Determination of lycopene in tissues and plasma of rats by normal-phase high-performance liquid chromatography with photometric detection

An analytical method for the determination of lycopene in tissues and plasma of rats is described. The method was validated for the determination of lycopene in liver and plasma with respect to selectivity, linearity, accuracy, recovery and precision. Following precipitation of proteins with water–ethanol plasma was extracted with hexane; tissues were extracted with acetone followed by precipitation of proteins with water–ethanol and extraction of lycopene with hexane. Separation and quantification of geometrical isomers of lycopene was achieved by normal-phase HPLC with UV/VIS detection at 471 nm. The method proved to be selective and specific for lycopene in plasma and liver. Detector response was linear in the range from 2 ng/g to 10 μg/g liver and 0.5 ng/ml to 2 μg/ml plasma, respectively. Average recoveries ranged from 96 to 101% in spiked liver samples and from 91 to 94% in spiked plasma samples. Intra-day variability (C.V.) was ≤6% and ≤5% in liver and plasma, respectively. Inter-day precision was ≤9% for liver samples and ≤6% for plasma samples. The procedures were successfully applied to the sample analysis of pharmacokinetic and metabolism studies.     

Lycopene and apo-10'-lycopenal do not alter DNA methylation of GSTP1 in LNCaP cells

DNA hypermethylation and silencing of tumor-suppressor genes are commonly seen in human cancers, and likely contribute to the process of carcinogenesis. A growing body of evidence suggests that dietary compounds may alter cancer risk through epigenetic modifications. Glutathione S-transferase P1 (GSTP1) is hypermethylated in >90% of prostate cancer cases making it one of the most common genome alterations in prostate cancer. LNCaP cells were treated either with lycopene or apo-10′-lycopenal for 7 days, and mRNA expression and DNA methylation of GSTP1 were evaluated. Neither compound significantly altered expression nor methylation of GSTP1 while treatment with 5-azacytidine decreased methylation by 78%. These findings demonstrate that lycopene and apo-10′-lycopenal are not effective demethylating agents of GSTP1 in the human LNCaP cell line.     

Tissue-specific accumulation of carotenoids in carrot roots

Raman spectroscopy can be used for sensitive detection of carotenoids in living tissue and Raman mapping provides further information about their spatial distribution in the measured plant sample. In this work, the relative content and distribution of the main carrot (Daucus carota L.) root carotenoids, α-, β-carotene, lutein and lycopene were assessed using near-infrared Fourier transform Raman spectroscopy. The pigments were measured simultaneously in situ in root sections without any preliminary sample preparation. The Raman spectra obtained from carrots of different origin and root colour had intensive bands of carotenoids that could be assigned to β-carotene (1,520 cm⁻¹), lycopene (1,510 cm⁻¹) and α-carotene/lutein (1,527 cm⁻¹). The Raman mapping technique revealed detailed information regarding the relative content and distribution of these carotenoids. The level of β-carotene was heterogeneous across root sections of orange, yellow, red and purple roots, and in the secondary phloem increased gradually from periderm towards the core, but declined fast in cells close to the vascular cambium. α-carotene/lutein were deposited in younger cells with a higher rate than β-carotene while lycopene in red carrots accumulated throughout the whole secondary phloem at the same level. The results indicate developmental regulation of carotenoid genes in carrot root and that Raman spectroscopy can supply essential information on carotenogenesis useful for molecular investigations on gene expression and regulation.     

大孔树脂对三孢布拉霉菌产番茄红素的吸附性能研究

为利用大孔吸附树脂分离纯化三孢布拉霉菌发酵液番茄红素,考察了4种大孔树脂(HP20, HP2MG, SP825, SP207)对番茄红素的吸附 解吸特性,筛选出最佳树脂。结果表明,HP20大孔树脂对番茄红素具有较好的吸附 解吸性能,且吸附迅速,能在1h内达到吸附平衡。用含55%和60%异丙醇的丙酮不连续梯度洗脱,回收率可达89.55%,重结晶后可获得纯度95%以上的番茄红素,经红外、质谱、核磁等鉴定为天然型番茄红素。该方法简单,可操作性强,极具工业应用价值。     

Expression profiling by whole-genome microarray hybridization reveals differential gene expression in breast cancer cell lines after lycopene exposure

The correlation between diet and variation in gene-expression is an important field which could be considered to approach cancer pathways comprehension. We examined the effects of lycopene on breast cancer cell lines using pangenomic arrays. Lycopene is derived predominantly from tomatoes and tomato products and there is some epidemiologic evidence for a preventive role in breast cancer. Previously, we investigated lycopene in breast cancer using a dedicated breast cancer microarray. To confirm these results and explore pathways other than those implicated in breast cancer, for this study we used pangenomic arrays containing 25,000 oligonucleotides. This in vitro study assayed two human mammary cancer cell lines (MCF-7 and MDA-MB-231), and a fibrocystic breast cell line (MCF-10a) treated or not with 10 μM lycopene for 48 h. A competitive hybridization was performed between Cy3-labeled lycopene treated RNA and Cy5-labeled untreated RNA to define differentially expressed genes. Using t-test analysis, a subset of 391 genes was found to be differentially modulated by lycopene between estrogen-positive cells (MCF-7) and estrogen-negative cells (MDA-MB-231, MCF-10a). Hierarchical clustering revealed 726 discriminatory genes between breast cancer cell lines (MCF-7, MDA-MB-231) and the fibrocystic breast cell line (MCF-10a). Modified gene expression was observed in various molecular pathways, such as apoptosis, cell communication, MAPK and cell cycle as well as xenobiotic metabolism, fatty acid biosynthesis and gap junctional intercellular communication.     

