Establisment of quantitative analysis of multi-components by singer marker for content determination of 4 carotenoids in Lycium barbarum / 中国药房

OBJECTIVE To establish a method for simultaneous determination of zeaxanthin ，β-carotene，β-cryptoxanthin palmitate and zeaxanthin dipalmitate in Lycium barbarum . METHODS L. barbarum was extracted with n-hexane-anhydrous ethanol-acetone-toluene（10∶6∶7∶7，V/V/V/V）by ultrasonic method. High performance liquid chromatography （HPLC）method was adopted. The determination was performed on YMC C 30 column with mobile phase A consisted of methanol-acetonitrile-water （81∶ 14 ∶ 5，V/V/V）and mobile phase B consisted of dichloromethane （gradient elution ）at the flow rate of 1.0 mL/min. The column temperature was set at 20 ℃. The detection wavelength was set at 450 nm，and sample size was 20 μL. Using zeaxanthin as control，the relative correction factors （RCFs）of β-carotene，β-cryptoxanthin palmitate and zeaxanthin dipalmitate were calculated ， and then the content of each component was calculated according to RCFs and compared with the results of external standard method（ESM）. RESULTS The linear range of zeaxanthin ，β-carotene，β-cryptoxanthin palmitate and zeaxanthin dipalmitate were 0.119 4-2.983 8，0.121 7-1.521 6，0.285 9-5.718 8，8.460 5-211.513 3 μg/mL（all R2＞0.999）. RSDs of precision ，repeatability and stability（16 h）tests were all less than 4％. The average recoveries were 103.34％，107.37％，105.64％，96.16％（RSD＜5％，n＝ 9）. The average RCFs of β-carotene，β-cryptoxanthin palmitate and zeaxanthin dipalmitate were 1.109，1.390，1.158. The relative errors of the content determination results by quantitative analysis of multi-components by singer marker （QAMS）and ESM were within ±1％. CONCLUSIONS The established HPLC-QAMS method is accurate and stable ，which can be used for the content determination and quality control of 4 carotenoids in L. barbarum .

Chemistry of California Lycium cooperi and Lycium andersonii

Aims: To examine the chemistry of two California Lycium species and evaluate the possible use of California Lycium species as dietary supplements especially for age related macular degeneration. Study Design: This exploratory analytical research used samples of Lycium andersonii and Lycium cooperi collected in the field and analyzed in the lab. Place and Duration of Study: University of Southern California School of Pharmacy, 1985 Zonal Avenue, Los Angeles, CA USA 90089. Methodology: Plant extracts were analyzed by high pressure liquid chromatography mass spectrometry with ultraviolet photodiode array detection in order to identify the chemical characteristics of compounds found in the plants. Results: Several known compounds were found in extracts of Lycium cooperi and Lycium andersonii foliage and fruit including: zeaxanthin, zeaxanthin monopalmitate and β-cryptoxanthin. The various California species of Lycium are discussed as possible alternatives to Chinese Lycium barbarum. Conclusion: California Lycium berries may be suitable substitutes for Chinese Lycium berries.

Zeaxanthin from Porphyridium purpureum induces apoptosis in human melanoma cells expressing the oncogenic BRAF V600E mutation and sensitizes them to the BRAF inhibitor vemurafenib

Abstract Zeaxanthin, an abundant carotenoid present in fruits, vegetables and algae was reported to exert antiproliferative activity and induce apoptosis in human uveal melanoma cells. It also inhibited uveal melanoma tumor growth and cell migration in nude mice xenograft models. Here we report that zeaxanthin purified from the rhodophyte Porphyridium purpureum (Bory) K.M.Drew & R.Ross, Porphyridiaceae, promotes apoptosis in the A2058 human melanoma cell line expressing the oncogenic BRAF V600E mutation. Zeaxanthin 40 µM (IC50) induced chromatin condensation, nuclear blebbing, hypodiploidy, accumulation of cells in sub-G1 phase, DNA internucleosomal fragmentation and activation of caspase-3. Western blot analysis revealed that zeaxanthin induced up-regulation of the pro-apoptotic factors Bim and Bid and inhibition of NF-κB transactivation. Additionally, zeaxanthin sensitized A2058 melanoma cells in vitro to the cytotoxic activity of vemurafenib, a BRAF inhibitor widely used for the clinical management of melanoma, suggesting its potential interest as dietary adjuvant increasing melanoma cells sensitivity to chemotherapy.

