Molecular cloning of the helokinestatin/CNP precursor from a venom-derived cDNA library of the Mexican beaded lizard, Heloderma horridum, and identification of a novel encoded bradykinin-antagonist peptide, helokinestatin-6

Helokinestatins 1–5 represent a novel family of bradykinin antagonist peptides originally isolated from the venom of the Gila Monster, Heloderma suspectum. We found that they were encoded in tandem along with a single copy of C-type natriuretic peptide (CNP), by two different but almost identical biosynthetic precursors that were cloned from a venom-derived cDNA library. Here we have applied the same strategy to the venom of a related species, the Mexican beaded lizard, Heloderma horridum. Lyophilised venom was used as a surrogate tissue to generate a cDNA library that was interrogated with primers from the previous study and for reverse phase HPLC fractionation. The structure of a single helokinestatin precursor was obtained following sequencing of 20 different clones. The open-reading frame contained 196 amino acid residues, somewhat greater than the 177–178 residues of the corresponding helokinestatin precursors in H. suspectum. The reason for this difference in size was the insertion of an additional domain of 18 amino acid residues encoding an additional copy of helokinestatin-3. Helokinestatin-6 (GPPFNPPPFVDYEPR) was a novel peptide from this precursor identified in venom HPLC fractions. A synthetic replicate of this peptide antagonised the relaxation effect of bradykinin on rat arterial smooth muscle. The novel peptide family, the helokinestatins, have been shown to be present in the venom of H. horridum and to be encoded by a single precursor of different structure to those from H. suspectum. Studies such as this reveal the naturally-selected structures of bioactive peptides that have been optimised for purpose and provide the scientist with a natural analogue library for pharmacological investigation.

Molecular cloning and deduced organization of the pHpG/CNP precursor from the venom of the African forest viper, Atheris squamigera

The discovery that the hypotensive sequela of envenomation by the South American viper, Bothrops jararaca, was mediated by peptides, represented a milestone in drug discovery research that led to the introduction of ACE inhibitors. These bradykinin-potentiating peptides (BPPs) have been found in the venoms of many species of viper and molecular cloning of biosynthetic precursors has revealed that each encodes several different BPPs in tandem with a single copy of a C-type natriuretic peptide (CNP) located at the C-terminus. Venoms of the African forest vipers (Atheris) have been poorly studied possibly because they do not represent a major danger to humans. However, initial studies have indicated that they contain some of the “classical” protein toxins of viper venoms and a novel class of peptide, the polyglycine/polyhistidine (pGpH) peptides. These peptides occur in several molecular forms with different numbers of repetitive glycine and histidine repeats. We have cloned the biosynthetic precursor of A. squamigera pGpH peptides from a venom-derived cDNA library and have confirmed that a single copy of CNP is located at the C-terminus and additionally that, like BPPs in other vipers, pGpH peptides are encoded in tandem within this single precursor. Solid phase peptide synthesis of pGpH peptides has proven to be extremely difficult but is progressing and acquisition of synthetic replicates of each peptide is a necessary prerequisite for systematic pharmacological characterisation as establishment of a biological function for these peptides remains elusive. pGpH peptides may prove to play a role as fundamental as that of the BPPs.

Extremely high conservation in the untranslated Region as well as the coding region of CNP mRNAs throughout elasmobranch species

C-type natriuretic peptide (CNP) is a crucial osmoregulatory hormone in elasmobranchs, participating in salt secretion and drinking. In contrast to teleosts and tetrapods in which the NP family is composed of a group of structurally related peptides, we have shown that CNP is the sole NP in sharks. In the present study, CNP cDNAs were cloned from four species of batoids, another group of elasmobranchs. The cloned batoid CNP precursors contained a plausible mature peptide of 22 amino acid residues that is identical to most shark CNP-22s, but five successive amino acids were consistently deleted in the prosegment compared with shark precursors, supporting the diphyletic classification of sharks and rays. In addition, molecular phylogenetic trees of CNP precursors were consistent with a diphyletic interpretation. Except for the deletion, the nucleotide and deduced amino acid sequences of the CNP cDNAs are extremely well-conserved among all elasmobranch species, even between sharks and rays. Surprisingly, high conservation is evident not only for the coding region, but also for the untranslated regions. It is most likely that the high conservation is due to the low nucleotide substitution rate in the elasmobranch genome, and high selection pressure. The 3′-untranslated region of the elasmobranch CNP cDNAs contained three to six repeats of the ATTTA motif that is associated with the regulation of mRNA stability and translation efficiency. Alternative polyadenylation sites were also found; the long 3′-untranslated region contains a core of ATTTA motifs while the short form has only one or no ATTTA motif, indicating that the post-transcriptional modification of mRNA is important for regulation of CNP synthesis. These characteristics in the 3′-untranslated region were conserved among all elasmobranch CNP cDNAs. Since CNP has been implicated as a fast-acting hormone to facilitate salt secretion from the rectal gland, the conserved 3′-untranslated region most likely contributes to rapid regulation of CNP synthesis in elasmobranchs in response to acute changes in internal and external environments.

