Construction and characterization of bacterial artificial chromosome library of black-handed spider monkey (Ateles geoffroyi)

The large-insert genomic DNA library is a critical resource for genome-wide genetic dissection of target species. We constructed a high-redundancy bacterial artificial chromosome (BAC) library of a New World monkey species, the black-handed spider monkey

Construction of a BAC library for Chinese amphioxus Branchiostoma belcheri and identification of clones containing Amphi-pax genes

Amphioxus is a crucial organism for the study of vertebrate evolution. Although a genomic BAC library of Branchiostoma floridae has been constructed, we report here another BAC library construction of its distant relative species Branchiostoma belcheri. The amphioxus BAC library established in present study consists of 45,312 clones arrayed in one hundred and eighteen 384-well plates. The average insert fragment size was 120 kb estimated by Pulsed Field Gel Electrophoresis (PFGE) analysis of 318 randomly selected clones. The representation of the library is about 12 equivalent to the genome, allowing a 99.9995% probability of recovering any specific sequence of interest. We further screened the library with 4 single copied Amphi-Pax genes and identified total of 26 positive clones with average of 6.5 clones for each gene. The result indicates this library is well suited for many applications and should also serve as a useful complemental resource for the scientific community.

Construction, characterization and chromosomal mapping of bacterial artificial chromosome (BAC) library of Yunnan snub-nosed monkey (Rhinopithecus bieti)

We constructed a high redundancy bacterial artificial chromosome library of a seriously endangered Old World Monkey, the Yunnan snub-nosed monkey (Rhinopithecus bieti) from China. This library contains a total of 136 320 BAC clones. The average insert size of BAC clones was estimated to be 148 kb. The percentage of small inserts (50-100 kb) is 2.74%, and only 2.67% non-recombinant clones were observed. Assuming a similar genome size with closely related primate species, the Yunnan snub-nosed monkey BAC library has at least six times the genome coverage. By end sequencing of randomly selected BAC clones, we generated 201 sequence tags for the library. A total of 139 end-sequenced BAC clones were mapped onto the chromosomes of Yunnan snub-nosed monkey by fluorescence in-situ hybridization, demonstrating a high degree of synteny conservation between humans and Yunnan snub-nosed monkeys. Blast search against human genome showed a good correlation between the number of hit clones and the size of the chromosomes, an indication of unbiased chromosomal distribution of the BAC library. This library and the mapped BAC clones will serve as a valuable resource in comparative genomics studies and large-scale genome sequencing of nonhuman primates. The DNA sequence data reported in this paper were deposited in GenBank and assigned the accession number CG891489-CG891703.

Coupled foundation-superstructure analysis and influence of building stiffness on foundation response

This paper proposes a simple method to include superstructure stiffness in foundation analyses. The method involves extracting a small "condensed structural matrix" from finite element models of the superstructure, which can then be incorporated into pile group or piled raft analyses using common approaches such as elastic continuum or load transfer methods. The matrix condensation method directly couples structural and geotechnical analyses, and eliminates the need for iterative analyses between structural and geotechnical engineers. Effectiveness of the approach is illustrated through analyses of several buildings designed with a typical floor plan but with varying heights. The parametric study illustrates that superstructure stiffness can have a significant influence on foundation settlement estimates, and the stiffening effects are dominated by the lower stories of the superstructure. The proposed method aims to bridge the gap between structural and geotechnical analyses. Also, being a computationally simple and accurate approach, it is applicable to parametric or optimization studies that would otherwise involve large amounts of analyses. © 2010 ASCE.

A NOVEL PLASMINOGEN-ACTIVATOR FROM SNAKE-VENOM - PURIFICATION, CHARACTERIZATION, AND MOLECULAR-CLONING

A novel plasminogen activator from Trimeresurus stejnegeri venom (TSV-PA) has been identified and purified to homogeneity. It is a single chain glycoprotein with an apparent molecular weight of 33,000 and an isoelectric point of pH 5.2. It specifically activates plasminogen through an enzymatic reaction. The activation of human native GIu-plasminogen by TSV-PA is due to a single cleavage of the molecule at the peptide bond Arg(561)-Val-(562). Purified TSV-PA, which catalyzes the hydrolysis of several tripeptide p-nitroanilide substrates, does not activate nor degrade prothrombin, factor X, or protein C and does not clot fibrinogen nor show fibrino(geno)lytic activity in the absence of plasminogen. The activity of TSV-PA was readily inhibited by phenylmethanesulfonyl fluoride and by p-nitrophenyl-p-guanidinobenzoate. Oligonucleotide primers designed on the basis of the N-terminal and the internal peptide sequences of TSV-PA were used for the amplification of cDNA fragments by polymerase chain reaction. This allowed the cloning of a full-length cDNA encoding TSV-PA from a cDNA library prepared from the venom glands. The deduced complete amino acid sequence of TSV-PA indicates that the mature TSV-PA protein is composed of 234 amino acids and contains a single potential N-gIycosylation site at Asn(1G1). The sequence of TSV-PA exhibits a high degree of sequence identity with other snake venom proteases: 66% with the protein C activator from Aghistrodon contortrix contortrix venom, 63% with batroxobin, and 60% with the factor V activator from Russell's viper venom. On the other hand, TSV-PA shows only 21-23% sequence similarity with the catalytic domains of u-PA and t-PA. Furthermore, TSV-PA lacks the sequence site that has been demonstrated to be responsible for the interaction of t-PA (KHRR) and u-PA (RRHR) with plasminogen activator inhibitor type 1.

