Automated determination of alpha-tocopherol in food and feed. Part 2. Continuous flow technique

The proposed determination of alpha-tocopherol is a continuous flow method with fluorometric detection. The only cleanup necessary is the usual saponification. A solution of the unsaponifiable matter in isooctane is automatically assayed. Isooctane is the carrier solvent and extractions are inserted between steps. These steps are selective reactions which render the method very specific. The natural homologs of alpha-tocopherol (alpha-T) do not interfere in the determination. A procedure for blank assays allows selective inhibition of alpha-T conversions and measurement of interfering fluorescence. The method is highly sensitive, which allows the determination of alpha-T in very dilute solutions. This in turn suppresses matrix effects and renders the results reliable. The time interval between 2 peaks is 6 min, washing included, and it is possible to carry out 50 determinations per day (sample preparation not included). The system is robust and maintenance is easy. Parallel determinations of foods and feeds have been carried out with a conventional thin layer chromatographic method.     

Free galactose concentrations in fresh and stored apples (Malus domestica) and processed apple products

Gas chromatography was used to quantitate free galactose in Braeburn, Fuji, Red Delicious, and Spartan apples during cold storage, after thermal processing of apple slices and in juice produced using clarification and/or liquifaction enzymes. Spartan had significantly higher galactose levels as compared to Red Delicious apples, but changes in galactose in all varieties during 9 months of cold storage were insignificant. Blanching and canning decreased galactose levels, but doubling the thermal processing during canning increased the free galactose concentration detected in plant tissue. An enzymatic liquefaction aid used to prepare apple juice dramatically increased the free galactose content while a clarification aid caused only a slight increase due to its selective action on soluble pectin. These findings provide useful information for dietitians to base diet recommendations for galactosemic patients.     

Middle Stone Age bedding construction and settlement patterns at Sibudu, South Africa

The Middle Stone Age (MSA) is associated with early behavioral innovations, expansions of modern humans within and out of Africa, and occasional population bottlenecks. Several innovations in the MSA are seen in an archaeological sequence in the rock shelter Sibudu (South Africa). At ~77,000 years ago, people constructed plant bedding from sedges and other monocotyledons topped with aromatic leaves containing insecticidal and larvicidal chemicals. Beginning at ~73,000 years ago, bedding was burned, presumably for site maintenance. By ~58,000 years ago, bedding construction, burning, and other forms of site use and maintenance intensified, suggesting that settlement strategies changed. Behavioral differences between ~77,000 and 58,000 years ago may coincide with population fluctuations in Africa.     

Ordering of quantum dots using genetically engineered viruses

A liquid crystal system was used for the fabrication of a highly ordered composite material from genetically engineered M13 bacteriophage and zinc sulfide (ZnS) nanocrystals. The bacteriophage, which formed the basis of the self-ordering system, were selected to have a specific recognition moiety for ZnS crystal surfaces. The bacteriophage were coupled with ZnS solution precursors and spontaneously evolved a self-supporting hybrid film material that was ordered at the nanoscale and at the micrometer scale into approximately 72-micrometer domains, which were continuous over a centimeter length scale. In addition, suspensions were prepared in which the lyotropic liquid crystalline phase behavior of the hybrid material was controlled by solvent concentration and by the use of a magnetic field.     

Forced unfolding of proteins within cells

To identify cytoskeletal proteins that change conformation or assembly within stressed cells, in situ labeling of sterically shielded cysteines with fluorophores was analyzed by fluorescence imaging, quantitative mass spectrometry, and sequential two-dye labeling. Within red blood cells, shotgun labeling showed that shielded cysteines in the two isoforms of the cytoskeletal protein spectrin were increasingly labeled as a function of shear stress and time, indicative of forced unfolding of specific domains. Within mesenchymal stem cells-as a prototypical adherent cell-nonmuscle myosin IIA and vimentin are just two of the cytoskeletal proteins identified that show differential labeling in tensed versus drug-relaxed cells. Cysteine labeling of proteins within live cells can thus be used to fluorescently map out sites of molecular-scale deformation, and the results also suggest means to colocalize signaling events such as phosphorylation with forced unfolding.     

Holocene deglaciation of Marie Byrd Land,West Antarctica

Surface exposure ages of glacial deposits in the Ford Ranges of western Marie Byrd Land indicate continuous thinning of the West Antarctic Ice Sheet by more than 700 meters near the coast throughout the past 10,000 years. Deglaciation lagged the disappearance of ice sheets in the Northern Hemisphere by thousands of years and may still be under way. These results provide further evidence that parts of the West Antarctic Ice Sheet are on a long-term trajectory of decline. West Antarctic melting contributed water to the oceans in the late Holocene and may continue to do so in the future.     

Accelerated uplift and magmatic intrusion of the Yellowstone caldera, 2004 to 2006

The Yellowstone caldera began a rapid episode of ground uplift in mid-2004, revealed by Global Positioning System and interferometric synthetic aperture radar measurements, at rates up to 7 centimeters per year, which is over three times faster than previously observed inflation rates. Source modeling of the deformation data suggests an expanding volcanic sill of approximately 1200 square kilometers at a 10-kilometer depth beneath the caldera, coincident with the top of a seismically imaged crustal magma chamber. The modeled rate of source volume increase is 0.1 cubic kilometer per year, similar to the amount of magma intrusion required to supply the observed high heat flow of the caldera. This evidence suggests magma recharge as the main mechanism for the accelerated uplift, although pressurization of magmatic fluids cannot be ruled out.     

Crystal structure of the malaria vaccine candidate apical membrane antigen 1

Apical membrane antigen 1 from Plasmodium is a leading malaria vaccine candidate. The protein is essential for host-cell invasion, but its molecular function is unknown. The crystal structure of the three domains comprising the ectoplasmic region of the antigen from P. vivax, solved at 1.8 angstrom resolution, shows that domains I and II belong to the PAN motif, which defines a superfamily of protein folds implicated in receptor binding. We also mapped the epitope of an invasion-inhibitory monoclonal antibody specific for the P. falciparum ortholog and modeled this to the structure. The location of the epitope and current knowledge on structure-function correlations for PAN domains together suggest a receptor-binding role during invasion in which domain II plays a critical part. These results are likely to aid vaccine and drug design.     

Regulation of cytokine receptors by Golgi N-glycan processing and endocytosis

The Golgi enzyme beta1,6 N-acetylglucosaminyltransferase V (Mgat5) is up-regulated in carcinomas and promotes the substitution of N-glycan with poly N-acetyllactosamine, the preferred ligand for galectin-3 (Gal-3). Here, we report that expression of Mgat5 sensitized mouse cells to multiple cytokines. Gal-3 cross-linked Mgat5-modified N-glycans on epidermal growth factor and transforming growth factor-beta receptors at the cell surface and delayed their removal by constitutive endocytosis. Mgat5 expression in mammary carcinoma was rate limiting for cytokine signaling and consequently for epithelial-mesenchymal transition, cell motility, and tumor metastasis. Mgat5 also promoted cytokine-mediated leukocyte signaling, phagocytosis, and extravasation in vivo. Thus, conditional regulation of N-glycan processing drives synchronous modification of cytokine receptors, which balances their surface retention against loss via endocytosis.