Determination of acylated anthocyanin in human urine after ingesting a purple-fleshed sweet potato beverage with various contents of anthocyanin by LC-ESI-MS/MS

Eighty-seven healthy volunteers ingested a purple-fleshed sweet potato beverage with various contents of anthocyanin (beverage A; 22.1 mg/250 ml, B; 107.8, C; 84.9). An acylated anthocyanin, peonidin 3-caffeoylsophoroside-5-glucoside, was detected in the urine 2 h after ingestion. The concentrations were 15.1+/-2.2 microg/l of urine (mean+/-SEM), 46.6+/-5.3, and 53.3+/-2.2 for beverages A, B, and C respectively.     

CysC,mALB和尿BGM联合检测在2型糖尿病早期肾损中的临床价值

目的:探讨CysC、mALB和尿BGM联合检测在2型糖尿病早期肾损中的临床价值。方法:选取2010年11月至2013年6月我院内分泌科收治的130名2型糠尿病患者,根据空腹血糖值的不同程度将其分为A、B两组,A组72例,B组58例,另选取同期在我院体检的年龄、性别等资料与之匹配的64名健康者设为C组。分别对其CysC、mALB和尿BGM进行测定,并进行对比分析。结果:通过分析可知,A组和B组的Cys C分别为(3.29±1.09)mg/L、(2.86±1.05)mg/L,相对于C组的(0.87±0.99)mg/L,均明显升高,但A组升高更加明显;A组、B组、C组的mALB分别为(37.36±4.82)mg/L、(36.55±4.31)mg/L、(6.61±3.84)mg/L,B组高于A组,C组高于A组和B组;三组的BGM中,以A组最高,为(634.15±55.24)μg/L,B组其次,为(626.92±48.18)μg/L,均高于C组的(57.12±11.75)μg/L;Cys C、mALB、BGM等阳性指标的检出率分别为76.15%、77.69%、72.31%,而Cys C、mALB、BGM联合检测的检出率为93.85%,明显高于其他单指标检测。且P0.05,差异具有统计学意义。结论:对CysC、mALB和尿BGM进行联合检测对2型糖尿病早期肾损患者具有较高的检出率,相对于单指标检测,敏感性更高。     

Anthocyanin biosynthetic genes in Brassica rapa

Background

Anthocyanins are a group of flavonoid compounds. As a group of important secondary metabolites, they perform several key biological functions in plants. Anthocyanins also play beneficial health roles as potentially protective factors against cancer and heart disease. To elucidate the anthocyanin biosynthetic pathway in Brassica rapa, we conducted comparative genomic analyses between Arabidopsis thaliana and B. rapa on a genome-wide level.

Results

In total, we identified 73 genes in B. rapa as orthologs of 41 anthocyanin biosynthetic genes in A. thaliana. In B. rapa, the anthocyanin biosynthetic genes (ABGs) have expanded and most genes exist in more than one copy. The anthocyanin biosynthetic structural genes have expanded through whole genome and tandem duplication in B. rapa. More structural genes located upstream of the anthocyanin biosynthetic pathway have been retained than downstream. More negative regulatory genes are retained in the anthocyanin biosynthesis regulatory system of B. rapa.

Conclusions

These results will promote an understanding of the genetic mechanism of anthocyanin biosynthesis, as well as help the improvement of the nutritional quality of B. rapa through the breeding of high anthocyanin content varieties.

Electronic supplementary material

The online version of this article (doi: 10.1186/1471-2164-15-426) contains supplementary material, which is available to authorized users.     