Descripción de las actividades de información del Consejo Nacional de Población del Perú (CNP)

Describe los objetivos y actividades del CNP, información utilizada y generada, su Centro de Documentación (CENDOC), la Red de Información sobre Población en el Perú (RIPOP), y sugerencias de cooperación interinstitucional y regional.

Formación física en los cuerpos de seguridad del Estado: especial atención al CNP

En este estudio se pretende investigar sobre la Formación Física en los Cuerpos de Seguridad del Estado centrándonos de manera especial en el Cuerpo Nacional de Policía. Para ello dividiremos los resultados de nuestro estudio en las exigencias de pruebas de acceso y en la formación física una vez dentro de los cuerpos policiales, en este caso el CNP. Para ello veremos el grado de aptitud física, el acceso a las unidades y finalmente la Agrupación Deportiva. Una vez obtenido los datos del estado actual de la cuestión se pretende discutir las razones de la formación y la exigencia existentes. Finalmente expondremos las conclusiones de todo el trabajo físico. Esperamos que la investigación tenga utilidad para reflexionar sobre todos estos aspectos en un ámbito tan importante en nuestra sociedad como es el ámbito policial.

Precursor Dependent Microstructure Evolution and Nonstoichiometry in Nanostructured Cerium Oxide Coatings Using the Solution Precursor Plasma Spray Technique

A solution precursor plasma spray (SPPS) technique has been used for direct deposition of cerium oxide nanoparticles (CNPs) from various cerium salt solutions as precursors. Solution precursors were injected into the hot zone of a plasma plume to deposit CNP coatings. A numerical study of the droplet injection model has been employed for microstructure development during SPPS. The decomposition of each precursor to cerium oxide was analyzed by thermogravimetric-differential thermal analysis and validated by thermodynamic calculations. The presence of the cerium oxide phase in the coatings was confirmed by X-ray diffraction studies. Transmission electron microscopy studies confirmed nanocrystalline (grain size <14 nm) characteristic of the coatings. X-ray photoelectron spectroscopy studies indicated the presence of a high concentration of Ce3+ (up to 0.32) in the coating prepared by SPPS. The processing and microstructure evolution of cerium oxide coatings with high nonstoichiometry are reported.

Report of the Study Group on “ Fisheries and Ecosystem Response to Recent Regime Shifts”

