Independency of anti-HIV-1 activity from ribosome-inactivating activity of trichosanthin

Trichosanthin (TCS) is a type I ribosome-inactivating (RI) protein possessing multiple biological and pharmacological activities. Its major action is inhibition of human immunodeficiency virus (HIV) replication but the mechanism is still elusive. All evidences showed that this action is related to its RI activity. Previous studies found that TCS mutants with reduced RI activity simultaneously lost some anti-HIV activity. In this study, an exception was demonstrated by two TCS mutants retaining almost all RI activity but were devoid of anti-HIV-1 activity. Five mutants were constructed by using site-directed mutagenesis with either deletion or addition of amino acids to the C-terminal sequence. Results showed that the RI activity of mutants with C-terminal deletion mutants (TCSC2, TCSC4, and TCSC14) decreased by 1.2-3.3-fold with parallel downshifting of its anti-HIV-1 activity (1.4-4.8-fold). Another two mutants, TCSC19aa and TCSKDEL having 19 amino acid extension and a KDEL signal sequence added to the C-terminal sequence, retained all RI activity but subsequently lost most of the anti-HIV-1 activity. These findings suggested that ribosome inactivation alone might not be adequate to explain the anti-HIV action of TCS. (C) 2003 Elsevier Science (USA). All rights reserved.

Tetranorclerodanes and clerodane-type diterpene glycosides from Dicranopteris dichotoma

The acetone extract of Dicranopteris dichotoma afforded two new tetranorclerodanes, 18-hydroxyaylthonic acid (1) and 18-oxo-aylthonic acid (2), and four new clerodane-type diterpene glycosides, (6S,13S)-6-O-[6-O-acetyl-beta-D-glucopyranosyl-(1 -> 4)-alpha

Periglaucines A-D, anti-HBV and -HIV-1 alkaloids from Pericampylus glaucus

Four new hasubanane-type alkaloids, periglaucines A-D (1-4), and three known alkaloids, norruffscine (5), (-)-8-oxotetrahydropalmatine (6), and (-)-8-oxocanadine (7), were isolated from the aerial parts of Pericampylus glaucus. Their structures were eluci

湖南尖吻蝮蛇毒出血毒素的圆二色性和化学修饰

用远紫外CD谱研究了湖南产尖吻蝮蛇毒的两个出血毒素(DaHT-1、DaHT-2)的溶液构象, 计算得DaHT-1的#alpha#螺旋、#beta#折叠、无规卷曲的含量分别为36.9%, 35.5%, 27.6%; DaHT-2的#alpha#螺旋、#beta#折叠、无规卷曲分别为23.4%, 31.3%, 45.3%。随pH的增大或减小。温度和pH对CD谱的影响相似。0.02mol/L EDTA便导致两个出血毒素呈极度的无序状态。胰蛋白酶不影响它们的出血活性。图4表4参10

乳酸脱氢酶C_(4)的精子免疫荧光定位特性

研究表明精子表面的乳酸脱氢酶C_(4)(LDH-C_(4))呈现多位点分区分布, 精子的 不同部位暴露的LDH-C_(4)的抗原决定簇不同。在被检动物中, LDH-C_(4)显示明显的同源性。图版1表4参12

银鲫卵黄原的诱导、分离纯化及其生化鉴定

以雄性银鲫为实验材料 ,通过雌二醇 (Estradiol- 1 7β,E2 )的多次诱导 ,使得 E2 诱导产物成为血清中的主要蛋白。而后 ,在快速蛋白液相色谱 (FPL C)系统上 ,利用高交换量的阴离子交换层析 Q柱 ,成功的从血清中提纯了与雌性特异蛋白相一致的 E2 诱导产物。糖、磷、脂蛋白分析表明 ,它是一类糖磷脂蛋白大分子。同时 ,它能被 Mg2 + - Ethylenediamine tetraaceticacid(Mg2 + - EDTA)部分沉淀 ,这进一步证明 ,与雌性特异蛋白相一致的

4种沼虾的SRFA指纹研究

本文用沼虾属（Ｍａｃｒｏｂｒａｃｈｉｕｍ）的４个主要种：罗氏沼虾（Ｍ．ｒｏｓｅｎｂｅｒｇｉ）、海南沼虾（Ｍ．ｈａｉｎａｎｅｎｓｅ）、日本沼虾（Ｍ．ｎｉｐｐｏｎｅｎｓｅ）、粗糙沼虾（Ｍ．ａｓｐｅｒｕｌｕｍ）作材料，构建了它们的ＳＲＦＡ－ＤＮＡ指纹，经比较得到这４个种的ＳＲＦＡ分子标记，并构建了它们之间的谱系关系图，发现日本沼虾和海南沼虾在遗传上比较接近，属于同一“支序”，粗糙沼虾与另外３者在遗传上都相距较远。