Multi-objective foundation optimization and its application to pile reuse

Pile reuse has become an increasingly popular option in foundation design, mainly due to its potential cost and environmental benefits and the problem of underground congestion in urban areas. However, key geotechnical concerns remain regarding the behavior of reused piles and the modeling of foundation systems involving old and new piles to support building loads of the new structure. In this paper, a design and analysis tool for pile reuse projects will be introduced. The tool allows coupling of superstructure stiffness with the foundation model, and includes an optimization algorithm to obtain the best configuration of new piles to work alongside reused piles. Under the concept of Pareto Optimality, multi-objective optimization analyses can also reveal the relationship between material usage and the corresponding foundation performance, providing a series of reuse options at various foundation costs. The components of this analysis tool will be discussed and illustrated through a case history in London, where 110 existing piles are reused at a site to support the proposed new development. The case history reveals the difficulties faced by foundation reuse in urban areas and demonstrates the application of the design tool to tackle these challenges. © ASCE 2011.

A horsefly saliva antigen 5-like protein containing RTS motif is an angiogenesis inhibitor

The Ag5 proteins are the most abundant and immunogenic proteins in the venom secretory ducts of stinging insects. An antigen 5-like protein (named tabRTS) composed of 221 amino acid residues was purified and characterized from the salivary glands of the horsefly, Tabanus yao (Diptera, Tabanidae). Its cDNA was cloned from the cDNA library of the horsefly's salivary gland. TabRTS containing the SCP domain (Sc7 family of extracellular protein domain) was found in insect antigen 5 proteins. More interestingly, there is an Arg-Thr-Ser (RTS) disintegrin motif at the C-terminus of tabRTS. The RTS motif is positioned in a loop bracketed by cysteine residues as those found in RTS-disintegrins of Crotalidae and Viperidae snake venoms, which act as angiogenesis inhibitors. Endothelial Cell Tube formation assay in vitro and chicken chorioallantoic membrane (CAM) angiogenesis assay in vivo were performed as to investigate the effect of tabRTS on angiogenesis. It was found that tabRTS could significantly inhibit angiogenesis in vitro and in vivo. Anti-alpha(1)beta(1) monoclonal antibody could dose-dependently inhibit the anti-angiogenic activity of tabRTS. This result indicated that tabRTS possibly targets the alpha(1)beta(1) integrin to exert the anti-angiogenic activity as snake venom RTS-/KTS-disintegrins do. The current work revealed the first angiogenesis inhibitor protein containing RTS motif from invertebrates, a possible novel type of RTS-disintegrin. (C) 2009 Elsevier Ltd. All rights reserved.

Five novel antimicrobial peptides from skin secretions of the frog, Amolops loloensis

While investigating antimicrobial peptide diversity of Amolops loloensis, five novel antimicrobial peptides belonging to two families were identified from skin secretions of this frog. The first family including two members is esculentin-2-AL (esculentin-2-ALa and -ALb): the second family including three members is temporin-AL (temporin-ALd to -ALf). The family of esculentin-2-AL is composed of 37 amino acid residues (aa); the family of temporin-AL is composed of 16, 13 and 10 aa, respectively. All of these antimicrobial peptides showed antimicrobial activities against tested microorganisms. cDNAs encoding precursors of esculentin-2-ALs and temporin-ALs were cloned from the skin cDNA library of A. loloensis. All the precursors share similar overall structures. There is a typical prohormone processing signal (Lys-Arg) located between the acidic propiece and the mature peptide. The antimicrobial peptide family of esculentin-2 is firstly reported in the genus of Amolops. Combined with previous reports, a total of four antimicrobial peptide families have been identified from the genus of Amolops; three of them are also found in the genus of Rana. These results suggest the possible evolutionary connection between the genera Amolops and Rana. (C) 2009 Elsevier Inc. All rights reserved.

