Impact of Waxy,Partial Waxy,and Wildtype Wheat Starch Fraction Properties on Hearth Bread Characteristics

Thirteen different wheat cultivars were selected to represent GBSS mutations: three each of wildtype, axnull, and bxnull, and two each of 2xnull and waxy. Starch and A‐ and B‐granules were purified from wheat flour. Hearth bread loaves were produced from the flours using a small‐scale baking method. A‐granules purified from wildtype and partial waxy (axnull, bxnull, and 2xnull) starches have significantly higher gelatinization enthalpy and peak viscosity compared with B‐granules. A‐ and B‐granules from waxy starch do not differ in gelatinization, pasting, and gelation properties. A‐ and B‐granules from waxy starch have the highest enthalpy, peak temperature, peak viscosity, breakdown, and lowest pasting peak time and pasting temperature compared with A‐ and B‐granules from partial waxy and wildtype starch. Waxy wheat flour has much higher water absorption compared with partial waxy and wildtype flour. No significant difference in hearth bread baking performance was observed between wildype and partial waxy wheat flour. Waxy wheat flour produced hearth bread with significantly lower form ratio, weight, a more open pore structure, and a bad overall appearance. Baking with waxy, partial waxy, and wildtype wheat flour had no significant effect on loaf volume.     

Photoinhibition, carotenoid composition and the co-regulation of photochemical and non-photochemical quenching in neotropical savanna trees

Plants in the neotropical savannas of central Brazil are exposed to high irradiances, high air temperatures and low relative humidities. These conditions impose a selection pressure on plants for strong stomatal regulation of transpiration to maintain water balance. Diurnal adjustments of non-photochemical energy dissipation in photosystem II (PSII) provide a dynamic mechanism to reduce the risk of photoinhibitory damage during the middle of the day when irradiances and leaf temperatures are high and partial closure of the stomata results in considerable reductions in internal CO(2) concentration. At the end of the dry season, we measured diurnal changes in gas exchange, chlorophyll fluorescence parameters and carotenoid composition in two savanna tree species differing in photosynthetic capacity and in the duration and extent of the midday depression of photosynthesis. Non-photochemical quenching and its quantum yield were tightly correlated with zeaxanthin concentrations on a total chlorophyll basis, indicating that the reversible de-epoxidation of violaxanthin to antheraxanthin and zeaxanthin within the xanthophyll cycle plays a key role in the regulation of thermal energy dissipation. In both cases, a single linear relationship fitted both species. Although efficient regulation of photochemical and non-photochemical quenching and adjustments in the partitioning of electron flow between assimilative and non-assimilative processes were operating, these trees could not fully cope with the rapid increase in irradiance after sunrise, suggesting high vulnerability to photoinhibitory damage in the morning. However, both species were able to recover quickly. The effects of photoinhibitory quenching were largely reversed by midday, and zeaxanthin rapidly converted back to violaxanthin as irradiance decreased in late afternoon, resulting in the maximal quantum yield of PSII of around 0.8 just before sunrise.     

Diagnosis of anatoxin-a poisoning in dogs from North America.

Anatoxin-a, a toxin produced by several genera of blue-green algae, is considered a potent neurotoxin. Ingestion of water contaminated with the toxin results in acute neurological signs and often death. This report describes fatal cases of anatoxin-a ingestion in 6 dogs, with confirmation of anatoxin-a exposure by liquid chromatography/tandem mass spectrometry (LC-MS/MS/MS). In 1 outbreak, 3 dogs developed seizures and died within an hour after swimming in a river in California, while the other outbreak involved 3 dogs that died within 1 hour after swimming in a pond in Ontario. Anatoxin-a poisoning is rarely reported in dogs as a cause of acute neurological signs and death. However, increased occurrences of blue-green algae blooms in North America make this neurotoxin an important consideration in the diagnosis of sudden death associated with environmental water exposure. This brief communication reports on the isolation and detection of anatoxin-a from environmental water sources and the stomach contents of North American dogs dying of acute neurotoxicosis. This demonstrates the first documented cases of anatoxin-a poisoning in dogs in North America and the importance of LC-MS/MS/MS in identifying neurotoxins responsible for sudden death in cases of suspected blue-green algae toxicosis; especially those cases showing no gross or histological lesions.     

