应用改进的多聚组氨酸标签T7表达载体构建肺癌cDNA文库

应用噬菌体C端展示系统构建的cDNA文库缺乏开放阅读框筛选机制,文库中多数噬菌体克隆展示框外非天然短肽,给后期蛋白质的筛选带来了不便. 为实现噬菌体的ORF筛选功能,利用PCR技术对已有载体T7Select10-3b进行改造,在MCS处外源cDNA插入位点的3′端引入6聚组氨酸筛选标签,经包装后挑取成功表达的单克隆构建肺癌cDNA文库. 经镍柱亲和层析后,收集文库中表达组氨酸的克隆,利用化学发光免疫试验进行筛选效果鉴定. 结果显示,改造的新型载体可成功表达组氨酸标签,以此构建的肺癌cDNA文库经筛选后,含ORF插入的克隆由筛前的6 %提高至70 %,本研究为提高cDNA文库的质量提供了一种简便可行的方法.     

Generalized Bohm’s criterion and negative anode voltage fall in electric discharges

The value of the voltage fall across the anode sheath is found as a function of the current density. Analytic solutions are obtained in a wide range of the ratio of the directed velocity of plasma electrons v ₀ to their thermal velocity v _T . It is shown that the voltage fall in a one-dimensional collisionless anode sheath is always negative. At the small values of v ₀/v _T , the obtained expression asymptotically transforms into the Langmuir formula. Generalized Bohm’s criterion for an electric discharge with allowance for the space charge density ρ(0), electric field E(0), ion velocity v _i (0), and ratio v ₀/v _T at the plasma-sheath interface is formulated. It is shown that the minimum value of the ion velocity v _i ^* (0) corresponds to the vanishing of the electric field at one point inside the sheath. The dependence of v _i ^* (0) on ρ(0), E(0), and v ₀/v _T determines the boundary of the existence domain of stationary solutions in the sheath. Using this criterion, the maximum possible degree of contraction of the electron current at the anode is determined for a short high-current vacuum arc discharge.     

Specific features of the structure of the Z-pinch emitting region formed during the implosion of a foam-wire load at the ANGARA-5-1 facility

Results are presented from experimental studies of the structure of the compressed plasma of a Z-pinch produced during the implosion of a foam-wire load at the current of up to 3 MA. The foam-wire load consisted of two nested cylindrical cascades, one of which was a solid or hollow cylinder made of low-density agar-agar foam, while the other was a wire array. The wall thickness of a hollow foam cylinder was 100–200 μm. The images of the pinch and its spectrum obtained with the help of multiframe X-ray cameras and a grazing incidence spectrograph with a spatial resolution were analyzed. Data on the spatial structure of the emitting regions and the soft X-ray (SXR) spectrum of the Z-pinch in the final stage of compression of a foam-wire load were obtained. The implosion modes characterized by the formation of hot regions during implosion of such loads were revealed. The characteristic scale lengths of the hot regions were determined. It is shown that the energy distribution of SXR photons in the energy range from 80 eV to 1 keV forms the spatial structure of Z-pinch images recorded during the implosion of foam-wire loads. It is revealed that the spectral density of SXR emission in the photon energy range of 300–600 eV from hot Z-pinch regions exceeds the spectral density of radiation from the neighboring Z-pinch regions by more than one order of magnitude. Groups of lines related to the absorption and emission of radiation by atoms and multicharged ions of carbon and oxygen in the outer foam cascade of a foam-wire load were recorded for the first time by analyzing the spatial distribution of the SXR spectra of multicharged ions of the Z-pinch. The groups of absorption lines of ions (C III, O III, O IV, and O VI) corresponding to absorption of SXR photons in the Z-pinch of a tungsten wire array, which served as the inner cascade of a foam-wire load, were identified. The plasma electron temperature measured from the charge composition of carbon and oxygen ions in the outer agar-agar foam cascade was 10–40 eV. During the implosion of foam-wire loads at currents of up to 3 MA, SXR pulses (hν > 100 eV) with a duration of 10 ns and peak power of 3 TW were detected. It is shown that the temporal profile of single-peak and double-peak SXR pulses can be controlled by varying the parameters of the outer and inner cascades of the foam-wire load.     

Formation of an internal transport barrier and magnetohydrodynamic activity in experiments with the controlled density of rational magnetic surfaces in the T-10 Tokamak

Results are presented from experiments on the formation of an internal electron transport barrier near the q = 1.5 rational surface in the T-10 tokamak. The experiments were carried out in the regime with off-axis electron cyclotron resonance (ECR) heating followed by a fast plasma current ramp-up. After suppressing sawtooth oscillations by off-axis ECR heating, an internal transport barrier began to form near the q = 1.5 rational surface. In the phase of the current ramp-up, the quality of the transport barrier improved; as a result, the plasma energy confinement time increased 2–2.5 times. The intentionally produced flattening of the profile of the safety factor q(r) insignificantly affected magnetohydrodynamic activity in the plasma column in spite of the theoretical possibility of formation of substantial m/n = 3/2 and 2/1 magnetic islands. Conditions are discussed under which the flattening of the profile of the safety factor q near low-order rational surfaces leads to the formation of either an internal transport barrier or the development of an island magnetic structure induced by tearing modes.     

