital uses food-grade aluminum foil as a primary support for storing cryo preserved human donor skin (511 donors). The possible release of heavy metals into the cryo preservation media (30% (v/v) glycerol in physiological water) and the possible impact this release could have on the quality of the cryo preserved donor skin was evaluated. Aluminum was the principal detection target. Possible contaminants of the aluminum foil as such (arsenic, cadmium, chromium and lead) were also investigated. The evaluation was set up after a Belgian Competent Authority inspection remark. Aluminum was detected at a concentration of 1.4 mg/l, arsenic and lead were not detected, while cadmium and chromium were detected in trace element quantities. An histological analysis revealed no differences between cryo preserved and fresh donor skin. No adverse reactions in patients, related to the presence of aluminum or heavy metal traces, were reported since the introduction of the cryo preserved donor skin in our burn wound centre

Feeder layer- and animal product-free culture of neonatal foreskin keratinocytes: improved performance, usability, quality and safety

Since 1987, keratinocytes have been cultured at the Queen Astrid Military Hospital. These keratinocytes have been used routinely as auto and allografts on more than 1,000 patients, primarily to accelerate the healing of burns and chronic wounds. Initially the method of Rheinwald and Green was used to prepare cultured epithelial autografts, starting from skin samples from burn patients and using animal-derived feeder layers and media containing animal-derived products. More recently we systematically optimised our production system to accommodate scientific advances and legal changes. An important step was the removal of the mouse fibroblast feeder layer from the cell culture system. Thereafter we introduced neonatal foreskin keratinocytes (NFK) as source of cultured epithelial allografts, which significantly increased the consistency and the reliability of our cell production. NFK master and working cell banks were established, which were extensively screened and characterised. An ISO 9001 certified Quality Management System (QMS) governs all aspects of testing, validation and traceability. Finally, as far as possible, animal components were systematically removed from the cell culture environment. Today, quality controlled allograft production batches are routine and, due to efficient cryopreservation, stocks are created for off-the-shelf use. These optimisations have significantly increased the performance, usability, quality and safety of our allografts. This paper describes, in detail, our current cryopreserved allograft production process

PRIMO-INFECTION HERPETIQUE GENERALISEE DE TYPE 2 CHEZ UN ADULTE 'SAIN'

A healthy 26-yr-old patient presented a primary herpetic type 2 infection. The infection disseminated to the skin, the mucosa and the liver. The clinical course of the infection had a favorable issue.SCOPUS: NotDefined.jinfo:eu-repo/semantics/publishe

PARALYSIE FACIALE PERIPHERIQUE ET SYPHILIS SECONDAIRE. PROBLEME DU TRAITEMENT DES MENINGITES SYPHILITIQUES

The authors are presenting a case of meningitis with facial nerve palsy due to early syphilis. The frequency, symptomatology and treatment of this meningeal invasion during the septicaemic phase of secundary syphilis are discussed.SCOPUS: NotDefined.jinfo:eu-repo/semantics/publishe

Treatment of epidermolysis bullosa with human cultured epidermal allografts

Junctional epidermolysis bullosa letalis type Herlitz Pearson is a genetically determined, life-threatening disease. Effective therapy has been lacking to date. Therefore any therapy that improves wound healing would be beneficial for these patients. Cultured epidermal grafts are known to enhance wound epithe- lialization and have been used with success in some epidermolysis bullosa disorders. Encouraged by these reports, we grafted cultured allogeneic keratino- cytes to an infant with a junctional epidermolysis bullosa letalis type. © 1994 S. Karger AG, Basel.SCOPUS: ar.jinfo:eu-repo/semantics/publishe

Granulome moniliasique chez l'adulte: Piège clinique et histologique

info:eu-repo/semantics/publishe