Cloning and expression analysis on full length CDS of zeaxanthin epoxidase gene in Pseudostellaria heterophylla / 中国中药杂志

Zeaxanthin epoxidase plays an important role in indirect pathway of plant abscisic acid biosynthesis. According to the data of Pseudostellaria heterophylla transcriptome, zeaxanthin epoxidase gene was isolated and named as PhZEP. The results of bioinformatics analysis showed that the coding sequence of PhZEP was 1 263 bp long and encoded 420 amino acids. The putative protein molecular weight was 47.34 kDa and its theoretical isoelectric point was 6.64. The characteristic structure domains were predicted, including binding site of lipoprotein and flavoprotein monooxyenase. A signal peptide was discovered at the N-terminal of amino acids. The Real-time PCR revealed that PhZEP had a higher expression level in leaves than other tissues of P.heterophylla. Highly expressed PhZEP was also observed at 10 d and 40 d tuberous root after flowering. PhZEP presented a different expression after treatment with ABA, fluridone and ABA +fluridone compared to the control. The expression of PhZEP in tuberous root after ABA treatment was close to that in control group, while PhZEP showed significant up-regulation in the fluridone treatment group. In this study, the PhZEP gene from P. heterophylla was cloned and this result has important significance for its functional identification. This research provides a basis for the further analysis on functional mechanism of ABA during development of P. heterophylla.

Multimodality imaging in macular telangiectasia 2: A clue to its pathogenesis.

Macular telangiectasia type 2 also known as idiopathic perifoveal telangiectasia and juxtafoveolar retinal telangiectasis type 2A is an acquired bilateral neurodegenerative macular disease that manifests itself during the fifth or sixth decades of life. It is characterized by minimal dilatation of the parafoveal capillaries with graying of the retinal area involved, a lack of lipid exudation, right‑angled retinal venules, refractile deposits in the superficial retina, hyperplasia of the retinal pigment epithelium, foveal atrophy, and subretinal neovascularization (SRNV). Our understanding of the disease has paralleled advances in multimodality imaging of the fundus. Optical coherence tomography (OCT) images typically demonstrate the presence of intraretinal hyporeflective spaces that are usually not related to retinal thickening or fluorescein leakage. The typical fluorescein angiographic (FA) finding is a deep intraretinal hyperfluorescent staining in the temporal parafoveal area. With time, the staining may involve the whole parafoveal area but does not extend to the center of the fovea. Long‑term prognosis for central vision is poor, because of the development of SRNV or macular atrophy. Its pathogenesis remains unclear but multimodality imaging with FA, spectral domain OCT, adaptive optics, confocal blue reflectance and short wave fundus autofluorescence implicate Müller cells and macular pigment. Currently, there is no known treatment for this condition.

Two Eggs per Day Increase Plasma Lutein and Zeaxanthin in a Pediatric Population Characterized by Low Intake of Fruits and Vegetables.

Aims: Eggs are a good source of lutein and zeaxanthin, carotenoids known for their antioxidant properties. Mexican children have been shown to consume limited amounts of fruit and vegetables. The purpose of the current study is to determine whether the inclusion of eggs in the diet increases plasma carotenoids in this population. Study Design: This study is a follow up on the effects of high egg intake on plasma lipids and atherogenic lipoproteins in children. Fifty four Mexican children (25 boys/29 girls) aged 8-12 y were randomly assigned to consume either 2 eggs/d (518 mg additional dietary cholesterol) (EGG period) or the equivalent amount of egg whites (SUB Period) in a cross-over design for 4 wk. After a 3 wk washout, children were crossed over to the alternate treatment. Methodology: 3-day dietary records, plasma carotenoids and apolipoproteins were measured at the end of the EGG and SUB Periods. Results: In agreement with the lack of effects of eggs in increasing atherogenic lipoprotein profiles, plasma apolipoprotein B concentrations did not change between periods indicating that increases in plasma cholesterol were not associated with higher number of LDL particles. Although the values for apo C-III were high compared to other pediatric populations, they were not affected by egg intake. Dietary records indicated low intake of carotenoids, especially during the SUB period. Plasma lutein and zeaxanthin were increased during the EGG period from 0.235 ± 0.071 to 0.280 ± 0.147 μmol/L (P<0.001) and 0.044 ± 0.019 to 0.051 ± 0.031 μmol/L (P<0.001), respectively. Conclusions: These results suggest that the eggs are a good source of lutein and zeaxanthin in this population and that the increases in LDL size during the egg period may also be related to a better transport of these carotenoids in plasma.