Instability of anthocyanin accumulation inVitis vinifera L. var. Gamay Fréaux suspension cultures

The inherent instability of metabolite production in plant cell culture-based bioprocessing is a major problem hindering its commercialization. To understand the extent and causes of this instability, this study was aimed at understanding the variability of anthocyanin accumulation during long-term subcultures, as well as within subculture batches, inVitis vinifera cell cultures. Therefore, four cell line suspensions ofVitis vinifera L. var. Gamay Fréaux, A, B, C and D, originated from the same callus by cell-aggregate cloning, were established with starting anthocyanin contents of 2.73±0.15, 1.45±0.04, 0.77±0.024 and 0.27±0.04 CV (Color Value)/g-FCW (fresh cell weight), respectively. During weekly subculturing of 33 batches over 8 months, the anthocyanin biosynthetic capacity was gradually lost at various rates, for all four cell lines, regardless of the significant difference in the starting anthocyanin content. Contrary to this general trend, a significant fluctuation in the anthocyanin content was observed, but with an irregular cyclic pattern. The variabilities in the anthocyanin content between the subcultures for the 33 batches, as represented by the variation coefficient (VC), were 58, 57, 54, and 84% forV. vinifera cell lines A, B, C and D, respectively. Within one subculture, the VCs from 12 replicate flasks for each of 12 independent subcultures were averaged, and found to be 9.7%, ranging from 4 to 17%. High- and low-producing cell lines, VV05 and VV06, with 1.8-fold differences in their basal anthocyanin contents, exhibited different inducibilities tol-phenylalanine feeding, methyl jasmonate and light irradiation. The low-producing cell line showed greater potential in enhanced the anthocyanin production.     

Anthocyanin production in selected cell lines of grape (Vitis vinifera L.)

Summary Anthocyanin production of two lines ofVitis vinifera cell cultures, i.e., 5.4 and 13.1, which were obtained from the same starting material after 20 and 37 mo. of clonal selection, respectively, was investigated. Cell suspension cultures of lines 5.4 and 13.1 maintained an anthocyanin content of 0.44 ± 0.15 and 1.02 ± 0.31 mg·g⁻¹ fresh weight during 50 and 32 weekly maintenance subcultures, respectively. Under anthocyanin-promoting culture conditions, both lines showed an enhancement of their anthocyanin level by approximately fourfold. While line 5.4 accumulated peonidin 3-glucoside and cyanidin 3-glucoside in decreasing order, line 13.1 accumulated primarily peonidin 3-p-coumaroylglucoside with lesser amounts of malvidin monoglucoside. Results show that while the anthocyanin content was improved during the course of repeated selections, the anthocyanin composition was modified markedly favoring the accumulation of more metabolically-advanced anthocyanins.     

Effects of Temperature on the Pattern of Anthocyanin Accumulation in Seedlings of Polygonum cuspidatum

Polygonum cuspidatum seedling. Anthocyanin accumulated first in the lower part of hypocotyls and then the site of accumulation gradually extended toward the upper part of hypocotyls when seedlings were irradiated with white light (WL) at 25 C. Etiolated seedlings accumulated anthocyanin only in the upper parts (hook and cotyledons) when the seedlings were irradiated with WL at 5 C. De-etiolated seedlings that had been pre-irradiated with WL for 1 day at 25 C accumulated anthocyanin both in upper and lower parts of the seedlings when the seedlings were irradiated with WL at 5 C. Spectral sensitivity was dependent on the temperature during irradiation. Red light (R), blue light (B), and near ultra-violet light (NUV) induced the accumulation of anthocyanin at 5 C but only NUV was effective in inducing the accumulation of anthocyanin at 25 C. Dichlorophenyl dimethylurea (DCMU) inhibited WL-induced anthocyanin accumulation but did not NUV-induced anthocyanin accumulation at 25 C. However, sucrose promoted NUV action at 25 C, indicating that photosynthesis can promote NUV-induced anthocyanin accumulation. Distribution of phytochrome in etiolated seedlings, that was examined by spectrophotometry, was similar to the distribution of anthocyanin at 5 C. Furthermore, phytochrome remained after 48 hr irradiation with WL at 5 C although phytochrome was rapidly degraded at 25 C. Received 12 July 1999/ Accepted in revised form 24 December 1999     

Anthocyanin Production in Cultured Euphorbia millii Cells Using Film Culture Vessels

The anthocyanin production of cultured Euphorbia millii cells using envelope-shaped film culture vessels increased with the decrease of film thickness. The optimum temperature for anthocyanin production under the illumination was at 22°C. The anthocyanin production was increased by 2 times with mild agitation.     

