Immunohistochemical localization of phospholipase D in strawberry (Fragaria ananassa Duch.) fruits

Phospholipase D, the key enzyme involved in membrane phospholipid catabolism, was localized in strawberry (Fragaria ananassa Duch.) fruits during development using immunohistochemical techniques. Typical structural features such as the nucleus, cell wall, plasma membrane, mitochondria and vacuole were noticeable in cells of young strawberry fruits. Cytoplasmic structural features were less pronounced in fully grown and turning red fruits, except cell wall, which was considerably enlarged. Also, extensive microvesiculation from the plasma membrane was noticeable in ripening fruits. The number of visible gold particles indicative of phospholipase D increased during development. Phospholipase D was found to be associated with membranous structures and vacuole. Interestingly, gold-particles could also be observed in the cell wall space in ripening fruits suggesting that cytosolic contents may have leaked into the cell wall space as a result of membrane deterioration and loss of compartmentalization. The role of phospholipase D during development and ripening of strawberry, a non-climacteric fruit, is discussed.     

Physico-chemical characteristics of nanovesicle-carbohydrate complexes in grape juice concentrate

It is generally assumed that polyphenols, such as anthocyanins in fruit juice, exist in a free soluble state and are readily available for absorption in the gastro-intestinal tract. In the present study, we have investigated the interaction of polyphenols with soluble carbohydrate polymers, such as pectin and lipid nanovesicles, that are generated during homogenization of the fruit tissue during juice extraction. A commercially available grape juice concentrate contained nearly 25% of polyphenol fraction bound to macromolecules that were nondialyzable. Treatment of dialyzed juice with cellulase, pectinase, and beta-galactosidase did not cause the release of bound polyphenols; however, treatment with triton X-100 caused an increased release of bound polyphenols. The dialyzate contained relatively more -3-O glucosides and -3-O-acetoyl glucosides in comparison to the bound fraction which was enriched in -3-O-coumaroyl glucosides, suggesting qualitative differences in the bound and the free anthocyanin composition. Electron microscopic analysis of the juice fractions revealed the presence of electron-dense nanovesicle-fiber complexes ranging from 10 to 200 nm in diameter. Such complexes were absent in the dialyzate fraction. Cellulase treatment did not change the morphology of the complexes; however, treatment with pectinase and beta-galactosidase disrupted the complexes, releasing vesicular structures, suggestive of the pectin nature of the fibrous matrix. The dialyzed and the dialyzate fractions also showed differences in their 1H NMR and fluorescence spectral characteristics. The dialyzed fraction containing polyphenol-pectin complexes showed no superoxide scavenging capacity, reduced hydroxyl radical scavenging activity, and high 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity, indicating potential changes in functionality because of the complex formation.     

Effects of phosphorus fertilizer supplementation on processing quality and functional food ingredients in tomato

Even though several types of phosphorus fertilizers are used in crop production, the influence of phosphorus on produce quality is not well understood. Several quality attributes of tomato juice were analyzed in relation to phosphorus supplementation during a three-year field study (2000-2002). In addition to the recommended phosphorus fertilization, phosphorus supplementations, either through soil (low and high) or through foliar spray (hydrophos, seniphos), were tested. In general, soil and foliar phosphorus supplementation did not provide a statistically significant increase in yield. Tomato juice was evaluated for various quality characteristics including pH, titratable acidity, precipitate weight ratio, total solids, serum viscosity, Brookfield viscosity, color, lycopene levels, vitamin C, and flavor volatiles. Changes observed in several quality parameters were marginal, statistically insignificant and influenced by the season. Therefore, it appears that phosphorus supplementation may not significantly affect the processing quality parameters in tomato fruits.     

Effect of processing conditions on phospholipase D activity of corn kernel subcellular fractions.

The influence of physicochemical conditions on the phospholipase D (PLD) activity of subcellular preparations of sweet corn (Zea mays L. cv. Peaches and Cream) kernels has been studied. The microsomal, mitochondrial, and cytosolic preparations of corn kernels possessed PLD activity albeit at varying proportions. The microsomal and cytosolic PLD activities were stimulated 2-fold between 5 and 15 degrees C. Ethanol had varying modulatory effects on PLD activity. By contrast, acetaldehyde was a potent inhibitor of PLD. As well, a naturally occurring C(6) aldehyde such as hexanal and an alcohol such as hexanol inhibited PLD activity efficiently. Divalent cations such as calcium chloride and magnesium chloride stimulated PLD activity at micromolar levels. Monovalent cations such as KCl and NaCl did not appear to affect PLD activity. Partial purification of PLD from the microsomal, mitochondrial, and cytosolic fractions separately revealed four major isoforms with relative molecular masses of 200, 140-150, 102-108, and 60-66 kDa. The importance of PLD in the maintenance of processed food quality is discussed.     

