Tissue distribution of a plasmid DNA encoding Hsp65 gene is dependent on the dose administered through intramuscular delivery

In order to assess a new strategy of DNA vaccine for a more complete understanding of its action in immune response, it is important to determine the in vivo biodistribution fate and antigen expression. In previous studies, our group focused on the prophylactic and therapeutic use of a plasmid DNA encoding the Mycobacterium leprae 65-kDa heat shock protein (Hsp65) and achieved an efficient immune response induction as well as protection against virulent M. tuberculosis challenge. In the present study, we examined in vivo tissue distribution of naked DNA-Hsp65 vaccine, the Hsp65 message, genome integration and methylation status of plasmid DNA. The DNA-Hsp65 was detectable in several tissue types, indicating that DNA-Hsp65 disseminates widely throughout the body. The biodistribution was dose-dependent. In contrast, RT-PCR detected the Hsp65 message for at least 15 days in muscle or liver tissue from immunized mice. We also analyzed the methylation status and integration of the injected plasmid DNA into the host cellular genome. The bacterial methylation pattern persisted for at least 6 months, indicating that the plasmid DNA-Hsp65 does not replicate in mammalian tissue, and Southern blot analysis showed that plasmid DNA was not integrated. These results have important implications for the use of DNA-Hsp65 vaccine in a clinical setting and open new perspectives for DNA vaccines and new considerations about the inoculation site and delivery system.     

Oxygen consumption in four species of teleosts from Greenland: no evidence of metabolic cold adaptation

Standard metabolic rate of Greenland cod or uvak, Gadus ogac, polar cod, Boreogadus saida, Atlantic cod, Gadus morhua, and sculpin, Myxocephalus scorpius, caught in the same geographical area on the west coast of Greenland was measured at 4.5°C, the temperature at which the fish were caught. The present data does not support the Metabolic Cold Adaptation theory in the traditional sense of the standard metabolic rate being 2–4 times higher for Arctic fishes than for temperate species. The standard metabolic rate of the two exclusively Arctic species of teleosts was only 10% and 26% higher, respectively, than the two species that occur in temperate as well as Arctic areas. The critical oxygen tension, with respect to oxygen consumption, of resting uvak was between 50 and 60 mmHg, and the lethal oxygen tension 20–25 mmHg at 4.5°C, which is considerably higher than for Atlantic cod from a temperate area measured at the same temperature.     

The role of adrenaline as a modulator of cardiac performance in two Antarctic fishes

The present work was performed to test the hypothesis that Antarctic teleosts rely mostly on cholinergic inhibition for autonomic modulation of the heart. The effects of adrenaline on the inotropic properties on paced, isometrically contracting muscle strips were examined in two distinct Antarctic teleosts, the haemoglobinless icefish Chaenocephalus aceratus and the red-blooded Notothenia coriiceps. All tissues examined revealed a negative force-frequency relationship. Under baseline conditions C. aceratus contracted with a force twice as great as that of N. coriiceps. While the degree to which ventricular tissues responded to adrenaline varied between species, adrenergic stimulation significantly increases myocyte contraction force in this group of fishes. Contraction and relaxation times were not significantly affected by adrenaline concentration while absolute rates of contraction were. Adrenergic stimulation does not enable tissues to achieve higher contraction frequencies, but is shown to be a potent modulator of contraction force.     

Partial trisomy of the long arm of human chromosome 1 as demonstrated by in situ hybridization with 5S ribosomal RNA

Summary In a newborn boy with multiple malformations, a tandem duplication was detected at the distal end of the long arm of one human chromosome 1. The Giemsa bands, 1q31 to 1q43–44, were repeated serially. Since 5S rRNA genes are located at 1q42–43, in situ hybridization of ¹²⁵I 5S rRNA with fixed chromosome preparations was used to confirm the chromosomal duplication. The infant exhibited numerous developmental and clinical abnormalities as might be expected with an abnormality of chromosome structure relating to a ribosome component.