Absence of both MGME1 and POLG EXO abolishes mtDNA whereas absence of either creates unique mtDNA duplications.

Both POLG and MGME1 are needed for mitochondrial DNA (mtDNA) maintenance in animal cells. POLG, the primary replicative polymerase of the mitochondria, has an exonuclease activity (3'→5') that corrects for the misincorporation of bases. MGME1 serves as an exonuclease (5'→3'), producing ligatable DNA ends. Although both have a critical role in mtDNA replication and elimination of linear fragments, these mechanisms are still not fully understood. Using digital PCR to evaluate and compare mtDNA integrity, we show that Mgme1 knock out (Mgme1 KK) tissue mtDNA is more fragmented than POLG exonuclease-deficient "Mutator" (Polg MM) or WT tissue. In addition, next generation sequencing of mutant hearts showed abundant duplications in/nearby the D-loop region and unique 100 bp duplications evenly spaced throughout the genome only in Mgme1 KK hearts. However, despite these unique mtDNA features at steady-state, we observed a similar delay in the degradation of mtDNA after an induced double strand DNA break in both Mgme1 KK and Polg MM models. Lastly, we characterized double mutant (Polg MM/Mgme1 KK) cells and show that mtDNA cannot be maintained without at least one of these enzymatic activities. We propose a model for the generation of these genomic abnormalities which suggests a role for MGME1 outside of nascent mtDNA end ligation. Our results highlight the role of MGME1 in and outside of the D-loop region during replication, support the involvement of MGME1 in dsDNA degradation, and demonstrate that POLG EXO and MGME1 can partially compensate for each other in maintaining mtDNA.

Bipolar radiofrequency lesion geometry: implications for palisade treatment of sacroiliac joint pain.

Ex vivo photographic temperature mapping of bipolar radiofrequency (RF) lesions in animal tissue is performed over a wide range of electrode tip spacings, tip lengths, tip diameters, tip temperatures, and lesion times. In vivo temperature measurements collected during clinical treatment of sacroiliac joint (SIJ) pain corroborate those collected ex vivo. Generation of a "strip lesion" connecting two separated bipolar electrode tips is demonstrated ex vivo for tip spacings as large as 20 mm. A rounded rectangular bipolar lesion with midline dimensions 12 mm × 15 mm × 8 mm (L × W × D) is demonstrated using 10 mm parallel tip spacing, 10 mm tip lengths, 20 gauge cannulae, 90°C tip temperature, and 3-minute lesion time. Lesion length can be increased to 18 mm by using 15 mm tip lengths. Lesion width can be increased to 17 mm by using 12 mm tip spacing. The size of conventional bipolar lesions can exceed the size of lesions produced both by conventional monopolar RF (12 mm × 7 mm × 7 mm ellipsoidal) and by cooled monopolar RF as used in spinal pain management (10 mm × 10 mm × 10 mm spherical). SIJ pain is treated by placing 5 to 7 straight RF cannulae perpendicular to the dorsal sacrum and producing 4 to 6 overlapping bipolar RF lesions between the dorsal sacral foramina and the ipsilateral SIJ. This bipolar "palisade" (a defensive fence) creates a continuous lesion spanning the region through which multiple sacral lateral branch nerves travel along irregular, branching paths to reach the SIJ.