Measurement of soil CO2 efflux using soda lime absorption: both quantitative and reliable

Measurement of soil CO₂ efflux using a non-flow-through steady-state (NFT-SS) chamber with alkali absorption of CO₂ by soda lime was tested and compared with a flow-through non-steady-state (FT-NSS) IRGA method to assess suitability of using soda lime for field monitoring over large spatial scales and integrated over a day. Potential errors and artifacts associated with the soda lime chamber method were investigated and improvements made. The following issues relating to quantification and reliable measurement of soil CO₂ efflux were evaluated: (i) absorption capacity of the soda lime, (ii) additional and thus artifactual absorption of CO₂ by soda lime during the experimental procedure, (iii) variation in the CO₂ concentration inside the chamber headspace, and (iv) effects of chamber closure on soil CO₂ efflux. Soil CO₂ efflux, as measured using soda lime (with a range of quantities: 50, 100, and 200 g per 0.082 m² ground area enclosed in chamber), was compared with transient IRGA measurements as a reference method that is based on well-established physical principles, using several forms of spatial and temporal comparisons. Natural variation in efflux rates ranged from 2 to 5.5 g C m⁻² day⁻¹ between different chambers and over different days. A comparison of the IRGA-based assay with measurement based on soda lime yielded an overall correlation coefficient of 0.82. The slope of the regression line was not significantly different from the 1:1 line, and the intercept was not significantly different from the origin. This result indicated that measurement of CO₂ efflux by soda lime absorption was quantitatively similar and unbiased in relation to the reference method. The soda lime method can be a highly practical method for field measurements if implemented with due care (in terms of drying and weighing soda lime, and in minimizing leakages), and validated for specific field conditions. A detailed protocol is presented for use of the soda lime method for measurement of CO₂ efflux from field soils.     

Lignans isolated from Campylotropis hirtella (Franch.) Schindl. decreased prostate specific antigen and androgen receptor expression in LNCaP cells

Accumulating epidemiological data suggest that Asian men have lower incidences of prostate cancer and benign prostate hyperplasia (BPH) compared with American and European populations and may have benefited from their higher intake of phytoestrogens in their diet. However, how these phytochemicals affect prostatic diseases is still unclear. In this study, we isolated six lignans from a plant, Campylotropis hirtella (Franch.) Schindl., which has been used as a folk medicine for treatment of BPH in China, through bioassay guided fractionation. They were dehydrodiconiferyl alcohol (C1), 4-[(-6-hydroxy-2,3-dihydro-1-benzofuran-3-yl)methyl]-5-methoxybenzene-1,3-diol (C2), erythro-guaiacylglycerol-beta-O-4'-coniferyl ether (C3), threo-guaiacylglycerol-beta-O-4'-coniferyl ether (C4), secoisolariciresinol (C5), and prupaside (C6), where C2 was identified as a new lignan analog. Their IC50 values for inhibition of prostate specific antigen (PSA) secretion were 19, 45, 110, 128, 137, and 186 microM, respectively, from C1 to C6 in LNCaP cells. Further study showed that C1-5 down-regulated cellular PSA expression and C1-4 also decreased androgen receptor (AR) expression in LNCaP cells. Furthermore, we investigated the proapoptotic effect of C1 on LNCaP cells. The active forms of caspase 3 associated with the specific proteolysis of poly (ADP-ribose) polymerase (PARP) were detected, and the antiapoptotic protein Bcl-2 was down-regulated after the treatment with C1. These results collectively indicated that these lignans may have chemopreventive or therapeutic actions for prostate cancer through suppressing AR signaling pathway and inducing apoptosis.     

Retention of provitamin A carotenoids in high beta-carotene maize (Zea mays) during traditional African household processing

High beta-carotene maize, biofortified with beta-carotene through plant breeding, is being developed as a cost-effective, sustainable agronomic approach to alleviating the problem of vitamin A deficiency in Africa. We used high beta-carotene maize (10.49+/-0.16 microg beta-carotene/g) to prepare traditional maize porridges and compared the carotenoid contents in the following: (1) whole kernels; (2) wet milled flour; (3) wet milled flour, fermented; (4) wet milled flour, cooked; (5) wet milled flour, fermented and cooked. The cumulative losses of beta-carotene in the final, cooked products were 24.5% (95% CI 22.8-26.2%) and 24.8% (95% CI 23.1-26.5%), for the fermented and unfermented porridges, respectively. Thus, fermentation, a traditional technology with documented nutritional and other health benefits, does not adversely affect the retention of beta-carotene in porridges prepared with high beta-carotene maize. The relatively good retention of beta-carotene during traditional maize processing provides additional experimental support for the feasibility of maize biofortification as a means to alleviate vitamin A deficiency.     

