Anti-tumor activity of a novel protein obtained from tartary buckwheat

TBWSP31 is a novel antitumor protein that was isolated from tartary buckwheat water-soluble extracts. The objective of this paper was to investigate the anti-proliferative effects of TBWSP31 on breast cancer Bcap37cells and to explore its possible mechanism. After treatment of Bcap37 cells with TBWSP31, typical apoptotic morphological changes were observed by inverted microscopy and scanning electron microscopy (SEM), such as detachment from the culture plate, change to a round shape, cell shrinkage, the absence of obvious microvilli, plasma membrane blebbing, and formation of apoptotic bodies. Cell-cycle analysis revealed that treatment with TBWSP31 resulted in a G(0)/G(1) arrest and prevented the cells from growing from G(0)/G(1) phase to S phase, which was most prominent at 48 h. The expression of bcl-2 and Fas were detected quantitatively by FCM, which showed that TBWSP31 induced-apoptosis may be involved with the participation of Fas and bcl-2. These results suggest that TBWSP31 is a potential antitumor compound and that apoptosis induced by TBWSP31 is a key antitumor mechanism.     

荞麦中黄酮类化合物的分析方法研究进展

荞麦中黄酮类化合物已被鉴定出的有40余种,定量荞麦总黄酮最好的方法是三氯化铝显色法;而高效液相色谱法是目前分离、定量荞麦黄酮类化合物最有效的方法。     

A rapid measurement of rutin-degrading enzyme activity in extract of tartary buckwheat seeds

In order to measure rutin-degrading enzyme activity conveniently, we have developed an isoabsorptive spectrophotometric method (ISM) for rapidly monitoring rutin-degrading enzyme (RDE) activity in extract of tartary buckwheat seeds. This technique uses discrepancy in absorbency at 372 nm and 344.5 nm, both of which are isoabsorptive wavelengths of rutin, that allow for calculations of quercetin concentrations, which is the only product of the reaction catalyzed by RDE. With this method, extracts containing RDE from buckwheat seeds were analyzed, and the measured data indicated that the sample contained quercetin concentration of 9.909 μg/ml, 8.04 times and 18.08 times greater than negative control (1.232 μg/ml) and positive control (0.548 μg/ml), respectively. ISM results of measuring rutin-degrading enzyme activity in tartary buckwheat seeds correlate with those of HPLC. However, it is beneficial to HPLC as it presents a more convenient and rapid method.     

中草药还原法制备银纳米颗粒及其抗菌性能表征

分别以中草药丁香、山茱萸、地榆和乌梅的提取液为还原剂和保护剂合成银纳米颗粒,利用紫外-可见吸收光谱(UV-vis)、透射电镜(TEM)以及X射线粉末衍射(XRD)对产物进行了表征。结果表明,所得银纳米颗粒呈近球形,提高提取液的pH有利于获得粒径较小、分散性好、稳定性高的银纳米颗粒。进一步考察制得的银纳米颗粒的抗菌性能,结果表明,其对大肠杆菌和金黄色葡萄球菌有很强的抑制作用,最小抑菌质量浓度(M IC)分别可达1.69和3.38 mg/L。     

The effect of primary antioxidants and synergists on the activity of plant extracts in lard

The antioxidant activities of combinations of plant extracts and primary antioxidants or citric acid were studied in lard stored at 75°C. Methanol extracts of oregano, thyme, marjoram, dittany, rosemary and sage were used. When combined with butylated hydroxyanisole, butylated hydroxytoluene or ascorbyl palmitate the extracts showed an additive antioxidant effect, while combinations with propyl gallate did not increase the induction period obtained by the phenolic antioxidant. Negative synergism was observed when the plant extracts were mixed with DL-α-tocopherol. Citric acid showed a high synergistic efficiency with marjoram extract and a minor one with thyme extract, but it had a negative effect with all the others. Ternary mixtures of the plant extracts with citric acid and ascorbyl palmitate had lower activity than the corresponding dual combinations with ascorbyl palmitate, except for the mixture of marjoram extract.     

