加工工艺对荞麦蛋白热性质和体外模拟消化过程的影响

本实验研究了加工工艺对荞麦蛋白(BWP)的热性质和体外模拟消化过程的影响。荞麦蛋白有两个变性温度,在80℃和102℃附近,分别对应8S和13S球蛋白的变性。脂肪的存在对荞麦蛋白的变性温度影响不大,但会降低其热焓。在模拟的胃蛋白酶消化过程中,BWP的氮释放量较大豆分离蛋白(SPI)先高后低;而在胰蛋白酶消化过程中,BWP的氮释放量较SPI高。这与其蛋白组成以及SPI中存在活性较高的胰蛋白酶抑制剂有关。荞麦蛋白的球蛋白(13S和8S)易被降解(胃蛋白酶消化阶段),而2S清蛋白的降解主要集中在胰蛋白酶消化阶段。经脱脂处理、超声协助提取的荞麦蛋白(DFU-BWP)和搅拌提取的荞麦蛋白(FM-BWP)的氮释放量在整个胃蛋白酶消化过程中类似,均明显高于超声提取的荞麦蛋白(FU-BWP)(p<0.05),说明胃蛋白酶对BWP的降解过程与其脂肪含量以及加工工艺有关。     

抗白色念珠菌海洋真菌的筛选及抑菌活性初探

实验以白色念珠菌为对象,对从渤海湾不同站位分离获得的55株海洋丝状真菌进行了抗真菌活性菌株的筛选。从中筛选得到了BH431、BH515、BH531和BH09721 4株抑菌活性较高的菌株,并进一步对不同浓度发酵液的抑菌活性及代谢物性质进行了检测。结果显示,4株海洋真菌对野生型白色念珠菌SC5314均具有较强抑菌活性,其最小抑菌浓度分别为 65 g/mL、19 g/mL、54 g/mL、23 g/mL;其中BH431和BH515菌株还显示出对基因敲除型菌株RM1000的抑菌活性,最小抑菌浓度分别为 32 g/mL、39 g/mL。4株海洋真菌所产生的活性代谢物具有较好的酸碱适应性,活性物质分子质量＜6 000 u;其中BH431菌株的活性代谢物还具有良好的热稳定性。     

加工工艺对荞麦蛋白功能特性的影响

探讨了加工工艺对荞麦蛋白(BWP)功能特性的影响。pH＜5．0和 pH＞6．0时,喷雾干燥制备的荞麦蛋白(SBWP)的溶解度高于商品大豆分离蛋白(SPI)(P＜0．05)。pH＜7．0时,超声协助提取荞麦蛋白(U- BWP)的溶解度较搅拌提取荞麦蛋白(M-BWP)和脱脂、超声协助提取荞麦蛋白(DU-BWP)好；pH＞7．0时,结果相反。总体来说,BWP 持油能力较 SPI 强,且冷冻干燥制备的荞麦蛋白(F-BWP)的持水、持油能力强于 S-BWP (P＜0．05)。BWP 的乳化活性与其溶解度呈正相关。pH 4．0～5．0时,BWP 的乳化活性指数(EAI)最小而乳化稳定性(ES)最高。脱脂处理明显提高了 BWP 的 EAI 和 ES。     

Preparation of tartary buckwheat protein product and its improving effect on cholesterol metabolism in rats and mice fed cholesterol-enriched diet

ABSTRACT:  Tartary buckwheat protein product (TBP) was prepared from buckwheat flour by alkali extraction and isoelectric precipitation. The protein content of TBP was 45.8%, and its amino acid composition of TBP was similar to that of common buckwheat protein product (BWP). SDS-PAGE analysis showed that the protein profile of TBP was partially different from that of BWP. TBP contained more quercetin (1710 mg/100 g) than BWP (5.4 mg/100 g), while there was a small difference in the contents of rutin between them. In experiment 1, the consumption of BWP and TBP at 20% net protein level for 13 d caused 32% and 25% reductions in serum cholesterol of rats fed cholesterol, respectively, when compared to the consumption of casein ( P < 0.05). The reduction of serum cholesterol by BWP and TBP was associated with enhanced excretion of fecal neutral sterols. In experiment 2, the consumption of BWP and TBP for 27 d caused 62% and 43% reductions in the lithogenic index in mice fed cholesterol, respectively ( P < 0.05). The reduction in lithogenic index was associated with enhanced excretion of fecal bile acids. Taken together, these results suggest a potential source of TBP as a functional food ingredient as well as BWP.     