Contribution to the Data on Copper Concentration in Blood and Urine in Patients with Wilson’s Disease and in Normal Subjects

Determination of copper in human tissues and body fluids may be crucial in the diagnosis of Wilson’s disease. In this study we evaluated urinary copper excretion and urine and blood concentration in 14 patients in whom Wilson’s disease was confirmed (group A) and in 21 subjects in whom the disease was only suspected (group B). The following values (mean ± SD) were found: 24-h urine (μg Cu/24 h), 152 ± 135 (A) and 31.8 ± 10.9 (B); urine (μg Cu/ml), 0.091 ± 0.087 (A) and 0.028 ± 0.011 (B); and blood (μg Cu/ml), 0.62 ± 0.25 (A) and 0.72 ± 0.09 (B). By comparison, urine copper concentration in the group of apparently healthy subjects was 0.035 ± 0.010 (n = 50), and blood copper concentration in autopsy cases of nonpoisoned people was 0.85 ± 0.19 (n = 73).     

Photoabatement by Anthocyanin Shields Photosynthetic Systems from Light Stress

Leaves and other chlorophyllous tissues of plants often show transient or permanent anthocyanin coloration. The question of whether anthocyanins can function as effective light screens to modulate photosynthesis in plants was addressed by comparing photosynthetic responses in reddish-purple pods with those in green pods of the ornamental leguminous tree Bauhinia variegata. For these comparisons the actinic radiation employed was either red radiation (RR) which was poorly absorbed by anthocyanin or blue-green radiation (BGR) which was strongly absorbed by anthocyanin. Photon yields of photosystem 2 (PS2) photochemistry and photochemical chlorophyll fluorescence quenching coefficients (q_p), measured over a range of photon flux densities (PFD) up to 1200 μmol m^-2 s^-1 at 23 °C and at five temperatures from 8 to 28 °C at a PFD of 260 μmol m^-2 s^-1, were almost identical in green pods irradiated with either RR or BGR and in purple pods irradiated with RR. However, q_p values remained much higher in purple pods irradiated with BGR, e.g., 0.80 in BGR versus 0.29 in RR at a PFD of 1200 μmol m-2 s^-1 at 23 °C, and 0.67 in BGR versus 0.28 in RR at a PFD of 260 μmol m^-2 s-1 at 8 °C. The higher values of q_p in BGR compared to RR indicated that photoabatement by anthocyanin allowed the first stable acceptor of PS2, Q_A, to be kept in a more oxidized state, thus decreasing the likelihood of photoinhibition. This was confirmed by demonstrating a lower susceptibility to photoinhibition in purple pods than in green pods in the sunlight, either naturally in pods on trees or in detached pods exposed to photoinhibitory conditions. We conclude that photoabatement by anthocyanin is a mechanism for allowing maintenance of higher oxidative levels of PS2 acceptor during episodes of high radiation stress, thereby minimizing photodamage to photosynthetic tissues. This revised version was published online in June 2006 with corrections to the Cover Date.     

Temperature Regulation of Anthocyanin Accumulation in Apple Skin

The regulation of anthocyanin accumulation in apple skin (cv.Jonathan) by temperature was studied. In the field the increasein anthocyanin in the skin before harvest coincided with decreasingtemperatures and with increasing ethylene production by thefruit. In detached apples held under continuous white light,the optimum temperatures for anthocyanin accumulation were 12°C in unripe apples and 16–24 °C in ripe apples.These effects were explained by corresponding changes in thelevel of phenylalanine ammonia-lyase (PAL), a key enzyme offlavonoid synthesis. PAL levels were higher at low than at hightemperatures and higher in ripe than in unripe apples. In intermittentlight the effects of temperature were similar but levels ofPAL and anthocyanin were lower, particularly in unripe apples.It is concluded that temperature, in conjunction with ripeningand light, is an important factor regulating anthocyanin accumulationand that its effects are mediated by effects on the level ofPAL activity. Key words: Apple, Anthocyanin, Phenylalanine ammonia-lyase     