RP-HPLC法测定超临界提取物中淫羊藿苷的含量

目的：建立淫羊藿叶超临界提取物中淫羊藿苷的含量测定方法。方法：采用RP-HPLC法,Hypersil ODS C18色谱柱（4.6 mm×250 mm,5μm）,甲醇-0.1%冰醋酸水溶液梯度洗脱,流速1 mL.min-1,检测波长270 nm。结果：淫羊藿苷在0.212～2.12μg进样量范围内线性关系良好,平均加样回收率98.7%,RSD为1.18%（n=6）。结论：该法简便、易操作,适用于淫羊藿超临界提取物中淫羊藿苷的含量测定,可有效地控制淫羊藿超临界提取物的质量。     

Catalytic adsorptive stripping voltammetric determination of Cr(VI) in EDTA extracts from solid samples

A catalytic adsorptive stripping voltammetric procedure which allows for Cr(VI) determination in EDTA extracts is presented. EDTA used for extraction can be exploited as a masking agent for Cr(III). The calibration graph for Cr(VI) for an accumulation time of 60 s was linear in the range from 2 × 10⁻¹⁰ to 3 × 10⁻⁸ mol L⁻¹. The relative standard deviation for a Cr(VI) concentration of 2 × 10⁻⁹ mol L⁻¹ was 5.1% (n = 5). The detection limit estimated from three times the standard deviation of low Cr(VI) concentration and accumulation time of 60 s was about 7 × 10⁻¹¹ mol L⁻¹. The influence of foreign ions commonly present in extracts from solid samples is presented. The proposed voltammetric procedure was applied to determine Cr(VI) in EDTA extractable chromium from soil certified reference materials CRM 483 and CRM 041. The performance of the method was also verified by studying the recovery of Cr(VI) from spiked river water.     

Antioxidant activity of tea extracts in lipids and correlation with polyphenol content

The present study examined the antioxidative activity of water and ethanol extracts of green and black tea leaves against the oxidation of heated sunflower oil and lard. Oxidation was conducted at 110 °C in the Rancimat test. Total polyphenols and catechin contents in tea extracts were measured. The research showed that the total polyphenol content in green and black tea leaves was 205.2 and 148.7 mg/g, respectively. In tea leaves extracts, it ranged between 245.9 mg/g and 837.7 mg/g and depended on the extraction solvent and the kind of tea used (p <0.001). The highest polyphenol content was observed in samples extracted with 95% ethanol, lower contents were found with the use of water. Results showed that the highest antioxidant activity, measured as an induction period, with 1000 ppm green tea ethanol extract, was comparable to á‐tocopherol activity in sunflower oil. In lard, the longest induction period was measured with 500 and 1000 ppm of green tea ethanol extract. Other tea extract concentrations were significantly less active. Statistical analysis of the tea extract antioxidant activity in lipids in the Rancimat test showed an essential influence of the catechin contents. Further statistical analysis also showed an influence of (?)‐epicatechin gallate (ECG), (?)‐epicatechin (EC), and (+)‐catechin (C) contents in the tea extracts on the antioxidant activity in lipids. It was stated that the antioxidant activity was higher in tea extracts containing high levels of ECG, EC, and C.     

Correlation of absolute temperature coefficients of ultrasonic velocity in solutions of dilute coal and lignite extracts with molecular masses

Absolute temperature coefficients of ultrasonic velocity b were determined from temperature dependences of the velocity of the longitudinal ultrasonic waves with frequency 2.5 MHz in solutions of pyridine and tetrahydrofuran extracts, obtained from lignite and coals of different ranks (54.1–85.6 wt% of carbon). The relationship between absolute temperature coefficients b of all extract solutions and extract molecular masses M was studied. It was stated that the dependence of b√M vs. M deviates considerably from the rule b√M=const, found empirically by Lagemann et al. for pure organic liquids. Experimental data for lignite and coal product solutions were best fitted by the equation b√M=AM+B, where constants A and B were determined from the fits having the correlation coefficients r within the range 0.99–0.999. This empirical formula was applied to determine the molecular mass of a sample, for which the Knauer osmometer method could not be used.     

