转生长激素基因鲤鱼几项血液指标的检测

以F2代转“全鱼”生长激素基因鲤鱼为材料,对其体重、体长和几项血液指标进行了检测。实验显示:转基因鱼体重和体长均极显著地高于对照鱼(P<0.01);转基因鱼每毫升血液中淋巴细胞总数为(25.2±6.7)×106个,对照鱼为(18.2±6.6)×106个,两者差异显著(P=0·015);转基因鱼红细胞压积值和白细胞吞噬百分率都高于对照鱼(P<0·05),但是吞噬指数没有统计学差异;转基因鱼的皮质醇水平为(63·8±16·8)ng/ml,对照鱼为(33·2±9·6)ng/ml,前者明显高于后者(P<0·01)。以上结果表明,转生长激素基因不但加速了鲤鱼的生长,还促进了其血液细胞的增殖和吞噬功能,并直接或间接地引起了血清皮质醇水平的升高。     

Profile of differentially expressed genes in blood vessels and blood cells of hyperlipidemia rats using suppression subtractive hybridization

Platelets, macrophages, endothelial cells, and smooth muscle cells can all contribute to atherosclerosis. To investigate the molecular events leading to atherosclerosis involving platelets, macrophages, endothelial cells, and smooth muscle cells, we carried out suppression subtractive hybridization (SSH) to generate a profile of genes overexpressed in the aorta and blood cells in high fat diet rats. From 85 random SSH-cDNA clones, we have screened 23 clones by Northern blotting, which were scored as overexpressed in the aorta and blood cells in high fat diet rats compared to normal diet rats. Sequencing showed that the gene corresponded to the known gene in the public databases, previously shown to be overexpressed in atherosclerosis, heparin-binding epidermal growth factor (EGF)-like growth factor (HB-EGF), and local production was seen in vascular smooth muscle and endothelial cells by RT-PCR and Western blotting. These results show that SSH provides a very efficient means to produce a profile of differentially expressed genes in atherosclerosis. The identified gene may provide novel points of therapeutic intervention and pathophysiological mechanisms in atherosclerosis.     

鲤鱼(Cyprinus Carpio) 促性腺激素基因的克隆、原核表达及多克隆抗体制备

采用逆转录-聚合酶链式反应(RT-PCR)方法,从鲤鱼脑垂体获得了两种GtH β亚基的cDNA, 克隆在pMD18-T载体.经测序确证后,将这两个基因克隆到原核表达载体pET -32(a)中,转化E.coli表达菌BL21(DE3),以IPTG诱导融合蛋白的高效表达.利用初步纯化后的抗原免疫新西兰大白兔,制备多克隆抗体.应用制备的兔抗血清与抽提的鲤鱼脑垂体的总蛋白分别进行Western-blot及ELISA分析,结果显示获得的多抗能特异识别各自的天然蛋白.该结果为纯化天然GtH蛋白提供了有效的检测手段,为进一步制备GtH单克隆抗体奠定了基础.     

Contribution of the cashew gum (Anacardium occidentale L.) for development of layer-by-layer films with potential application in nanobiomedical devices

The search for bioactive molecules to be employed as recognition elements in biosensors has stimulated researchers to pore over the rich Brazilian biodiversity. In this sense, we introduce the use of natural cashew gum (Anacardium occidentale L.) as an active biomaterial to be used in the form of layer-by-layer films, in conjunction with phthalocyanines, which were tested as electrochemical sensors for dopamine detection. We investigated the effects of chemical composition of cashew gum from two different regions of Brazil (Piauí and Ceará states) on the physico-chemical characteristics of these nanostructures. The morphology of the nanostructures containing cashew gum was studied by atomic force microscopy which indicates that smooth films punctuated by globular features were formed that showed low roughness values. The results indicate that, independent of the origin, cashew gum stands out as an excellent film forming material with potential application in nanobiomedical devices as electrochemical sensors.     

Distribution of the residual tritium content in organs of <Emphasis Type="Italic">Carassius gibelio</Emphasis> (Prussian carp) freshwater ray-finned fish

The distribution of residual tritium in organs of Carassius gibelio (Prussian carp) freshwater rayfinned fish as a representative of the Carassius phytophagous genus of Cyprinidae family was studied for determining the ³Н accumulation in the aquatic plants–fishes food chain in water bodies contaminated with radionuclides. In the first step, tritium was accumulated by 3000 g (wet weight) of the aquatic plant biomass in aquaria of 100 L volume with the specific water activity of 100 kBq L^–1. The aquatic plants were shown to actively take up the introduced tritium, accumulating up to 70% of the radionuclide. The content of tritium bound to organics in the biomass increased to 21% of the whole accumulated tritium. After that, the plants (Ceratophyllum, Lemna) were dried and mixed with dry fish fodder (1: 1), and the mixture was pressed in granules. The activity of each granule was up to 50 Bq. These granules were used for forced feeding of fishes, so that each individual took up in the course of the 550-day experiment up to 330 kBq of tritium. According to the analysis results, about 75% of the whole amount of the accumulated tritium was present in muscle tissues. Determination of the tritium content of proteins and lipids in the experimental systems revealed no differences in the use of Ceratophyllum and Lemna as fodder.     

