Activation of antifreeze proteins from larvae of the beetle Dendroides canadensis

Summary Purified antifreeze proteins (AFPs) from the larvae of the beetle Dendroides canadensis do not produce the high levels of antifreeze activity seen in the hemolymph of overwintering larvae, even when the purified AFPs are assayed at very high concentrations. However, addition of certain proteins or agar (at concentrations sufficiently low that the gel state does not result) to the Dendroides AFP resulted in a 2–3-fold increase in activity. A 70-kDa protein with AFP-activating capabilities was purified from Dendroides larvae. Addition of this endogenous activator protein to a 4 mg·ml^-1 solution of AFP increased the activity of the AFPs to values comparable to those of the hemolymph of overwintering larvae. Data derived from a modified immunoblot technique demonstrate that the activators bind to the AFP, or vice versa. Formation of this association must allow the AFP to block ice crystal growth by binding to the surface of potential seed crystals in the normal fashion. However, because the AFP-activator complex is much larger than the AFP alone, the complex probably blocks a greater surface area of the crystal and is thus a more efficient antifreeze.Abbreviations AFP antifreeze protein - BSA bovine serum albumine - DEAE diethylaminoethyl - Ig immunoglubolin - LPIN lipoprotein ice nucleator - PIN protein ice nucleator - SDS sodium dodecyl sulfate - PAGE polyacrylamide gel electrophoresis - TH thermal hysteresis     

Further investigation on nuclear transplantation in different orders of teleost: the combination of the nucleus of Tilapia (Oreochromis nilotica) and the cytoplasm of Loach (Paramisgurnus dabryanus).

The nucleus of a blastula cell from Tilapia (Oreochromis nilotica, family Cichlidae, order Perciformes) was transplanted into an enucleated egg of Loach (Paramisgurnus dabryanus, family Cobitidae, order Cypriniformes). From among 3747 nucleo-cytoplasmic hybrid (NCH) eggs two NCH larval fish (0.05%) were obtained; one died on the 6th day and the other died on the 12th day after the operation. Morphological examinations showed that both NCH larval fish had developed normally with an opened mouth except they could not take food after complete utilization of their egg yolk on the 5th day of development. The possible mechanisms for obtaining such inter-order NCH larval fish are discussed. This is the first report indicating that inter-order NCH larval fish can be obtained in spite of their evolutionary divergence.     

烙铁头蛇毒纤维蛋白原溶酶研究 Ⅰ分离纯化及理化性质(英文)

通过DEAE-SephadexA-50,DEAE-SepharoseCL-6B,MonoQ （FPLC）三步离子交换柱层析,纯化得到一新的纤维蛋白原溶酶,在碱性聚丙烯酰胺凝胶电泳和SDS-聚丙烯酰胺凝胶电泳上均呈单一的蛋白带。分子量为2600,等电点pl4.7;它是一个糖蛋白,含糖量6.4%,其中中性糖0.3%,已糖胺4.9%,唾液酸1.2%。烙铁头蛇毒纤维蛋白原溶酶TMVFg由187个氨基酸组成,含有较高的酸性氨基酸,此外甘氨酸含量也较高。TMVFg热稳定性强,而酸不稳定,在280 nm处具有典型的蛋白吸收峰,在无离子水中紫外消光系数E0.1%/280 nm=1.558。纯化的TMVFg具有较强的精氨酸酯酶活力,对苯甲酰-L-精氨酸乙酯（BAEE）的Km值为1.4×10[-3]M。TMVFg的活性受苯甲基丹磺酰氟（PMSF）抑制;乙二胺四乙酸（EDTA）对活性无影响。TMVFg不能使纤维蛋白原凝固,但能水解纤维蛋白原α、β链。纤溶实验表明TMVFg具有激活纤溶作用。纯化的烙铁头蛇毒纤维蛋白原溶酶对酪蛋白无作用,无出血活性,因而与凝血酶样酶、出血毒素及β-纤维蛋白原溶酶（OUYANG et al.,1977）明显不同。     