Growth‐promoting properties of yeast extracts produced at different pH values,with different autolysis promoters and bacterial populations

The effect of the autolysis conditions of bakers' yeast on the growth‐promoting properties of the resulting yeast extracts (YE) were examined on cultures of Lactococcus lactis ssp lactis D25 and of Lactococcus lactis ssp cremoris R2. The various YE were produced by conducting autolysis at four pH levels (pH 4.0, 5.5, 7.0 or 8.5) and with or without the autolysis promoters (AP) ethyl acetate or chitosan. To determine if the presence of contaminating bacteria during autolysis affected the YE characteristics, one series of YE was also prepared with the bakers' yeast cell suspension contaminated with 10⁸ CFU/cm^?3 of typical yeast plant bacteria (mostly of the Bacillus genus) at the beginning of autolysis. The growth curves of the lactococci in media supplemented with the various YE were analyzed to determine the maximum optical density (OD_max), the maximum growth rate (µ_max) and the fermentation time (F‐time) at which OD_max was reached. The addition of bacterial contaminants (at 10⁸ CFUcm^?3) did not significantly influence the growth‐promoting properties of YE. The lactococci had higher OD_max and µ_max values when the media contained YE produced at pH 4.0 or 5.5, and these YE contained 40% more total nitrogen than those obtained at pH 7.0 or 8.5. There was no major effect of AP on the biological value of the resulting YE, but a significant interaction between pH and AP was found for L lactis D25. Thus the OD_max of YE produced at pH 7.0 and 8.5 were highest when ethyl acetate was used as autolysis promoter, as compared with control and chitosan at pH 7.0–8.5, while the opposite was obtained with products produced at pH 4.0 and 5.5. This study shows that parameters used to produce YE influence their biological value with respect to the growth‐promoting properties of lactococci. © 2001 Society of Chemical Industry     

公、母丁香乙醇提取物离体抑菌活性比较

采用生长速率法和孢子萌发法测定了公丁香和母丁香的乙醇提取物对5种病原菌（番茄灰霉病菌、水稻稻瘟病菌、黄瓜炭疽病菌、苹果轮纹病菌和玉米小斑病菌）的菌丝生长和3种病原菌（番茄灰霉病菌、稻瘟病菌和玉米小斑病菌）孢子萌发的影响。结果表明,提取物对两者均有一定的抑制作用：对于同一种病原菌来说,丁香的乙醇提取物对其孢子萌发的抑制效果好于对其菌丝生长的抑制效果;在相同的供试浓度下,公丁香的抑菌效果优于母丁香。     

体外消化对蓝莓提取物抗氧化、抗癌及组分的影响

以蓝莓为原料,考察了体外消化前、后蓝莓提取物(总酚和花色苷)含量、抗氧化性、抗癌作用及花色苷组成的变化,并推测了花色苷的降解途径。分别采用福林-酚法和pH示差法测定总酚和花色苷含量;以脂质体的抑制率、DPPH和ABTS+自由基清除能力评价体外消化前后提取物的抗氧化能力;利用高效液相色谱-电喷雾二级质谱联用技术分析花色苷组成。结果表明,与未经消化的提取物相比,经肠道消化后样品总酚含量增加47.21%,花色苷含量降低71.82%,脂质体的抑制率、DPPH和ABTS+自由基清除率分别提高38.45%、29.41%和29.12%;对Hep G2肝癌细胞、A549肺癌细胞和Hela人宫颈癌细胞的生长抑制作用显著增加,蓝莓花色苷组分由12种降为9种。对胃肠消化花色苷的降解过程推测发现,矢车菊素-3-葡萄糖苷降解形成槲皮素的过程与天竺葵素-3-葡萄糖苷降解成山柰酚的过程机制相同。     

Determination of aromatic and aliphatic CH groups in coal by FT-i.r.: 1. Studies of coal extracts

A set of coal extracts have been characterized by FT-i.r. and proton n.m.r. spectroscopy. The n.m.r. measurements of aliphatic and aromatic hydrogen allow a direct calibration of the absorption coefficients of appropriate i.r. bands. This allows methods for calculating these coefficients based on relating i.r.-band intensities to elemental hydrogen to be tested directly. Reasonable agreement was obtained.     

