The efficacy of commercial tooth storage media for maintaining the viability of human periodontal ligament fibroblasts.

AIM: To evaluate Save-A-Tooth (SAT), EMT Toothsaver (EMT) and Hank's Balanced Salt Solution (HBSS) for their influence on the viability and proliferative capacity of human periodontal ligament fibroblasts (HPDLFs). METHODOLOGY: Primary HPDLFs were seeded into 96-well cell culture plates and exposed to SAT, EMT, HBSS and water (negative control) for 0.5, 1, 3, 6, 12 and 24 h at room temperature (22 °C). After each exposure time, cell viability was measured through quantifying adenosine triphosphate (ATP) using a luminescent dye. The proliferative capacity was also quantified using the PrestoBlue assay after 12 or 24 h storage in each medium. The data were analysed statistically by two-way anova and post hoc Least Significant Difference (LSD) test (P < 0.05). The morphology of the cells after 12 h storage was also investigated through live/dead viability/cytotoxicity kit together with fluorescence microscopy. RESULTS: There was no significant difference in cell viability amongst HBSS, SAT and EMT groups up to 6 h. SAT was effective in maintaining cell viability only up to 12 h and then became detrimental to HPDLF; after 24 h, the effectiveness of SAT in maintaining cell viability was similar to that of water (P > 0.05). Amongst all the media, only EMT could maintain the proliferative capacity of HPDLFs significantly higher than the negative control, that is water (P < 0.05) after 24 h storage. CONCLUSION: EMT maintained the proliferative capacity of HPDLFs after 24 h storage.

First record of a new member of Anopheles Hyrcanus Group from Iran: molecular identification, diagnosis, phylogeny, status of kdr resistance and Plasmodium infection.

The current study aimed to provide further evidence on the status of species composition, insecticide resistance, and vectorial capacity within the members of Anopheles (Anopheles) Hyrcanus Group in Ardebil, Giulan, and Khuzestan provinces of Iran. Sequencing the internal transcribed spacer 2 (ITS2) of ribosomal DNA gene led to identification of two members of Hyrcanus complex: Anopheles hyrcanus Pallas and a new species/form, hereafter called Anopheles hyrcanus sp(IR) as a world record. Furthermore, we identified and compared partial sequences of exons I and II and the whole intron I region of insecticide resistance-related voltage-gated sodium channel (vgsc) gene in populations of Hyrcanus Group and other main old world Anopheles species. The ITS2 and vgsc sequences in members of Hyrcanus Group and other Anopheles species were used for construction of phylogenetic tree, which demonstrated the evolutionary relatedness among Western and Eastern Palearctic taxa within the Hyrcanus Group. A nested polymerase chain reaction assay for detection of Plasmodium species revealed the infection of Plasmodium falciparum within An. hyrcanus collected from Fooman district in Guilan province. The data from this study led to the introduction of a new member/form within the Hyrcanus Group, identification and definition of the status of knockdown resistance related to pyrethroids and DDT in their vgsc gene, detection of Plasmodium infection, and further evidence on genetic relatedness within these taxa. The overall results may suggest reconsidering the role ofAn. hyrcanus in malaria transmission, which would be useful for implementation and evaluation of malaria control programs in Western Palearctic region.