Effects of oxidizing adulterants on detection of 11-nor-delta9-THC-9-carboxylic acid in urine. (1/14)
Bleach, nitrite, chromate, and hydrogen peroxide-peroxidase are effective urine adulterants used by the illicit drug users to conceal marijuana-positive results. Methods for detecting nitrite and chromate are available. Effects of other oxidizing agents that could possibly be used as adulterants and are difficult to detect or measure are presented in this report. Urine samples containing 40 ng/mL of 11-nor-delta9-THC-9-carboxylic acid (THC-acid) were treated with 10 mmol/L of commonly available oxidizing agents. Effects of horseradish peroxidase of activity 10 unit/mL and extracts from 2.5 g of red radish (Raphanus sativus, Radicula group), horseradish (Armoracia rusticana), Japanese radish (Raphanus sativus, Daikon group), and black mustard seeds (Brassica nigra), all with 10 mmol/L of hydrogen peroxide, were also examined. After 5 min, 16 h and 48 h of exposure at room temperature (23 degrees C) the specimens were tested by a gas chromatographic-mass spectrometric method for THC-acid. A control group treated with sodium hydrosulfite to reduce the oxidants, was also tested to investigate the effect of oxidizing agents on THC-acid in the extraction method. THC-acid was lost completely in the extraction method when treated with chromate, nitrite, oxone, and hydrogen peroxide/ferrous ammonium sulfate (Fenton's reagent). Some losses were also observed with persulfate and periodate (up to 25%). These oxidants, and other oxidizing agents like permanganate, periodate, peroxidase, and extracts from red radish, horseradish, Japanese radish and black mustard seeds destroyed most of the THC-acid (> 94%) within 48 h of exposure. Chlorate, perchlorate, iodate, and oxychloride under these conditions showed little or no effect. Complete loss was observed when THC-acid was exposed to 50 mmol/L of oxychloride for 48 h. Several oxidizing adulterants that are difficult to test by the present urine adulterant testing methods showed considerable effects on the destruction of THC-acid. The time and temperature for these effects were similar to those used by most laboratories to collect and test specimens. In several cases, the loss of THC-acid was > 94%. (+info)An N-terminal peptide extension results in efficient expression, but not secretion, of a synthetic horseradish peroxidase gene in transgenic tobacco. (2/14)
BACKGROUND AND AIMS: Native horseradish (Armoracia rusticana) peroxidase, HRP (EC 1.11.1.7), isoenzyme C is synthesized with N-terminal and C-terminal peptide extensions, believed to be associated with protein targeting. This study aimed to explore the specific functions of these extensions, and to generate transgenic plants with expression patterns suitable for exploring the role of peroxidase in plant development and defence. METHODS: Transgenic Nicotiana tabacum (tobacco) plants expressing different versions of a synthetic horseradish peroxidase, HRP, isoenzyme C gene were constructed. The gene was engineered to include additional sequences coding for either the natural N-terminal or the C-terminal extension or both. These constructs were placed under the control of a constitutive promoter (CaMV-35S) or the tobacco RUBISCO-SSU light inducible promoter (SSU) and introduced into tobacco using Agrobacterium-mediated transformation. To study the effects of the N- and C-terminal extensions, the localization of recombinant peroxidase was determined using biochemical and molecular techniques. KEY RESULTS: Transgenic tobacco plants can exhibit a ten-fold increase in peroxidase activity compared with wild-type tobacco levels, and the majority of this activity is located in the symplast. The N-terminal extension is essential for the production of high levels of recombinant protein, while the C-terminal extension has little effect. Differences in levels of enzyme activity and recombinant protein are reflected in transcript levels. CONCLUSIONS: There is no evidence to support either preferential secretion or vacuolar targeting of recombinant peroxidase in this heterologous expression system. This leads us to question the postulated targeting roles of these peptide extensions. The N-terminal extension is essential for high level expression and appears to influence transcript stability or translational efficiency. Plants have been generated with greatly elevated cytosolic peroxidase activity, and smaller increases in apoplastic activity. These will be valuable for exploring the role of these enzymes in stress amelioration and plant development. (+info)Distribution and Translocation of 141Ce (III) in Horseradish. (3/14)
BACKGROUND AND AIMS: Rare earth elements (REEs) are used in agriculture and a large amount of them contaminate the environment and enter foods. The distribution and translocation of (141)Ce (III) in horseradish was investigated in order to help understand the biochemical behaviour and toxic mechanism of REEs in plants. METHODS: The distribution and translocation of (141)Ce (III) in horseradish were investigated using autoradiography, liquid scintillation counting (LSC) and electron microscopic autoradiography (EMARG) techniques. The contents of (141)Ce (III) and nutrient elements were analysed using an inductively coupled plasma-atomic emission spectrometer (ICP-AES). RESULTS: The results from autoradiography and LSC indicated that (141)Ce (III) could be absorbed by horseradish and transferred from the leaf to the leaf-stalk and then to the root. The content of (141)Ce (III) in different parts of horseradish was as follows: root > leaf-stalk > leaf. The uptake rates of (141)Ce (III) in horseradish changed with the different organs and time. The content of (141)Ce (III) in developing leaves was greater than that in mature leaves. The results from EMARG indicated that (141)Ce (III) could penetrate through the cell membrane and enter the mesophyll cells, being present in both extra- and intra-cellular deposits. The contents of macronutrients in horseradish were decreased by (141)Ce (III) treatment. CONCLUSIONS: (141)Ce (III) can be absorbed and transferred between organs of horseradish with time, and the distribution was found to be different at different growth stages. (141)Ce (III) can enter the mesophyll cells via apoplast and symplast channels or via plasmodesmata. (141)Ce (III) can disturb the metabolism of macronutrients in horseradish. (+info)Thermal unfolding studies of a phytocyanin. (4/14)
(+info)The structure of horseradish peroxidase C characterized as a molten globule state after Ca(2+) depletion. (5/14)
(+info)Permeability change of arterial endothelium is an age-dependent function of lesion size in apolipoprotein E-null mice. (6/14)
(+info)Stabilization of enzymes in silk films. (7/14)
(+info)Biphenyl-metabolizing bacteria in the rhizosphere of horseradish and bulk soil contaminated by polychlorinated biphenyls as revealed by stable isotope probing. (8/14)
(+info)'Armoracia' is the genus name for a type of plant commonly known as horseradish. It belongs to the family Brassicaceae, which also includes vegetables such as broccoli, cabbage, and mustard greens. The root of the horseradish plant is used as a spice or condiment due to its pungent flavor, which is caused by the release of volatile oils when the root is grated or crushed.
Horseradish has been used in traditional medicine for various purposes, such as treating respiratory infections and promoting digestion. However, it is important to note that while some studies suggest potential health benefits of horseradish, more research is needed to confirm its effectiveness and safety. As with any medication or supplement, it is recommended to consult with a healthcare professional before using horseradish for medicinal purposes.