Distortion of genetically modified organism quantification in processed foods: influence of particle size compositions and heat-induced DNA degradation

Milling fractions from conventional and transgenic corn were prepared at laboratory scale and used to study the influence of sample composition and heat-induced DNA degradation on the relative quantification of genetically modified organisms (GMO) in food products. Particle size distributions of the obtained fractions (coarse grits, regular grits, meal, and flour) were characterized using a laser diffraction system. The application of two DNA isolation protocols revealed a strong correlation between the degree of comminution of the milling fractions and the DNA yield in the extracts. Mixtures of milling fractions from conventional and transgenic material (1%) were prepared and analyzed via real-time polymerase chain reaction. Accurate quantification of the adjusted GMO content was only possible in mixtures containing conventional and transgenic material in the form of analogous milling fractions, whereas mixtures of fractions exhibiting different particle size distributions delivered significantly over- and underestimated GMO contents depending on their compositions. The process of heat-induced nucleic acid degradation was followed by applying two established quantitative assays showing differences between the lengths of the recombinant and reference target sequences (A, deltal(A) = -25 bp; B, deltal(B) = +16 bp; values related to the amplicon length of the reference gene). Data obtained by the application of method A resulted in underestimated recoveries of GMO contents in the samples of heat-treated products, reflecting the favored degradation of the longer target sequence used for the detection of the transgene. In contrast, data yielded by the application of method B resulted in increasingly overestimated recoveries of GMO contents. The results show how commonly used food technological processes may lead to distortions in the results of quantitative GMO analyses.     

Enantioselective analysis of methyl-branched alcohols and acids in rhubarb (Rheum rhabarbarum L.) stalks

The enantiomeric compositions of 2-methylbutanol (1), 4-methylhexanol (2), 2-methylbutanoic acid (3), and 4-methylhexanoic acid (4) present in rhubarb (Rheum rhabarbarum L.) stalks were determined. Enantiodifferentiation was achieved via multidimensional gas chromatography using heptakis(2,3,6-tri-O-ethyl)-beta-cyclodextrin as a chiral stationary phase. For all compounds the enantiomeric ratios were in favor of the (R)-enantiomers. The alcohols (1 and 2) exhibited generally high excesses of the (R)-enantiomers, the ratios varying slightly from batch to batch. For the acid (3) a rather narrow range averaging 65% (R):35% (S) was observed. The procedure applied to isolate the volatiles (vacuum headspace technique, simultaneous distillation--extraction, liquid--liquid extraction) had no significant impact on the enantiomeric ratios. The study describes for the first time a plant used as food material in which 2-methyl-branched volatiles are not nearly exclusively present as (S)-enantiomers. This information enlarges the current regulatory knowledge regarding the classification of these important flavor compounds as "natural" on the basis of their enantiomeric ratios.     

Alga-like forms in onverwacht series, South Africa: oldest recognized lifelike forms on Earth

Spheroidal and cupshaped, carbonaceous alga-like bodies, as well as filamentous structures and amorphous carbonaceous matter occur in sedimentary rocks of the Onverwacht Series (Swaziland System) in South Africa. The Onverwacht sediments are older than 3.2 eons, and they are probably the oldest, littlealtered sedimentary rocks on Earth. The basal Onverwacht sediments lie approximately 10,000 meters stratigraphically below the Fig Tree sedimentary rocks, from which similar organic microstructures have been interpreted as alga-like microfossils. The Onverwacht spheroids and filaments are best preserved in black, carbonrich cherts and siliceous argillites interlayered with thick sequences of lavas. These lifelike forms and the associated carbonaceous substances are probably biological in origin. If so, the origins of unicellular life on Earth are buried in older rocks now obliterated by igneous and metamorphic events.     

Autosomal deletion mapping in man

Two families were observed in which morphologically similar dele tions involving a G-group chromosome were associated in the propositi with con spicuous abnormalities in ossification of the skull. The findings suggest that specific genetic information relating to morphogenesis of the skull may be located on a G-group chromosome.     

The V-antigen of Yersinia forms a distinct structure at the tip of injectisome needles

Many pathogenic bacteria use injectisomes to deliver effector proteins into host cells through type III secretion. Injectisomes consist of a basal body embedded in the bacterial membranes and a needle. In Yersinia, translocation of effectors requires the YopB and YopD proteins, which form a pore in the target cell membrane, and the LcrV protein, which assists the assembly of the pore. Here we report that LcrV forms a distinct structure at the tip of the needle, the tip complex. This unique localization of LcrV may explain its crucial role in the translocation process and its efficacy as the main protective antigen against plague.     

Methylene-C14-dioxyphenyl compounds: metabolism in relation to their synergistic action

The methylene-C(14) group is hydroxylated yielding formate-C(14) in the microsome-reduced nicotinamide-adenine dinucleotide phosphate system in vitro and yielding expired C(14)O(2), in living mice and houseflies. Methylenedioxyphenyl compounds apparently serve as alternate substrates for this enzymatic hydroxylation system of microsomes, and thus reduce the rate of metabolism and prolong the action of certain drugs and insecticide chemicals.