Association between Gene Polymorphism of Manganese Superoxide Dismutase and Prostate Cancer Risk

Manganese superoxide dismutase (MnSOD) is the most effective antioxidant enzyme in mitochondria and protects cells from reactive oxygen species‐induced oxidative damage. The aim of this study was to investigate the association between MnSOD Ala‐9Val gene polymorphism and prostate cancer (PCa) risk in Turkish men with prostate cancer. 33 patients with PCa and 81 control individuals were included in the study. We observed an association between MnSOD Ala/Ala frequency and a higher PCa risk. In addition, we found that the increased risk of early‐onset PCa (under age of 65) in the men homozygous for Ala allele was higher than the men homozygous for Val allele. However, we determined that MnSOD Ala‐9Val genotype was not associated with the aggressiveness of the disease. The results of our study suggest that MnSOD Ala/Ala genotype may influence on early‐onset of PCa patients, but no effect on subsequent development of the disease in Turkish men. However, our study has a limitation that is small numbers of individuals for cases and controls. Therefore, the presented study limited our statistical power to fully investigate the gene polymorphism on cancer risk. © 2013 Wiley Periodicals, Inc. J BiochemMol Toxicol 27:213‐218, 2013; View this article online at wileyonlinelibrary.com . DOI 10.1002/jbt.21472     

Antitumor activity of amides of dihydrobetulonic acid in vitro and in vivo

Amides containing homopiperidine and piperazine cycles were synthesized from dihydrobetulonic acid obtained by the oxidation of dihydrobetulin. All substances were shown to have a high antitumor activity (CCID₅₀ = 3.5–36.2 μM) in vitro in lymphoid (CEM-13, U-937) and monocytic (MT-4) human cell lines. Amides containing the methyl- and ethylpiperazine residues do not influence the viability of Lung Lewis Carcinoma cells in culture and do not have a significant effect on its transplants in C57BL/6 mice. However, these amides efficiently inhibit the development of metastases in lungs of these mice. The antimetastatic activity of the studied amides increases with changing the methyl by ethyl aliphatic residue in the piperazine cycle.     

High prevalence of serum folate receptor autoantibodies in children with autism spectrum disorders

Abstract

Background: Supplementation of folic acid by pregnant mothers is thought to lower the risk of autism spectrum disorders (ASDs) in the offspring. Folic acid is taken up by cells via receptors with high affinity for folate and reduced folic acid derivatives. However, this is blocked by the presence of folate receptor autoantibodies (FRAA). Cerebral FRAA have been detected with high frequency in children with ASDs, suggesting the existence of a link between folic acid uptake and disease aetiology.

Methods: We investigated the frequency of FRAA in serum samples from 40 children with ASDs and 42 gender- and age-matched children with typical development (TD). Serum FRAA concentrations were measured by enzyme-linked immunosorbent assay.

Results: We found a significant difference in the frequency of serum FRAA in the two study cohorts. Serum FRAA were present in 77.5% (31/40) of children with ASDs compared with 54.8% (23/42) of TD children (p?=?0.03746, Fischer’s exact test). Thus, serum FRAA are more prevalent in children with ASDs than in TD children.

Conclusions: Our data suggest that children with ASDs may have defects in folic acid absorption that play a role in the onset of ASDs.     

Glycolipids of Pseudomonas and Rhodococcus oil-degrading bacteria used in bioremediation preparations: Formation and structure

We studied formation and structural features of biosurfactants produced by five oil-degrading Pseudomonas and Rhodococcus strains. These bacteria were found to be capable of intense formation of extracellular glycolipid biosurfactants when grown on mineral salts medium with 2% hexadecane. Under these conditions, the surface tension of the cultures decreased from 77 mN/m to 31–34 mN/m. The strain Rhodococcus sp. S26 forming up to 780 mg glycolipids/l of culture medium proved the most efficient biosurfactant producer. Extracellular glycolipids were purified from the crude extracts by column chromatography. Their structural features were determined by thin layer chromatography and electrospray ionization mass spectrometry. Strains Pseudomonas putida BS3701 and Pseudomonas fluorescens 142NF synthesized a number of glycolipids identified as rhamnolipid B and its homologues. Glycolipids produced by Rhodococcus sp. X5 and Rhodococcus sp. S26 were assigned to trehalose tetraesters.