Formation of prolamellar-body-like ultrastructures in etiolated cyanobacterial cells overexpressing light-dependent protochlorophyllide oxidoreductase in Leptolyngbya boryana.

Protochlorophyllide (Pchlide) reduction is the penultimate step of chlorophyll (Chl) biosynthesis, and is catalyzed by two evolutionarily unrelated enzymes: dark-operative Pchlide oxidoreductase (DPOR) and light-dependent Pchlide oxidoreductase (LPOR). Because LPOR is the sole Pchlide reductase in angiosperms, dark-grown seedlings of angiosperms become etiolated. LPOR exists as a ternary complex of Pchlide-NADPH-LPOR to form paracrystalline prolamellar bodies (PLBs) in etioplasts. Because LPOR is distributed ubiquitously across oxygenic phototrophs including cyanobacteria, it would be important to determine whether cyanobacterial LPOR has the ability to form PLBs. We isolated a DPOR-less transformant ΔchlL/LPORox, carrying a plasmid to overexpress cyanobacterial LPOR in the cyanobacterium Leptolyngbya boryana. The transformant did not produce Chl in the dark and became etiolated with an accumulation of Pchlide and LPOR. Novel PLB-like ultrastructures were observed in etiolated cells, which disappeared during the early stage of the light-dependent greening process. However, the rate of Chl production in the greening process of ΔchlL/LPORox was almost the same as that observed in the control cells, which carried an empty vector. An in vitro LPOR assay of extracts of dark-grown ΔchlL/LPORox cells suggested that the PLB-like structures are deficient in NADPH. Low-temperature fluorescence emission spectra of membrane fractions of the etiolated cells indicated the absence of the photoactive form of Pchlide, which was consistent with the inefficiency of the greening process. Cyanobacterial LPOR exhibited an intrinsic ability to form PLB-like ultrastructures in the presence of the co-accumulation of Pchlide; however, the PLB-like structure differed from the authentic PLB regarding NADPH deficiency.

Anaerobic respiration coupled with mitochondrial fatty acid synthesis in wax ester fermentation by Euglena gracilis.

In Euglena gracilis, wax ester fermentation produces ATP during anaerobiosis. Here, we report that anaerobic wax ester production is suppressed when the mitochondrial electron transport chain complex I is inhibited by rotenone, whereas it is increased by the uncoupler carbonyl cyanide m-chlorophenylhydrazone (CCCP). The ADP/ATP ratio in anaerobic cells is elevated by treatment with either rotenone or CCCP. Gene silencing experiments indicate that acyl-CoA dehydrogenase, electron transfer flavoprotein (ETF), and rhodoquinone (RQ) participate in wax ester production. These results suggest that fatty acids are synthesized in mitochondria by the reversal of ß-oxidation, where trans-2-enoyl-CoA is reduced mainly by acyl-CoA dehydrogenase using the electrons provided by NADH via the electron transport chain complex I, RQ, and ETF, and that ATP production is highly supported by anaerobic respiration utilizing trans-2-enoyl-CoA as a terminal electron acceptor.

PFG-NMR study for evaluating freezing damage to onion tissue.

We assessed the damage to onion tissue due to freeze-thawing as the water permeability determined by using PFG-NMR and light microscopy. The water diffusion in fresh onion tissue was restricted due to cellular barriers, and the estimated water permeability was 6.99 x 10(-6)m/s. The water diffusion became considerably less restricted after freeze-thawing; the convergent value for the restricted diffusion coefficient increased and the water permeability significantly increased to 2.85 x 10(-5) m/s. While NMR could detect a distinct change in the diffusion behavior of water molecules in freeze-thawed tissue, light microscopy revealed no significant tissue damage. These results suggest that freeze-thawing damaged the vegetable tissues primarily through destruction of the cell membrane rather than the cell wall.

Oral polio vaccines have not yet covered West Africa: survey of immunization coverage conducted in Niger.

Following the re-emergence of polio in West Africa, an investigation was conducted on the occasion when transients gather for the festival in Niger, where the oral polio vaccine (OPV) and BCG coverage among children under the age of 5 years can be evaluated. A total of 259 children were investigated, including 186 from settled families and 73 from unsettled families. OPV coverage was found to be as low as 32.4%, and 61.8% of all participants in the study had not received both OPV and BCG. There were more children who had not received the OPV in unsettled families than in settled families. As there are still unvaccinated children in Niger, polio continues to occur among them. Moreover, outbreaks can transfer to more densely-populated areas, causing much larger outbreaks. To stop the chain of transmission, it is essential to reconsider the strategy of mass vaccination in order to cover all children thoroughly, including transients.