Determination of alpha-amanitin in serum and liver by multistage linear ion trap mass spectrometry

This paper describes a rapid LC-MS/MS/MS method for the analysis of alpha-amanitin in serum and liver. Serum was initially prepared by precipitation of proteins with acetonitrile and subsequent removal of acetonitrile with methylene chloride. Liver was prepared by homogenization with aqueous acetonitrile and subsequent removal of acetonitrile using methylene chloride. For both matrices, the aqueous phase was then extracted using mixed-mode C18/cation exchange SPE cartridges and analyzed on a linear ion trap LC-MS system. Standards were prepared in extracts of control matrix. Seven replicate fortifications of serum at 0.001 mug/g (1 ng/g) of alpha-amanitin gave a mean recovery of 95% with 8.8% CV (relative standard deviation) and a calculated method detection limit of 0.26 ng/g. Seven replicates of control liver fortified at 1 ng/g gave a mean recovery of 98% with 17% CV and a calculated method detection limit of 0.50 ng/g. This is the first report of a positive mass spectrometric identification and quantitation of alpha-amanitin in serum and liver from suspect human and animal intoxications.     

Conformational switches modulate protein interactions in peptide antibiotic synthetases

Protein dynamics plays an important role in protein function. Many functionally important motions occur on the microsecond and low millisecond time scale and can be characterized by nuclear magnetic resonance relaxation experiments. We describe the different states of a peptidyl carrier protein (PCP) that play a crucial role in its function as a peptide shuttle in the nonribosomal peptide synthetases of the tyrocidine A system. Both apo-PCP (without the bound 4'-phosphopantetheine cofactor) and holo-PCP exist in two different stable conformations. We show that one of the apo conformations and one of the holo conformations are identical, whereas the two remaining conformations are only detectable by nuclear magnetic resonance spectroscopy in either the apo or holo form. We further demonstrate that this conformational diversity is an essential prerequisite for the directed movement of the 4'-PP cofactor and its interaction with externally acting proteins such as thioesterases and 4'-PP transferase.     

Integrated standardization concept for Angelica botanicals using quantitative NMR

Despite numerous in vitro/vivo and phytochemical studies, the active constituents of Angelica sinensis (AS) have not been conclusively identified for the standardization to bioactive markers. Phytochemical analyses of AS extracts and fractions that demonstrate activity in a panel of in vitro bioassays, have repeatedly pointed to ligustilide as being (associated with) the active principle(s). Due to the chemical instability of ligustilide and related issues in GC/LC analyses, new methods capable of quantifying ligustilide in mixtures that do not rely on an identical reference standard are in high demand. This study demonstrates how NMR can satisfy the requirement for simultaneous, multi-target quantification and qualitative identification. First, the AS activity was concentrated into a single fraction by RP-solid-phase extraction, as confirmed by an alkaline phosphatase, (anti-)estrogenicity and cytotoxicity assay. Next, a quantitative ¹H NMR (qHNMR) method was established and validated using standard compounds and comparing processing methods. Subsequent 1D/2D NMR and qHNMR analysis led to the identification and quantification of ligustilide and other minor components in the active fraction, and to the development of quality criteria for authentic AS preparations. The absolute and relative quantities of ligustilide, six minor alkyl phthalides, and groups of phenylpropanoids, polyynes, and poly-unsaturated fatty acids were measured by a combination of qHNMR and 2D COSY. The qNMR approach enables multi-target quality control of the bioactive fraction, and enables the integrated biological and chemical standardization of AS botanicals. This methodology can potentially be transferred to other botanicals with active principles that act synergistically, or that contain closely related and/or constituents, which have not been conclusively identified as the active principles.