Inhibitory effects of Serjania erecta on the development of Chrysodeixis includens.

The soybean looper, Chrysodeixis includens, is a primary soybean pest that reduces crop productivity. This work examined control of C. includens populations with methanolic extract of Serjania erecta, a native Cerrado plant, while minimizing risks to pollinators, natural enemies and the environment. Serjania erecta specimens were collected, identified, and subjected to methanol extraction. Bioassays were performed using newly hatched and second-instar caterpillars and different extract concentrations on the diet surface to obtain IC50 values. Two replicates, containing 10 caterpillars, were established in triplicate. The IC50 values were 4.15 and 6.24 mg of extract mL-1 for first-instar and second-instar caterpillars, respectively. These growth inhibition results informed the extract concentrations assessed in subsequent development inhibition assays, in which the pupal weight was higher under the control than under the treatments. Extract treatments increased the duration of the larval, pupal and total development. The potential of different concentrations of S. erecta extract to inhibit the enzymes carboxylesterases was also evaluated. Carboxylesterases activity decreased by 41.96 and 43.43% at 7.8 and 15.6 µg mL-1 extract, respectively. At 31.3 µg mL-1 extract, enzymatic activity was not detected. Overall, S. erecta leaf methanolic extract showed inhibitory potential against carboxylesterases.

Isolation and characterization of an arginine ester hydrolase from Bothrops jararacussu venom which induces contractions of the isolated rat uterus.

The isolation and partial characterization of a serine protease with arginine ester hydrolase activity from Bothrops jararacussu snake venom are described. The purification procedure consisted of a gel filtration of the crude venom on Sephadex G-75 followed by an ion-exchange chromatography of the active fraction on DEAE-cellulose and a rechromatography on Bio-Rex 70 resin. The esterase fraction (DI-III), M(r) = 25,000 by SDS-PAGE, showed proteolytic activity on fibrinogen and casein. After 2 hr incubation, the A alpha and B beta chains of fibrinogen were intensely hydrolysed, while the gamma chain kept apparently intact, even after 20 hr of incubation. In spite of that, DI-III did not clot fibrinogen. DI-III induced edema in the rat paw. Although unable to release bradykinin, it induced contractions of the isolated rat uterus. DI-III did not catalyse the hydrolysis of bradykinin. Its arginine ester hydrolase activity was completely inhibited by diisopropyl fluorophosphate after 1 hr incubation, but not by phenylmethylsulfonyl fluoride under the same conditions.

Regulation of protein synthesis by minigene expression.

Peptidyl-tRNA hydrolase (Pth), an enzyme essential for Escherichia coli viability, scavenges peptidyl-tRNA released during abortive polypeptide chain elongation. Bacterial strains of E coli partially defective in Pth activity are unable to maintain bacteriophage lambda growth. Phage mutations that overcome the bacterial defect have been located to several regions in the lambda genome named bar. Plasmid constructs expressing just the bar region are toxic and cause a general arrest of protein synthesis in Pth-defective cells. Inspection of the nucleotide sequence from two bar regions reveals the short coding sequence AUG AUA Stop, spaced by an AT-rich segment from a Shine Dalgarno-like sequence (S-D). These sequences have been named minigenes. Base changes altering the putative S-D, the two sense codons, or the stop codon have been found to reduce Bar-toxicity. Transcripts containing bar function as mRNA. Upon expression in pth mutants, wild-type (bar+) transcripts are found associated with ribosomes. In addition, bar+ RNA forms ternary complexes with the 30S ribosomal subunit and the initiator tRNA and can be released upon run-off translation in the same way as an authentic mRNA. A cell free system for protein synthesis reproduces the in vivo effects: bar+ expression inhibits protein synthesis, bar+ RNA sequences are associated with ribosomes in the inhibited extracts, addition of purified Pth restores synthesis, and excess of tRNA(Lys), specific for the last sense codon in a mutant toxic minigene, prevents protein synthesis inhibition. Also, bar expression promotes association of methionine with ribosomes possibly in a translation complex. These results are consistent with a model proposing tRNA starvation to explain the behaviour of a pth mutant, thermosensitive for protein synthesis.