Using vibrational molecular spectroscopy to detect moist heating induced carbohydrates structure changes in cool-climate adapted barley grain

There is limited research to study how moist heating affects internal structure of barley grain on a molecular basis. The objectives of this study were to use vibrational molecular spectroscopy: 1) to determine the moist heating induced changes of barley carbohydrate (CHO) structure on a molecular basis, 2) to study the effects of moist heating on CHO chemical profiles, Cornell Net Carbohydrate and Protein System (CNCPS) subfractions, in situ rumen degradation, and predicted intestinal carbohydrate supply of barley grain; and 3) to reveal the association between molecular structure spectral features and CHO related metabolic characteristics. Barley samples (CDC cowboy) were collected from Kernen Crop Research Farm (Saskatoon, Canada) during two consecutive years. Half of each sample was kept as raw barley and the other half underwent moist heating (autoclaving at 120 °C for 60 min). The molecular spectroscopy (attenuated total reflectance-fourier transform infrared, ATR-FTIR) was used to detect the barley CHO related molecular structure spectral features. Moist heating did not affect carbohydrate related chemical profiles and CNCPS subfractions but it decreased rumen degradable carbohydrate. Rumen undegradable and intestinal digestion of CHO subfractions were not affected by moist heating. The advanced vibrational molecular spectroscopy can be used to detect carbohydrate molecular spectral features. Nutrient utilization prediction using molecular spectral characteristics is warranted and further investigation is encouraged.     

Soluble Klotho protein suppresses THP-1-derived foam cell formation

AIMTo investigate the role of soluble Klotho protein in THP-1-derived foam cell formation. METHODSTHP-1 monocytes were induced into macrophages by treatment with 160 nmol/L phorbol myristate acetate for 48 h, and then were divided into 6 groups: negative control group (THP-1-derived macrophages), positive control group [THP-1-derived foam cells induced by oxidized low-density lipoprotein (ox-LDL) for 48 h], and 25, 50, 100 and 200 μg/L soluble Klotho protein groups (THP-1-derived macrophages pretreated with soluble Klotho protein at the indicat?ed concentraions for 2 h and then induced by ox-LDL for 48 h). Lipid droplets in cytoplasm were observed by oil red O staining. The cholesterol outflow rate was detected by scintillation counting technique. The content of intracellular total cholesterol, free cholesterol and cholesterol ester was detected by enzyme fluorescence analysis. The expression of acyl-coenzyme A:cholesterol acyltransferase 1 （ACAT1） and ATP-binding cassette transport?er A1 (ABCA1) at mRNA and protein levels was determined by RT-qPCR and Western blot, respectively. RESULTSOil red O staining and lipid mass quantification showed that THP-1-derived foam cell formation was dose-dependently suppressed by soluble Klotho protein. The cholesterol efflux rate of THP-1-derived foam cells was increased by soluble Klotho protein in a dose-dependent manner (P<0.05). In addition, soluble Klotho protein decreased the expression of ACAT1 and increased the expression of ABCA1 in a dose-dependent manner (P<0.05). CONCLUSION The soluble Klotho protein inhibits THP-1-derived foam cell formation in a dose-dependent manner by down-regulating the expression of ACAT1 and up-regulating the expression of ABCA1.     

Effects of calpain-2 and autophagy-related protein 5 on hepatocyte apoptosis induced by endoplasmic reticulum stress

AIMTo investigate the effects of calpain-2 and autophagy-related protein 5 (Atg5) on apoptosis of BRL-3A rat normal liver cells during endoplasmic reticulum stress (ERS) induced by dithiothreitol (DTT). METH?ODS: BRL-3A cells were treated with DTT at 2.0 mmol/L for 0, 6, 12 and 24 h to induce ERS. Real-time cell analysis (RTCA) was used to measure the effect of DTT on BRL-3A cell proliferation. Apoptosis and cell cycle distribution were analyzed by flow cytometry. The mRNA expression of calpain-2 and Atg5 was detected by real-time PCR. The protein levels of calpain-2, Atg5, Atg7, Atg12 and microtubule-associated protein 1 light chain 3 (LC3) were determined by Western blot. The interaction between calpain-2 and Atg5 was investigated by co-immunoprecipitation (Co-IP). RESULTSThe proliferation of BRL-3A cells treated with DTT was significantly inhibited. The apoptosis of BRL-3A cells was significantly increased after DTT treatment for 6, 12 and 24 h as compared with 0 h group (P<0.05). The cell cycle was arrested in G₁ phase after DTT treatment (P<0.05). After DTT treatment for 6, 12 and 24 h, the mRNA expression of calpain-2 and Atg5 in the BRL-3A cells was significantly increased as compared with 0 h group (P<0.05). The protein levels of calpain-2, Atg12 and Atg7 in the cells treated with DTT for 6, 12 and 24 h were significantly higher than those in 0 h group, and the ratio of LC3-II/LC3-I was also significantly higher than that in 0 h group, while Atg5 expression was significantly lower than that in 0 h group (P<0.05). The results of Co-IP found that the anti-calpain-2 antibody precipitated Atg5 protein from the cell lysates, and the anti-Atg5 antibody also precipitated calpain-2 from the cell lysates, which confirmed the interaction between calpain-2 and Atg5. CONCLUSION Calpain-2 may participate in ERS-induced hepatocyte apoptosis by interacting with Atg5.     