Zea mays rhizosphere respiration, but not soil organic matter decomposition was stable across a temperature gradient

In a greenhouse experiment, we grew maize plants at different densities. We added fertilizer to half of the pots and created a temperature gradient. After 10 weeks of plant growth, we measured soil CO₂ efflux (SCE) and determined rhizosphere respiration (R_rhizo) and the decomposition rate of soil organic matter (R_SOM) using the different δ¹³C of the C₃ soil and C₄ plants. Whereas R_rhizo remained stable across the temperature gradient, R_SOM significantly increased with growth temperature. Neither plant density, nor the fertilizer treatment affected the relation between R_rhizo or R_SOM and growth temperature. Although R_rhizo might still increase with temperature in the short term, long term exposure to higher temperatures revealed full thermal acclimation of R_rhizo, but not of R_SOM.     

Microassay for rapid screening of alpha-amylase activity

A microassay was developed for measuring the activity of alpha-amylases in the nanogram enzyme concentration range, based on the use of dye-labeled cross-linked starch as the substrate, and the release of soluble colored fragments formed in enzyme hydrolysis. Reaction conditions were optimized to generate a linear correlation between the increase in absorbance and a reaction time of 0-10 min, as well as enzyme concentrations in the range of 0-50 ng. A standard curve for the conversion of absorbance to enzyme activity units was constructed. The protocol developed was applied to monitoring the production of ultralow concentrations of recombinant barley alpha-amylase in yeast cells.     

Application of isotope dilution gas chromatography-mass spectrometry in analysis of organochlorine pesticide residues in ginseng root

A highly accurate and precise method based on isotope dilution gas chromatography-mass spectrometry was developed for the determination of five matrix-bound organochlorine pesticides, namely, hexachlorobenzene and hexachlorocyclohexanes (alpha-, beta-, delta-, and gamma- isomers), in a reference sample of Panax gingseng. Identification of the analytes was confirmed under selective ion monitoring mode by the presence of two dominant ion fragments within the specific time windows (+/-1% of the relative retention time with respect to the calibration standards) and matching of relative ion intensities of the concerned ions in samples and calibration standards (within +/-5%). Quantification was based on the measurement of concentration ratios of the natural and isotopic analogues in the sample and calibration blends. To circumvent the tedious iterative process of exact isotope matching that is often used in isotope dilution mass spectrometry analysis, a single-point calibration procedure was adopted with the isotopic amount ratios in the sample and calibration blends close to unity (0.9-1.1). Under the described approach, intraday and interday repeatability of replicate analyses of organochlorine pesticides in the ginseng root sample were below 1.4%. The expanded relative uncertainty ranging from 4.0 to 6.5% at a coverage factor of 2 was significantly lower than those of conventional gas chromatographic methods using other calibration techniques (internal or external standards). A deviation of less than 2.0% from the certified values was achieved when applying the developed method to determine hexachlorobenzene, alpha-, and beta-hexachlorocyclohexane in a certified reference material (CRM), BCR-CRM 115. Because of the unavailability of relevant CRMs of herbal origins, the concerned ginseng root sample, after verification of the "true values" of the concerned organochlorine pesticides by the valid primary method, is suitable for serving as an in-house reference material for quality assurance and method validation purposes.     

The Accumulation of Boron in Agropyron elongatum Grown in Coal Fly Ash and Sewage Sludge Mixture

A greenhouse pot experiment was conducted to investigate the boron (B) release capacity of coal fly ash and sewage sludge mixtures, and the accumulation of B in Agropyron elongatum after two consecutive growing seasons. Sludge was amended with fly ash at application rates of 0, 5, 10, 35, and 50% (w/w), and each mixture was then mixed with a loamy soil at either 1:1 or 1:5 (v/v). Both water soluble B (WS-B) and hot water soluble B (HWS-B) increased with increasing fly ash amendment rate. Shoot B concentrations also increased significantly according to the rate of ash amendment. The ash-sludge mixture improved plant growth with the highest total dry weight yield at 10% ash amendment rate. Boron toxicity symptoms in leaf tips were observed at 35% and 50% ash amendment rate at both soil mixing ratios. Hot water soluble B and WS-B decreased significantly after consecutive cropping of Agropyron especially at low ratio of mixure with soil i.e., 1:5 (v/v). However, soil available B contents at ≥ 35% ash application rate and 1:1 (v/v) soil mixing ratio were still excessive for normal plant growth, suggesting that deleterious effects on plant growth would be experienced in later seasons owing to the high amounts of residual B.     

