Der p 1 is the primary activator of Der p 3, Der p 6 and Der p 9 the proteolytic allergens produced by the house dust mite Dermatophagoides pteronyssinus

Background

The enzymatic activity of the four proteases found in the house dust mite Dermatophagoides pteronyssinus is involved in the pathogenesis of allergy. Our aim was to elucidate the activation cascade of their corresponding precursor forms and particularly to highlight the interconnection between proteases during this cascade.

Methods

The cleavage of the four peptides corresponding to the mite zymogen activation sites was studied on the basis of the Förster Resonance Energy Transfer method. The proDer p 6 zymogen was then produced in Pichia pastoris to elucidate its activation mechanism by mite proteases, especially Der p 1. The role of the propeptide in the inhibition of the enzymatic activity of Der p 6 was also examined. Finally, the Der p 1 and Der p 6 proteases were localised via immunolocalisation in D. pteronyssinus.

Results

All peptides were specifically cleaved by Der p 1, such as proDer p 6. The propeptide of proDer p 6 inhibited the proteolytic activity of Der p 6, but once cleaved, it was degraded by the protease. The Der p 1 and Der p 6 proteases were both localised to the midgut of the mite.

Conclusions

Der p 1 in either its recombinant form or in the natural context of house dust mite extracts specifically cleaves all zymogens, thus establishing its role as a major activator of both mite cysteine and serine proteases.

General significance

This finding suggests that Der p 1 may be valuable target against mites.     

Real‐time PCR for identification of the soybean aphid,Aphis glycines Matsumura

The soybean aphid (Aphis glycines Matsumura) is an economically significant pest in North America, causing extensive damage to soybean crops through direct feeding damage and disease transmission. If unchecked, this pest could cause billions of dollars of damage to soybean crops. Identification of the soybean aphid can be difficult due to its small size, complex life cycle and morphological plasticity. Generally, an expert is required to identify a specimen. Additionally, identification of some life stages, such as eggs, is impossible. DNA barcoding has been successfully used to differentiate aphid species, including A. glycines, based on sequencing of a standardized gene region. Although this method represents an important step towards accurate identification, samples must still be sent to specialized facilities for analysis. Using existing DNA barcode sequences in the publically accessible Barcode of Life Data System (BOLD; www.boldsystems.org ), species‐specific differences were identified and used to develop a real‐time PCR assay to identify soybean aphids. This assay can be run on portable systems for rapid, accurate and simple identification in the field. The use of a non‐destructive DNA extraction protocol allows the original insect to be vouchered and therefore available for further study if necessary. This work represents an important step in soybean aphid management.     

六价铬对水绵生长的毒性效应

为研究六价铬（Cr^6＋）对水绵（Spirogyrasp．）的毒性效应和水绵对六价铬的毒性响应,实验设置5个暴露组（4、6、8、10和12mg／L）和1个对照组。结果表明：96h后各暴露组对水绵的生长均有抑制作用,铬浓度越高,藻细胞叶绿素a越低,显示出较明显的剂量一效应关系;六价铬对水绵的96h的Ec。值为7．25mg／L。细胞浸出液电导率在低浓度（4～6mg／L）组上升较缓慢,在高浓度组（8～12mg／L）上升显著;丙二醛（MDA）累积含量在Cr6＋≥4mg／L时,累计含量上升较高（P〈0．01）,当Cr^6＋≥8mg／L时,累积含量上升较少。水绵细胞浸出液电导率与MDA存在显著正相关（P〈0．05,r=0．891）。水绵细胞浸出液电导率和MDA含量与六价铬浓度分别存在显著（P〈0．05,r=0．951）和极显著（P〈0．01,r=0．977）的非线性的毒性响应关系。     

红树植物桐花树叶片多酚提取物对酪氨酸酶活性抑制及抗自由基和抗菌活性分析

采用超声波辅助-乙酸乙酯提取方法获得桐花树〔Aegiceras corniculatum(Linn.)Blanco〕叶片多酚提取物,研究了该提取物对酪氨酸酶活性的抑制作用及其动力学特征,并分析了该提取物对DPPH·自由基的清除作用及其抗菌活性。结果显示:多酚提取物得率约为(122.0±31.4)mg·g-1,提取物中多酚含量约为(521.8±17.2)mg·g-1。该提取物对酪氨酸酶活性的抑制作用呈明显正相关的量效关系,IC50为0.650 g·L-1,与阳性对照槲皮素对酪氨酸酶活性的抑制能力相近;该提取物通过降低酶活性实现对酪氨酸酶活性的抑制,且该抑制作用具有可逆性;随提取物质量浓度的提高酶促反应Km值增大、vm值减小,其动力学特征符合混合Ⅰ型抑制类型;对游离酶的抑制常数Ki为0.833 g·L-1,对酶-底物络合物的抑制常数Kis为1.823 g·L-1,表明该提取物与游离酶的亲和力大于其与酶-底物络合物的亲和力。随质量浓度提高,该提取物对DPPH·自由基的清除率逐渐增大并在0.000~0.600 g·L-1范围内呈明显量效关系,且IC50为0.304 g·L-1,与0.036 g·L-1槲皮素等效,显示该多酚提取物对自由基的清除能力较强。随质量浓度提高,该提取物对大肠杆菌(Escherichia coli)、金黄色葡萄球菌(Staphylococcus aureus)和枯草芽孢杆菌(Bacillus subtilis)的抑菌圈直径均逐渐增大,显示该提取物对3种供试菌均有抑菌作用,最小抑菌浓度均为25 g·L-1。结果表明:通过深入的开发研究,桐花树叶片多酚提取物可作为兼具辅助防腐抑菌功能的新型酪氨酸酶抑制剂。     

