Kinetic Study of Prolidase Activity in Erythrocytes against Different Substrates Using Capillary Electrophoresis with Electrochemiluminescence Detection

Capillary electrophoresis (CE) with electrochemiluminescence (ECL) detection was used to explore the kinetics ofthe enzymatic reaction. The different effects ofreaction conditions including the concentration of Mn2l, incubation temperature and pH on PFOlidase (PLD, EC 3.4.13.9) activity in erythrocyte lysates against three different substrates, Gly-Pro, Val-Pro and Leu-Pro were investigated. Also, the effects of colchicine which can prevent or delay cancer ofliver on the PLD activity were studied.

Purification, sequencing and biological activity of polypeptide from velvet antler

The velvet antler polypeptide CNT14 was extracted and purified by gel filtration, ion exchange chromatography and RP C, which showed a single peak in HPLC chromatography and a single band in SDS-PAGE. The molecular weight measured by MALDI/TOF/MS spectrum was 1479. 9028. The polypeptide consisted mostly of Glu, Leu, Val, Pro. The amino acid sequence of the polypeptide was detected with ESI-MS/ MS, and the sequence was E-P-T-V-L-D-E-V-C-L-A-H-G-P. The experiments of biological activity of polypeptide CNT14 in vivo were carried out, and the results show that CNT14 has stimulant effects on the growth of rat HT22 cells. Then we produced the polypeptide CNT14 according the amino acid sequence by solid phase synthesis, confirmed the sequence of the polypeptide to be consistent with the amino acid sequence of polypeptide CNT14 which was separated from the velvet antler.

Systematic study of the transfer of amino acids across the water/1,2-dichloroethane interface facilitated by dibenzo-18-crown-6

Facilitated ion transfer reactions of 20 amino acids with di.benzo-18-crown-6 (DB18C6) at the water/1,2-dichloroethane (W/DCE) interfaces supported at the tips of micro- and nano-pipets were investigated systematically using cyclic voltammetry. It was found that there were only 10 amino acids, that is, Leu, Val, Ile, Phe, Trp, Met, Ala, Gly, Cys, Gln (in brief), whose protonated forms as cations can give well-defined facilitated ion transfer voltammograms within the potential window, and the reaction pathway was proven to be consistent with the transfer by interfacial complexation/dissociation (TIC/TID) mechanisms. The association constants of DB 18C6 with different amino acids in the DCE (beta(0)), and the kinetic parameters of reaction were evaluated based on the steady-state voltammetry of micro- or nano-pipets, respectively The experimental results demonstrated that the selectivity of complexation of protonated amino acid by DB18C6 compared with that of alkali metal cations was low, which can be attributed to the vicinal effect arising from steric hindrance introduced by their side group and the steric bulk effect by lipophilic stabilization.

PUMA760切割作业力与位置混合控制

本文介绍在 PUMA760 上实现了的切割作业控制系统。我们在 PUMA760 控制系统中引进了力传感器信号,加入了力控制算法,通过调整机器人手部位置,控制机器人与环境物体间的相互作用力。在 VAL 系统中加入了根据传感器信号动作的指令,实现了一种机器人力与位置的混合控制。用扩充后的 VAL 系统编程,成功地使 PUMA760完成了多种材料的切割作业。

机器人规划与编程系统的研究

机器人的程序设计问题是智能机器人研究的重要方面。也是自动程序设计的一个重要方面。给定一个机器人的任务描述。应该能够得一个完整的程序代码（用某种程序设计语言书写）。本文描述了一个这样的系统－LRPS。LRPS用一种比较简单的任务描述语言TOL（Task-Oriented Language）来接收用户的任务描述，然后经过分析。规划，最后生成一段用VAL语言编写的程序代码，系统能够给出机器人的宏动作序列－－机器人规划。并能对若干知识库进行操作。在规划器中采用了一种新的称为插入法的方法来进行规划，整个系统在IBM－PC机上用PASCAL语言实现。

A gueroba (Syagrus oleracea Becc.) nas comunidades rurais II: sistema de producao e avaliacao economica.

