Validation study of the in vitro skin irritation test with the LabCyte EPI-MODEL24.

A validation study on an in vitro skin irritation assay was performed with the reconstructed human epidermis (RhE) LabCyte EPI-MODEL24, developed by Japan Tissue Engineering Co. Ltd (Gamagori, Japan). The protocol that was followed in the current study was an optimised version of the EpiSkin protocol (LabCyte assay). According to the United Nations Globally Harmonised System (UN GHS) of classification for assessing the skin irritation potential of a chemical, 12 irritants and 13 non-irritants were validated by a minimum of six laboratories from the Japanese Society for Alternatives to Animal Experiments (JSAAE) skin irritation assay validation study management team (VMT). The 25 chemicals were listed in the European Centre for the Validation of Alternative Methods (ECVAM) performance standards. The reconstructed tissues were exposed to the chemicals for 15 minutes and incubated for 42 hours in fresh culture medium. Subsequently, the level of interleukin-1 alpha (IL-1 α) present in the conditioned medium was measured, and tissue viability was assessed by using the MTT assay. The results of the MTT assay obtained with the LabCyte EPI-MODEL24 (LabCyte MTT assay) demonstrated high within-laboratory and between-laboratory reproducibility, as well as high accuracy for use as a stand-alone assay to distinguish skin irritants from non-irritants. In addition, the IL-1α release measurements in the LabCyte assay were clearly unnecessary for the success of this model in the classification of chemicals for skin irritation potential.

Inactivation of antineoplastics in clinical wastewater by electrolysis.

Wastewater from clinical institutions contains a considerable amount of toxic substances. Among the toxic substances, antineoplastics may induce carcinogenesis, teratogenesis, and the emergence of mutant microorganisms in the environment. Although the incineration or chemical treatments of disposed antineoplastics are recommended, a high energy during incineration and a careful quality control during chemical treatment are required. In this study, we determined the conditions for the electrolytic treatment of an antineoplastic, epirubicin hydrochloride (EH), using two platinum electrodes with a constant current of 100 mA. We analyzed the cytotoxicity, mutagenicity and antibacterial activity of electrolyzed EH and compared them with those of unelectrolyzed EH. Nearly 100% cytotoxicity, mutagenicity and antibacterial activity were eliminated and HPLC did not detect an EH molecule, in the case of electrolysis for 6 h. We also examined the biological cytotoxicities of electrolyzed irinotecan hydrochloride, vincristine sulfate, mitomycin C, paclitaxel, methotrexate and cisplatin, and found that 72.1-99.999% toxicity was eliminated by electrolysis under the same conditions. The biological toxicity of a mixture of these drugs was determined to be decreased by approximately 99% by electrolysis under the same conditions.

Tryptophan pyrrole ring cleavage enzymes in placenta.

Tryptophan pyrrole ring cleavage enzymes were assayed in pregnant uterus of mouse. The highest activity was observed at 6.5 days post-coitus (dpc) with a small activity shoulder at 9.5 to 12.5 dpc concepti and placenta. The highest peak at early concepti of 6.5 dpc was coincided with gene expression of tryptophan 2,3-dioxygenase (TDO). And the shoulder from 9.5 dpc was coincided with the expression of indoleamine 2,3-dioxygenase (IDO). These enzymes showed a different inhibitory behavior to a proper IDO inhibitor, 1-methyl tryptophan. In situ hybridization analysis revealed that TDO was expressed in the decidua of the early concepti. This is the first report of the extra hepatic TDO.