安徽省SMV株系的鉴定及其抗源筛选

为明确安徽省大豆花叶病毒（soybean mosaic virus,SMV）株系的最新组成、分布及消长动态,以及筛选对流行株系的抗性种质,2009-2010年广泛采集安徽省各大豆主产区疑似SMV病样461份,采用生物学及血清学方法鉴定得到83个SMV分离物。用全国统一的10个鉴别寄主对这些分离物进行鉴定,共鉴定出14个SMV株系,如SC3~SC9、SC11、SC13~SC15、SC17、SC19和SC22,其中SC5、SC11、SC14、SC15、SC17、SC19和SC22是本研究在大豆主产区新发现的。进一步比较分析发现,SMV株系SC3和SC7仍为目前安徽省大豆产区的流行株系,分别占分离物总数的16.9%和25.3%;其次为新发现的强毒株系SC15,在安徽省也有较为广泛的分布,比率为12.1%。针对流行株系SC3和SC7的抗源筛选结果表明,来自参加安徽省区试的149份资源中有中作J8023、中黄45、L85-2308、中涡47号和濉科13等5份材料对SC3表现高抗,中黄45、宿03-4-1-4和鲁06-7等9份材料对SC7表现高抗;参加我国南方区试的95份种质资源中有13份对强毒株系SC15表现抗病,占鉴定品种的13.7%。本研究明确了安徽省大豆产区的流行株系及变化趋势,筛选出针对SMV流行株系的抗源,为培育抗病品种和抗病品种布局提供了依据。     

Phenotypic consequences of LYS4 gene disruption in Candida albicans

A BLAST search of the Candida Genome Database with the Saccharomyces cerevisiae LYS4 sequence known to encode homoaconitase (HA) revealed ORFs 19.3846 and 19.11327. Both alleles of the LYS4 gene were sequentially disrupted in Candida albicans BWP17 cells using PCR‐based methodology. The null lys4Δ mutant exhibited lysine auxotrophy in minimal medium but was able to grow in the presence of l ‐Lys and α‐aminoadipate, an intermediate of the α‐aminoadipate pathway, at millimolar concentrations. The presence of d ‐Lys and pipecolic acid did not trigger lys4Δ growth. The C. albicans lys4Δ mutant cells demonstrated diminished germination ability. However, their virulence in vivo in a murine model of disseminated neonatal candidiasis appeared identical to that of the wild‐type strain. Moreover, there was no statistically significant difference in fungal burden of infected tissues between the strains. Copyright © 2014 John Wiley & Sons, Ltd.     

Distinct locational differences observable in biophysical functions of the facial skin: with special emphasis on the poor functional properties of the stratum corneum of the perioral region

The face is composed of complicated anatomical components, presenting unique portions, such as the eyes, nose and mouth in a relatively narrow area. Moreover, the facial skin is densely populated by the pilosebaceous units and sweat glands, and its stratum corneum (SC) is much thinner than that of the trunk and limbs, although it is always exposed to the environment. Among various portions of the facial skin, some are more easily irritated than others by environmental stimuli, or are more often affected by certain dermatoses. However, the functional aspects of the different portions of the facial skin have not been studied in detail under a strictly controlled environment in sufficiently large numbers of subjects covering different age groups. Thus, we conducted studies in winter with various biophysical techniques, such as transepidermal water loss (TEWL), as a parameter for SC barrier function, high-frequency conductance as that for skin surface hydration state, skin surface lipids, pH, blood flow and skin surface temperature on the forehead, mid-portion of the cheek (cheek in short), nasal tip (nose in short), nasolabial fold and chin of 20 healthy Japanese females aged 22-37 years (average 25 years) in a climate chamber adjusted to 21 degrees C and 50% relative humidity. Thereafter, we studied the influence of ageing on these biophysical parameters by collecting data of TEWL, high-frequency conductance and size of superficial corneocytes on the cheek, nasolabial fold and chin of 303 healthy Japanese female volunteers of different ages. The obtained results showed that the barrier function of the SC was best on the cheek, presenting the lowest TEWL, which was significantly higher on the nasolabial fold and chin than on the cheek. TEWL showed a decrease with age. In contrast, skin hydration state was higher on the nose, but it tended to be lower on the nasolabial fold, showing a mild age-related increase. The corneocytes on the nasolabial fold and chin were smaller than those on the cheek. They revealed a clear increase in size with age. Skin surface lipids were richest on the nose, whereas the superficial pH on the nose was the lowest among the regions tested. The skin temperature was lowest on the cheek than on other areas of the face; although, together with the nose, its blood flow was higher than that of the others. These data indicate great regional differences observable in SC functions on the face. In general, the SC barrier function increases with age, probably because of a decreased epidermal turnover rate as recognized by the increase in corneocyte size. Among the various sites, the skin of the nasolabial fold and chin, whose SC consisted of the smallest corneocytes, showed poorest SC properties in barrier function, suggesting the presence of mild invisible inflammation. It is understandable that this area easily develops not only the complaint of sensitive skin to cosmetics but also dermatitis because of various external agents.     

