Determination of oleandrin in tissues and biological fluids by liquid chromatography-electrospray tandem mass spectrometry

A rapid LC-MS/MS method, using a triple-quadrupole/linear ion trap mass spectrometer, was developed for the quantitative determination of oleandrin in serum, urine, and tissue samples. Oleandrin, the major cardiac glycoside of oleander (Nerium oleander L.), was extracted from serum and urine samples with methylene chloride and from tissues with acetonitrile. The tissue extracts were cleaned up using Florisil solid-phase extraction columns. Six replicate fortifications of serum and urine at 0.001 microg/g (1 ppb) oleandrin gave average recoveries of 97% with 5% CV (relative standard deviation) and 107% with 7% CV, respectively. Six replicate fortifications of liver at 0.005 microg/g (5 ppb) oleandrin gave average recoveries of 98% with 6% CV. This is the first report of a positive mass spectrometric identification and quantitation of oleandrin in tissue samples from oleander intoxication cases. The sensitivity and specificity of the LC-MS/MS analysis enables it to be the method of choice for toxicological investigations of oleander poisoning.     

Creaming in black tea

Tea cream is the precipitate formed as tea cools. Its formation has been studied by X-ray scattering, and it is shown that a higher tea concentration leads to earlier onset of creaming and larger particles and that addition of theaflavin and calcium promotes creaming. Association constants between the major components of black tea have been obtained using NMR and show that calcium and glucose enhance the self-association of caffeine, polyphenols, and theaflavin but have little effect on hetero-association. Glycosylation of a polyphenol reduced self-association and reduced binding to caffeine. We conclude that theaflavin is important for the initiation of creaming, forming nanoclusters of typically 3 nm diameter, whereas caffeine acts more to fill in the gaps within the clusters and thus adds to the bulk of tea cream without being necessary for its initiation. Tea creaming may be reduced by increasing the solubility of the polyphenols (i.e., by glycosylation) or by removing calcium. Tea cream; theaflavin; caffeine; small-angle X-ray scattering; NMR; colloid.     

Effects of urbanization on California’s fish diversity: Differentiation, homogenization and the influence of spatial scale

Human development of freshwater ecosystems has lead to drastic changes in freshwater fish faunas, including the loss of many native species and the gain of non-natives. Typically conservation ecologists view these two opposing forces as contributing to biological homogenization, and consider homogenization as one of the principle negative consequences of urbanization. However, homogenization is only one outcome out of many that can result from the loss and gain of species. In particular, it is possible for invasions and extinctions to lead to differentiation; a process whereby two (or more) regions become less similar to one another through time. Using the freshwater fishes of California, we show that urbanization is highly positively correlated to both the endangerment of native fish and the invasion of non-native fish within watersheds. Despite this, the fish faunas of California’s watersheds have differentiated from one another through time. Furthermore, the degree of differentiation is positively correlated with measures of urbanization, which is contrary to expectation. We suggest that this result reflects: (1) the haphazard manner in which non-native fishes have been introduced into California watersheds, (2) the difficulty that both native and non-native fishes have in expanding their geographical ranges, and (3) the continued presence of vestiges of formerly distinct regional faunas. This pattern of differentiation among watersheds is likely a matter of scale, as previous work on freshwater fishes has demonstrated homogenization at both larger and smaller spatial scales. In addition the observed pattern is probably a short-term (temporal) phenomena and will disappear with continued invasion and extinction. We suggest that similar patterns may occur for other taxa that have limited natural dispersal abilities and that are idiosyncratically released as non-natives via human activities (e.g. herptiles).     

Effect of fermentation on Sweetpotato (Ipomoea batatas) toxicity in mice

Unfortunate bovine fatalities occurring after ingestion of mold-damaged sweetpotatoes preclude the use of the culled tubers in livestock feed. In cattle, mold-damaged sweetpotatoes induce an acute respiratory distress syndrome resulting in asphyxiation. Because of this potential toxicity and the general abundance of culled sweetpotatoes, the detoxification efficacy of ensiling was explored since it is an easy and economically viable technique often applied to preserve livestock feed. Sweetpotato slices with or without mold damage were stored either frozen (to represent unfermented samples) or fermented for 6 weeks at room temperature. Following fermentation, organic extracts were generated for administration to mice. Thirty hours following administration of the extracts, mice were evaluated for gross and microscopic lesions affecting the lungs, liver, and kidneys. Fermentation of 6 weeks duration was observed to inadequately eliminate the lung, liver, and kidney toxicity caused by mold-damaged sweetpotatoes. In fact, fermentation exacerbated the hepatotoxicity of mold-damaged sweetpotatoes. This is also the first demonstration that sweetpotato regions lacking visible mold damage can induce lung and kidney injury, which, however, is preventable by fermentation.     