EXECUTIVE SUMMARY 1. DECADAL-SCALE CLIMATE EVENTS 1.1 Introduction 1.2 Basin-scale Patterns 1.3 Long Time Series in the North Pacific 1.4 Decadal Climate Variability in Ecological Regions of the North Pacific 1.5 Mechanisms 1.6 References 2. COHERENT REGIONAL RESPONSES 2.1 Introduction 2.2 Central North Pacific (CNP) 2.3 California Current System (CCS) 2.4 Gulf of Alaska (GOA) 2.5 Bering Sea and Aleutian Islands 2.6 Western North Pacific (WNP) 2.7 Coherence in Regional Responses to the 1998 Regime Shift 2.8 Climate Indicators for Detecting Regime Shifts 2.9 References 3. IMPLICATIONS FOR THE MANAGEMENT OF MARINE RESOURCES 3.1 Introduction 3.2 Response Time of Biota to Regime Shifts 3.3 Response Time of Management to Regime Shifts 3.4 Provision of Stock Assessment Advice 3.5 Decision Rules 3.6 References 4. SUGGESTED LITERATURE 4.1 Climate Regimes 4.2 Impacts on Lower Trophic Levels 4.3 Impacts on Fish and Higher Trophic Levels 4.4 Impacts on Ecosystems and Possible Mechanisms 4.5 Regimes and Fisheries Management APPENDIX 1: RECENT ECOSYSTEM CHANGES IN THE CENTRAL NORTH PACIFIC A1.1 Introduction A1.2 Physical Oceanography A1.3 Lower Trophic Levels A1.4 Invertebrates A1.5 Fishes A1.6 References APPENDIX 2: RECENT ECOSYSTEM CHANGES IN THE CALIFORNIA CURRENT SYSTEM A2.1 Introduction A2.2 Physical Oceanography A2.3 Lower Trophic Levels A2.4 Invertebrates A2.5 Fishes A2.6 References APPENDIX 3: RECENT ECOSYSTEM CHANGES IN THE GULF OF ALASKA A3.1 Introduction A3.2 Physical Oceanography A3.3 Lower Trophic Levels A3.4 Invertebrates A3.5 Fishes A3.6 Higher Trophic Levels A3.7 Coherence in Gulf of Alaska Fish A3.8 Combined Standardized Indices of Recruitment and Survival Rate A3.9 References APPENDIX 4: RECENT ECOSYSTEM CHANGES IN THE BERING SEA AND ALEUTIAN ISLANDS A4.1 Introduction A4.2 Bering Sea Environmental Variables and Physical Oceanography A4.3 Bering Sea Lower Trophic Levels A4.4 Bering Sea Invertebrates A4.5 Bering Sea Fishes A4.6 Bering Sea Higher Trophic Levels A4.7 Coherence in Bering Sea Fish Responses A4.8 Combined Standardized Indices of Bering Fish Recruitment and Survival Rate A4.9 Aleutian Islands A4.10 References APPENDIX 5: RECENT ECOSYSTEM CHANGES IN THE WESTERN NORTH PACIFIC A5.1 Introduction A5.2 Sea of Okhotsk A5.3 Tsushima Current Region and Kuroshio/Oyashio Current Region A5.4 Bohai Sea, Yellow Sea, and East China Sea A5.5 References (168 page document)

黑土微生物活力与组成对养分添加的响应

东北黑土是我国重要的商品粮生产基地，在保证我国的粮食安全中起着重要作用。鉴于东北黑土所具有的重要战略地位及其面临的严峻形势，如何通过化肥合理配施培肥黑土已经成为一个亟待解决的关键问题。 本文以东北黑土为供试土壤，设置CK、C、N、P、NP、CN、CP、CNP，8个养分组合，进行室内恒温模拟培养。通过对黑土呼吸强度的监测，研究黑土微生物活力对不同养分响应的规律；同时采用磷脂脂肪酸（Phospholipid Fatty Acids, PLFAs）和中性脂肪酸（Neutral Lipid Fatty Acids, NLFAs）为标识物，研究黑土微生物群落结构以及生理生态特性在不同养分条件下的动态变化，揭示黑土微生物活力与群落组成对不同养分响应的规律。 研究结果表明：黑土微生物活力首先受到可利用C源的限制，单加葡萄糖能够使土壤呼吸释放的CO2-C累积量达到对照的10倍以上，使黑土微生物活力显著提高；而在碳限制的状况被缓解后，氮便成为黑土微生物生长的主要限制因素，可利用碳底物充足的条件下，N的加入能够显著提高土壤呼吸强度，使释放出的CO2-C累积量达到对照处理的13倍以上，CN交互作用非常显著；在碳氮养分都充足的条件下，P的施入能更好地刺激微生物生长，提高黑土微生物活力，CNP处理，黑土微生物活力在培养的第3 d即达到峰值。CN和CNP组合能够迅速刺激真菌、细菌生长，使PLFA和NLFA含量、真菌/细菌比值均增加；并且CNP组合优于CN组合。单独加入C和CP组合，不能在短时间内使真菌、细菌的PLFA和NLFA含量显著增加，真菌/细菌比值也处于较低的水平。G+/G-比值在CN条件下，达到最大之后逐渐趋于平稳；而在单加C、CP和CNP条件下，G+/G-比值均在培养时间内达到两次峰值，显示出不同的养分条件下，微生物群落结构发生着内在的动态变化。

基于合同网协议交互投标的动态调度方法研究

柔性制造系统使生产加工路径有很多可选性,所以调度系统必须考虑机器调度问题。分配规则调度是一种最基本、最具影响力的动态调度方法。然而,分配规则调度方法很少考虑机器顺序选择。兼顾工件选择和机器选择两方面,本文运用交互投标过程,构建基于合同网协议调度的协商规则。研究作业车间动态调度问题,提出并构建了5种合同网规则调度方法。通过实验分析结果表明,基于合同网交互投标模式的规则调度能够大大改善调度系统性能,提高设备的利用率和设备负荷平衡指标。