高静水压和热处理对荞麦蛋白功能性质的影响

高静水压（high hydrostatic pressure,HHP）处理技术作为一种新型的非热食品加工技术,在食品加工中具有对食物营养和感官品质破坏较小等多种优势。本实验通过HHP处理与传统热处理法（heat treatment,HT）处理荞麦蛋白（buckwheat protein,BWP）,并测定BWP溶解性、起泡性和乳化性等性质,此外通过BWP羰基含量、巯基含量和表面疏水性测定以及傅里叶变换红外光谱分析,研究不同加工处理方式对BWP功能性质的影响。结果表明：随着HHP处理压力的增加,BWP的羰基含量、表面疏水性显著升高（P＜0.05）,而巯基含量显著降低（P＜0.05）;在500 MPa、30 min时,BWP乳化性、起泡性均达到最大值（67.3 m2/g和91.5%）,较空白对照组（未经HHP处理）分别提高33.0%、16.1%,较HT处理组分别提高61.5%、52.8%。傅里叶变换红外光谱结果表明,HHP处理后BWP的α-螺旋和β-折叠相对含量减少,β-转角和无规卷曲相对含量增加。经相关性分析发现,BWP的溶解性、乳化性、起泡性、羰基含量与巯基含量之间均存在相关性。其中,BWP的乳化性与起泡性、表面疏水性呈极显著正相关（P＜0.01）,并且与巯基含量呈极显著负相关（P＜0.01）。综上,HHP处理可以改变BWP的功能性质。可为进一步研究BWP在功能食品的应用提供理论和实验依据。     

New modules for PCR-based gene targeting in Candida albicans: rapid and efficient gene targeting using 100 bp of flanking homology region

The use of PCR-based techniques for directed gene alterations has become a standard tool in Saccharomyces cerevisiae. In our efforts to increase the speed of functional analysis of Candida albicans genes, we constructed a modular system of plasmid vectors and successfully applied PCR-amplified functional analysis (FA)-cassettes in the transformation of C. albicans. These cassettes facilitate: (a) gene disruptions; (b) tagging of 3'-ends of genes with green fluorescent protein (GFP); and (c) replacements of endogenous promoters to achieve regulated expression. The modules consists of a core of three selectable marker genes, CaURA3, CaHIS1 and CaARG4. Modules for C-terminal GFP-tagging were generated by adding GFP-sequences flanked at the 5'-end by a (Gly-Ala)3-linker and at the 3'-end by the S. cerevisiae URA3-terminator to these selection markers. Promoter exchange modules consist of the respective marker genes followed by the regulatable CaMAL2 or CaMET3 promoters at their 3'-ends. In order to ensure a reliably high rate of homologous gene targeting, the flanking homology regions required a size of 100 bp of gene-specific sequences, which were provided with the oligonucleotide primers. The use of shorter flanking homology regions produced unsatisfactory results with C. albicans strain BWP17. With these new modules only a minimal set of primers is required to achieve the functional analysis of C. albicans genes and, therefore, provides a basic tool to increase the number of functionally characterized C. albicans genes of this human pathogen in the near future.     