Sequencing, annotation and comparative analysis of nine BACs of giant panda (Ailuropoda melanoleuca)

A 10-fold BAC library for giant panda was constructed and nine BACs were selected to generate finish sequences. These BACs could be used as a validation resource for the de novo assembly accuracy of the whole genome shotgun sequencing reads of giant panda newly generated by the Illumina GA sequencing technology. Complete sanger sequencing, assembly, annotation and comparative analysis were carried out on the selected BACs of a joint length 878 kb. Homologue search and de novo prediction methods were used to annotate genes and repeats. Twelve protein coding genes were predicted, seven of which could be functionally annotated. The seven genes have an average gene size of about 41 kb, an average coding size of about 1.2 kb and an average exon number of 6 per gene. Besides, seven tRNA genes were found. About 27 percent of the BAC sequence is composed of repeats. A phylogenetic tree was constructed using neighbor-join algorithm across five species, including giant panda, human, dog, cat and mouse, which reconfirms dog as the most related species to giant panda. Our results provide detailed sequence and structure information for new genes and repeats of giant panda, which will be helpful for further studies on the giant panda.

Natural frequency and damping ratio of a vertically vibrated surface foundation

It is possible and common to obtain equivalent natural frequency and damping for a soil-foundation system from results of experimental or numerical analysis assuming the system has a single degree of freedom. Three approaches to extract natural frequency and damping were applied to the vertically vibrated soil-foundation system. The sensitivity of the computed natural frequency and damping to the soil properties was evaluated through parametric studies. About 10-20% of discrepancy in values of natural frequency was observed due to different approaches. The results help to assess the reliability of equivalent soil properties determined from the reported natural frequency of the system. Finally the results obtained using theoretical predictions with linear soil properties measured in situ were compared to those calculated from experimental data. The prediction and experimental results showed good agreements if the embedment of the foundation is neglected with stepped sine test but considered with impulse test. © 2010 Elsevier Ltd.

Wave propagation velocity under a vertically vibrated surface foundation

The ultimate objective of the research conducted by the authors is to explore the feasibility of determining reliable in situ values of soil modulus as a function of strain. In field experiments, an excitation is applied on the ground surface using large-scale shakers, and the response of the soil deposit is recorded through receivers embedded in the soil. The focus of this paper is on the simulation and observation of signals that would be recorded at the receiver locations under idealized conditions to provide guidelines on the interpretation of the field measurements. Discrete models are used to reproduce one-dimensional and three-dimensional geometries. When the first times of arrival are detected by receivers under the vertical impulse, they coincide with the arrival of the P wave; therefore related to the constrained modulus of the material. If one considers, on the other hand, phase differences between the motions at two receivers, the picture is far more complicated and one would obtain propagation velocities, function of frequency and measuring location, which do not correspond to either the constrained modulus or Young's modulus. It is necessary then to conduct more rigorous and complicated analyses in order to interpret the data. This paper discusses and illustrates these points. Copyright © 2008 John Wiley & Sons, Ltd.

Wave propagation under vertically excited surface foundation

Recently developed equipment allows measurement of the shear modulus of soil in situ as a function of level of strain. In these field experiments, the excitation is applied on the ground surface using large scale shakers, and the response of the soil deposit is recorded through embedded receivers. The focus of this paper is on the simulation of signals which would be recorded at the receiver locations in idealized conditions to provide guidelines on the interpretation of field measurements. Discrete and finite element methods are employed to model one dimensional and three dimensional geometries, respectively, under various lateral boundary conditions. When the first times of arrival are detected by receivers under the vertical impulse, they coincide with the arrival of the P wave, related to the constrained modulus of the material, regardless of lateral boundary conditions. If one considers, on the other hand, phase differences between the motions at two receivers the picture is far more complicated and one would obtain propagation velocities, function of frequency and depth, which do not correspond to either the constrained modulus or Young's modulus. It is thus necessary to apply some care when interpreting the data from field tests based on vertical steady state vibrations. The use of inverse analysis can be considered as a way of extracting the shear modulus of soil from the field test measurements. © 2008 ASCE.

Characterization of the porcine differentially expressed PDK4 gene and association with meat quality

To investigate the differential expression of genes in the skeletal muscle between Yorkshire and Chinese indigenous breed Meishan pigs, suppression subtractive hybridization was carried out and many genes were proved to be expressed significantly different in the two breeds. One gene highly expressed in Meishan but lowly expressed in Yorkshire specific library, shared strong homology with human pyruvate dehydrogenase kinase 4 (PDK4). Using semi-quantity and quantity PCR, We confirmed its differential expression between the two breeds. Temporal and spatial expression analysis indicated that porcine PDK4 gene is highly expressed in skeletal muscle and the highest in neonatal pigs. Complete cDNA cloning and sequence analysis revealed that porcine PDK4 gene contains an open reading frame of 1,221 bp. The deduced amino acid sequence showed conservation in evolution. A G/A mutation in intron 9 was identified and association analysis showed that it was significantly associated with intramuscular fat, muscle water content.