Light Control of Anthocyanin Biosynthesis in Zea Seedlings

Evidence for involvement of two non-photosynthetic pigments in photoinduction of anthocyanin biosynthesis in the roots and mesocotyls of Zea mays L. seedlings is presented. Short (5 min), low energy (4.5 × 10³ J m^?2) fluences of red light neither induced anthocyanin synthesis nor enhanced phenylalanine ammonia-lyase activity in dark-grown maize seedlings. Little anthocyanin synthesis and no enhancement of phenylalanine ammonia-lyase activity was induced by continuous far-red light. Continuous white or blue light induced both anthocyanin synthesis and enhanced phenylalanine ammonia-lyase activity. These results show that phytochrome alone cannot induce anthocyanin synthesis in maize seedlings. However, a strong phytochrome mediation of white light induced pigment synthesis was demonstrated. This effect was not demonstrable with white light enhanced phenylalanine ammonia-lyase activity, indicating that phytochrome controls another step in anthocyanin biosynthesis.     

中间产物对玫瑰茄培养细胞合成花青苷的影响

用Ｂ５培养基悬浮培养产色素的玫瑰茄培养细胞,培养１３天时,花青苷产量最高,为０．２５ｇ／Ｌ。培养基中添加终浓度为１０＾－６ｍｏｌ／Ｌ的外源Ｌ－Ｐｈｅ能够显著地增加产色素细胞花青苷的积累量。浓度为１０＾－７ｍｏｌ／Ｌ的槲皮素,可使悬浮培养的玫瑰茄细胞花青苷产量提高１．３倍,无论是Ｌ－Ｐｈｅ还是槲皮素均不能启动不产色素的细胞系产花青苷。     

Effect of storage time and temperature on steroid and protein hormone concentrations in porcine plasma and serum

The effect of storage time and temperature of porcine blood prior to quantitation of hormone concentrations by radioimmunoassay (RIA) was evaluated. Blood from each of four luteal phase gilts was used to determine cortisol (CS) and progesterone (P) concentrations, while blood from each of four ovariectomized gilts and each of four lactating sows was used to determine luteinizing hormore (LH) and prolactin (PRL) concentrations, respectively. Blood was collected via jugular puncture from each animal within a 30-sec time period and placed into 18 heparinized and 18 nonheparinized tubes. One sample with and without heparin was stored in ice water (4°C) or at 28°C for 0.25, 2, 4, 6, 8, 12, 24, 36 or 48 hours. After storage, blood was centrifuged at 4°C and plasma or serum was collected and stored at ?20°C until quantitated by RIA. There were no differences (P>0.05) between plasma and serum concentrations (X ± SE, ng/ml) of CS (26.9 ± 0.8 vs 28.5 ± 0.8), P (24.7 ± 0.7 vs 24.8 ± 0.8), LH (2.1 ± 0.1 vs 2.2 ± 0.1) or PRL (53.2 ± 2.3 vs 52.6 ± 2.1). Similarly, storage temperature (4 vs 28°C) did not affect the concentrations of P (25.7 ± 0.8 vs 23.9 ± 0.7), LH (2.2 ± 0.1 vs 2.2 ± 0.1) or PRL (53.7 ± 2.1 vs 53.2 ± 2.3). Howver, CS concentrations decreased (P<0.05) from 28.5 ± 0.5 (4°C) to 26.9 ± 0.8 ng/ml (28°C). There was an animla x time interaction for CS concentration when plasma and serum were stored at both 4°C (P<0.001) and 28°C (P<0.003). There was also and animal x time interaction (P<0.03) for LH concentrations. The P and PRL concentrations decreased linearly by 0.0615 ng/hr (P<0.001) and 0.0625 ng/hr (P<0.004), respectively, with increased storage time.     

