Induction of sexual reproduction in Opalina sudafricana by injecting its host Bufo regularis with gibberellic acid

Induction of sexual reproduction in Opalina sudafricana by injecting its host Bufo regularis with gibberellic acid. International Journal for Parasitology4, 203–206. Opalina sudafricana parasitic in the rectum of Bufo regularis was induced to reproduce sexually when its host was injected subcutaneously with 0·3 mg of gibberellin-A₃. This plant growth substance had no effect on the induction of encystation in the parasites in vitro. Urine of toads injected with gibberellin-A₃ induced sexual reproduction (encystation) in the opalinids in vitro. It is speculated that the plant hormone must either be broken down into an active substance by the toad or cause the toad to excrete its own gonadal hormones (or other hormones) into the urine. This active substance or the excreted hormones may induce division in the parasites resulting in the formation of small forms which encyst.     

Puromycin aminonucleoside increases the synthesis of ribonuclease inhibitor and other proteins in kidney

The incorporation of labelled amino acids into proteins was measured in vivo in the kidney of control rats and rats that received puromycin aminonucleoside. There was an increase in the synthesis of kidney proteins after the aminonucleoside that was similar to the increased synthesis previously observed in cell-free and slice preparations. The increased synthesis in vivo and in vitro especially involved proteins of the prealbumin fraction of average molecular weights of approx. 50 000, 35 000, 25000, 18 000, and 10 000. The largest of these proteins was identified as kidney ribonuclease inhibitor and additional evidence was obtained for the increased synthesis of the kidney inhibitor after aminonucleoside.     

Some novel flavin-nicotinamide biscoenzymes and their reactivities

The present study was undertaken to investigate flavin-nicotinamide reactions and interactions. A series of novel flavin-nicotinamide biscoenzymes have been synthesized by a general three-step procedure. The structures of these compounds were confirmed by nuclear magnetic resonance spectra, absorption spectra and elemental analysis. These compounds consist of short linear hydrocarbon chains interconnecting the N-1 of nicotinamide and the N-10 of the 7,8-dimethyl-isoalloxazine ring. The compounds were reduced with sodium dithionite (Na₂S₂O₄) and the flavin portion was reoxidized with ferricyanide. Re-reduction of the flavin portion by the nicotinamide portion of the molecule was followed anaerobically at 442 nm. When the interconnecting hydrocarbon chain was unsaturated, a second order reaction was observed with a rate equal to that of lumiflavin and 1-propyl-1,4-dihydronicotinamide (NprNicH₂) under the same conditions. When the two halves of the biscoenzymes were connected by saturated three- and four-carbon chains, the expected unimolecular reaction was not observed. Instead, the reduced biscoenzyme, after separation from excess sodium dithionite, was shown to have a strong absorption at 298 nm. This absorption is characteristic of hydration of dihydronicotinamides at the 5,6-double bond.In further studies, the C₃-biscoenzyme exhibited an absorption at 600 nm due to a complex between the reduced flavin and oxidized nicotinamide portions of the molecule. Absorbance at 600 nm increased linearly with the C₃-biscoenzyme concentration, clearly indicating that this is an intramolecular complex. When the C₃-biscoenzyme was at 0°C in 60–75% dimethylformamide buffer solution, no absorption at 600 nm was observed. When excess dithionite was removed, the spectrum under these conditions showed definite peaks at 297 and 357 nm. These respective peaks were attributed to hydrated dihydronicotinamide and dihydronicotinamide species present in the reaction mixture.The reduced flavin was postulated to be a catalyst for the hydration of dihydronicotinamide. This hypothesis was tested by incubating 1-propyl-1,4-dihydronicotinamide alone and with several concentrations of reduced riboflavin under basic anaerobic conditions. The results show that the reduced flavin increases the rate of disappearance of the dihydronicotinamide species and that the product shows an absorption near 298 nm. These results indicate that a reduced form of the flavin nucleus catalyzes the hydration of dihydronicotinamides.     

On the carbohydrate—protein linkage group in vitreous humor hyaluronate

Fractional molar ratios of serine, threonine and aspartic acid to neutral sugars in the purified bovine vitreous humor hyaluronate, and a 4–5-fold increase in the percentage of these amino acids and the absence of sugar alditols in hyaluronate reduced with NaBH₄---PdCl₂ after alkali treatment indicated the absence of a carbohydrate—protein linkage. Gel filtration behavior, a decrease in intrinsic viscosity of reduced hyaluronate to about one-half and a significant decrease in its specific rotation suggested that the two antiparallel chains of the hyaluronate double helix may come apart upo reduction. The vitreous humor hyaluronate contained 109.2 ppm of “bound” silicon. It is suggested that the bound silicon may bridge the two antiparallel chains through the neutral sugars and/or through the hydroxyl group of the uronic acid moiety.     

Isolation and characterization of a 3-chlorobenzoate degrading pseudomonad

A pseudomonad has been isolated from sewage, which can utilize 3-chlorobenzoic acid as a sole carbon source. In cells grown on benzoate the enzymes of the -ketoadipic acid pathway are present. Considerable enzymic activities for chlorinated substrates were found in benzoate grown cells only for the oxygenation of 3-chlorobenzoate and the dehydrogenation of 3- and 5-chloro-3,5-cyclohexadiene-1,2-diol-1-carboxylic acid. 3-Chlorobenzoate grown cells show additional high activities for the turnover of 3- and 4-chlorocatechols and chloromuconic acids.Abbreviations Used DHB (-)-3,5-cyclohexadiene-1,2-diol-1-carboxylic acid (derived from the trivial name, dihydrodihydroxybenzoate) - 3- and 5-Cl-DHB correspondingly 3- and 5-chloro-3,5-cyclohexadiene-1,2-diol-1-carboxylic acid     

Isolation of ouabain-resistant human diploid fibroblasts

Seventeen clones resistant to the cytotoxic action of ouabain were isolated in culture by direct selection from 5 independent strains of diploid human fibroblasts. Resistant clones were recovered at frequencies on the order of 10^?7 per wild type cell selected from populations treated with the mutagen EMS, but no resistant cells were detected among 10⁸ unmutagenized cells. Most selected clones remained ouabain-resistant following further propagation in the absence of drug. The growth of wild type cells was inhibited by 50% at ouabain concentrations of 2–5 × 10^?8 M, while resistant clones required 15–180 fold higher drug concentrations to cause equivalent inhibition. Ouabain-resistant clones showed increased resistance of K+ transport function to ouabain inhibition that paralleled their increased resistance to growth inhibition. Initial experiments suggest that under selective conditions the resistant diploid fibroblasts differ significantly from wild type in binding of ³H-ouabain per unit surface area. The ouabain-resistant cells were similar to wild type in transport properties unrelated to ouabain inhibition. Resistant cells had normal karyotypes and senesced with a lifespan similar to control clones. The ouabain-resistant phenotypes of these diploid human fibroblast isolates apparently reflect point mutations that specifically affect the Na+/K+ transport ATPase with respect to ouabain-binding and/or response to bound ouabain.