A gueroba, guariroba ou gariroba e uma palmeira importante para a populacao do Cerrado, sendo extraido dela um palmito de sabor amargo muito apreciado na culinaria regional. Esse produto e comercializado, geralmente, in natura e sua industrializacao ainda e artesanal. Seu principal mercado localiza-se nos Estados de Goias, Tocantins, Mato Grosso, Mato Grosso do Sul, algumas regoes da Bahia e de Minas Gerais e no Distrito Federal. Com a exploracao predatoria e a acelerada destruicao da vegetacao nativa por meio da expansao da fronteira agricola, a oferta do produto, oriunda do extrativismo, vem sendo reduzida substancialmente. No entanto, tem-se observado o cultivo dessa palmeira em alguns Estados, principalmente em Goias que, em 1999, cultivava 4.499 hectares. Neste estudo, encontra-se descrito o sistema de producao da cultura de gueroba, consorciada com milho e feijao, nos dois primeiro anos, bem como a matriz dos coeficientes tecnicos, as estimativas do Custo Operacional Efetivo (COE) e os fluxos liquidos de caixa, referentes a quatro alternativas de producao (gueroba-milho-feijao, laraja-pera-rio, milho e arroz de sequeiro) para um periodo de oito anos. Para comparar as diferentes alternativas de investimento, foi utilizado como principal indicador o Valor Atual Liquido (VAL) que considerando o nivel de preço medio, o valor estimado por hectare foi de R$ 35.599,60 para o consorcio queroba-milho-feijao; de R$ 20.994,74 para a cultura de laranja-pera-rio; de R$ 609,33 para o milho e de R$ (-810,36) para o arroz de sequeiro. Como indicadores-auxiliar, foram utilizados: a relacao beneficio/custo, o preco de equilibrio e a margem bruta. Os resultados indicam que, para as condicoes vigentes de mercado e os niveis de tecnologia considerados, o sistema gueroba proporciona, em todos os niveis de precos, o maior retorno do capital investido e maior relacao beneficio/custo.

Evaluation of microbial adjuncts and their effect on the ripening of cheddar cheese

A bacteriocin-producing strain of Lactobacillus paracasei DPC 4715 was used as an adjunct culture in Cheddar cheese in order to control the growth of “wild” nonstarter lactic acid bacteria. No suppression of growth of the indicator strain was observed in the experimental cheese. The bacteriocin produced by Lactobacillus paracasei DPC 4715 was sensitive to chymosin and cathepsin D and it may have been cleaved by the rennet used for the cheese manufactured or by indigenous milk proteases. A series of studies were performed using various microbial adjuncts to influence cheese ripening. Microbacterium casei DPC 5281, Corynebacterium casei DPC 5293 and Corynebacterium variabile DPC 5305 were added to the cheesemilk at level of 109 cfu/ml resulting in a final concentration of 108 cfu/g in Cheddar cheese. The strains significantly increased the level of pH 4.6-soluble nitrogen, total free amino acids after 60 and 180 d of ripening and some individual free amino acids after 180 d. Yarrowia lipolytica DPC 6266, Yarrowia lipolytica DPC 6268 and Candida intermedia DPC 6271 were used to accelerate the ripening of Cheddar cheese. Strains were grown in YG broth to a final concentration of 107 cfu/ml, microfluidized, freeze-dried and added to the curd during salting at level of 2% w/w. The yeasts positively affected the primary, secondary proteolysis and lipolysis of cheeses and had aminopeptidase, dipeptidase, esterase and 5’ phosphodiestere activities that contributed to accelerate the ripening and improve the flavor of cheese. Hafia alvei was added to Cheddar cheesemilk at levels of 107 cfu/ml and 108 cfu/ml and its contribution during ripening was evaluated. The strain significantly increased the level of pH 4.6-soluble nitrogen, total free amino-acids, and some individual free amino-acids of Cheddar cheese, whereas no differences in the urea-polyacrylamide gel electrophoresis (urea-PAGE) electrophoretograms of the cheeses were detected. Hafia alvei also significantly increased the level of some biogenic amines. A low-fat Cheddar cheese was made with Bifidobacterium animalis subsp. lactis, strain BB-12® at level of 108 cfu/ml, as a probiotic adjunct culture and Hi-Maize® 260 (resistant high amylose maize starch) at level of 2% and 4% w/v, as a prebiotic fiber which also played the role of fat replacer. Bifidobacterium BB-12 decreased by 1 log cycle after 60 d of ripening and remained steady at level of ~107 cfu/g during ripening. The Young’s modulus also increased proportionally with increasing levels of Hi-maize. Hencky strain at fracture decreased over ripening and increased with increasing in fat replacer. A cheese based medium (CBM) was developed with the purpose of mimicking the cheese environment at an early ripening stage. The strains grown in CBM showed aminopeptidase activity against Gly-, Arg-, Pro- and Phe-p-nitroanalide, whereas, when grown in MRS they were active against all the substrates tested. Both Lb. danicus strains grown in MRS and in CBM had aminotransferase activity towards aromatic amino acids (Phe and Trp) and also branched-chain amino acids (Leu and Val). Esterase activity was expressed against p-nitrophenyl-acetate (C2), pnitrophenyl- butyrate (C4) and p-nitrophenyl-palmitate (C16) and was significantly higher in CBM than in MRS.