草莓果实磷脂酶D活性的测定

以草莓果实为试材,建立了适合草莓果实组织磷脂酶D制备及活性测定的技术体系。以放射标记的磷脂酰胆碱为底物,在1mL反应体系中,膜蛋白的含量以5-10μg为宜;25℃反应温度下,反应时间以5-10min为宜;反应体系的适宜pH为6.5-7。对草莓果实不同组分磷脂酶D酶学特性的进一步分析认为,该技术体系的建立有助于进一步探讨磷脂酶D在草莓果实发育成熟和衰老进程中的作用。     

Hexanal and 1-MCP treatments for enhancing the shelf life and quality of sweet cherry (Prunus avium L.)

Sweet cherry has a short post-harvest shelf life and this greatly affects the consumer preference and export of fresh fruits. In this study, the effects of pre-harvest application of a hexanal formulation (enhanced freshness formulation, EFF) and post-harvest application of hexanal vapour and 1-MCP on quality parameters and shelf life of sweet cherry were investigated. Cherries subjected to pre-harvest spray with EFF had better color, brightness and firmness than unsprayed cherries even after 30 days of storage at 4 °C. These EFF-treated cherries also showed higher chroma values indicative of enhanced red color. Post-harvest application of either, hexanal vapour, 1-MCP, or a combination of both, enhanced the firmness of cherries. These treatments also resulted in higher levels of superoxide dismutase and ascorbate peroxidase activities. The levels of anthocyanins and phenolic components were either enhanced or maintained during the 30-day storage period. Our results suggest that a pre-harvest application of EFF combined with post-harvest treatment of hexanal and 1-MCP may enhance the quality and shelf life of sweet cherry.     

Effects of phosphorus fertilizer supplementation on antioxidant enzyme activities in tomato fruits

The effects of soil and foliar phosphorus supplementation on the activities and levels of superoxide dismutase (SOD), guaiacol peroxidase (POX), and ascorbate peroxidase (APX) in tomato fruits were evaluated by determining enzyme activities and isoenzyme analysis. Both protein levels and enzyme activities varied depending on the variety and season. In general, phosphorus supplementation did not alter SOD, POX, and APX activities significantly;however, some treatments showed season- and stage-specific enhancement in activities as noticed with hydrophos and seniphos supplementation. Three different SOD isozymes were observed, and these isozymes showed very similar staining intensities in response to P application and during the three developmental stages studied. Two major isozymes of POX and two different APX isozymes were observed at all the developmental stages. The results suggest that antioxidant enzyme activities may be influenced by the availability of phosphorus, but are subject to considerable variation depending on the developmental stage and the season.     

Treatment of mcf-7 breast cancer cells with a red grape wine polyphenol fraction results in disruption of calcium homeostasis and cell cycle arrest causing selective cytotoxicity

Food components influence the physiology by modulating gene expression and biochemical pathways within the human body. The disease-preventive roles of several fruit and vegetable components have been related to such properties. Polyphenolic components such as flavonoids are strong antioxidants and induce the expression of several xenobiotic-detoxifying enzymes. The mechanism of selective cytotoxicity induced by red grape wine polyphenols against MCF-7 breast cancer cells was investigated in relation to their interference with calcium homeostasis. MCF-7 cells showed an increase in cytosolic calcium levels within 10 min of treatment with the polyphenols. Immunohistochemical localization of calmodulin with secondary gold-labeled antibodies showed similar levels of gold labeling in both MCF-7 cells and the spontaneously immortalized, normal MCF-10A cell line. MCF-7 cells treated with the red wine polyphenol fraction (RWPF) showed swelling of endoplasmic reticulum, dissolution of the nucleus, and loss of plasma membrane integrity as well as reduced mitochondrial membrane potential. These cells were arrested at the G2/M interphase. By contrast, MCF-10A cells did not show such changes after RWPF treatment. The results suggest that polyphenol-induced calcium release may disrupt mitochondrial function and cause membrane damage, resulting in selective cytotoxicity toward MCF-7 cells. This property could further be developed toward breast cancer prevention strategies either independently or in conjunction with conventional prevention therapies where a positive drug-nutrient interaction can be demonstrated.