Elemental concentrations in fishes from the Salton Sea,Southeastern California

The Salton Sea is a 93 000 ha saline lake fed by drainage water from more than 283 000 ha of irrigated lands in the Imperial and Coachella valleys of California. A total of 21 composite samples of four recreationally important fish species — bairdiella (Bairdiella icistia), orangemouth corvina (Cynoscion xanthulus), sargo (Anisotremus davidsoni), and Mozambique tilapia (Tilapia mossambica) —collected there were analyzed for 14 elements. Twelve of these elements were detected in one or more of the samples: As, B, Co, Cu, Fe, Hg, Mo, Ni, Pb, Se, V, and Zn. Cadmium and Tl were not detected. The ranges in concentrations of elements in the skinless fillets of bairdiella, corvina, and sargo, and in whole bodies of all four fishes were comparable to levels that are typically measured in saltwater fishes. Only Se concentrations were elevated (as much as 14 µg g^?1 dry weight in both fillets and whole bodies) in this series of samples. Elevated concentrations of Se have already led to public health advisories concerning the consumption of fish and might eventually cause the demise of fish populations from toxic effects.     

Utilization of the genetic resources of wild species to create a nontransgenic high flavonoid tomato

Flavonoids represent a large and important group of plant natural products that are ubiquitous in the plant kingdom. Epidemiological studies have shown the health benefits of a diet high in flavonoids. However, the dietary intake of flavonoids in most western populations is limited, creating a need to find alternative food sources for these polyphenolic secondary metabolites. The domestication of many of our cultivated food crops has resulted in alterations in the biosynthetic pathways of many essential micronutrients and vitamins through inadvertent counterselection against nutritional traits in favor of agronomic ones. Flavonoids are nearly absent from fruits of cultivated tomato (Lycopersicon esculentum Mill.), a major vegetable in human diets. Previous attempts to restore the flavonoid pathway in tomato fruits have been limited to transgenic strategies, suggesting that the problem was intractable through traditional methods. Here, we describe for the first time a nontransgenic metabolic engineering approach to developing a high flavonoid tomato using a wild tomato species (Lycopersicon pennelliiv. puberulum) and demonstrate the opportunities for restoring functional pathways using the genetic resources of wild species, resulting in production of healthier foods.     

Cotton lint yield variability in a heterogeneous soil at a landscape scale

Landscape variability associated with topographic features affects the spatial pattern of soil water and N redistribution, and thus N uptake and crop yield. A landscape-scale study was conducted in a center pivot irrigated field on the southern High Plains of Texas in 1999 to assess soil water, soil NO₃-N, cotton (Gossypium hirsutum L.) lint yield, and N uptake variability in the landscape, and to determine the spatial correlation between these landscape variables using a state-space approach. The treatments were irrigation at 50 and 75% cotton potential evapotranspiration (ET). Neutron access tubes were placed at a 15-m interval along a 710 m (50% ET) and 820 m (75% ET) transect across the field. Soil NO₃-N in early spring was autocorrelated at a distance varying between 60 and 80 m. Measured soil volumetric water content (WC), total N uptake, and lint yield were generally higher on lower landscape positions. Cotton lint yield was significantly correlated to soil WC (r=0.76), soil NO₃-N (r=0.35), and site elevation (r=−0.54). Differences of site elevation between local neighboring points explained the soil water, NO₃-N and lint yield variability at the micro-scale level in the landscape. Soil WC, cotton lint yield, N uptake, and clay content were crosscorrelated with site elevation across a lag distance of ±30–40 m. The state-space analysis showed that cotton lint yield was positively weighted on soil WC availability and negatively weighted on site elevation. Cotton lint yield state-space models give insights on the association of soil physical and chemical properties, lint yield, and landscape processes, and have the potential to improve water and N management at the landscape-scale.     

Thermal oxidation of 9'-cis-neoxanthin in a model system containing peroxyacetic acid leads to the potent odorant beta-damascenone

The potent odorant beta-damascenone was formed directly from 9'-cis-neoxanthin in a model system by peroxyacetic acid oxidation and two-phase thermal degradation without the involvement of enzymatic activity. Beta-damascenone formation was heavily dependent on pH (optimum at 5.0) and temperature, occurring over the two sequential phases. The first was incubation with peroxyacetic acid at 60 degrees C for 90 min, and the second was at above 90 degrees C for 20 min. Only traces of beta-damascenone were formed on application of only one of the two phases. Formate and citrate solutions produced a much better environment for beta-damascenone formation than acetate and phosphate. About 7 microg/L beta-damascenone was formed from 5.8 mg/L 9'-cis-neoxanthin under optimal experimental condition. The detailed pathway by which beta-damascenone is formed remains to be elucidated.     

Development of a continuous process to make casein films

A semicontinuous pilot plant process was developed to make films from calcium caseinate plasticized with glycerol. Calcium caseinate/glycerol solutions were wetted and spread on polyethylene or Mylar belts and were readily removed after forming films. A Meier rod was used to spread the solution onto the belt. Feed rates were 3.3 and 5 g/cm. The resulting films using the polyethylene belt had a tensile strength of 5 MPa and 30% elongation with thickness of 0.14 mm. When the glycerol concentration was adjusted, the process produced films with properties similar to those previously reported for a batch process. It is anticipated that this process will apply to other proteins.