Development of an Oligidic Diet forin VitroRearing ofTrichogramma galloiZucchi andTrichogramma pretiosumRiley

The goal of this research was to develop an artificial diet for rearingTrichogramma galloiandTrichogramma pretiosum,which are important egg parasitoids of several pests in Brazil. The diet was based on different proportions of hemolymph ofHelicoverpa zealarvae, chicken egg yolk, powdered milk solutions, bovine fetal serum, and an extract fromHeliothis virescenseggs. The ideal volume of diet for rearingT. galloiwas also evaluated. The parasitoids showed different nutritional requirements for their development. The best proportion of diet components forT. galloiwas 70% hemolymph, 20% egg yolk, 10% fetal serum, and 0.2% streptomycin, and forT. pretiosumit was 70% hemolymph, 20% egg yolk, 5% fetal serum, 5% egg extract, and 0.2% streptomycin. Egg yolk was found to be essential for the development of both parasitoids, and the powdered milk solution was not an important component in the diet. Development was not obtained in diets with hemolymph concentrations lower than 40%. A 3- to 4-μl amount of diet was the ideal volume on which to rearT. galloiwith the techniques reported.     

An optimized Boc synthesis of indolicidin

Summary Indolicidin, an antimicrobial peptide from bovine neutrophils containing five tryptophan out of a total of 13 residues, has the highest molar proportion of tryptophan of any known peptide sequence and is thus considered a difficult synthetic target. Conventional Boc chemistry can be applied to the synthesis of indolicidin with an appropriate choice of scavenger mixtures, reaction times and temperatures at the crucial acidolytic cleavage and deprotection step. In particular, treatment with HF/p-cresol/p-thiocresol (90:7:3) for 40 min at −8°C results in a crude product containing ca. 90% indolicidin, from which the target compound can be isolated in satisfactory yields and purity after reverse-phase purification. The main byproducts arising during the synthesis and cleavage steps have been identified by HPLC with on-line electrospray mass spectrometric detection.     

棉铃虫一种新的实用人工饲料(英文)

本文报道饲养棉铃虫的一种新的含有番茄酱的实用人工饲料。取食这种饲料时,绝大部分幼虫只有5个龄期。因此,幼虫历期大为缩短。在该饲料中加入适量绿豆粉取代麦胚能在一定程度上提高成虫的繁殖力。在连续12代饲养期间,幼虫期存活率在87．5％－98．7％之间,蛹期死亡率不到5％;第11代成虫平均产卵千粒左右,孵化率达86．5％。这些结果表明,经过2年连续饲养后,棉铃虫的活力并没有明显的降低。     

A NEW AND PRACTICAL ARTIFICIAL DIET FOR THE COTTON BOLL-WORM*

Abstract A new and practical artificial diet for the cotton bollworm, Helicoverpa armigera (Hübner), containing canned tomato paste has been developed. The developmental duration of larvae fed on the diet was greatly shortened because most of them only went through 5 stadia before pupation. To add certain amount of mung bean flour to partly substitute for wheat germ in the diet was beneficial to the adult reproduction. During rearing period of 12 successive generations, 87. 5%-98. 7% of the larvae survived and only less than 5 % of the pupae failed to emerge. The number of egg laid by the adults of the 11th generation averaged about 1 000/female, with a hatching rate of 86. 5%. These results suggested that the vitality of the cotton bollworm did not decline obviously after reared with the diet for two years.     

Synthesis and spectroscopic characterization of protected N-phosphonomethylglycine dipeptides.

A series of terminally blocked dipeptides containing C-terminal N-phosphonomethylglycine (glyphosate, an extremely effective non-selective post-emergence herbicide) have been synthesized by a solution method. The presence of their two conformers, cis (syn) and trans (anti), was shown in solutions by NMR spectroscopy. Molecular structures of the peptides were also determined in the solid state by X-ray diffraction. The attempts for the selective and total removal of the groups protecting amino, carboxylic and phosphonate functions were in many cases unsuccessful due to the formation of cyclic structures and breakage of the phosphorus-to-carbon bond.     

Conformational analysis of the Galpha(s) protein C-terminal region.

