Practical application of seminal plasma

Natural mating deposits a large number of sperm in a high volume of seminal plasma. Semen processed for AI has the seminal plasma greatly diluted, and processing of boar sperm for cryopreservation removes all seminal plasma. Boar sperm were evaluated for the impact of seminal plasma on the functional status of boar sperm. Seminal plasma prevented or reversed capacitation in frozen-thawed boar sperm. However, supplementing frozen-thawed semen doses with 10% seminal plasma did not significantly affect the number of sperm found in the utero-tubal sperm reservoir, nor did it affect farrowing rates.     

现阶段中国生态农业可持续发展面临的实践和理论问题探讨

随着全球经济一体化的发展以及中国实施可持续发展和全面建设小康社会的需要,中国生态农业面临着新的机遇和挑战。我国生态农业经过20多年的实践,现已取得了一定进展,并具备了一定的发展基础。目前,全国2000多个县、乡镇先后实施了生态农业建设。然而,当前我国生态农业可持续发展面临着一系列实践和理论问题。在实践方面,存在着思想认识、技术、资金、建设与管理等问题。在科学理论方面则需要解决如生态农业的基本内涵、分类、生态模式的内在过程与机理及其生态服务功能、模式的尺度转换以及生态农业模式变化规律、生态农业安全等问题。因此,加快开展全国生态农业建设规划,加大资金投入和政策扶持力度,加强农业基础设施和社会化服务体系建设,加快农业产业结构调整和优化升级,发展生态产业,全面推动生态农业的产业化、标准化、信息化和现代化发展。是当前生态农业建设面临的重要任务。     

Practical application of fundamental concepts in exercise physiology

The collection of primary data in laboratory classes enhances undergraduate practical and critical thinking skills. The present article describes the use of a lecture program, running in parallel with a series of linked practical classes, that emphasizes classical or standard concepts in exercise physiology. The academic and practical program ran under the title of a particular year II module named Sports Performance: Physiology and Assessment, and results are presented over a 3-yr period (2004-2006), based on an undergraduate population of 31 men and 34 women. The module compared laboratory-based indexes of endurance (e.g., ventilatory threshold and exercise economy) and high-intensity exercise (e.g., anaerobic power), respectively, with measures of human performance (based on 20-m shuttle run tests). The specific experimental protocols reinforced the lecture content to improve student understanding of the physiological and metabolic responses (and later adaptations) to exercise. In the present study, the strongest relationship with endurance performance was the treadmill velocity at maximal aerobic power (r = +0.88, P < 0.01, n = 51); in contrast, the strongest relationship with high-intensity exercise performance was the mean power output (in W/kg) measured during a 30-s all-out cycle ergometer sprint (r = +0.80, P < 0.01, n = 48). In class student data analysis improved undergraduate indepth or critical thinking during seminars and enhanced computer and data presentation skills. The endurance-based laboratories are particularly useful for examining the underlying scientific principles that determine aerobic performance but could equally well be adapted to investigate other topics, e.g., differences in the exercise response between men and women.     

Expanding chemical biology of 2-oxoglutarate oxygenases

Beyond established roles in collagen biosynthesis, hypoxic signaling and fatty acid metabolism, recent reports have now revealed roles for human 2-oxoglutarate-dependent oxygenases in histone and nucleic acid demethylation and in signaling protein hydroxylation. The emerging role of these oxygenases in enabling a multiplicity of histone modifications has some analogy with their role in enabling structural diversity in secondary metabolism.     

Cytochrome P450 oxygenases of monoterpene metabolism

The cytochrome P450 monoterpene oxygenases are largely responsible for imparting structural and functional diversity to this family of natural products. In most cases, cytochrome P450-mediated allylic hydroxylation of a parental monoterpene olefin leads to a series of redox transformations and conjugation reactions which yield a family of structurally related derivatives and isomers. An overview is provided of the extant monoterpene oxygenases, with examples mainly from the mint (Lamiaceae) family of essential oil plants, and, where possible, information on the structure, mechanism, localization and regulation of these enzymes is described. The review concludes with a brief assessment of biotechnological applications and a view to future research in this area.     

