Antimalarial compounds from Parinari capensis

The antimalarial activity of the raw petroleum ether and dichloromethane extracts of the stems of Parinari capensis (Chrysobalanceae) was determined. Phytochemical investigation of these extracts led to the isolation of three diterpene lactones that possess antimalarial activity with IC(50) values of 0.54, 0.67, and 1.57 microg/mL. Although their antimalarial activity is promising, the toxicity profiles of these diterpene lactones prevent further biological evaluation. They could however be used effectively as lead compounds in the synthesis of novel antimalarial agents.     

A recently acquired host plant provides an oligophagous insect herbivore with enemy-free space

Enemy-free space (EFS) is a potentially important factor affecting host plant use by phytophagous insects. Yet only a few field studies have demonstrated that natural enemy activity is the sole mechanism underlying use of novel host plants by herbivorous insects. This may be due to the fact that in earlier studies, both herbivores and natural enemies had the opportunity to adapt to the new host plant. Here we studied the possibility that EFS underlies the recently recorded increase in Phthorimaea operculella densities on tomato plants in a few areas within its geographical range. Through field experiments in Ethiopia, we show that all three conditions proposed by Berdegue et al. to demonstrate EFS are fulfilled. First, a significantly higher proportion of larvae survive on caged than on exposed potato plants, showing that natural enemies are an important mortality factor on the original host, potato. Second, larval survival was significantly higher on exposed tomato than potato plants, implying greater protection for the herbivore from its natural enemies on tomato than on potato plants. Thus tomato plants provide P. operculella with an EFS. Finally, larval survival was significantly higher on caged potato than on caged tomato plants at the preblossom stage, indicating that, in the absence of natural enemies, there is a fitness cost when larvae feed on the sub-optimal tomato plants. Fulfillment of this third condition points to the importance of natural enemy activity relative to that of other unidentified factors, such as food quality and competition. An intensive field survey provides further support for this conclusion.     

Structural divergence among cannabinoids influences membrane dynamics: A H Solid-State NMR analysis

Cannabinoids are compounds that can modulate neuronal functions and immune responses via their activity at the CB₁ receptor. We used ²H NMR order parameters and relaxation rate determination to delineate the behavior of magnetically aligned phospholipid bilayers in the presence of several structurally distinct cannabinoid ligands. THC (Δ⁹-Tetrahydrocannabinol) and WIN-55,212-2 were found to lower the phase transition temperature of the DMPC and to destabilize their acyl chains leading to a lower average S_CD (≈ 0.13), while methanandamide and CP-55,940 exhibited unusual properties within the lipid bilayer resulting in a greater average S_CD (≈ 0.14) at the top of the phospholipid upper chain. The CB₁ antagonist AM281 had average S_CD values that were higher than the pure DMPC lipids, indicating a stabilization of the lipid bilayer. R_1Z versus |S_CD|² plots indicated that the membrane fluidity is increased in the presence of THC and WIN-55,212-2. The interaction of CP-55,940 with a variety of zwitterionic and charged membranes was also assessed. The unusual effect of CP-55,940 was present only in bicelles composed of DMPC. These studies strongly suggest that cannabinoid action on the membrane depends upon membrane composition as well as the structure of the cannabinoid ligands.     

Co-evolving residues in membrane proteins

MOTIVATION: The analysis of co-evolving residues has been exhaustively evaluated for the prediction of intramolecular amino acid contacts in soluble proteins. Although a variety of different methods for the detection of these co-evolving residues have been developed, the fraction of correctly predicted contacts remained insufficient for their reliable application in the construction of structural models. Membrane proteins, which constitute between one-fourth and one-third of all proteins in an organism, were only considered in few individual case studies. RESULTS: We present the first general study of correlated mutations in alpha-helical membrane proteins. Using seven different prediction algorithms, we extracted co-evolving residues for 14 membrane proteins having a solved 3D structure. On average, distances between correlated pairs of residues lying on different transmembrane segments were found to be significantly smaller compared to a random prediction. Covariation of residues was frequently found in direct sequence neighborhood to helix-helix contacts. Based on the results obtained from individual prediction methods, we constructed a consensus prediction for every protein in the dataset that combines obtained correlations from different prediction algorithms and simultaneously removes likely false positives. Using this consensus prediction, 53% of all predicted residue pairs were found within one helix turn of an observed helix-helix contact. Based on the combination of co-evolving residues detected with the four best prediction algorithms, interacting helices could be predicted with a specificity of 83% and sensitivity of 42%. AVAILABILITY: http://webclu.bio.wzw.tum.de/helixcorr/     

Inhibition of monoamine oxidase B by selected benzimidazole and caffeine analogues