Twentieth Century Society Spring lecture series: second lives for Twentieth Century masterpieces

The Twentieth Century Society’s Spring lecture series (six in total) looks at the restoration and refurbishment of key C20 buildings in Britain and the US. Buildings covered: BBC Broadcasting House in London (G Val Meyer 1930-32, MacCormac Jamieson Prichard 2000-09). Speaker: Mark Hines (Mark Hines Architects), was the project architect and is the author of The Story of Broadcasting House: Home of the BBC. 5 February 2009. Crown Hall, Chicago (Mies van der Rohe 1952), the Art and Architecture Building, Yale University, New Haven (Paul Rudolf 1961-63) and the former Wills head office in Bristol (SOM with YRM 1970-75). Speaker: Patrick Bellew (Atelier 10 Engineers), 12 February 2009. Center for British Art, Yale University, New Haven (Louis Kahn 1969-77). Speaker: Peter Inskip (Inskip and Jenkins Architects), 17 February 2009. Brunswick Centre London (Patrick Hodgkinson 1967-72; Levitt Bernstein with Patrick Hodgkinson 2006). Speaker: Stuart Tappin (Stand Consulting Engineers Ltd), 26 February 2009. De La Warr Pavilion, Bexhill-on-Sea (Mendelsohn and Chermayeff 1934-5, John McAslan and Partners 2000-05). Speaker: Mark Cannata (HOK Architects), 5 March 2009. Finsbury Health Centre London (Lubetkin & Tecton 1938, first phase of conservation work Avanti Architects 1995.). Speaker: John Allan of Avanti Architects, 12 March 2009.

Notes and memoranda

Notes and observations on: Polyprion cerniu, Val. Scomber scomber Linn (the mackerel). Year-old pilchards. Muggiaea atlantica. Hippoglossus vulgaris, Linn. (the halibut). Rhombus maximus, Linn (the turbot).

Sentinel-3 Mission Performance Centre: Ensuring a high-quality altimetric dataset

Sentinel-3A is scheduled for launch in Oct. 2015, with Sentinel-3B to follow 18 months later. Together these missions are to take oceanographic remote-sensing into a new operational realm. To achieve this a large number of processing, calibration and validation tasks have to be applied to their data in order to assess for quality, absolute bias, short-term changes and long-term drifts. ESA has funded the Sentinel-3 Mission Performance Centre (S3MPC) to carry out this evaluation on behalf of ESA and EUMETSAT. The S3MPC is run by a consortium led by ACRI [1] and this paper describes the work on the calibration/validation (cal/val) of the Surface Topography Mission (STM), which is co-ordinated by CLS and PML.

Creación de una filmoteca de prácticas para la Facultad de Ciencias de la Información de la UCM: Adecuación de espacio y estrategias de catalogación

En el presente trabajo se da el primer paso para la creación de una Filmoteca de Prácticas en la facultad de Ciencias de la Información de la Universidad Complutense de Madrid. El Patrimonio Fílmico se ve gravemente expuesto al paso del tiempo y el abandono por lo que se necesita un rescate urgente. Mediante un riguroso trabajo de investigación, en el que se estudian diferentes propuestas y normativas expuestas por organismos especializados, se redacta un manual de inicio. Se han investigado, diseñado y propuesto las herramientas básicas y útiles para poner en funcionamiento la primera Filmoteca de Prácticas en el ámbito universitario español.

Novel dipeptidyl peptidase IV resistant analogues of glucagon-like peptide-1(7-36)amide have preserved biological activities in vitro conferring improved glucose-lowering action in vivo