Absorption of acylated anthocyanins in rats and humans after ingesting an extract of Ipomoea batatas purple sweet potato tuber

We evaluated the absorbability of anthocyanins in humans and rats administered with a beverage prepared from an extract of the tuber of purple sweet potato (Ipomoea batatas Cultivar Ayamurasaki), or with an anthocyanin concentrate. Two major anthocyanin components, cyanidin 3-O-(2-O-(6-O-(E)-caffeoyl-beta-D-glucopyranosyl)-beta-D-glucopyranoside)-5-O-beta-D-glucopyranoside) and peonidin 3-O-(2-O-(6-O-(E)-caffeoyl-beta-D-glucopyranosyl)-beta-D-glucopyranoside)-5-O-beta-D-glucopyranoside), were detected in the plasma and urine of both rats and humans by HPLC or liquid chromatography/mass spectrometry (LC/MS). The plasma concentration of anthocyanins in humans reached a maximum 90 minutes after ingestion, and the recovery of anthocyanins in the urine was estimated as 0.01-0.03%. These results indicate that acylated anthocyanins could be selectively absorbed after ingesting food.     

Effects of luteinizing hormone releasing hormone on gonadotrophins in the hamster

The changes in serum gonadotrophins in male hamsters following one injection of 15 μg luteinizing hormone releasing hormone (LHRH) (Group A) were compared with those following the last injection of LHRH in animals receiving an injection approximately every 12 hr for 4 days (Group B) or 12 days (Group C). Peak follicle stimulating hormone (FSH) levels (ng/ml) were 1776±218 (Group A), 2904±346 (Group B), and 4336±449 (Group C). Peak luteinizing hormone (LH) values (ng/ml) were 1352±80 (Group A), 410±12 (Group B), and 498±53 (Group C). Serum FSH:LH ratios, calculated from the concentrations measured 16 hr after the last LHRH injections, were higher in Groups B and C than in Group A. Similar injections of LHRH (100 ng or 15 μg/injection) for 6 days elevated the serum FSH:LH ratio in intact males. Five such LHRH injections (100 ng/injection) blunted the rise in serum LH in orchidectomized hamsters. Direct effects of LHRH on gonadotrophin secretory dynamics or altered brain-pituitary-testicular interactions may alter the ratio of FSH to LH in the hamster.     

Effect of treatment of cow’s urine “Gomutra” and antioxidants in alleviating the lindane-induced oxidative stress in kidney of Swiss mice (Mus musculus)

The study aimed to evaluate the effect of cow urine and combination of antioxidants against lindane induced oxidative stress in Swiss mice. Male healthy mice, 8–10 weeks old, weighing 30 ± 5 g were randomly selected and divided into eight groups, namely, control (C); lindane (L); antioxidant (A), antioxidant+lindane (A+L), cow urine (U), cow urine+lindane (U+L), cow urine+antioxidants (U+A) and cow urine+antioxidants+lindane (U+A+L). Group C animals were administered only the vehicle (olive oil); doses selected for other treatments were: lindane: 40 mg/kg b.w.; antioxidants: 125 mg/kg b.w. (vitamin C: 50 mg/kg b.w., vitamin E: 50 mg/kg b.w., α-lipoic acid: 25 mg/kg b.w.) and cow urine: 0.25 ml/kg b.w. In group A+L and U+L antioxidants and cow urine were administered 1 h prior to lindane administration and in group U+A and U+A+L cow urine was administered 10 min before antioxidants. All treatments were administered orally continuously for 60 days. Lindane treated group showed increased lipid peroxidation, whereas glutathione, glutathione peroxidase, superoxide dismutase, catalase, protein and endogenous levels of vitamin C and E were significantly decreased compared to control. Administration of cow urine and antioxidants alleviated the levels of these biochemical parameters.     