We have recently reported that a series of (E)-8-styrylcaffeines and (E)-2-styrylbenzimidazoles are moderate to very potent competitive inhibitors of monoamine oxidase B (MAO-B). The most potent member of the series was found to be (E)-8-(3-chlorostyryl)caffeine (CSC) with an enzyme-inhibitor dissociation constant (K(i) value) of 128 nM. In the present study, we have prepared additional caffeine and benzimidazole analogues in an attempt to identify compounds with improved MAO-B inhibition potency while still acting reversibly. The most potent inhibitor among the caffeine analogues was (E)-8-(3,4-dichlorostyryl)caffeine with a K(i) value of 36 nM, approximately 3.5 times more potent than CSC. The most potent inhibitor among the benzimidazole analogues was (E)-2-(4-trifluoromethylstyryl)-1-methylbenzimidazole with a K(i) value of 430 nM. An SAR analysis indicated that the potency of MAO-B inhibition by (E)-2-styryl-1-methylbenzimidazole analogues depended upon the Taft steric parameter (E(s)) of the substituents attached to C-4 of the styryl phenyl ring. Substituents with a large degree of steric hindrance appear to enhance inhibition potency. The proposal that potent MAO-B inhibition by (E)-8-styrylcaffeines and (E)-2-styrylbenzimidazoles can be explained by a mode of binding that involves traversing both the entrance and substrate cavities was supported by the finding that 1-methylbenzimidazole only weakly inhibited MAO-B with a K(i) value of 2084 microM. Without the styryl side chain, 1-methylbenzimidazole is not expected to be able to bind simultaneously to both the entrance and substrate cavities.     

Single-parent segregant pools for allocation of markers to a specified chromosomal region in outcrossing species

Bulked co-segregant analysis is a method of rapidly allocating unmapped genetic markers to a specific chromosomal region. Although originally developed for utilization in populations derived from crosses between fully inbred lines, it has been proposed that co-segregant pools could also serve the same purpose in outbreeding populations, if individuals from only a single large family are pooled. Large, fully mapped, single-sire backcross and half-sib families are presently available as part of the international chicken and bovine reference family panels respectively. In this study, power and tests of significance for single-parent co-segregant analysis are derived for full-sib, single-parent back-cross and single-parent half-sib families, as a function of proportion of recombination between index marker and linked marker, proportion of single-parent alleles among the mates, number of individuals in each segregant pool and technical error variance. Power was found to be greater than 0–80 for many reasonable parameter combinations. The method is illustrated using microsatellite markers and a large single-sire bovine family, part of the international bovine reference family panel.     

Quantitative trait locus mapping in dairy cattle by means of selective milk DNA pooling using dinucleotide microsatellite markers: analysis of milk protein percentage.

"Selective DNA pooling" accomplishes quantitative trait locus (QTL) mapping through densitometric estimates of marker allele frequencies in pooled DNA samples of phenotypically extreme individuals. With poly(TG) microsatellites, such estimates are confounded by "shadow" ("stutter") bands. A correction procedure was developed on the basis of an observed linear regression between shadow band intensity and allele TG repeat number. Using this procedure, a selective DNA pooling study with respect to milk protein percentage was implemented in Israel-Holstein dairy cattle. Pools were prepared from milk samples of high and low daughters of each of seven sires and genotyped with respect to 11 markers. Highly significant associations with milk protein percentage were found for 5 of the markers; 4 of these markers confirmed previous reports. Selective DNA pooling accessed 80.6 and 48.3%, respectively, of the information that would have been available through individual selective genotyping or total population genotyping. In effect, the statistical power of 45,600 individual genotypings was obtained from 328 pool genotypings. This methodology can make genome-wide mapping of QTL accessible to moderately sized breeding organizations.     

Developmental regulation of juvenile hormone biosynthesis by the ring gland ofDrosophila melanogaster

Summary The synthesis in vitro of the putative dipteran juvenile hormone (JHB₃) by ring glands isolated from third instarDrosophila melanogaster larvae was quantified by a radiochemical assay. The data indicate that JHB₃ synthesis is developmentally regulated during the period prior to wandering until after puparium formation. The highest level of basal production occurred during the postfeeding stage, and synthesis declined after pupariation. Similar relative profiles of synthesis were obtained upon the addition of the JHB₃ precursor, farnesoic acid, although the absolute levels of production were elevated considerably. Basal JHB₃ production by brain-ring gland complexes in vitro was also highest during the postfeeding stage, although the synthetic rates were much lower than displayed by isolated ring glands. Further analysis revealed that methyl farnesoate, a JHB₃ precursor, was synthesized by brain-ring gland complexes in significant quantity. A dual mechanism of brain-centered control of JHB₃ biosynthesis is proposed.To whom offprint requests should be sent