Peptidoglycan recognition protein of Chlamys farreri (CfPGRP-S1) mediates immune defenses against bacterial infection

Peptidoglycan recognition protein (PGRP) is an essential molecule in innate immunity for both invertebrates and vertebrates, owing to its prominent ability in detecting and eliminating the invading bacteria. Several PGRPs have been identified from mollusk, but their functions and the underlined mechanism are still unclear. In the present study, the mRNA expression profiles, location, and possible functions of PGRP-S1 from Zhikong scallop Chlamys farreri (CfPG RP-St) were analyzed. The CfPGRP-S1 protein located in the mantle, gill, kidney and gonad of the scallops. Its mRNA expression in hemocytes was up-regulated extremely after PGN stimulation (P < 0.01), while moderately after the stimulations of LPS (P < 0.01) and beta-glucan (P < 0.05). The recombinant protein of CfPGRP-S1 (designated as rCfPGRP-S1) exhibited high affinity to PGN and moderate affinity to LPS, but it did not bind beta-glucan. Meanwhile, rCfPGRP-S1 also exhibited strong agglutination activity to Gram-positive bacteria Micrococcus luteus and Bacillus subtilis and weak activity to Gram-negative bacteria Escherichia coli. More importantly, rCfPGRP-S1 functioned as a bactericidal amidase to degrade PGN and strongly inhibit the growth of E. coli and Staphyloccocus aureus in the presence of Zn2+. These results indicated that CfPGRP-S1 could not only serve as a pattern recognition receptor recognizing bacterial PGN and LPS, but also function as a scavenger involved in eliminating response against the invaders. (C) 2010 Elsevier Ltd. All rights reserved.

Cflec-5, a pattern recognition receptor in scallop Chlamys farreri agglutinating yeast Pichia pastoris

C-type lectins are a superfamily of carbohydrate-recognition proteins which play crucial roles as pattern recognition receptors (PRRs) in the innate immunity. In this study, the full-length cDNA of a C-type lectin was cloned from scallop Chlamys farreri (designated as Cflec-5) by expression sequence tag (EST) analysis and rapid amplification of cDNA ends (RACE) approach The full-length cDNA of Cflec-5 was of 1412 bp. The open reading frame encoded a polypeptide of 153 amino acids, including a signal sequence and a conserved carbohydrate-recognition domain with the EPN motif determining the mannose-binding specificity The deduced amino acid sequence of Cflec-5 showed high similarity to members of C-type lectin superfamily. The quantitative real-time PCR was performed to investigate the tissue distribution of Cflec-5 mRNA and its temporal expression profiles in hemocytes post pathogen-associated molecular patterns (PAMPs) stimulation. In healthy scallops, the Cflec-5 mRNA was mainly detected in gill and mantle, and marginally in other tissues The mRNA expression of Cflec-5 could be significantly induced by lipopolysaccharide (LPS) and glucan stimulation and reached the maximum level at 6 h and 12 h, respectively But its expression level did not change significantly during peptidoglycan (PGN) stimulation The function of Cflec-5 was investigated by recombination and expression of the cDNA fragment encoding its mature peptide in Escherichia coli Rosetta Gami (DE3) The recombinant Cflec-5 agglutinated Pichia pastoris in a calcium-independent way The agglutinating activity could be inhibited by D-mannose. LPS and glucan, but not by D-galactose or PGN. These results collectively suggested that Cflec-5 was involved in the innate Immune response of scallops and might contribute to nonself-recognition through its interaction with various PAMPs (C) 2010 Elsevier Ltd All rights reserved

三维VSP技术的研究及应用

Today, because of high petroleum consumption of our country, society steady development and difficulty increase in new resources exploration, deep exploitation of the existing oilfield is needed. More delicate reservoir imaging and description, such as thin layer identification, interlayer exploitation monitoring, subtle structure imaging, reservoir anisotropy recognition, can provide more detail evidence for new development adjustment scheme and enhanced oil recovery. Now, the people have already realized the 3D VSP technique more effective than the general methods in solving these aspects. But VSP technique especially 3D VSP develop slowly due to some reasons. Carrying out the research of VSP technique, it will be very useful to the EOR service. 3D VSP techniques include acquisition、data processing and interpretation. In this paper, the author carried out some researches around acquisition and processing. The key point of acquisition is the survey design, it is critical to the quality of the data and it will influence the reservoir recognition as follows. The author did detailed researches on the layout pattern of shot point and geophone. Some attributes relate to survey design such as reflectivity, incidence angle, observation area, reflection points distribution, fold, minimum well source distance, azimuth angle and so on are studied seriously. In this geometry design of 3D-VSP exploration in deviated wells, the main problems to be solved are: determining the center position of shots distribution, the effect of shots missing on coverage areas and coverage times，locating the shots and receivers of multi-wells. Through simulating and analyzing, the above problems are discussed and some beneficial conclusions are drawn. These will provide valuable references to actual survey design. In data processing, researches emphasize on those relatively key techniques such as wavefield separation, VSP-CDP imaging, the author carried out deep researches around these two aspects. As a result, variant apparent slowness wavefield separation method developed in this article suit the underground variant velocity field and make wavefield separation well, it can overcome reflection bending shortage aroused by conventional imaging method. The attenuateion range of underground seismic wave is very important for amplitude compensation and oil/gas identification.In this paper, seismic wave attenuateion mechanism is studied by 3D-VSP simulateion and Q-inversion technique. By testing with seismic data, the method of VSP data attenuateion and relationship of attenuateion attribute variant with depth is researched. Also the software of survey design and data processing is developed, it fill the gap of VSP area in our country. The technique developed applied successfully in SZXX-A Oilfield、QKYY-B Oilfield、A area and B area. The good results show that this research is valuable, and it is meaningful to the VSP technique development and application of offshore oil industry and other areas in our country.

Computer-assisted automatic peak recognition and result evaluation for analysis of chlorinated hydrocarbons in environmental samples

A data manipulation method has been developed for automatic peak recognition and result evaluation in the analysis of organic chlorinated hydrocarbons with dual-column gas chromatography. Based on the retention times of two internal standards, pentachlorotoluene and decachlorobiphenyl, the retention times of chlorinated hydrocarbons can be calibrated automatically and accurately. It is very convenient to identify the peaks by comparing the retention times of samples with the calibrated retention times calculated from the relative retention indices of standards. Meanwhile, with a suggested two-step evaluation method the evaluation coefficients and the suitable quantitative results of each component can be automatically achieved for practical samples in an analytical system using two columns with different polarities and two internal standards. (C) 2002 Elsevier Science B.V. All rights reserved.

Recognition interactions of metal-complexing imprinted polymer

Molecularly imprinted polymer, exhibiting considerable enantioselectivity for L-mandelic acid, was prepared using metal coordination-chelation interaction. By evaluating the recognition characteristics in the chromatographic mode, the recognition interactions were proposed: specific and nonspecific metal coordination-chelation interaction and hydrophobic interaction were responsible for substrate binding on metal-complexing imprinted polymer; while the selective recognition only came from specific metal coordination-chelation interaction and specific hydrophobic interaction.