Electroporation-mediated delivery of the FER gene in the resolution of trauma-related fatal pneumonia.

Injured patients with lung contusion (LC) are at risk of developing bacterial pneumonia (PNA) followed by sepsis and death. A recent genome-wide association study (GWAS) showed FER gene expression positively correlating with survival rates among individuals with above conditions. We sought to determine whether electroporation (EP)-mediated delivery of FER gene could indeed improve survival, in a lethal model of combined LC and PNA. C57BL/6 mice sustained unilateral LC, which preceded a 500 Klebsiella colony forming unit (CFU) inoculation by 6 h. In-between these insults, human FER plasmid (pFER) was introduced into the lungs followed by eight EP pulses applied externally (10 ms at 200 V cm-1). Control groups included EP of empty vector (pcDNA3) or Na+/K+-ATPase genes (pPump) and no treatment (LC+PNA). We recorded survival, histology, lung mechanics, bronchial alveolar lavage (BAL) fluid, FER and inflammatory gene expression and bacteriology. The data show that 7-day survival was significantly improved by pFER compared with control groups. pFER increased BAL monocytes and activated antibacterial response genes (nitric oxide synthase (NOS), Fizz). pFER treatment showed decreased lung and blood Klebsiella counts reaching, in some cases, complete sterilization. In conclusion, FER gene delivery promoted survival in LC+PNA mice via recruitment of activated immune cells, improving efficiency of bacterial clearance within contused lung.

[The role of intracellular Ca(2+) pools in the regulation of protoplast volume. Effect of red light on the Ca(2+) mobilization in cytoplasm of Arabidopsis cells]. / Rol' vnutrikletochnykh pulov Ca(2+) v reguliatsii ob''ema protoplastov. Vliianie krasnogo sveta na mobilizatsiiu Ca(2+) v tsitoplazme kletok Arabidopsis.

The changes in cytosol Ca2+ concentration associated with the shrinkage of Arabidopsis cells induced by the inhibitor of Ca(2+)-ATPase, cyclopiazonic acid and the Ca2+ ionophore ionomycin were monitored using the fluorescence of Ca(2+)-sensitive probe chlortetracycline hydrochloride. It was found that these compounds elicited a substantial decrease in fluorescence intensity closely associated with Ca(2+)-release from the intracellular stores to the cytoplasm. The release of Ca2+ from the intracellular depots was accompanied by decrease of plant cell volume. Thapsigargin and 2,5'-ditert-butyl-1,4-benzohydroquinone (highly specific inhibitors of Ca(2+)-ATPase of endoplasmic reticulum) resulted in much weaker changes than cyclopiazonic acid did. It was also found with the help of the same technique that red light (lambda = 660 nm) illumination induced a similar Ca2+ release from the intracellular stores. Moreover, the amplitudes of light-induced fluorescence responses registered in mutant plants differing in the content of phytochrome A (phyAOX) and phytochrome B (phyBOX) were much higher than those registered in wild-type of Arabidopsis.

[Activation of poly(ADP-ribose)-polymerase inhibits apoptosis of thymocytes, caused by short-wave ultraviolet radiation]. / Aktivatsiia poli(ADF-ribozo)-polimerazy ingibiruet apoptoz timotsitov, vyzvannyi korotkovolnovym ul'trafioletovym izlucheniem.

The effect of (i) aphidicolin, a specific inhibitor of delta- and epsilon-polymerases, and nucleotide excision repair; (ii) 3-aminobenzamide, an inhibitor of poly(ADP-ribose) polymerase and base excision repair; and (iii) actinomycin D and cycloheximide, inhibitors of protein and RNA synthesis, respectively, on the induction of suppression of apoptosis of rat thymocytes by different doses of short-wavelength ultraviolet radiation was studied by flow cytometry. 3-Aminobenzamide suppressed the inhibition of apoptosis induced by the doses of short-wavelength ultraviolet radiation higher than 20 J/m2, increasing the cell death to a maximum. Thus, the inhibition of apoptosis by high short-wavelength ultraviolet radiation doses depends on the status of poly(ADP-ribose) polymerase and is prevented by 3-aminobenzamide. As opposed to 3-aminobenzamide, aphidicolin did not affect the cell death at short-wavelength radiation doses higher than 10 J/m2 but induced the apoptosis of unirradiated cells and cells irradiated with short-wavelength ultraviolet radiation doses lower than 10 J/m2. The inhibitors of protein and RNA synthesis cycloheximide and actinomycin D prevented the induction of apoptosis caused by low and medium doses but did not abolish the apoptosis-inhibiting activity of high doses of short-wavelength ultraviolet radiation.

[Effect of structural analogs of platelet activating factor on the intracellular signal transduction in murine peritoneal neutrophils and macrophages of P388D1 line]. / Deistvie strukturnykh analogov factora aktivatsii trombotsitov na peredachu vnutrikletochnykh signalov v peritoneal'nykh neitrofilakh myshi i makrofagakh linii P388D1.

Direct and modulate effects of platelet activating factor (PAF), its structural analogues and ATP on primary and second processes at peritoneal neutrophils and P388D1 cells activation has been studied. The effect of compounds was evaluated on changes in Ca2+ transport and generation of reactive oxygen species. It was shown, that the synthetic analogues of MS series interact with PAF receptor, mobilize Ca2+ from thapsigargin-dependent intracellular stores and inhibit Ca2+ response on PAF in both types of cells. Unlike PAF the analogues do not induce the formations of reactive oxygen species in neutrophils and inhibit the PMA-induced respiratory burst. The activation of pyrinoreceptor of P388D1 cells by exogenous ATP does not inhibit PAF induced Ca2+ rise in cytoplasm, though partly releases Ca2+ from the same store.