Analysis of gamma-irradiated melon, pumpkin, and sunflower seeds by electron paramagnetic resonance spectroscopy and gas chromatography-mass spectrometry

Seeds of melon (Citrullus lanatus var. sp.), pumpkin (Cucurbita moschata), and sunflower (Heliantus annus) were gamma-irradiated at 1, 3, 5, and 10 kGy and analyzed by electron paramagnetic resonance (EPR) and gas chromatography-mass spectrometry (GC-MS) according to EN1787:2000 and EN1785:2003, respectively. Distinguishable triplet signals due to the presence of induced cellulose radicals were found at 2.0010-2.0047 g in the EPR spectra. The gamma-irradiated radiolytic markers of 2-dodecylcyclobutanone (2-DCB) and 2-tetradecylcyclobutanone (2-TCB) were identified in all irradiated seed samples. Both the free radicals and the alkylcyclobutanones were found to increase with irradiation dose. In general, linear relationships between the amount of radicals and irradiation dosage could be established. Studies at an ambient temperature (20-25 degrees C) in a humidity-controlled environment showed a complete disappearance of the cellulosic peaks for irradiated samples upon 60 days of storage. Such instability behavior was considered to render the usefulness of using EPR alone in the determination of irradiated seed samples. On the other hand, 2-DCB and 2-TCB were also found to decompose rapidly (>85% loss after 120 days of storage), but the radiolytic markers remained quantifiable after 120 days of postirradiation storage. These results suggest that GC-MS is a versatile and complimentary technique for the confirmation of irradiation treatment to seeds.     

Balance and turnover of nutrients in a bamboo plantation (Bambusa bambos) of different ages

The accumulation of nutrients by a bamboo plantation and their rate of uptake and release through litter fall was determined. The nutrients in the above-ground living biomass increased linearly with plant age. The balance of nutrients showed an increase in loss by litter fall, and annual nutrient accumulation and enrichment ratio were enhanced with age. Annual percentage turnover showed no consistent trend. Enrichment ratios were in the order of K>N>Mg>Ca>P. In conclusion, precautions are necessary during the exploitation of bamboo plantations on a large scale in order to prevent nutrient depletion of the soil. Received: 7 January 1996     

Antidiabetic activity of red wine polyphenolic extract, ethanol, or both in streptozotocin-treated rats

A polyphenol extract from a Corbières (France) red wine (P, 200 mg/kg), ethanol (E, 1 mL/kg), or a combination of both (PE) was administered by daily gavage for 6 weeks to healthy control or streptozotocin (60 mg/kg i.v.)-induced diabetic rats (180-200 g). Treatment groups included C or D (untreated control or diabetic) and CP, CE, or CPE (treated control) or DP, DE, or DPE (treated diabetic). P treatment induced a reduction in body growth, food intake, and glycemia in both CP and DP groups. In DP, hyperglycemia was reduced when measured 1 h after daily treatment but not at sacrifice (no treatment on that day). The hyperglycemic response to the oral glucose tolerance test (OGTT) and plasma insulin at sacrifice were impaired similarly in DP and D groups. In contrast, in DE or DPE, body growth was partially restored while hyperglycemia was reduced both during treatment and at sacrifice. In addition, hyperglycemia response to OGTT was reduced and plasma insulin was higher in DE or DPE than in D animals, indicating a long-term correction of diabetes in ethanol-treated animals. Morphometric studies showed that ethanol partially reversed the enlarging effect of diabetes on the mesenteric arterial system while the polyphenolic treatment enhanced it in the absence of ethanol. In summary, our study shows that (i). a polyphenol extract from red wine ("used at a pharmacological" dose) reduces glycemia and decreases food intake and body growth in diabetic and nondiabetic animals and (ii). ethanol ("nutritional" dose) administered alone or in combination with polyphenols is able to correct the diabetic state. Some of the effects of polyphenols were masked by the effects of ethanol, notably in diabetic animals. Further studies will determine the effect of "nutritional" doses of polyphenols as well as their mechanism of action.