High-level expression of biologically active glycoprotein hormones in Pichia pastoris strains—selection of strain GS115, and not X-33, for the production of biologically active N-glycosylated 15N-labeled phCG

The methylotrophic yeast Pichia pastoris is widely used for the production of recombinant glycoproteins. With the aim to generate biologically active 15N-labeled glycohormones for conformational studies focused on the unravelling of the NMR structures in solution, the P. pastoris strains GS115 and X-33 were explored for the expression of human chorionic gonadotropin (phCG) and human follicle-stimulating hormone (phFSH). In agreement with recent investigations on the N-glycosylation of phCG, produced in P. pastoris GS115, using ammonia/glycerol-methanol as nitrogen/carbon sources, the N-glycosylation pattern of phCG, synthesized using NH4Cl/glucose–glycerol–methanol, comprised neutral and charged, phosphorylated high-mannose-type N-glycans (Man8–15GlcNAc2). However, the changed culturing protocol led to much higher amounts of glycoprotein material, which is of importance for an economical realistic approach of the aimed NMR research. In the context of these studies, attention was also paid to the site specific N-glycosylation in phCG produced in P. pastoris GS115. In contrast to the rather simple N-glycosylation pattern of phCG expressed in the GS115 strain, phCG and phFSH expressed in the X-33 strain revealed, besides neutral high-mannose-type N-glycans, also high concentrations of neutral hypermannose-type N-glycans (Manup-to-30GlcNAc2). The latter finding made the X-33 strain not very suitable for generating 15N-labeled material. Therefore, 15N-phCG was expressed in the GS115 strain using the new optimized protocol. The 15N-enrichment was evaluated by 15N-HSQC NMR spectroscopy and GLC-EI/MS. Circular dichroism studies indicated that 15N-phCG/GS115 had the same folding as urinary hCG. Furthermore, 15N-phCG/GS115 was found to be similar to the unlabeled protein in every respect as judged by radioimmunoassay, radioreceptor assays, and in vitro bioassays.

Translational fusion of two beta-subunits of human chorionic gonadotropin results in production of a novel antagonist of

The strategy of translationally fusing the alpha-and beta-subunits of human chorionic gonadotropin (hCG) into a single-chain molecule has been used to produce novel analogs of hCG. Previously we reported expression of a biologically active singlechain analog hCG alpha beta expressed using Pichia expression system. Using the same expression system, another analog, in which the alpha-subunit was replaced with the second beta-subunit, was expressed (hCG beta beta) and purified. hCG beta beta could bind to LH receptor with an affinity three times lower than that of hCG but failed to elicit any response. However, it could inhibit response to the hormone in vitro in a dose- dependent manner. Furthermore, it inhibited response to hCG in vivo indicating the antagonistic nature of the analog. However, it was unable inhibit human FSH binding or response to human FSH, indicating the specificity of the effect. Characterization of hCG alpha beta and hCG beta beta using immunological tools showed alterations in the conformation of some of the epitopes, whereas others were unaltered. Unlike hCG, hCG beta beta interacts with two LH receptor molecules. These studies demonstrate that the presence of the second beta-subunit in the single-chain molecule generated a structure that can be recognized by the receptor. However, due to the absence of alpha-subunit, the molecule is unable to elicit response. The strategy of fusing two beta-subunits of glycoprotein hormones can be used to produce antagonists of these hormones.

Production of Dirac particles due to riccion coupling

It has been noted that at high energy the Ricci scalar is manifested in two different ways, as a matter field as well as a geometrical field (which is its usual nature even at low energy). Here, using the material aspect of the Ricci scalar, its interaction with Dirac spinors is considered in four-dimensional curved spacetime. We find that a large number of fermion-antifermion pairs can be produced by the exponential expansion of the early universe.