不同光源对4种蔬菜幼苗叶绿素荧光及生理指标的影响

为了解不同光源或光源组合对茄子、辣椒、番茄、黄瓜幼苗叶绿素荧光及生理指标的影响,在4种基本光源的基础上,比较了不同光源处理下不同蔬菜植株叶片的叶绿素荧光参数、可溶性蛋白和可溶性糖含量。结果表明:红蓝光源不利于4种蔬菜幼苗叶片光合色素的形成,茄子、辣椒、番茄和黄瓜叶片中的光合色素含量分别在2白4红2黄、3白3红2黄、8黄或8红、8白光源下最高;茄子和番茄幼苗叶片在红蓝光源下,辣椒和黄瓜在3白3红2黄光源下,最大光化学效率和实际光化学效率都最大;茄子和辣椒幼苗在红蓝光源下所受非生物胁迫最小,叶片中可溶性蛋白含量最低,而番茄和黄瓜分别在3白3红2黄和8白光源下的可溶性蛋白含量最低;红蓝光源可明显增加4种蔬菜幼苗叶片的可溶性糖含量,对幼苗的形态建成有积极的影响。在冬春季蔬菜育苗生产中,可根据不同光源优势为不同种类蔬菜组配光源,以促进幼苗生长发育。     

无机碳源对零价铁介导的自养脱氮序批式反应器处理高氨氮废水的影响

无机碳源作为自养微生物的能量来源,是影响自养脱氮细菌富集的重要因素。文章通过添加不同量的KHCO3作为ZVI介导的自养脱氮体系中无机碳源,研究反应体系的脱氮效果影响趋势以及适宜的无机碳源添加量。结果如下:1)KHCO3作为无机碳源可以显著提高NH^+4-N去除率,KHCO3添加量越多,其NH^+4-N去除率越高。KHCO3添加量分别为2 g,1 g,0.5 g的SBR反应器R2,R1,R0.5的NH^+4-N去除率分别为98.39%,65.18%,44.56%,相较不添加KHCO3的R0分别提高86.5%,53.29%,32.67%。ZVI的添加会降低KHCO3对NH^+4-N氧化的促进作用。2)KHCO3可明显提高TIN去除效果,促进总氮脱除的高低顺序是R1>R0.5>R2,SBR反应器R2,R1,R0.5的平均TIN去除率分别为19.01%,32.04%,27.62%,相较于空白组R0分别提高了10.60%,23.63%,19.21%。3)KHCO3可中和硝化反应产生的H^+,对反应体系具有缓冲作用,可使微生物处于较适宜的酸碱环境中,更有利于反应器的稳定运行。4)适量的无机碳源KHCO3可以促进氨氧化细菌和厌氧氨氧化细菌的活性。该研究探讨了无机碳源在零价铁脱氮体系中的影响趋势,为铁型脱氮技术提供了理论支撑。     

Identification of resistance to Aspergillus flavus infection in cotton germplasm

Natural resistance in cottonseed to Aspergillus flavus infection has not been explored to date. A green fluorescent protein (GFP) expressing A. flavus strain was used to assess the resistance of seed from 35 cotton varieties from Gossypium arboreum, G. barbadense, and G. hirsutum. Mature cotyledons devoid of seed coat were wounded, inoculated, and assessed for innate resistance to A. flavus infection. Of the initial 35 varieties tested, we observed a range of resistance to infection in representatives of all three species. A subset of 15 representatives was further analyzed. Within this group, G. arboreum cultivar A2 186 and G. hirsutum cultivar SA 1582 were most resistant to fungal infection. The most susceptible cultivar in this group was G. hirsutum SA 1595. The remaining 12 representatives tested in the secondary screen (3 G. arboreum, 3 G. barbadense, and 6 G. hirsutum lines) exhibited intermediate resistance. We did not observe any relationship between species and resistance. Within each species, there was a range of responses to fungal infection. Future studies using this methodology to screen additional diploid and tetraploid cotton lines may enable us to identify naturally resistant germplasm that can be used to develop cotton with enhanced resistance to fungal infection.     

