Digital PCR methods improve detection sensitivity and measurement precision of low abundance mtDNA deletions

Mitochondrial DNA (mtDNA) mutations are a common cause of primary mitochondrial disorders, and have also been implicated in a broad collection of conditions, including aging, neurodegeneration, and cancer. Prevalent among these pathogenic variants are mtDNA deletions, which show a strong bias for the loss of sequence in the major arc between, but not including, the heavy and light strand origins of replication. Because individual mtDNA deletions can accumulate focally, occur with multiple mixed breakpoints, and in the presence of normal mtDNA sequences, methods that detect broad-spectrum mutations with enhanced sensitivity and limited costs have both research and clinical applications. In this study, we evaluated semi-quantitative and digital PCR-based methods of mtDNA deletion detection using double-stranded reference templates or biological samples. Our aim was to describe key experimental assay parameters that will enable the analysis of low levels or small differences in mtDNA deletion load during disease progression, with limited false-positive detection. We determined that the digital PCR method significantly improved mtDNA deletion detection sensitivity through absolute quantitation, improved precision and reduced assay standard error

The effect of low level laser therapy in different wavelengths in the treatment of oral mucositis-proposal for extra-oral implementation

The oral mucositis is the most frequent acute oral complication resulting from antineoplastic treatment and may worsen the clinical condition of the patient and interfere with his/her quality of life. This study aimed to comparatively evaluate, from a clinical point of view, the effect of Laser Therapy lambda 660 nm (wavelength of the red Laser) and lambda 830 nm (wavelength of the infrared Laser), at extra-oral points, in remission of severity of oral mucositis and pain associated with it in pediatric oncological patients undergoing chemotherapy with the anticancer drug methotrexate, noting which of the two wavelength is the most appropriate to this new technique. The sample consisted of 13 patients placed at random in each group and subjected to sessions of Low Level Laser Therapy, at pre-determined extra-oral points for five consecutive days, starting at the beginning of the observation of mucositis injuries. It became possible to note that from the group of patients in the group of Laser lambda 830 nm (n = 6; 46.15%), four (n = 4; 66.67%) of these patients had remission of injuries to grade 0 (WHO), and as for pain, five patients (n = 5; 83.33%) showed no painful symptoms for mucositis injuries. In the Laser lambda 660 nm group (n = 7; 53.85%), only two patients (n = 2; 28.57%) achieved a regression of lesions to grade 0 (WHO), while four patients (n = 4; 57.14%) had no pain. So, the extra-oral application of Laser Therapy was effective in treating injuries of oral mucositis in the patients treated; and Laser Therapy in the infrared spectrum (lambda 830 nm) was more effective in the treatment of oral mucositis injuries compared to the red spectrum (lambda 660 nm), which can be explained by the greater power of penetration of infrared rays, acting in a more expressive way in deeper places.1991912191