白杨素及其衍生物在Caco-2细胞模型中的转运规律

目的 研究白杨素（chrysin,ChR）及其衍生物6,8-二-三氟甲基-7-乙酰氧基白杨素（dFMAChR）在人源结肠腺癌细胞系Caco-2细胞单层模型的转运规律.方法 建立精确灵敏、重复性好的ChR及dFMAChR的高效液相色谱（HPLC）检测方法;MTT（四甲基偶氮唑盐）法测定ChR和dFMAChR对Caco-2细胞的毒性作用.体外培养Caco-2细胞,用Transwell建立单层细胞模型;确定ChR和dFMAChR转运的最佳pH条件.评价时间、浓度对dFMAChR和ChR的双向转运的影响,计算转运速率及表观渗透系数（Papp）,并与阳性对照药普萘洛尔进行比较.结果 ChR和dFMAChR的转运最佳介质pH分别为6.0和6.5,两者的转运无论是Apical侧（A侧）到Basolateral侧（B侧）还是B侧到A侧,均具有时间依赖性和浓度依赖性.ChR和dFMAChR的平均Papp （A-B）分别是（1.60±0.15）×10-6 cm/s和（10.63±0.35）×10-6 cm/s,平均Papp （B-A）分别是（1.22±0.17）×10-6 cm/s和（10.43±0.28）×10-6 cm/s,两者Papp （A-B） ＞Papp （B-A）,且Papp （A-B） /Papp（B-A） ＜2.结论 在Caco-2模型中,dFMAChR和ChR均由被动扩散方式转运,dFMAChR的转运速率明显高于ChR,吸收接近普萘洛尔,属于吸收良好的化合物.     

Effect of milk protein glycation and gastrointestinal digestion on the growth of bifidobacteria and lactic acid bacteria

In this paper, β-lactoglobulin (β-Lg) and sodium caseinate (SC) have been glycated via Maillard reaction with galactose and lactose and, subsequently, the effect of glycoconjugates hydrolyzed under simulated gastrointestinal digestion on the growth of pure culture of Lactobacillus, Streptococcus and Bifidobacterium has been investigated. Glycopeptides were added to the growth media as the sole carbon source. None of the bacterial strains was able to grow in hydrolysates of native and control heated β-Lg and SC. However, glycopeptides were fermented, in different degree, by Lactobacillus and Bifidobacterium and hardly any effect was detected on the growth of Streptococcus. Digested β-Lg glycoconjugates showed a strain-dependent effect whereas growth profiles of bacteria when hydrolysates of SC glycoconjugates were used as substrates were very similar, regardless of the strain. A general preference towards peptides from β-Lg/SC glycated with galactose, particularly at the state of the reaction in which the highest content in the Amadori compound tagatosyl-lysine is present, was observed. SC glycoconjugates were quickly fermented by some strains, promoting their growth in a greater extent than β-Lg complexes or even glucose. Therefore, from the results obtained in this work it can be concluded that conjugation of both milk proteins with galactose and lactose via the Maillard reaction could be an efficient method to obtain novel food ingredients with a potential prebiotic character.     

仿刺参卵和体壁多肽的制备及免疫活性

以仿刺参卵和体壁为原料制备多肽,测定其基本营养成分和分子质量分布,并研究其对淋巴细胞增殖作用和小鼠免疫功能的影响。采用生物酶解技术和切向流超滤技术分离和制备仿刺参卵和体壁多肽,磺酰罗丹明B比色分析法测定了多肽质量浓度（10、50、100、500 μg/mL）对小鼠淋巴细胞增殖（spleen lymphocyte proliferation,SLP）能力的影响,设定水对照组和多肽低、中、高（日剂量83.3、166.7、500.0 mg/kg）剂量组进行30 d小鼠灌胃实验,检测了小鼠脏器/体质量、迟发型变态反应能力、伴刀豆球蛋白A诱导的小鼠脾淋巴细胞转化能力、抗体生成细胞数、小鼠血清溶血素水平、小鼠碳廓清能力、小鼠腹腔巨噬细胞吞噬鸡红细胞能力及自然杀伤细胞活力等指标。结果表明,仿刺参卵和体壁1～10 kDa多肽（EP1和BWP1）在500 μg/mL时均对SLP具有显著的促进作用,其粗蛋白含量分别为64.74、70.25 g/100 g,氨基酸总量分别为45.69、63.26 g/100 g,分子质量分别分布在130～1 600 Da及130～2 500 Da之间。经口服灌胃给予小鼠不同剂量的多肽30 d,EP1可提高小鼠的细胞免疫功能和单核-巨噬细胞吞噬功能,BWP1可提高小鼠的体液免疫功能和单核-巨噬细胞吞噬功能。结果提示EP1和BWP1均具有增强免疫力的功能,可开发为新的免疫调节产品。     

荞麦蛋白的功能特性研究

采用碱萃取酸沉析法提取制备了荞麦蛋白，并较为系统地研究了蛋白质浓度、温度、pH值、氯化钠和蔗糖对荞麦蛋白的三种功能特性（溶解性质、乳化特性和泡沫特性）的影响。结果表明，本实验条件下制备的荞麦蛋白具有优良的溶解性质，其乳化性质与对照样品大豆分离蛋白相当，但泡沫特性较差。     

