Identification, characterization, epitope mapping, and three-dimensional modeling of the alpha-subunit of beta-conglycinin of soybean, a potential allergen for young pigs

Soybean meal (SBM), the major byproduct of soybean oil extraction, is the main protein source for swine diets globally. In the United States, 8.6 million metric tons of SBM was used in swine rations in 2004. The pathological effect and immunological response of SBM feeding have been demonstrated in swine. In this study, we have utilized plasma collected from piglet feed with SBM in immunoblot analysis to detect proteins that elicited antigenic responses. We have identified soybean beta-conglycinin alpha-subunit as being a potential allergen for young piglets. Characterization of this protein indicated that deglycosylation and pepsin digestion did not eliminate immunoreactivity of this protein. Epitope mapping utilizing planar cellulose supports technology (SPOT) showed that three peptides spanning amino acids S185-R231 were critical for the allergenicity. A computer-generated three-dimensional structure model of the alpha-subunit of beta-conglycinin indicated that the antigenic epitopes were located on the surface of the protein. Information from this study may assist in the construction of recombinant nonallergenic soybean protein useable for both immunotherapy and the potential production of hypoallergenic soybean plants.     

Proteome-level investigation of Brassica carinata-derived resistance to Leptosphaeria maculans

Plants resistant to the fungal pathogen Leptosphaeria maculans were generated by an interspecific cross between the highly susceptible Brassica napus (canola) and the highly resistant Brassica carinata. Changes in the leaf protein profiles of these lines were investigated in order to understand the biochemical basis for the observed resistance. Two-dimensional electrophoresis followed by tandem mass spectrometry led to the identification of proteins unique to the susceptible (5 proteins) and resistant genotypes (7 proteins) as well those that were differentially expressed in the resistant genotype 48 h after challenge with the pathogen (28 proteins). Proteins identified as being unique in the resistant plant material included superoxide dismutase, nitrate reductase, and carbonic anhydrase. Photosynthetic enzymes (fructose bisphosphate aldolase, triose phosphate isomerase, sedoheptulose bisphosphatase), dehydroascorbate reductase, peroxiredoxin, malate dehydrogenase, glutamine synthetase, N-glyceraldehyde-2-phosphotransferase, and peptidyl-prolyl cis-trans isomerase were observed to be elevated in the resistant genotype upon pathogen challenge. Increased levels of the antioxidant enzyme superoxide dismutase were further validated and supported by spectrophotometric and in-gel activity assays. Other proteins identified in this study such as nitrate reductase and peptidylprolyl isomerase have not been previously described in this plant-pathogen system, and their potential involvement in an incompatible interaction is discussed.     

Accumulation of Cadmium and Antioxidant and Hormonal Responses in the Indian Major Carp <Emphasis Type="Italic">Cirrhinus mrigala</Emphasis> During Acute and Sublethal Exposure

Acute (24 h) and sublethal (35 days) effects of cadmium chloride (CdCl₂) were examined in Cirrhinus mrigala using various endpoints (accumulation pattern, thyroid hormones (THs), and antioxidants). The mean concentrations of CdCl₂ for 24 and 96 h were found to be 35.974 and 22.387 mg L^?l, respectively. LC50 concentration of CdCl₂ for 24 h (35.97 mg L^?l) was used for the acute study. For the sublethal studies, fish were exposed to 3.59 mg L^?1 (Treatment I) and 7.19 mg L^?1 (Treatment II) corresponding to 1/10th and 1/5th of 24 h LC50 of the CdCl₂. During acute exposure, higher accumulation of CdCl₂ was noticed in the gill, liver, and kidney of C. mrigala, which is found in the order gill > liver > kidney tissues. Similarly, in sublethal treatments (Treatment I and II), a concentration and time-dependent increase of CdCl₂ accumulation was noticed in the order of gill > liver > kidney. GSH, GST, and GPx activities were found to be relatively lower from the treated groups in both acute and sublethal treatments. However, LPO activity was significantly increased in CdCl₂-treated fish C. mrigala. Further, plasma T₃ reduction was more pronounced than T₄ in acute study. During sublethal treatments, both T₄ and T₃ levels showed a continuous decrease as the exposure period extended. All the values in this study were statically significant (P < 0.01 and P < 0.05).     

Resveratrol metabolites do not elicit early pro-apoptotic mechanisms in neuroblastoma cells

Resveratrol, a nontoxic polyphenol, has been shown to inhibit tumor growth in a xenograft mouse model of neuroblasoma. However, resveratrol is rapidly metabolized, mainly to its glucuronidated and sulfated derivatives. This study demonstrates that resveratrol alone, and not the glucuronidated or sulfated metabolites, is taken up into tumor cells, induces a rise in [Ca(2+)](i), and ultimately leads to a decrease in tumor cell viability. A new water-soluble resveratrol formulation was delivered directly at the site of the tumor in a neuroblastoma mouse model. The amount of unmodified resveratrol associated with the tumor increased more than 1000-fold. The increase of unmodified resveratrol associated with the tumor resulted in tumor regression. The number of residual tumor cells that remained viable also decreased as the ratio of the metabolites relative to unmodified resveratrol declined.     

