Mitochondrial Disease: A Historical,Biochemical, and London Perspective

Roland Luft is credited with describing the first truly mitochondrial disorder in the late 1950s and early 1960s. Cases such as his have proven to be exceptionally rare. Some years later, methods of mitochondrial analysis--enzymatic, polarographic, and spectroscopic, which had been developed primarily by groups in Philadelphia--were applied to the study of mitochondria isolated from skeletal muscle biopsies of patients thought to have defects of oxidative phosphorylation. In the vanguard of these investigations were groups in New York and London. John Clark led the latter group. Application of biochemical studies, more recently supplemented by molecular mtDNA and nuclear DNA studies, have revealed that mitochondrial disorders are among the most common of all metabolic disorders.     

Effect of epinephrine on muscle glycogenolysis during exercise in trained men

Febbraio, M. A., D. L. Lambert, R. L. Starkie, J. Proietto,and M. Hargreaves. Effect of epinephrine on muscle glycogenolysis during exercise in trained men. J. Appl.Physiol. 84(2): 465-470, 1998.To test thehypothesis that an elevation in circulating epinephrine increasesintramuscular glycogen utilization, six endurance-trained men performedtwo 40-min cycling trials at 71 ± 2% of peak oxygen uptake in20-22°C conditions. On the first occasion, subjects wereinfused with saline throughout exercise (Con). One week later, afterdetermination of plasma epinephrine levels in Con, subjects performedthe second trial (Epi) with an epinephrine infusion, which resulted ina twofold higher (P < 0.01) plasmaepinephrine concentration in Epi compared with Con. Although oxygenuptake was not different when the two trials were compared, respiratoryexchange ratio was higher throughout exercise in Epi compared with Con(0.93 ± 0.01 vs. 0.89 ± 0.01; P < 0.05). Muscle glycogenconcentration was not different when the trials were comparedpreexercise, but the postexercise value was lower(P < 0.01) in Epi compared with Con.Thus net muscle glycogen utilization was greater during exercise withepinephrine infusion (224 ± 37 vs. 303 ± 30 mmol/kg for Con andEpi, respectively; P < 0.01). Inaddition, both muscle and plasma lactate and plasma glucoseconcentrations were higher (P < 0.05) in Epi compared with Con. These data indicate that intramuscularglycogen utilization, glycolysis, and carbohydrate oxidation areaugmented by elevated epinephrine during submaximal exercise in trainedmen.

     

The recovery of Arcobacter butzleri NCTC 12481 from various temperature treatments

AIMS: The aim of this study was to determine the growth and survival characteristics for Arcobacter butzleri NCTC 12481. METHODS AND RESULTS: The temperature and pH growth ranges were 15-39 degrees C and pH 6.0-8.0, as determined using impedance microbiology. The maximum specific growth rate was 00.57 h(-1) at 30 degrees C, pH 7.0. Arcobacter butzleri harvested from the exponential phase was more resistant to heat treatment than stationary phase cells (D55 1.1 and 0.4 min, respectively). Fluorescent dye uptake, and the release of UV-absorbing material, increased in heat-treated cells. After 21 d storage at 4 and -20 degrees C, A. butzleri was recovered on blood agar, but not on the isolation media CAT or CCDA. CONCLUSION: Arcobacter butzleri cells from the exponential phase were less heat sensitive than those from the stationary phase. The organism was able to survive cold storage for at least 3 weeks. SIGNIFICANCE AND IMPACT OF THE STUDY: The growth and survival characteristics have been quantified thus providing a greater understanding of this newly emerging pathogen.     

Effect of pH on tolerance of Hormidium rivulare to zinc and copper

Summary The toxicity of zinc to a population of Hormidium rivulare isolated from an acid mine drainage was shown to be least at the optimum pH range for the growth of the alga, pH 3.5–4.0; toxicity increases markedly at higher pH values. Calcium clearly antagonizes the toxicity of zinc. Populations of H. rivulare isolated from higher pH values and which are resistant to zinc, are also especially resistant to low pH values, although they are unlikely ever to encounter such values naturally. Nevertheless raised levels of calcium bring about only a slight improvement of growth at very low pH values in the absence of zinc, so the mechanisms of pH and zinc tolerance are not the same. Although the acid stream population grows in the field in an environment with rather similar levels of zinc and copper, copper is less toxic than zinc at pH 3.5, but much more toxic than zinc at pH 6.0.     

Chlorimipramine: a novel anticancer agent with a mitochondrial target

Mitochondria have been suggested to be a potential intracellular target for cancer chemotherapy. In this report, we demonstrate the ability of the tricyclic antidepressant chlorimipramine to kill human glioma cells in vitro by a molecular mechanism resulting in an increase in caspase 3 activity following inhibition of glioma oxygen consumption. Studies with isolated rat mitochondria showed that chlorimipramine specifically inhibited mitochondrial complex III activity, which causes decreased mitochondrial membrane potential as well as mitochondrial swelling and vacuolation. The use of chlorimipramine in human as an effective, non-toxic cancer therapeutic having a strong selectivity between cancer cells and normal cells on the basis of their mitochondrial function is discussed.     

Effects of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine on differentiating mouse N2a neuroblastoma cells

The effect of the neurotoxin 1-methyl-4-phenyl-1,2,3, 6-tetrahydropyridine (MPTP) was investigated in mouse N2a neuroblastoma cells, induced to differentiate by serum withdrawal and addition of dibutyryl cyclic AMP, over a 24-h period. Addition of MPTP (10 microM) during differentiation caused a change in cell morphology characterised by an inhibition of axon outgrowth, in the absence of cell death. Biochemical characterisation by western blotting revealed that MPTP had no significant effects on the levels of actin, alpha-tubulin, or total heavy-chain neurofilament (NF-H). However, NF-H phosphorylation appeared to increase following MPTP treatment when blots were probed with the phosphorylation state-specific antibodies RMd09 and Ta51. In addition, indirect immunofluorescence analysis revealed an accumulation of phosphorylated NF-H in the cell perikaryon, suggesting that altered NF-H distribution was associated with the observed effects of MPTP on cell morphology. These changes may represent a useful in vitro marker of MPTP neurotoxicity within a simple differentiating neuronal cell model system.     

CRISPR Diversity in E. coli Isolates from Australian Animals,Humans and Environmental Waters

Seventy four SNP genotypes and 54 E. coli genomes from kangaroo, Tasmanian devil, reptile, cattle, dog, horse, duck, bird, fish, rodent, human and environmental water sources were screened for the presence of the CRISPR 2.1 loci flanked by cas2 and iap genes. CRISPR 2.1 regions were found in 49% of the strains analysed. The majority of human E. coli isolates lacked the CRISPR 2.1 locus. We described 76 CRISPR 2.1 positive isolates originating from Australian animals and humans, which contained a total of 764 spacer sequences. CRISPR arrays demonstrated a long history of phage attacks especially in isolates from birds (up to 40 spacers). The most prevalent spacer (1.6%) was an ancient spacer found mainly in human, horse, duck, rodent, reptile and environmental water sources. The sequence of this spacer matched the intestinal P7 phage and the pO111 plasmid of E. coli.