Mitochondria are subcellular organelles composed of two discrete membranes in the cytoplasm of eukaryotic cells. They have long been recognized as the generators of energy for the cell and also have been known to associate with several metabolic pathways that are crucial for cellular function. Mitochondria have their own genome, mitochondrial DNA (mtDNA), that is completely separated and independent from the much larger nuclear genome, and even have their own system for making proteins from the genes in this mtDNA genome. The human mtDNA is a small (~16.5 kb) circular DNA and defects in this genome can cause a wide range of inherited human diseases. Despite of the significant advances in discovering the mtDNA defects, however, there are currently no effective therapies for these clinically devastating diseases due to the lack of technology for introducing specific modifications into the mitochondrial genomes and for generating accurate mtDNA disease models. The ability to engineer the mitochondrial genomes would provide a powerful tool to create mutants with which many crucial experiments can be performed in the basic mammalian mitochondrial genetic studies as well as in the treatment of human mtDNA diseases. In this review we summarize the current approaches associated with the correction of mtDNA mutations in cells and describe our own efforts for introducing engineered mtDNA constructs into the mitochondria of living cells through bacterial conjugation.
Conjugation, Genetic ; Cytoplasm ; DNA ; DNA, Circular ; DNA, Mitochondrial ; Eukaryotic Cells ; Genome ; Genome, Mitochondrial ; Humans ; Membranes ; Metabolic Networks and Pathways ; Mitochondria ; Organelles ; Proteins

OBJECTIVEDust sample mass gain is too smaller to satisfy the limit of detection (LOD) even in most cases during dust sampling at workplaces nowdays, especially for respirable fraction. Therefore, it is aimed to solve the problem by increasing sample load with high flow rate samplers.

METHODSIn A and B two shipyards respirable welding fume was sampled by high flow rate cyclone samplers of FSP-10 (10 L/min) for 2-2.5 hours and normal flow rate FSP-2 (2 L/min) for 3-4 hours with a stratigy of parallele sampling at the same workpalce, in order to compare their mass gain, coincidence rate with LOD, and airborn dust concentration.

RESULTSSample mass gain of 0.97±0.40 mg and 1.61±0.86 mg respectively in the two factories by FSP-10 was significantly higher than that of 0.29±0.12 mg and 0.51±0.27 mg by FSP-2 (t-test, P<0.05 in both cases) , increasing herewith the coincidence rate with LOD from 26.8% (when sampling with FSP-2, calculated together with samples of the two factories) to 89.7%. However there was no significant difference in dust concentrations by the two different samplers, 0.53±1.88 vs 0.73±1.61 mg/m(3) by FSP-2 and FSP-10 in the shipyard A and 1.14±1.78 vs 1.01±1.63 mg/m(3) in the factory B (t-test, P>0.05 in every case) . In addtion, sample loading by FSP-2 was found to be correlated to sampling time (R(2)=0.7906, y=0.002 6x) , therefore, it has to sample for ≥192.3 min to meet the LOD (0.5 mg) in case of normal flow rate.

CONCLUSIONBy using of high flow rate cyclone FSP-10 the problem of LOD could be solved, along with increased sample mass and similar respirable dust concentration by the two samplers. Some techincal improvements of FSP-10 and increasing of LOD coincidence rate by other methods was also disscussed.

Air Pollutants, Occupational ; analysis ; Construction Industry ; Dust ; analysis ; Environmental Monitoring ; instrumentation ; Occupational Exposure ; Ships ; Workplace