应用改进Canny法检测工业零件含噪图像边缘

针对工业零件含噪图像边缘检测,根据Canny算法原理,提出了一些改进策略,形成了一种矩形透镜最大梯度模边缘检测算法。采用中值滤波完成图像平滑,有效抑制了图像噪声;采用5×5邻域一阶偏导有限差分计算图像的梯度幅值,提高了边缘定位的精度;采用最大类间方差法(OTSU)求解了最优区域分割阈值,实现了边缘的自动检测。以磁环和极片工业零件图像边缘检测为例进行了实验,结果表明,该算法具有较好的去噪和边缘检测效果。     

Hesperidin inhibits development of atopic dermatitis-like skin lesions in NC/Nga mice by suppressing Th17 activity

Hesperidin (previously called vitamin P) is a predominant flavanone present in citrus fruits, and is presumed to have a role in their beneficial effect for human health because it possesses various physiological activities. In this study, we investigated the anti-allergic and anti-inflammatory effects of hesperidin and α-glucopyranosyl (αG)-hesperidin, its derivative with enhanced water-solubility, in NC/Nga mice, a human-like mouse model of atopic dermatitis. NC/Nga mice were fed a 0.1% αG-hesperidin or hesperidin diet for 8 weeks. αG-hesperidin and hesperidin feeding effectively inhibited skin lesions and immunoglobulin E (IgE) elevation. At the end of the 8-week-experimental period, the production of inflammatory cytokine interleukin (IL)-17 and interferon-gamma (IFN-γ) from splenocytes was lower in the αG-hesperidin/hesperidin-fed group than in the control group. Changes in mRNA expression in splenocytes are also examined using DNA microarray and real-time RT-PCR. It was revealed that cytotoxic T-lymphocyte antigen 4 (CTLA4), a regulatory T-cell (Treg) marker, was markedly upregulated in splenocytes, particularly by αG-hesperidin feeding. These results suggest that αG-hesperidin attenuated exacerbation of AD-like symptoms, decreased systemic immune hyper-responsiveness in part through the reduction of IgE, IL-17 and IFN-γ, and also modulated Th17/Treg balance in NC/Nga mice. Therefore, αG-hesperidin may be useful in the management of Th17-mediated allergic disorders.     

Effects of different dairy ingredients on the rheological behaviour and stability of hot cheese emulsions

The influence of sodium caseinate (SC), butter milk powder (BMP) and their combinations on particle size, rheological properties, emulsion stability and microstructure of hot cheese emulsions made from mixtures of Cheddar and soft white cheese was studied. All emulsions exhibited shear-thinning flow behaviour and increasing SC concentration (1–4%) led to an increase in particle size and a decrease in apparent viscosity. In contrast, increasing BMP concentration caused significant decrease in particle size and slightly reduced the apparent viscosity. Stability against creaming and precipitation increased with increasing concentration of SC, whereas BMP destabilised the emulsions resulting in extensive precipitation. Confocal laser scanning microscopy images showed that SC exerted markedly better emulsification ability than BMP. Emulsions containing equal amounts of SC and BMP presented better stability against creaming and precipitation and this could be developed into a novel strategy to replace emulsifying salts in production of cheese powder.     

Characterisation of a Bifidobacterium strain with acquired resistance to cholate--a preliminary study

The susceptibility levels against bile salts (ox gall) and sodium cholate of 19 Bifidobacterium strains from different origins ranged between 0.125% and 2.0% and 0.05% and 0.4%, respectively. Seven of these strains were subsequently selected for the isolation of sodium cholate resistant derivatives by exposure to gradually increasing concentrations of this compound. Derivatives resistant to at least 1.2% of cholate were obtained from Bifidobacterium bifidum CECT 4549 and B. bifidum M6. Further analysis of the CECT 4549 derivative indicated that the phenotype induced was stable; the enzymatic activities analysed remained unchanged and no major rearrangements of chromosomal DNA were produced. However, the cells of the resistant derivative were smaller and more regular in average size than those of the original strain. In addition, the derivative had lost the ability to use lactose, fructose and galactose, and showed reduced expression levels of two membrane proteins of 78 and 114 kDa. Finally, the resistant strain displayed higher survival and lower growth inhibition in the presence of cholate than the original strain.