Influence of thermal power station effluents on hydrobiology of seawater

The hydrobiological parameters of the waters in the vicinity of ash slurry and cooling water outfall from Ennore Thermal Power Station located on the shore of Bay of Bengal, Madras were determined. In the outfall site, an increase in mean temperature (35 °C), salinity (3.5%), nitrite ? N (25 μg L^?1), ammonia ? N (185 μg L^?1), phosphate ? P (1 mg L^?1), silicate ? SiO₂ (1.65 mg L^?1), chlorophyll a (21.1 mg m^?3), respiration (76% of gross production), suspended solids (3.84 g L^?1), BOD (3.65 mg O₂) and decrease in pH (8.1), dissolved oxygen (DO) (5.5 mg L^?1), nitrate ? N (15 jig L^?1), gross production (16.6 Mg C m^?3 hr^?1) were recorded. Phaeopigment and respiration were inversely related with chlorophyll a and net productivity, respectively. Microbial biomass varied spatially, but was generally low (114.5 jig L^?1), at the outfall site. The hydrobiological characteristics of water with high turbidity indicate that the ash slurry and coolant water play a crucial role in a localized area resulting in environmental disturbance. The effect was profoundly influenced by an initial dilution, dispersion, wave mixing and current direction. The ecological relationships between these parameters and their implications in coastal pollution are discussed.     

Arsenic Removal from Drinking Water using Iron Oxide-Coated Sand

This article describes experiments in which iron oxide-coated sand(IOCS) was used to study the removal of both As(V) and As(III) to a level less than 5 μg L^-1 in drinking water. Iron oxide-coated sand 2 (IOCS-2) prepared through high temperature coating process was used in batch and column studies to assess the effectiveness and suitability. The isotherm study results showed that the observed data fitted well with the Langmuir model, and the adsorption maximum for IOCS-2 at pH 7.6 was estimated to be 42.6 and 41.1 μg As g^-1 IOCS-2 for As(V) and As(III), respectively. In the fixed bed column tests to study arsenic removal from the tap water, good performance ofIOCS-2 was observed in respect of bed volumes achieved and arsenic removal capacity. Five cycles of column tests were conducted to evaluate the performance of IOCS-2, and arsenic wassuccessfully recovered from the media through regeneration and backwash operations. High bed volumes (860 to 1403) up to a breakthrough concentration of 5 μg L^-1 were achieved inthe column studies with tap water, and the bed volumes achievedin the studies with natural water (containing arsenic) were 1520.The results of both the batch and column studies showed that ironoxide-coated sand filtration could be effectively used to achieveless than 5 μg L^-1 As in drinking water.     

Nitrogen lowers the sulfur amino acid content of soybean (Glycine max [L.] Merr.) by regulating the accumulation of Bowman-Birk protease inhibitor

Soybeans in general contain 35-40% protein. Efforts are underway to increase further this protein content, thus enhancing their nutritive value. Even though higher protein is a desirable characteristic, whether such an increase will be accompanied by enhanced protein quality is not known. Soybean protein quality could be significantly improved by increasing the concentration of the sulfur-containing amino acids, cysteine and methionine. To ascertain if a correlation existed between protein quantity and quality, a comparison of the amino acids of soybeans differing in protein content was made. Soybeans with higher protein content had a significantly lower percentage of sulfur amino acids, while those with lower protein exhibited a higher content of cysteine and methionine. Nitrogen application elevated the protein content but lowered that of the sulfur amino acids. Transmission electron microscopy examination of thin sections of low protein soybean seeds revealed several protein storage vacuoles that were partially filled with storage proteins. Fluorescence two-dimensional difference gel electrophoresis of soybean seed proteins revealed that nitrogen application favored the accumulation of the beta-subunit of beta-conglycinin while decreasing the accumulation of Bowman-Birk protease inhibitor (BBI), a protein rich in cysteine. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of 60% 2-propanol-extracted proteins showed a drastic reduction in the accumulation of BBI with increasing protein content. Northern blot analysis indicated that nitrogen had a negative influence on the expression of the BBI gene. Our results indicate that the negative correlation between total protein and sulfur amino acid content is mostly mediated by the differential accumulation of BBI.     

Determination of spore inactivation during thermal and pressure-assisted thermal processing using FT-IR spectroscopy

The efficacy of microbial inactivation techniques is currently tested using time-consuming and labor-intensive plate count methods, which are the principal rate-limiting steps in developing inactivation kinetic parameters for alternative food processing technologies. Fourier transform infrared (FT-IR) spectroscopy combined with multivariate analysis was used to quantify viable spores and identify some biochemical changes in samples treated by autoclaving, pressure-assisted thermal processing (PATP), and thermal processing (TP). Spore suspensions ( approximately 109 CFU/mL) of Bacillus amyloliquefaciens TMW 2.479 Fad 82, B. amyloliquefaciens TMW 2.482 Fad 11/2, B. sphaericus NZ 14, B. amyloliquefaciens ATCC 49764, and Clostridium tyrobutyricum ATCC 25755 were treated by PATP (121 degrees C and 700 MPa) for 0, 10, 20, and 30 s and by TP (121 degrees C) for 0, 10, 20, and 30 s. The concentrations of spores in treated samples were determined by plating (reference method). Models developed using partial least-squares regression (PLSR) for predicting spore levels in treated samples had correlation coefficients (r) of >0.99 and standard errors of cross-validation ranging between 100.2 and 100.5 CFU/mL. Changes in dipicolinic acid (DPA) and secondary structure of proteins were found to occur during inactivation of spores by PATP and TP. FT-IR spectroscopy could rapidly estimate viable bacterial spore levels in PATP- and TP-treated spore suspensions, providing an accurate analytical tool for monitoring the efficacy of sterilization techniques in inactivating spore-forming microorganisms.