Development of a point-of-care-device for fast detection of periodontal pathogens.

BACKGROUND: A number of pathogens can cause severe destruction of the periodontal apparatus during the course of periodontitis. The aim of this work was the development of a diagnostic device for the use at the point-of-need for the detection of periodontal pathogens to enable a personalized therapy for treatment of periodontitis. METHODS: This test system is based on the polymerase chain reaction of DNA isolated from periodontal pathogens and was examined to precisely detect species-specific sequences on a rotating chip with lyophilized reagents for polymerase chain reaction. The preservation of the reagents was optimized to ensure their stability during the storage. RESULTS: In the current work, we have developed a model point-of-care device and showed a proof of concept. It requires low sample volume, is timesaving and can therefore facilitate early diagnosis and treatment of periodontal diseases. CONCLUSIONS: The developed device can provide fast diagnosis of the composition and amount of patients' oral flora and might help to assess the stage of periodontitis infection. This can facilitate an optimization of therapeutic approaches in order to prevent some of the more serious consequences of the disease.

GiFRD encodes a protein involved in anaerobic growth in the arbuscular mycorrhizal fungus Glomus intraradices.

Fumarate reductase is a protein involved in the maintenance of redox balance during oxygen deficiency. This enzyme irreversibly catalyzes the reduction of fumarate to succinate and requires flavin cofactors as electron donors. Two examples are the soluble mitochondrial and the cytosolic fumarate reductases of Saccharomyces cerevisiae encoded by the OSM1 and FRDS1 genes, respectively. This work reports the identification and characterization of the gene encoding cytosolic fumarate reductase enzyme in the arbuscular mycorrhizal fungus, Glomus intraradices and the establishment of its physiological role. Using a yeast expression system, we demonstrate that G. intraradices GiFRD encodes a protein that has fumarate reductase activity which can functionally substitute for the S. cerevisiae fumarate reductases. Additionally, we showed that GiFRD transformants are not affected by presence of salt in medium, indicating that the presence of this gene has no effect on yeast behavior under osmotic stress. The fact that GiFRD expression and enzymatic activity was present only in asymbiotic stage confirmed existence of at least one anaerobic metabolic pathway in this phase of fungus life cycle. This suggests that the AMF behave as facultative anaerobes in the asymbiotic stage.