The C-terminal domain of the heterotrimeric G protein a-subunits plays a key role in selective activation of G proteins by their cognate receptors. Several C-terminal fragments of Galpha(s) (from 11 to 21 residues) were recently synthesized. The ability of these peptides to stimulate agonist binding was found to be related to their size. Galpha(s)(380-394) is a 15-mer peptide of intermediate length among those synthesized and tested that displays a biological activity surprisingly weak compared with that of the corresponding 21-mer peptide, shown to be the most active. In the present investigation, Galpha(s)(380-394) was subjected to a conformational NMR analysis in a fluorinated isotropic environment. An NMR structure, calculated on the basis of the data derived from conventional 1D and 2D homonuclear experiments, shows that the C-terminal residues of Galpha(s)(380-394) are involved in a helical arrangement whose length is comparable to that of the most active 21 -mer peptide. A comparative structural refinement of the NMR structures of Galpha(s)(380-394) and Galpha(s)(374-394)C379A was performed using molecular dynamics calculations. The results give structural elements to interpret the role played by both the backbone conformation and the side chain arrangement in determining the activity of the G protein C-terminal fragments. The orientation of the side chains allows the peptides to assume contacts crucial for the G protein/receptor interaction. In the 15-mer peptide the lack as well as the disorder of some N-terminal residues could explain the low biological activity observed.     

Amorphastilbol,an antimicrobial agent from Amorpha nana

Bioassay-directed fractionation of Amorpha nana demonstrated that the anti Gram positive/antitubercular constituent is amorphastilbol. The chemical structure of amorphastilbol was confirmed by chemical transformation to known (Radula variabilis) dihydroamorphastilbol and its methyl ether. Inactive 3,5-dihydroxy-7,4′-dimethoxyflavone was detected in this plant for the first time and identified by interconversion with kaempferol. All compounds were evaluated in vitro for antimicrobial spectrum and potency.     

Biochemical and biophysical properties of flacherie virus of the silkworm

Flacherie virus of the silkworm (FVS) was extracted from diseased silkworms, both larvae and pupae, and purified by 15 to 30% sucrose density gradient centrifugation. FVS III and FVS IV, in addition to the FVS I and FVS II described in the previous paper (Himeno et al., 1974), were found. The FVS I, FVS III, and FVS IV showed the same mobility in 2.4% polyacrylamide gel electrophoresis and could not be distinguished from each other in the gel. However, the purified FVS II was separated into two bands, FVS IIa and FVS IIb, in 2.4% gel. FVS III was a spherical particle with a diameter of 28 ± 1 nm and showed a sedimentation coefficient of about 90 S. FVS III was easily decomposed into FVS IV which sedimented at about 30 S in sucrose gradient centrifugation. FVS I and FVS II each contained a single molecule of RNA which showed the same molecular weight. FVS I consisted of three polypeptides with molecular weights of 67,000, 50,000, and 33,000. FVS II consisted of 10 polypeptides; among them 2 polypeptides with molecular weights of 50,000 and 33,000 were also found. Labeling experiments with [³²P]orthophosphate revealed that FVS II was found at an early stage of infection and FVS I at a late stage. FVS II was also isolated at an early stage from silkworms infected with FVS II, and FVS I was found at a late stage in these silkworms. The correlation among FVS I, FVS II, FVS III, and FVS IV was discussed and it was suggested that they might be closely related to one another and that few particles in them were immature. It is possible that FVS II changes to FVS I via FVS III by cleavage of large polypeptides.     

Activity and stability of human kallikrein-2-specific linear and cyclic peptide inhibitors.

Human glandular kallikrein (KLK2) is a highly prostate-specific serine protease, which is mainly excreted into the seminal fluid, but part of which is also secreted into circulation from prostatic tumors. Since the expression level of KLK2 is elevated in aggressive tumors and it has been suggested to mediate the metastasis of prostate cancer, inhibition of the proteolytic activity of KLK2 is of potential therapeutic value. We have previously identified several KLK2-specific linear peptides by phage display technology. Two of its synthetic analogs, A R R P A P A P G (KLK2a) and G A A R F K V W W A A G (KLK2b), show specific inhibition of KLK2 but their sensitivity to proteolysis in vivo may restrict their potential use as therapeutic agents. In order to improve the stability of the linear peptides for in vivo use, we have prepared cyclic analogs and compared their biological activity and their structural stability. A series of cyclic variants with cysteine bridges were synthesized. Cyclization inactivated one peptide (KLK2a) and its derivatives, while the other peptide (KLK2b) and its derivatives remained active. Furthermore, backbone cyclization of KLK2b improved significantly the resistance against proteolysis by trypsin and human plasma. Nuclear magnetic resonance studies showed that cyclization of the KLK2b peptides does not make the structures more rigid. In conclusion, we have shown that backbone cyclization of KLK2 inhibitory peptides can be used to increase stability without losing biological activity. This should render the peptides more useful for in vivo applications, such as tumor imaging and prostate cancer targeting.