Effects of organic solvent extracts from herbage on feeding behavior in goats

The effects of organic solvent extracts from perennial ryegrass (Lolium perenne L. cv. Friend), orchardgrass (Dactylis glomerata L. cv. Akimidori) andLolium ×Festuca hybrid on the hay consumption of goats were measured by simple two-choice feeding assays. Orchardgrass hay treated with methanol extracts (resuspended in water) of perennial ryegrass showed an increased intake by goats compared to untreated hay (P < 0.01). The palatability of hay tended to be slightly decreased by spraying it with pentane extract of perennial ryegrass and orchardgrass. Furthermore, extracts of hybrid with pentane inhibited goats from feeding (P < 0.05). Methanol extract stimulated hay consumption whereas the pentane extracts did not change or reduce intake.     

Earthworm extracts utilized in the green synthesis of gold nanoparticles capable of reinforcing the anticoagulant activities of heparin

Gold nanoparticles were obtained using a green synthesis approach with aqueous earthworm extracts without any additional reducing or capping agents. The gold nanoparticles were characterized using UV-visible spectrophotometry, high-resolution transmission electron microscopy, atomic force microscopy, field emission scanning electron microscopy, X-ray diffraction, Fourier transform infrared spectroscopy, and inductively coupled plasma mass spectrometry. The anticoagulant activity of the gold nanoparticles was assessed using the activated partial thromboplastin time and was mildly enhanced by combining the gold nanoparticles with heparin. In addition to the generation of spherical nanoparticles with an average diameter of 6.13 ± 2.13 nm, cubic and block-shaped nanoparticles with an average aspect ratio, defined as the length divided by width, of 1.47 were also observed.     

Processing of soybean (Glycine max) extracts in aqueous two‐phase systems as a first step for the potential recovery of recombinant proteins

BACKGROUND: The potential use of plants as production systems to establish bioprocesses has been established over the past decade. However, the lack of efficient initial concentration and separation procedures affect the generic acceptance of plants as economically viable systems. In this context the use of aqueous two‐phase systems (ATPS) can provide strategies to facilitate the adoption of plants as a base for bioprocesses. Among the crops, soybeans (Glycine max) represent an attractive alternative since potentially they can produce high levels of recombinant protein. In this paper the processing of fractionated soybean extracts using ATPS is evaluated as a first step to recover recombinant proteins expressed in plants, using β‐glucuronidase (GUS; E.C. 3.2.1.31) as a model protein. RESULTS: The evaluation of the effect of system parameters provided the conditions under which the contaminant proteins from fractionated soybean extracts and GUS concentrated in opposite phases. A PEG 600/phosphate system comprising 14.5% (w/w) polyethylene‐glycol (PEG), 17.5% (w/w) phosphate, a volume ratio (Vr) equal to 1.0, and a system pH of 7.0 resulted in the potential 83% recovery of GUS from the complex mixture and an increase in purity of 4.5‐fold after ATPS. CONCLUSIONS: The findings reported here demonstrate the potential of ATPS to process fractionated soybean extract as a first step to isolate and purify a recombinant protein expressed in soybeans. The proposed approach can simplify the way in which recombinant proteins expressed in plants can be recovered. Copyright © 2007 Society of Chemical Industry     

苦荞提取物的关键香气成分分析

采用固相微萃取(SPME)结合气相色谱-质谱联用技术(GC-MS)分析苦荞提取物中的挥发性成分;同时采用气相色谱-嗅闻仪(GC-O)结合香气提取稀释分析(AEDA)确定其香气活性成分,通过对香气活性成分定量并计算其香气活性值(OAV)最终确定苦荞提取物的关键性香气成分。结果显示,SPME-GC-MS在苦荞提取物中共鉴定出92种挥发性成分,包括酯类28种、醛类7种、醇类9种、酮类7种、酸类6种、烯烃8种、烷烃20种、其他类7种;经GC-O嗅闻仪共鉴定出17种香气活性化合物,定量结果显示,苯甲醇(8637.40μg/kg),苯乙醛(3150.47μg/kg)、D-柠檬烯(3080.00μg/kg)含量较高;结合稀释因子(FD)和香气活性值(OAV)推测,苯甲醛(FD=81,OAV=5)、二甲基三硫(FD=243,OAV=48)、苯甲醇(FD=81,OAV=9)、苯乙醛(FD=729,OAV=787)、丙二酸二乙酯(FD=81,OAV=1.2)5种物质对苦荞提取物的风味贡献最大,是苦荞提取物的关键香气成分。     