Rapid quantification of total microcystins in cyanobacterial samples by periodate-permanganate oxidation and reversed-phase liquid chromatography

Microcystins (MCs) comprise a family of more than 80 related cyclic hepatotoxic heptapeptides. Oxidation of MCs causes cleavage of the chemically unique C-20 beta-amino acid (2S, 3S, 8S, 9S)-3-amino-9-methoxy-2,6,8-trimethyl-10-phenyldeca-4,6-dienoic acid (Adda) amino to form 2-methyl-3-methoxy-4-phenylbutanoic acid (MMPB), which has been exploited to enable analysis of the entire family. In the present study, the reaction conditions (e.g. concentration of the reactants. temperature and pH) used in the production of MMPB by oxidation of cyanobacterial samples with permanganate-periodate were optimized through a series of well-controlled batch experiments. The oxidation product (MMPB) was then directly analyzed by high-performance liquid chromatography with diode array detection. The results of this study provided insight into the influence of reaction conditions on the yield of MMPB. Specifically, the optimal conditions, including a high dose of permanganate (>= 50 mM) in saturated periodate solution at ambient temperature under alkaline conditions (pH similar to 9) over 1-4 h were proposed, as indicated by a MMPB yield of greater than 85%. The technique developed here was applied to determine the total concentration of MCs in cyanobacterial bloom samples, and indicated that the MMPB technique was a highly sensitive and accurate method of quantifying total MCs. Additionally, these results will aid in development of a highly effective analytical method for detection of MMPB as an oxidation product for evaluation of total MCs in a wide range of environmental sample matrices, including natural waters, soils (sediments) and animal tissues. (C) 2009 Elsevier B.V. All rights reserved.

Antisense for gonadotropin-releasing hormone reduces gonadotropin synthesis and gonadal development in transgenic common carp (Cyprinus carpio)

Generating transgenic fish with desirable traits (e.g., rapid growth, larger size, etc.) for commercial use has been hampered by concerns for biosafety and competition if these fish are released into the environment. These obstacles may be overcome by producing transgenic fish that are sterile, possibly by inhibiting hormones related to reproduction. In vertebrates, synthesis and release of gonadotropin (GtH) and other reproductive hormones is mediated by gonadotropin-releasing hormone (GnRH). Recently two cDNA sequences encoding salmon-type GnRH (sGnRH) decapeptides were cloned from common carp (Cyprinus carpio). This study analyzed the expression of these two genes using real-time polymerase chain reaction (RT-PCR) in different tissues carp at varying developmental stages. Transcripts of both genes were detected in ovary and testis in mature and regressed, but not in juvenile carp. To evaluate the effects of sGnRH inhibition, the recombinant gene CAsGnRHpc-antisense, expressing antisense sGnRH RNA driven by a carp beta-actin promoter, was constructed. Blocking sGnRH expression using antisense sGnRH significantly decreased GtH in the blood of male transgenic carp. Furthermore, some antisense transgenic fish had no gonadal development and were completely sterile. These data demonstrate that sGnRH is important for GtH synthesis and development of reproductive organs in carp. Also, the antisense sGnRH strategy may prove effective in generating sterile transgenic fish, eliminating environmental concerns these fish may raise. (c) 2007 Published by Elsevier B.V.

Effects of 2,3,7,8-TCDD and benzo[a]pyrene on modulating vitellogenin expression in primary culture of crucian carp (Carassius auratus) hepatocytes

Vitellogenin (Vtg) is the precursor of yolk protein. Its expression and secretion are estrogen-regulated and are crucial for oocyte maturation. An in vitro xenoestrogen screening model was established by measuring Vtg induction in cultured primary hepatocytes from crucian carp. Vtg production was detected by biotin-avidin sandwich ELISA method while Vtg and cytochrome P4501A1 (CYP1A1) mRNA induction were measured by semi- quantitative PCR-primer dropping technique. Vtg and Vtg mRNA were dose-dependently induced by diethylstilbestrol (DES, 0.2-200 ng/mL) in hepatocytes of crucian carp. Co-treatment of the DES-induced hepatocytes with either 2,3,7,8-TCDD (TCDD, 0.1-4 pg/mL) or benzo[a]pyrene (B[a]P, 5-1000 ng/mL) resulted in a reduction of Vtg production and an increment of CYP1A1 mRNA expression both in a dose dependent manner, indicating the anti-estrogenic effects of the compounds. However, at lower tested concentrations, TCDD (0.1, 0.2 pg/mL), B[a]P (5 ng/mL) seemed to have a potentiating effect on Vtg expression and secretion, although by their own these compounds had no observable estrogenic effect on Vtg induction. Tamoxifen (a selective estrogen receptor modulators, 1 nmol/L-1 mumol/L), and P-naphtho-flavone (beta-NF, an aryl hydrocarbon receptor inducing compounds, 2.5-1000 ng/mL) also were employed to study the possible interactions in DES-induced Vtg expression. In co-treatment of the DES-induced hepatocytes with beta-NF or tamoxifen, the decrease in Vtg production did parallel induction of CYP1A1 for beta-NF, but tamoxifen inhibited Vtg induction did not parallel induced CYP1A1 expression in all test concentrations. On the contrary, it was found that in co-treatment of the TCDD-induced hepatocytes with DES, TCDD induced CYP1A1 mRNA production was inhibited by DES also. These results implicated a possible cross talk between estrogen receptor- and aryl hydrocarbon receptor-mediated pathways in the hepatocytes.