Although the incretin hormone glucagon-like peptide-1 (GLP-1) is a potent stimulator of insulin release, its rapid degradation in vivo by the enzyme dipeptidyl peptidase IV (DPP IV) greatly limits its potential for treatment of type 2 diabetes. Here, we report two novel Ala(8)-substituted analogues of GLP-1, (Abu(8))GLP-1 and (Val(8) GLP-1 which were completely resistant to inactivation by DPP IV or human plasma. (Abu(8))GLP-1 and (Val(8))GLP-1 exhibited moderate affinities (IC50: 4.76 and 81.1 nM, respectively) for the human GLP-1 receptor compared with native GLP-1 (IC50: 0.37 nM). (Abu(8))GLP-1 and (Val(8))GLP-1 dose-dependently stimulated cAMP in insulin-secreting BRIN BD11 cells with reduced potency compared with native GLP-1 (1.5- and 3.5-fold, respectively). Consistent with other mechanisms of action, the analogues showed similar, or in the case of (Val(8))GLP-1 slightly impaired insulin releasing activity in BRIN BD11 cells. Using adult obese (ob/ob) mice, (Abu(8))GLP-1 had similar glucose-lowering potency to native GLP-1 whereas the action of (Val(8))GLP-1 was enhanced by 37%. The in vivo insulin-releasing activities were similar. These data indicate that substitution of Ala(8) in GLP-1 with Abu or Val confers resistance to DPP IV inactivation and that (Val(8))GLP-1 is a particularly potent N-terminally modified GLP-1 analogue of possible use in type 2 diabetes.

Pachymedusa dacnicolor tryptophyllin-1 (PdT-1): structural characterization, pharmacological activity and cloning of precursor cDNA.

Tryptophyllins are a heterogenous group of amphibian skin peptides originally identified in skin extracts of Neotropical leaf frogs, Phyllomedusa sp., by chemical means. Until now, biosynthetic precursor structure and biological activity remain unreported. Here we describe the isolation of a novel, post-translationally modified tryptophyllin, Lys-Pro-Hyp-Ala-Trp-Val-Pro.amide (PdT-1), from the skin secretion of the Mexican leaf frog, Pachymedusa dacnicolor. Using a 3'- and 5'-RACE strategy and an in vitro skin cDNA library, the PdT-1-encoding precursor was cloned and found to consist of an open-reading frame of 62 amino acids with a single copy of PdT-1 located towards the C-terminus. A synthetic replicate of PdT-1 was found to be a potent myoactive agent, relaxing mammalian arterial smooth muscle and contracting small intestinal smooth muscle at nanomolar concentrations. PdT-1 is thus the first amphibian skin tryptophyllin to be pharmacologically characterized and the first whose precursor cDNA has been cloned.

Mutations affecting the Rossman fold of isoleucyl-tRNA synthetase are correlated with low-level mupirocin resistance in Staphylococcus aureus.

The isoleucyl-tRNA synthetase (ileS) gene was sequenced in toto from 9 and in part from 31 Staphylococcus aureus strains with various degrees of susceptibility to mupirocin. All strains for which the mupirocin MIC was greater than 8 µg/ml contained point mutations affecting the Rossman fold via Val-to-Phe changes at either residue 588 (V588F) or residue 631 (V631F). The importance of the V588F mutation was confirmed by an allele-specific PCR survey of 32 additional strains. Additional mutations of uncertain significance were found in residues clustered on the surface of the IleS protein.

Effects of the aurora kinase inhibitors AZD1152-HQPA and ZM447439 on growth arrest and polyploidy in acute myeloid leukemia cell lines and primary blasts.

BACKGROUND:
Aurora kinases play an essential role in the orchestration of chromosome separation and cytokinesis during mitosis. Small-molecule inhibition of the aurora kinases has been shown to result in inhibition of cell division, phosphorylation of histone H3 and the induction of apoptosis in a number of cell systems. These characteristics have led aurora kinase inhibitors to be considered as potential therapeutic agents.
DESIGN AND METHODS:
Aurora kinase gene expression profiles were assessed in 101 samples from patients with acute myeloid leukemia. Subsequently, aurora kinase inhibitors were investigated for their in vitro effects on cell viability, histone H3 phosphorylation, cell cycle and morphology in acute myeloid leukemia cell lines and primary acute myeloid leukemia samples.
RESULTS:
The aurora kinase inhibitors AZD1152-HQPA and ZM447439 induced growth arrest and the accumulation of hyperploid cells in acute myeloid leukemia cell lines and primary acute myeloid leukemia cultures. Furthermore, both agents inhibited histone H3 phosphorylation and this preceded perturbations in cell cycle and the induction of apoptosis. Single cell cloning assays were performed on diploid and polyploid cells to investigate their colony-forming capacities. Although the polyploid cells showed a reduced capacity for colony formation when compared with their diploid counterparts, they were consistently able to form colonies.
CONCLUSIONS:
AZD1152-HQPA- and ZM447439 are effective apoptosis-inducing agents in acute myeloid leukemia cell lines and primary acute myeloid leukemia cultures. However, their propensity to induce polyploidy does not inevitably result in apoptosis.