Effect of culture pH on the antioxidant properties of Antrodia camphorata in submerged culture

Effects of culture pH ranging from pH 3.0 to 6.0 on yields of bioactive compounds, antioxidant activity, and free radical scavenging ability of methanolic extracts from mycelia and filtrates of Antrodia camphorata in a submerged culture were investigated. Culture pH significantly affected antioxidant and scavenging free radical activities of both the methanolic extracts of the mycelia and the filtrates. A low culture pH (3.0) favored cell growth, but higher culture pH (5.0) favored antioxidant properties. Antioxidant activities of the methanolic extracts of the mycelia and the filtrates were found to be correlated with total phenolic content, polysaccharide content, and protein/polysaccharide ratios. Antioxidant production was maximum at pH 5.0 at which the total phenolics and polysaccharide/protein ratio in the methanolic extracts of the mycelia were 152.2 mg/g and 0.335, respectively. The total phenolics might be the compounds responsible for the antioxidant activities of the methanolic extracts of the mycelia and the filtrates on lipid peroxidation and scavenging effects of ferrous ions. The maximum polysaccharide content of the methanolic extracts of the filtrates was 55.3 mg/g at pH 5.0. Higher yields of antioxidants were obtained in the submerged culture as compared with that of fruiting bodies. An alternative approach for producing antioxidants of A. camphorata in a submerged culture instead of solid state cultures was demonstrated in this study.     

Effects of dietary glucosylceramide on dermatitis in atopic dermatitis model mice

The effects of dietary plant and yeast cerebroside (glucosylceramide), a major sphingolipid in plants and yeast, on atopic dermatitis (AD) like symptoms were investigated in a mouse model. After 7 wk of feeding with a diet containing maize glucosylceramide, plasma IgE levels became significantly lower and in contrast, the levels of interleukin (IL)‐12, which induces cellular immunity, became significantly higher in the AD mice than in the controls. However, the sphingolipid constituents of the skin fraction in the maize glucosylceramide fed group did not contain sphingoid bases of plant origin, such as 8‐unsaturated sphingoid bases. The results of the present study indicated that dietary plant glucosylceramide prevented AD‐like symptoms in AD model mice via regulation of Th1/Th2 balance. Practical applications: Dietary plant and yeast glucosylceramides have been shown to suppress AD‐like symptoms in AD model mice via regulation of helper T‐cell Th1/Th2 balance. Glucosylceramide are capable of preventing AD and may be useful in skincare products.     

葡萄球菌S-35对小鼠的毒性作用

[目的]对葡萄球菌S-35进行动物安全性评价。[方法]采用Ames体外试验、小鼠急性毒性试验、小鼠精子畸形试验、小鼠骨髓嗜多染红细胞微核试验及小鼠30 d喂养慢性毒性检测等方法,测定了该菌株对小鼠的毒性、血液生化指标、血项指标及脏器的影响,并做组织切片观察。[结果]在1.6×10^10~2×10^12 cfu/皿剂量范围内,葡萄球菌S-35 Ames体外试验结果为阴性;葡萄球菌S-35对小鼠的急性经口毒性为“低毒”级(LD50>4640 mg/kg);S-35在0.78~12.5 mg/L试验剂量范围内,小鼠精子畸形和骨髓细胞微核试验结果为阴性。小鼠30 d喂养试验,葡萄球菌S-35在125 mg/L的剂量下,小鼠血液生化和血项各项指标均无异常。[结论]葡萄球菌S-35对动物机体安全,该菌株有进一步研究开发为生防菌的价值。