Role of lutein in preventing/slowing down age-related macular degeneration: a Meta-analysis / 中华临床营养杂志

Objective To evaluate the role of lutein in preventing and slowing down age-related macular degeneration (AMD). Methods Literatures concerning lutein and AMD were collected by computerized search, manual search, expert consultations. The quality of included studies was evaluated. RevMan software 4.2was used for meta-analysis. Results Totally 9 studies were included, with a total OR of 0. 81 (range: 0. 71-0.92). Conclusion Lutein can, to some extent, prevent or slow down AMD.

Transgenic Tobacco Expressing the Crocus CSzCD Gene Showed the Production of Crocetin / 中国生物工程杂志

To investigate the feasibility to produce crocetin in tobacco plants.The coding region of zeaxanthin cleavage dioxygenase (CSzCD) gene from Crocus sativus L. was inserted into the downstream of the cauliflower mosaic virus (CaMV) 35S promoter of a binary vector pBI121, and integrated into the genome of Nicotiana tabacum L. Twenty-one transgenic lines were identified by genomic southern blotting analysis. Western blotting and HPLC analysis of the leaf extracts of transgenic tobacco showed that crocetin was produced in CSzCD gene-transformed plants, while no crocetin was found in the untransformed tobacco.

Factors Effecting the Bioavailability of Carotenoid in Elderly Korean Women / 대한지역사회영양학회지

Carotenoid-rich foods focus one's attention on the prevention age-related diseases. This study was conducted to investigate the carotenoid status and look into the factors that affect the bioavailability of carotenoid in 121 elderly nonsmoking Korean women. Carotenoids and lipids in plasma, and nutrient intakes including carotenoid were studied. The mean plasma total-cholesterol, HDL-cholesterol, LDL-cholesterol and triacylglycerol concentrations were 220.0 mg/dl, 49.5 mg/dl, 139.2 mg/dl and 157.4 mg/dl, respectively. Significantly positive correlations were found between the plasma lutein + zeaxanthin, lycopene and beta-carotene concentrations and the intake of fruits (r = 0.17, r = 0.20, r = 0.19). However, significantly negative correlations were found between the plasma lutein+zeaxanthin, and beta-carotene concentrations that adjusted for carotenoid intakes and intakes of vegetables (r = - 0.21, r = - 0.19), and between plasma lutein+zeaxanthin, lycopene and beta-carotene concentrations that adjusted for carotenoid intakes and intakes of fruits (r = - 0.21, r = - 0.18, r = - 0.24). After the adjustment for plasma lipids, there was no correlation between the plasma carotenoid concentrations and the carotenoid-rich foods. However, after adjustment for fiber intake, significantly strong positive correlations were found between the plasma carotenoid concentrations and carotenoid-rich foods. The plasma levels of carotenoid biomarkers (plasma carotenoid concentrations adjusted for dietary fiber intakes) decreased with age, and the plasma levels of lycopene biomarkers (plasma lycopene concentrations adjusted for dietary fiber intakes) increased with regular exercise. However alcohol drinking had no impact. These results suggested that age, physical activity, and dietary fiber intake affected the bioavailability of carotenoid. Therefore, when the elderly have carotenoid-rich foods, they should consider ways of increasing the bioavailability of carotenoid through cooking methods and physical activity.

THE DETERMINATION AND CONTENTS OF LUTEIN,ZEAXANTHIN AND ?-CAROTENE IN VEGETABLES IN BEIJING REGION / 营养学报

Objective To determine the contents of lutein,zeaxanthin and ?-carotene in 31 vegetables.Method High performance liquid chromatography method with Develosil C30 column and ultraviolet detector was used to detect lutein,zeaxanthin and ?-carotene at wavelength of 480nm.A gradient elution method with elution A (acetonitrile:methol=75:25) and B (methyl-tert-butyl-ether) was used to separate the carotenoids in both raw and cooked vegetables.Results The retention time of lutein,zeaxanthin and ?-carotene was 10.355,10.732 and 20.789 min respectively.The contents of lutein,zeaxanthin and ?-carotene were linearly related with the peak area.Analysis showed that dark leafy vegetable such as amaranth,Chinese celery leaf,parsley,spinach and pakchoi petiole contained the highest amount of lutein.Zeaxanthin was detected only in dehydrated daylily.There was little or no loss of lutein,zeaxanthin and ?-carotene but an average increase in lutein and ?-carotene in vegetables after cooking.Conclusion Vegetables,especially dark leafy vegetables are major sources of dietary lutein,and ?-carotene.Consuming cooked vegetables tends to increase the intakes of lutein,zeaxanthin and their bioavailability.