昆虫嗅觉相关蛋白的研究进展

昆虫对自然环境中复杂化学信号的识别多依赖于其灵敏的嗅觉系统,选择寄主、觅食、寻找配偶等行为的发生都以嗅觉识别为基础,而完成嗅觉识别还需要多种嗅觉相关蛋白的参与。嗅觉相关蛋白主要包括6种,即气味结合蛋白、化学感受蛋白、气味受体、感觉神经元膜蛋白、离子型受体和气味降解酶。不同种类和性别的昆虫中,嗅觉蛋白的种类、数量和分布各不相同。由于嗅觉蛋白在昆虫识别外界气味分子中的重要作用,国内外近年来对其展开了广泛、深入的研究。本文从几种嗅觉相关蛋白的生化特性、分子结构、生理功能、分布表达部位和研究概况等角度,较详细地综述了近年来国内外昆虫嗅觉相关蛋白的研究进展。     

大豆RING/U-box蛋白Glyma.13G115900的克隆及其对非生物胁迫的应答

课题组前期对干旱胁迫下大豆转录组的数据分析发现大豆Glyma.13G115900基因编码一个RING/U-box蛋白,其表达水平受干旱胁迫影响显著。本研究以垦丰16大豆为试验材料,克隆Glyma.13G115900基因。氨基酸多重序列比对表明其编码的蛋白与其它物种都具有高度保守的RING/U-box结构域。构建原核表达载体pET-29b-Glyma.13G115900转化到大肠杆菌中,Glyma.13G115900蛋白在大肠杆菌中能够表达。荧光定量PCR分析表明Glyma.13G115900基因的表达量受PEG、NaCl和ABA的影响显著,但基本不受冷胁迫的诱导。经PEG和NaCl处理后,该基因表达量与CK相比呈现出显著下降的趋势,PEG处理的表达量变化比NaCl下调的更明显;在ABA诱导下与CK相比该基因的mRNA丰度呈现出先上升后下降的趋势,在4 h表达量出现峰值,推测该基因可能通过依赖于ABA途径参与非生物胁迫应答。以上结果为进一步深入研究该基因的调控途径奠定基础。     

水稻齿叶矮缩病毒Pns10蛋白在水稻原生质体内的表达

【目的】水稻齿叶矮缩病毒(Rice ragged stunt virus,RRSV)Pns10蛋白在介体昆虫细胞内可形成类似病毒原质(viroplasm)的内含体,是RRSV侵染介体所必需。然而Pns10蛋白在水稻寄主中是否具有类似功能及其表达情况如何未见报道。【方法】利用大肠杆菌系统表达Pns10蛋白,免疫家兔制备多克隆抗体;通过水稻原生质体病毒侵染体系,利用免疫荧光技术分析Pns10蛋白在水稻原生质体内的分布情况,利用实时定量PCR技术和Western blot技术分别检测Pns10 RNA和Pns10蛋白在水稻原生质体内的积累情况。【结果】将Pns10基因克隆到Gateway系统原核表达载体p DEST17中,IPTG诱导表达成功后,制备融合蛋白抗血清。Western blot检测显示,该抗血清可检测感病水稻叶片中的Pns10蛋白。病毒侵染水稻原生质体后,Pns10蛋白可形成类似病毒原质的内含体;Pns10 RNA在病毒接种8 h后开始积累,24 h后达到最大值,随后开始下降;Pns10蛋白在24 h后开始表达,之后维持较高水平,60 h后略有下降。【结论】成功获得了Pns10抗血清;Pns10在水稻原生质体内成功表达,可形成类似病毒原质的内含体,并且Pns10 RNA的表达先于其蛋白的表达。     

Characterization of Spray-Dried Hydrolyzed Proteins from Pink Perch Meat Added with Maltodextrin and Gum Arabic

In the present study, spray-dried pink perch meat protein hydrolysate (PPH) and hydrolysate with maltodextrin and gum arabic (PPHMG) were characterized in relation to their physical, rheological, functional, antioxidant, thermal, and sensory properties. The Herschel–Bulkley model was suitable to explain the flow behavior of PPH and PPHMG emulsions. Both the samples exhibited functional and antioxidant properties. Antioxidant activities were significantly higher for PPH compared with PPHMG (p<0.05). PPH and PPHMG alone did not exhibit antimicrobial activity but enhanced the activity of chitosan against pathogens. Sensory acceptability of the samples in soup revealed that PPHMG of up to 4% was highly scored without any traces of bitterness, whereas for PPH, a concentration of up to 3% was acceptable.