A lung type prostaglandin F synthase is expressed in bovine liver: cDNA sequence and expression in E. coli.

Using the cDNA of bovine lung prostaglandin F synthase (EC 1.1.1.2) as a probe, we isolated a clone from a bovine liver cDNA library which differed in only eleven nucleotides from the probe. The corresponding protein contained three amino acid substitutions, including a leucine residue which is conserved throughout all aldo-keto reductases. We inserted the liver cDNA into expression vector pUC19 and expressed the recombinant liver enzyme in E.coli. The purified liver enzyme reduced prostaglandin H2 as well as prostaglandin D2 and various carbonyl compounds. The high relative activity against prostaglandin H2 in combination with a high Km value for prostaglandin D2 identified this liver enzyme as a lung type prostaglandin F synthase. However, the binding constant for NADPH of the liver enzyme was 3.5 fold higher than that of lung prostaglandin F synthase.     

The influence of peripheral nerve graft's predegeneration stage on the regrowth of hippocampal injured neurites and concomitant changes in submicrosomal fraction proteins of grafts.

The present work has a twofold aim: 1. To ascertain whether the stimulative influence of peripheral nerve grafts on injured hippocampal neurons depends on the time lapse after transection and; 2. To examine whether the mentioned effect runs parallel to the time-dependent changes of proteins contents and composition in the submicrosomal fraction from transected rat sciatic nerves. Fluorescence microscope examination revealed that FITC-HRP labeled cells extending their neurites into the implanted peripheral nerve segments were particularly numerous among the hippocampal neurons when 7- and 35-day-old predegenegated distal stumps were used as grafts. Discontinuous SDS-slab polyacrylamide gel electrophoresis of submicrosomal fraction proteins obtained from distal stumps of rat sciatic nerves was performed at the 7, 14, 21 and 35 days after transection. Among the obtained protein fractions the most interesting seem to be the ones of 47 and 54 kDa, which reached maximal levels at the 7th day and the 50 kDa fraction with a maximum at the 35th experimental day. It is possible that the growth promoting power of the employed grafts depends on the presence of proper proteins.     

Photosynthetic oxygen evolution and CO2 photoassimilation by cyanobacteria that form water-bloom spots in a sulfur spring with a high sulfide content

Cyanobacteria belonging mainly to the genera Anabaena and Oscillatoria were isolated from water-bloom spots of a sulfur spring in Staraya Matsesta. Their suspensions evolved O2 at a rate of 6--8 nM/min per 1 mg of dry cell weight at an intensity of solar radiation being 60--75 mV/cm2 per 1 sec. The cells were also capable of CO2 photoassimilation in the presence of solfide at a rate of 10(-4) mg C per mg per hour. DCMU at a concentration of 10(-5) M completely inhibited O2 evolution and inhibited CO2 fixation by 80%. Oxygen assimilation in dark by the suspensions did not depend on the addition of cyanide and was caused apparently by nonenzymatic reduction of O2 with sulfide dissolved in the spring water. Oxygen assimilation by the suspensions in light in the presence of DCMU was by 20--30% greater than in dark. Therefore, the cells of cyanobacteria are characterized by photorespiration at the level of photosystem I. Presumably, sulfide at a concentration of 9 mM cannot significantly inhibit the photosynthetic processes in cyanobacteria producing water-bloom spots in the sulfur spring.