Structural diversity and functional novelty of new carotenoid biosynthesis genes

Many new carotenoid synthesis genes have recently been identified through genomic sequencing or functional cloning. Some of them exhibit novel structures and/or novel functions. This review describes such examples in the families of lycopene β-cyclases, putative homologues of phytoene dehydrogenases and new carotenoid hydroxylases. Both the functionally novel lycopene β-monocyclases and structurally novel fusion-type of lycopene β-cyclases were described. Another newly discovered sequence of lycopene β-cyclase described might represent a new class of lycopene β-cyclases previously not identified in several cyanobacteria. Three examples of putative homologues of phytoene dehydrogenases were described, however, they were confirmed to encode different and/or new functions such as β-carotene ketolase, 4,4′-diapolycopene oxygenase or prolycopene isomerase. Two new carotenoid hydroxylase genes were described that encoded the new function of 2,2′-β-ionone ring hydroxylase or 3,3′-isorenieratene hydroxylase. Phylogenetic analysis of these genes shed light on their possible evolutionary origins. These new genes also provide tools for synthesis of novel and desirable carotenoids by genetic engineering.     

Improvement of antioxidant and defense properties of Tomato (var. Pusa Rohini) by application of bioaugmented compost

Nutrient management practices play a significant role in improving the nutritional quality of tomato. The present study deals with the evaluation of compost prepared using Effective Microorganisms (EM), on antioxidant and defense enzyme activities of Tomato (Lycopersicon esculentum). A field experiment with five treatments (control, chemical fertilizer and EM compost alone and in combination) was conducted in randomized block design. An increment of 31.83% in tomato yield was recorded with the combined use of EM compost and half recommended dose of chemical fertilizers (N₅₀P₃₀K₂₅ + EM compost at the rate of 5 t ha⁻¹). Similarly, fruit quality was improved in terms of lycopene content (35.52%), antioxidant activity (24–63%) and defense enzymes activity (11–54%), in tomatoes in this treatment as compared to the application of recommended dose of fertilizers. Soil microbiological parameters also exhibited an increase of 7–31% in the enzyme activities in this treatment. Significant correlation among fruit quality parameters with soil microbiological activities reveals the positive impact of EM compost which may be adopted as an eco-friendly strategy for production of high quality edible products.     

Effects of lycopene supplementation on antioxidant status,oxidative stress,performance and carcass characteristics in heat-stressed Japanese quail

Lycopene, a carotenoid present predominantly in tomatoes, is one of the most efficient antioxidants. This experiment was conducted to evaluate the effects of dietary lycopene supplementation on performance, carcass characteristics, biomarkers of oxidative stress (malondialdehyde (MDA) and homocysteine), and concentrations of vitamins C, E, A, cholesterol, triglyceride, and glucose in Japanese quails (Coturnix coturnix Japonica) exposed to high-ambient temperature of 34 °C. Two hundred and forty Japanese quails (10 day-old) were randomly assigned to eight treatment groups consisting of 10 replicates of three birds. The birds were kept at a temperature-controlled room at 22 °C (Thermoneutral, TN groups) or 34 °C (for 12 h/day; 09.00 am–05.00 pm; Heat stress, HS groups). Birds were fed either a basal (control) diet (TN and HS) or the basal diet supplemented with 50, 100 or 200 mg of lycopene/kg of diet. Lycopene supplementation linearly increased feed intake (P=0.05$P=0.05$), live weight gain (P=0.01$P=0.01$), feed efficiency (P=0.01$P=0.01$) and cold carcass weight (P=0.01$P=0.01$) and yield (P=0.05$P=0.05$) under heat stress conditions but did not show the same effect at thermoneutral conditions (P>0.05$P>0.05$). The interaction Serum vitamin C, E, and A (P=0.01$P=0.01$) concentrations increased linearly in birds reared at high temperature while non-significant changes occurred at TN groups. Homocysteine level in serum and malondialdehyde (MDA) levels in serum, liver, and heart (P=0.001$P=0.001$) linearly decreased in all birds of both TN and HS groups as dietary lycopene supplementation increased. Heat stress-induced increase in serum cholesterol (P=0.01$P=0.01$), triglycerides (P=0.05$P=0.05$) and glucose (P=0.01$P=0.01$) concentrations were linearly reversed by lycopene supplementation. Supplementation of lycopene increased the HDL concentration whereas, the VLDL and LDL concentrations reduced with lycopene supplementation (P=0.01$P=0.01$, linear), particularly at a dietary concentration of 200 mg/kg. Lycopene could not be detected in control birds while a linear increase was observed in the sera of lycopene supplemented birds The results of the study indicate that lycopene supplementation attenuated the increase in oxidative stress and depletion in antioxidants caused by heat stress in Japanese quails.     

Prolonged root hypoxia induces ammonium accumulation and decreases the nutritional quality of tomato fruits

Here we examined the effects of root hypoxia (1-2% oxygen) on the physiology of the plant and on the biochemical composition of fruits in tomato (Solanum lycopersicum cv. Micro-Tom) plants submitted to gradual root hypoxia at first flower anthesis. Root hypoxia enhanced nitrate absorption with a concomitant release of nitrite and ammonium into the medium, a reduction of leaf photosynthetic activity and chlorophyll content, and an acceleration of fruit maturation, but did not affect final fruit size. Quantitative metabolic profiling of mature pericarp extracts by (1)H NMR showed that levels of major metabolites including sugars, organic acids and amino acids were not modified. However, ammonium concentration increased dramatically in fruit flesh, and ascorbate and lycopene concentrations decreased. Our data indicate that the unfavorable effects of root hypoxia on fruit quality cannot be explained by two of the well-known effects of root hypoxia on the plant, namely a decrease in photosynthesis or an excess in ethylene production, but may instead result from disturbances in the supply of either growth regulators or ammonium, by the roots.