常用萃取剂提取福寿螺卵中类胡萝卜素的初步研究

【背景】福寿螺为世界性恶性入侵水生动物,也是我国公布的第一批外来人侵物种之一。福寿螺大量啃食为害水稻、茭白、白莲等重要农作物,对我国南方各省农业生产造成了巨大威胁,因此,预防和控制福寿螺灾害显得尤为重要。合理利用福寿螺能有效控制福寿螺的数量和危害,是生物防治的一个重要部分。福寿螺卵中含丰富的类胡萝卜素,充分利用螺卵中的类胡萝卜素能拓展福寿螺的利用途径和方法。【方法】为探寻福寿螺卵中类胡萝卜素的提取方法,本研究采用甲醇、无水乙醇、丙酮、乙酸乙酯、二氯甲烷、石油醚等6种常用萃取剂提取福寿螺卵中类胡萝卜素,用紫外可见光分光光度计测定其含量。【结果】结果显示,不同萃取剂中类胡萝卜素的提取量不同,醇类为较适合的提取液（甲醇〉无水乙醇〉丙酮）。【结论与意义】本研究对福寿螺卵中类胡萝卜素的提取方法进行了探索和研究,找出了合适的提取液,为拓展福寿螺的利用途径,以及福寿螺的综合防治提供了参考。     

Carboxymethylcellulose-montmorillonite nanocomposite films activated with murta (Ugni molinae Turcz) leaves extract

The functionality of nanocomposite films based on carboxymethylcellulose (CMC) and montmorillonite (MMT) activated with murta (Ugni molinae Turcz) leaves extract was studied. Films were prepared by casting from film-forming dispersions containing CMC, glycerol (used as plasticizer) and different concentrations of MMT, using water or murta extract as solvent. The addition of MMT increased the tensile strength and the elasticity modulus of the films, and decreased their permeabilities to water vapor, oxygen and carbon dioxide. Besides the antioxidants properties provided to the films, the addition of murta leaves extract changed the gas permeability in different forms according to the MMT content, and plasticized the nanocomposite matrix.     

Metabolic engineering of soybean affords improved phytosterol seed traits

Different combinations of three rate‐limiting enzymes in phytosterol biosynthesis, the Arabidopsis thaliana hydroxyl methylglutaryl CoA1 (HMGR1) catalytic subunit linked to either constitutive or seed‐specific β‐conglycinin promoter, and the Glycine max sterol methyltransferase1 (SMT1) and sterol methyltransferase2‐2 (SMT2‐2) genes, under the control of seed‐specific Glycinin‐1 and Beta‐phaseolin promoters, respectively, were engineered in soybean plants. Mature seeds of transgenic plants displayed modest increases in total sterol content, which points towards a tight control of phytosterol biosynthesis. However, in contrast to wild‐type seeds that accumulated about 35% of the total sterol in the form of intermediates, in the engineered seeds driven by a seed‐specific promoter, metabolic flux was directed to Δ⁵‐24‐alkyl sterol formation (99% of total sterol). The engineered effect of end‐product sterol (sitosterol, campesterol, and stigmasterol) over‐production in soybean seeds resulted in an approximately 30% increase in overall sitosterol synthesis, a desirable trait for oilseeds and human health. In contradistinction, increased accumulation of cycloartenol and 24(28)‐methylencylartanol (55% of the total sterol) was detected in plants harbouring the constitutive t‐HMGR1 gene, consistent with the previous studies. Our results support the possibility that metabolic flux of the phytosterol family pathway is differentially regulated in leaves and seeds.     

Anti-inflammatory and anti-tumor-promoting effects of 5-deprenyllupulonol C and other compounds from Hop (Humulus lupulus L.)

A new phloroglucinol derivative, 5‐deprenyllupulonol C ( 1 ), along with four other phloroglucinol derivatives, 2 – 5 , five chalcones, 6 – 10 , four flavanones, 11 – 14 , two flavonol glycosides, 15 and 16 , and five triterpenoids, 17 – 21 , were isolated from the female inflorescence pellet extracts of hop (Humulus lupulus L.). Upon evaluation of these compounds against the Epstein? Barr virus early antigen (EBV‐EA) activation induced by 12‐O‐tetradecanoylphorbol 13‐acetate (TPA) in Raji cells, twelve compounds, i.e., 1 – 4, 11 – 14, 17 – 19 , and 21 , showed potent inhibitory effects on EBV‐EA induction, with IC₅₀ values in the range of 215–393 mol ratio/32 pmol TPA. In addition, eleven compounds, i.e., 1 – 4, 6, 11, 12, 14, 17, 18 , and 20 , were found to inhibit TPA‐induced inflammation (1 μg/ear) in mice, with ID₅₀ values in the range of 0.13–1.06 μmol per ear. Further, lupulone C ( 2 ) and 6‐prenylnaringenin ( 14 ) exhibited inhibitory effects on skin‐tumor promotion in an in vivo two‐stage mouse‐skin carcinogenesis test based on 7,12‐dimethylbenz[a]anthracene (DMBA) as initiator and with TPA as promoter.