DNA origami with complex curvatures in three-dimensional space

We present a strategy to design and construct self-assembling DNA nanostructures that define intricate curved surfaces in three-dimensional (3D) space using the DNA origami folding technique. Double-helical DNA is bent to follow the rounded contours of the target object, and potential strand crossovers are subsequently identified. Concentric rings of DNA are used to generate in-plane curvature, constrained to 2D by rationally designed geometries and crossover networks. Out-of-plane curvature is introduced by adjusting the particular position and pattern of crossovers between adjacent DNA double helices, whose conformation often deviates from the natural, B-form twist density. A series of DNA nanostructures with high curvature--such as 2D arrangements of concentric rings and 3D spherical shells, ellipsoidal shells, and a nanoflask--were assembled.     

Geographic barriers isolate endemic populations of hyperthermophilic archaea

Barriers to dispersal between populations allow them to diverge through local adaptation or random genetic drift. High-resolution multilocus sequence analysis revealed that, on a global scale, populations of hyperthermophilic microorganisms are isolated from one another by geographic barriers and have diverged over the course of their recent evolutionary history. The identification of a biogeographic pattern in the archaeon Sulfolobus challenges the current model of microbial biodiversity in which unrestricted dispersal constrains the development of global species richness.     

Temperature-dependent autoxidation of conjugated trienols from apple peel yields 6-methyl-5-hepten-2-one, a volatile implicated in induction of scald

Conjugated triene (CT) oxidation products of alpha-farnesene have long been thought to be involved in development of superficial scald in apple fruit. Early studies found that CT hydroperoxides and the volatile 6-methyl-5-hepten-2-one (MHO) are major in vitro autoxidation products of alpha-farnesene. However, it was recently shown that > or =99% of the oxidation products of alpha-farnesene that accumulate in apple peel are conjugated trienols (CTols), isomers of 2,6,10-trimethyldodeca-2,7,9,11-tetraene-6-ol. HPLC-purified CTols from fruit of two scald-susceptible cultivars, Granny Smith (GS) and Red Delicious (RD), were used to study autoxidation of these compounds in vitro. Incubation of CTols in sealed glass vials under air resulted in accumulation of MHO. Oxygen enrichment did not increase the amount of MHO produced. Regardless of which cultivar CTols were derived from, at 0 degrees C autoxidation yielding MHO was quite slow and linear, whereas at 20 degrees C MHO production was much more rapid, and after several hours the rate increased abruptly. However, CTols isolated from GS and RD fruit differed in the duration of the initial lag phase and the overall level of MHO generated at 20 degrees C. The sharp increase in MHO production occurred after 3 h with GS CTols and at about 12 h with RD CTols. Also, the yield of MHO from GS CTols after 6 h at 20 degrees C was nearly 6-fold greater than that from RD CTols after 20 h at 20 degrees C. The antioxidants butylated hydroxytoluene and diphenyamine reduced the yield of MHO by about 97%. Recent work has shown that MHO can induce scald-like symptoms in apple peel and that tissue sensitivity increases with time in storage. This may explain the correlation between high CTol levels and scald development, and why symptoms rapidly intensify when fruits are removed from cold storage.     

Effects of methanol concentration and solvent:peanut ratio on extraction of aflatoxin from raw peanuts

Aflatoxin B1 was extracted by a water slurry process using methanol concentrations of 55, 60, 65, and 70% in water and solvent:peanut ratios of 3, 4, 5, and 6 mL/g. Results failed to show that methanol concentration had an effect on amount of B1 extracted; however, the amount of B1 extracted increased with an increase in solvent:peanut ratio. Aflatoxin B1 was also extracted by the official AOAC method II, using methanol concentrations of 55, 60, 65, and 70% in water and solvent:peanut ratios of 2, 3, 4, and 5 mL/g. Results showed that the amount of B1 extracted increased with percent methanol at low solvent:peanut ratios but not at high ratios. Also, the amount of B1 extracted increased with solvent:peanut ratios at all methanol concentrations.     

Simulation of aflatoxin testing plans for shelled peanuts in the United States and in the export market

The 1987 United States aflatoxin testing plan for shelled peanuts was designed with a final accept level of 25 parts per billion (ppb) total aflatoxin. Some of the importers of U.S. peanuts use aflatoxin testing plans with accept levels lower than 25 ppb. For example, the accept level of a testing plan used in The Netherlands is 5 ppb B1 or 10 ppb total aflatoxin. Whenever export lots are re-tested for aflatoxin by an importing country, some lots accepted in the United States will be rejected by the importing country's aflatoxin testing plan. Computer models were developed to determine the effects of decreasing the final accept level of the U.S. testing plan on the number of lots accepted and rejected in the United States and the number of exported lots accepted and rejected by The Netherlands testing plan. Decreasing the final accept level of the U.S. testing plan from 25 to 5 ppb increased the number of lots rejected in the United States by 371% while reducing the number of exported lots rejected by 51%. For every additional 8.3 lots rejected in the United States, one less export lot will be rejected.     

Two new antioxidant malonated caffeoylquinic acid isomers in fruits of wild eggplant relatives

Fruits of the cultivated eggplant species Solanum melongena and its wild relative Solanum incanum have a high content of hydroxycinnamic acid conjugates, which are implicated in the human health benefits of various fruits and vegetables. Monocaffeoylquinic acid esters, in particular 5-O-(E)-caffeoylquinic acid, are usually predominant in solanaceous fruits and tubers. Two closely related caffeoylquinic acid derivatives with longer C(18) HPLC retention times than those of monocaffeoylquinic acids are minor constituents in cultivated eggplant fruit. In a prior study, the two compounds were tentatively identified as 3-O-acetyl- and 4-O-acetyl-5-O-(E)-caffeoylquinic acids and composed ≤2% of the total hydroxycinnamic acid conjugates in fruit of most S. melongena accessions. It was recently found that the pair of these caffeoylquinic acid derivatives can compose 15-25% of the total hydroxycinnamic acid conjugates in fruits of S. incanum and wild S. melongena. This facilitated C(18) HPLC isolation and structural elucidation using (1)H and (13)C NMR techniques and HR-ToF-MS. The isomeric compounds were identified as 3-O-malonyl-5-O-(E)-caffeoylquinic acid (isomer 1) and 4-O-(E)-caffeoyl-5-O-malonylquinic acid (isomer 2). Both exhibited free radical scavenging activity, albeit about 4-fold lower than that of the flavonol quercetin dihydrate. By contrast, the iron chelation activities of isomers 1 and 2, respectively, were about 3- and 6-fold greater than that of quercetin dihydrate. Reports of malonylhydroxycinnamoylquinic acids are rare, and only a few of these compounds have been structurally elucidated using both NMR and MS techniques. To the authors' knowledge, these two malonylcaffeoylquinic acid isomers have not previously been reported.