Adsorption of Zn(II) in aqueous solution by activated carbons prepared from evergreen oak (Quercus rotundifolia L.)

In the present work activated carbons have been prepared from evergreen oak wood. Different samples have been prepared varying the concentration of the activating agent (H(3)PO(4)) and the treatment temperature. The yield of the process decreases with increasing phosphoric acid concentrations. Furthermore, high concentrations of activating agent lead to mainly mesoporous activated carbons to the detriment of the microporous texture. Treatment temperatures up to 450 degrees C lead to a progressive increase of the micro- and mesopore volumes. Values of specific surface area (S(BET)) as high as 1723 m(2) g(-1)have been obtained using appropriate phosphoric acid concentrations and treatment temperatures. The samples prepared have been successfully used in the removal of Zn(II) from aqueous solutions. From the adsorption kinetic data it may be stated that the equilibrium time is, in all cases, below 170 h. The adsorption process as a rule becomes faster as the mesopore volume and specific surface area of the samples increase. The adsorption isotherms in liquid phase point out that the adsorption capacity (n(0)(s)) and the affinity towards the solute (K(ci)) are higher for the sample showing the most developed mesoporous texture and surface area as well.     

Rapid quantification of carotenoids and fat in Atlantic Salmon (Salmo salar L.) by Raman spectroscopy and chemometrics

Raman spectroscopy (785 nm excitation) was used to determine the overall carotenoid (astaxanthin and cantaxanthin) and fat content in 49 samples of ground muscle tissue from farmed Atlantic salmon (Salmo salar L.). Chemically determined contents ranged from 1.0 to 6.8 mg/kg carotenoids and 36 to 205 g/kg fat. In addition to the raw Raman spectra, three types of spectral preprocessing were evaluated: the first derivative, subtraction of the fitted fourth-order polynomial (POLY), and the intensity normalized versions of POLY (POLY-SNV). Further, variable selection based on significance testing by use of jack-knifing was performed on each spectral data set. Partial least-squares regression resulted in a root mean square error of prediction of 0.33 mg/kg (R = 0.97) for carotenoids for the variable selected versions of all the preprocessed spectral data sets. The fat content was best estimated by the variable selected POLYSNV, resulting in a root mean square error of prediction of 15.5 g/kg (R = 0.95). Both preprocessing and variable selection improved the regression models significantly. The results demonstrate that Raman spectroscopy is a suitable method for simultaneous, rapid, and nondestructive quantification of both pigments and fat in ground salmon muscle tissue.     

Phytofiltration of cadmium from water by Limnocharis flava (L.) Buchenau grown in free-floating culture system

A hydroponics experiment was conducted to examine the phytofiltration of Cd by Limnocharis flava (L.) Buchenau grown in low-level Cd-contaminated water. For this, 45 d old seedlings of L .flava were transferred to a floating-support culture system containing nutrient solution spiked with four levels of Cd (0.5, 1, 2 and 4 mg l⁻¹) and were separately harvested after 3, 7, 21 and 30 d. After 30 d harvesting, the percentage removal of Cd from the above four treatments reached up to 98, 96, 95 and 93%, respectively. Interestingly, all treatments had higher growth rate than control at 95% confidence level and plants still remained healthy at 4 mg l⁻¹ Cd exposure. The bioaccumulation study showed a linear relationship of Cd (R² = 0.896–0.999) in all plant parts with the exposure time (3–30 d) and Cd concentrations in hydroponics system (0.5–4 mg l⁻¹). Although, the root of L. flava had higher Cd concentration than leaf and peduncles, the total Cd concentrations in aerial plant parts were higher than the roots. The maximum bioconcentration factor (BCF) and translocation factor (TF) value of L. flava were calculated as 984.42 and 1.43, respectively. Estimated Cd accumulation capacity of L. flava per unit area (m²) was found to be in the range of 218. 35–1698.92 mg m⁻².The experimental results demonstrated that L. flava is a suitable candidate for the phytofiltartion (>93%) of Cd from low-level Cd-contaminated water.