Role of oxygenases in pisatin biosynthesis and in the fungal degradation of maackiain

Some isolates of the plant pathogen Nectria haematococca detoxify the isoflavonoid phytoalexin (−)maackiain by hydroxylation at carbon 6a. Precursor feeding studies strongly suggest that the penultimate step in (+)pisatin biosynthesis by Pisum sativum is 6a-hydroxylation of (+)maackiain. We have used ¹⁸O labeling to test the involvement of oxygenases in these two reactions. When fungal metabolism of maackiain took place under ¹⁸O₂, the product was labeled with 99% efficiency; no label was incorporated by metabolism in H₂¹⁸O. Pisatin synthesized by pea pods in the presence of ¹⁸O₂ or H₂¹⁸O was a mixture of molecules containing up to three labeled oxygen atoms. Primary mass spectra of such mixtures were complex but were greatly simplified by tandem MS. This analysis indicated that the 6a oxygen of pisatin was derived from H₂O and not from O₂. Labeling patterns for the other five oxygen atoms were consistent with the proposed pathway for biosynthesis of pisatin and related isoflavonoids. We conclude that the fungal hydroxylation of maackiain is catalyzed by an oxygenase, but the biosynthetic route to the 6a hydroxyl of pisatin is unknown.     

Role of the jelly-roll fold in substrate binding by 2-oxoglutarate oxygenases

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Highlights? Structural features of 2OG oxygenases involved in substrate recognition are analyzed. ? Crystallographic studies reveal the versatility of the jelly roll fold in substrate binding. ? Defined structural regions that interact with substrate(s) are biased by fold topology. ? The utility of the enzyme–substrate structures for engineering and selective inhibition are discussed.     

Key issues in bioheat transfer simulations for the application of cryosurgery planning

The bioheat transfer simulation is undoubtedly the foundation for developing computerized tools for cryosurgery planning and analysis. While a large variety of techniques for bioheat transfer simulations are available in the literature of the past several decades, it is only their integration with clinical criteria and constraints which can make computerized planning a practical reality. This brief communication outlines (in the opinion of this author) the key issues that must be addressed in the application of bioheat transfer to cryosurgery planning and analysis, while drawing attention to recent and relevant publications in other journals, with reference to the most recent publication on the topic in the Journal of Cryobiology [Z. Magalov, A. Shitzer, D. Degani, Isothermal volume contours generated in a freezing gel by embedded cryo-needles with applications to cryo-surgery, Cryobiology 55 (2) (2007) 127–137].     

Practical application of ligand efficiency metrics in lead optimisation

The use of composite metrics that normalise biological potency values in relation to markers of physicochemical properties, such as size or lipophilicity, has gained a significant amount of traction with many medicinal chemists in recent years. However, there is no consensus on best practice in the area and their application has attracted some criticism. Here we present our approach to their application in lead optimisation projects, provide an objective discussion of the principles we consider important and illustrate how our use of lipophilic ligand efficiency enabled the progression of a number of our successful drug discovery projects. We derive, from this and some recent literature highlights, a set of heuristic guidelines for lipophilicity based optimisation that we believe are generally applicable across chemical series and protein targets.     

Variable composition of heme oxygenases with different regiospecificities in Pseudomonas species

Heme oxygenases (HO) degrade heme yielding iron, carbon monoxide and one of four possible biliverdin (BV) isomers. Pseudomonas aeruginosa PAO1 is thus far the only organism to contain two HOs with different regiospecificities: BphO and PigA. While BphO cleaves heme to exclusively yield BV IXα, PigA produces the BV isomers IXβ and IXδ. We bioinformatically identified putative HOs in diverse Pseudomonas strains, tested their enzymatic functionality and determined their regiospecificity. Surprisingly, even high amino acid sequence identities to the P. aeruginosa HOs were not sufficient to correctly predict the HO regiospecificity in all cases. Based on our results, Pseudomonas strains differ in their HO composition containing either BphO or PigA or both HO types. Concomitantly with the existence of bphO is the occurrence of at least one gene encoding a bacterial phytochrome implying that only BV IXα is the sufficient phytochrome chromophore. In contrast, pigA genes are organized in gene clusters associated with iron utilization implying a role of PigA in iron acquisition. However, at least in strains containing no PigA this function maybe fulfilled by BphO. Only a combination of homology searches and analyses of genetic environments is appropriate for a reliable prediction of the regiospecificity of Pseudomonas HOs.