Variation of Kernel Anthocyanin and Carotenoid Pigment Content in USA/Mexico Borderland Land Races of Maize

Variation of Kernel Anthocyanin and Carotenoid Pigment Content in USA/Mexico Borderland Land Races of Maize. Maize is the only major cereal crop that displays abundant variation for health-promoting carotenoid and anthocyanin pigments. Traditional farmers in the USA/Mexico Borderland region utilize many land race varieties with diverse kernel characteristics reflecting enculturated preferences, including color. Food prepared using these varieties may provide benefits to human health, but the kernel pigment content, and grain physical and compositional traits, have not been characterized. Seed from 48 diverse accessions representing 18 races of maize originating from the Borderland region were obtained from Native Seeds/SEARCH and planted in replicated nurseries at two locations (Ohio and Arizona) in 2008. We visually determined kernel color and quantified total carotenoid and anthocyanin pigment content of samples obtained from these nurseries using spectrophotometric analysis. Nonpigmented (white) followed by yellow kernel colors were most abundant. Populations with high carotenoid pigment content (i.e., above 40 μg/g) were not observed, whereas many accessions produced ears with mixtures of red, purple, and blue kernels containing anthocyanin pigments. A wide range in anthocyanin pigment content was observed across and within populations—some kernels displayed concentrations above 50 mg/100 g. Kernel hardness was determined visually, and protein and oil content were determined by near-infrared spectrometric analysis. Flinty (hard) followed by floury (soft) kernel types were most abundant. Carotenoid content was highest in orange- and yellow-colored pop-type kernels. Anthocyanin content was highest in blue- and purple-colored floury and flint-type kernels. Kernel weight, protein, oil, and carotenoid content were significantly affected by location. Preservation of culturally-adapted varieties with diverse kernel pigments is important not only because of their genetic diversity—they also may contribute to enhanced human health and nutrition.     

Comparison of three protocols for superovulation of brown brocket deer (Mazama gouazoubira)

This study aims to evaluate the ovulation rate and the presence of functional corpora lutea after treatment by three different protocols designed to cause superovulation in brown brocket deer. Six female received an intravaginal device containing 0.33 g of progesterone (CIDR®) for 8 days, followed by 0.5 mg injection of estradiol benzoate at the time of insertion and 265 µg of cloprostenol at the time of removal. Afterwards, the hinds were divided into three groups (n = 2): Treatment A received injection of 600 IU eCG on Day 4 after CIDR® insertion; Treatment B received injection of 300 IU eCG at the same time; and Treatment C received injection of 250 IU FSH dissolved in PVP, also on Day 4 post‐insertion. The treatments were crossed over with 44–48 day intervals after CIDR® removal, such that all the deer were submitted to all three treatments. The mean ovulation rate (Treatment A = 3.40 ± 0.68, Treatment B = 1.40 ± 0.24, Treatment C = 0.80 ± 0.49), total ovarian stimulation (Treatment A = 4.80 ± 1.02, Treatment B = 1.80 ± 0.37, Treatment C = 1.40 ± 0.60), and mean CL diameter (Treatment A = 7.33 ± 0.76 mm, Treatment B = 3.94 ± 0.19 mm, Treatment C = 2.18 ± 0.49 mm) in Treatment A were significantly higher than the mean ovulation rates, total ovarian stimulation, and mean CL diameter in Treatments B and C. The mean fecal progesterone metabolites at the luteal phase in Treatment A (6,277.94±2,232.47 ng/g feces) was significantly different from Treatment C (1,374.82±401.77 ng/g feces). Thus, although fertility was not evaluated directly, Treatment A proved capable of induce superovulation in the species Mazama gouazoubira, presenting the greatest mean ovulation rates, with the formation of functional corpora lutea. lutea. Zoo Biol 31:642‐655, 2012. © 2011 Wiley Periodicals, Inc.