Role of sulphated tyrosine residue in influencing the biologic activity of human cholecystokinin-33

To evaluate the role of the sulphated tyrosine residue in position 27 in human cholecystokinin-33, parallel bioassay of the sulphated form of human cholecystokinin-33 and the unsulphated form of human cholecystokinin-33 was performed on the pancreatic protein secretion. Both peptides increased the protein output in a dose-related manner. However, the sulphated form possessed a considerably higher activity than the sulphated form. The relative potency of the unsulphated human cholecystokinin-33 compared to that of the sulphated human cholecystokinin-33 (taken as 1.0) was 0.08. From the results, it was suggested that the sulphated tyrosine may play an important role in controlling the activity of the longer molecular forms as well as that of the smaller forms of cholecystokinin.     

Nascent high density lipoproteins from liver perfusates of orotic acid-fed rats

Uniformly fatty livers from orotic acid-fed rats secreted almost no very low density lipoproteins (VLDL) but normal amounts of nascent high density lipoproteins (HDL) accumulated in perfusates. When lecithin:cholesterol acyltransferase (LCAT) was inhibited, nascent HDL were uniformly discoidal and lacked cholesteryl esters. Lipid and apoprotein compositions of nascent HDL from normal and fatty livers were similar whether LCAT was inhibited or not. Apolipoprotein B-100 was not detected in perfusates of uniformly fatty livers, but small amounts of apolipoprotein B-48 were present in HDL2 fractions. Nascent lipoproteins were not seen in Golgi compartments, but lipid-rich particles were clearly evident in endoplasmic reticulum cisternae adjacent to the cis face of the Golgi complex, suggesting that orotic acid blocks VLDL secretion by preventing translocation of nascent particles from the endoplasmic reticulum to the cis Golgi compartment. The accumulation of normal amounts of discoidal HDL in liver perfusates despite virtual absence of triglyceride-rich lipoproteins in Golgi secretory compartments, the space of Disse, and the perfusate is inconsistent with the concept that nascent HDL are exclusively a product of surface remnants cast off during lipolysis of chylomicrons and VLDL.     

营养膜无土栽培技术

前言营养膜技术(Nutrient Film Technique简称NFT)是国外广泛采用的一种特殊的无土栽培方法。它是将植物栽植在一个狭长的不透水的营养槽中,槽中有缓慢流动的营养液。植物根系的一部分浸没在营养液中,一部分露在空气中,植物从这种小环境中吸收水、营养物和氧气。由于这种方法采用很薄一层营养液在根际周围流动,裸露的根     

单面针的生物碱研究

自芸香科(Rutaceae)花椒属植物单面针(Zanthoxylum nitidum var. fastuosum How ex Huang)的根皮中分得五种已知生物碱:乙氧基白屈菜红碱(ethoxychelerythrine)(Ⅰ);氯化光花椒碱(nitidine chloride)(Ⅱ);去甲基白屈菜红碱(des-N-methychelerythrine)(Ⅲ);α—别隐品碱(α-allocryptopine)(Ⅳ);鹅掌揪宁(liriodenine)(Ⅴ).     

流行性感冒病毒重组株京生75-29 R2 T1及冷适应株31-广的基因分析

用聚丙烯酰胺凝胶电泳方法分析了流行性感冒病毒重组株京生75-29R2 T1(H3N2)及冷适应株31-广(H3N2)的RNA及多肽。重组株京生75-29R2 T1的HA及M基因系来自流行病毒亲本株/甲/北京/29/75(H3N2),而P_2、NA、NP及NS基因则来自温度敏感母株福R3(H2N2)。流行病毒株甲/穗/03/68(H3N2)在低温条件下经鸡胚尿囊腔传递24代而获得的冷适应疫苗毒株31-广(H3N2)其基因型与野毒株一致。