The molecular characterization of RNA segment S9 of grass carp hemorrhage virus (GCHV), an aquareovirus

Although reovirus infection is one of the major virus diseases of grass carp in China, the available knowledge on the structure and function of genes and proteins of the virus is limited. The complete sequence of the S9 genome segment of grass carp hemorrhage virus (GCHV) was determined. The segment consists of 1130 nucleotides and has a large open reading frame (ORF) encoding a protein of 352 amino acids with predicted molecular mass of 37.7 kDa. Amino acid sequence comparison revealed that the deduced protein encoded by GCHV S9 is closely related to the sigma NS proteins of mammalian reovirus (MRV) and avian reovirus (ARV). Secondary structure analysis displayed that the form of alpha -helices (40.1%) and beta -sheets (49.4%) are the richest two contents in the protein encoded by S9, and this protein is predicted to be a nonstructural protein. (C) 2001 Elsevier Science B.V. All rights reserved.

High-power operation of uncoated strain-compensated quantum cascade lasers at 4.8 mu m

High-power operation of uncoated 22-mu m-wide quantum cascade lasers (QCLs) emitting at lambda approximate to 4.8 mu m is reported. The emitting region of the QCL structure consists of a 30-period strain-compensated In0.68Ga0.32As/In0.37Al0.63As superlattice. For a 4-mm-long laser in pulsed mode, a peak output power is achieved in excess of 2240mW per facet at 81K with a threshold current density of 0.64kA/cm(2). The effects of varying the cavity lengths from 1 to 4mm on the performances of the QCLs are analysed in detail and the low waveguide loss of only about 1.4 cm(-1) is extracted.

High responsivity resonant-cavity-enhanced InGaAs/GaAs quantum-dot photodetector for wavelength of similar to 1 mu m at room temperature

The characteristics of a resonant cavity-enhanced InGaAs/GaAs quantum-dot n-i-n photodiode with only a bottom distributed Bragg reflector used as the cavity mirror, are reported. To suppress the dark current, an AlAs layer is inserted into the device structure as the blocking layer. It turns out that the structure still possesses the resonant coupling nature, and makes Rabi splitting discernible in the photoluminescence spectra. The measured responsivity spectrum of the photocurrent shows a peak at lambda = 1030 nm, and increases rapidly as the bias voltage increases. A peak responsivity of 0.75 A/W, or equivalently an external quantum efficiency of 90.3%, is obtained at V-bias = -1.4 V.

Molecular beam epitaxy growth and photoluminescence study of room temperature 1.31 mu m (InyGa1-yAs/GaAs1-x Sb-x)/ GaAs bilayer quantum wells

Molecular beam epitaxy (MBE) growth of (InyGa1-yAs/GaAs1-xSbx)/GaAs bilayer quantum well (BQW) structures has been investigated. It is evidenced by photo luminescence (PL) that a strong blue shift of the PL peak energy of 47 meV with increasing PL excitation power from 0.63 to 20 mW was observed, indicating type II band alignment of the BQW. The emission wavelength at room temperature from (InyGa1-yAs/GaAs1-xSbx)/GaAs BQW is longer (above 1.2 μ m) than that from InGaAs/GaAs and GaAsSb/GaAs SQW structures (1.1 μ m range), while the emission efficiency from the BQW structures is comparable to that of the SQW. Through optimizing growth conditions, we have obtained room temperature 1.31 μ m wavelength emission from the (InyGa1-yAs/GaAs1-xSbx)/GaAs BQW. Our results have proved experimentally that the GaAs-based bilayer (InyGa1-yAs/GaAs1-xSbx)/GaAs quantum well is a useful structure for the fabrication of near-infrared wavelength optoelectronic devices. © 2005 Elsevier B.V. All rights reserved.