Production of biosurfactant “Biosur-Pm” by Pseudomonas maltophila CSV89: characterization and role in hydrocarbon uptake

Pseudomonas maltophilia CSV89, a soil bacterium, produces an extracellular biosurfactant, ''Biosur-Pm''. The partially purified product is nondialyzable and chemically composed of 50% protein and 12-15% sugar, which indicates the complex nature of Biosur-Pm. It reduces the surface tension of water from 73 to 53 x 10(-3) N m(-1) and has a critical micellar concentration of 80 mg/l. Compared to aliphatic hydrocarbons, Biosur-Pm shows good activity against aromatic hydrocarbons. The emulsion formed is stable and does not require any metal ions for emulsification. The kinetics of Biosur-Pm production suggest that its synthesis isa growth-associated and pH-dependent process. At pH 7.0, cells produced more Biosur-Pm with less cell surface hydrophobicity. At pH 8.0, however, the cells produced less Biosur-Pm with more cell surface hydrophobicity and showed a twofold higher affinity for aromatic hydrocarbons compared to the cells grown at pH 7.0. The Biosur-Pm showed a pH-dependent release, stimulated growth of the producer strain on mineral salts medium with 1-naphthoic acid when added externally, and facilitated the conversion of salicylate to catechol. All these results suggest that Biosur-Pm is probably a cell-wall component and helps in hydrocarbon assimilation/uptake.

The production of AlN-rich matrix composites by the reactive infiltration of Al alloys in nitrogen

Aluminium nitride (AlN)-Al matrices reinforced with Al2O3 particulate have been fabricated by reactive infiltration of Al-2% Mg alloy into Al2O3 preforms in N-2 in the temperature range of 900-1075 degreesC. The growth of composites of useful thickness was facilitated by the presence of a Mg-rich external getter, in the absence of which composite growth is self-limiting and terminates prematurely. Successful growth of composites has been attributed to the reduction in residual oxygen partial pressure brought about by the reaction with oxygen of highly volatile Mg in the getter alloy. The microstructure of the matrix consists of AlN-rich regions contiguous with the particulate with metal-rich channels in-between, thereby suggesting that nitridation initiates by preferential wicking of alloy along the particle surfaces. The increase in nitride content of the matrix with temperature is consistent with hardness values that vary between similar to3 and 10 GPa. (C) 2002 Acta Materialia Inc. Published by Elsevier Science Ltd. All rights reserved.

Production of niobium powder by electronically mediated reaction (EMR) using calcium as a reductant

Explored in this study is an electronically mediated reaction (EMR) route for the production of niobium powder using calcium as a reductant for niobium oxide (Nb2O5). Feed material, Nb2O5, and reductant calcium alloy containing aluminum and nickel were charged into electronically isolated locations in a molten salt (e.g. CaCl2) at 1173 K. The current flow through an external path between the feed and reductant locations was monitored. A current approximately 0.4 A was measured during the reaction in the external circuit connecting cathode and anode location. Niobium powder with low aluminum and nickel content was obtained although liquid Ca–Al–Ni alloy was used as the reductant. This clearly demonstrates that niobium metal powder can be produced by an electronically mediated reaction (EMR), without direct physical contact between feed (Nb2O5) and reductant (calcium). Mechanism of calciothermic reduction of Nb2O5 in the molten salt is discussed using an isothermal chemical potential diagram.

Forecasting Climate Impacts on Future Production of Commercially Exploited Fish and Shellfish

Foreword [pdf, < 0.1 MB] Acknowledgements PHASE 1 [pdf, 0.2 MB] Summary of the PICES/NPRB Workshop on Forecasting Climate Impacts on Future Production of Commercially Exploited Fish and Shellfish (July 19–20, 2007, Seattle, U.S.A.) Background Links to Other Programs Workshop Format Session I. Status of climate change scenarios in the PICES region Session II. What are the expected impacts of climate change on regional oceanography and what are some scenarios for these drivers for the next 10 years? Session III. Recruitment forecasting Session IV. What models are out there? How is climate linked to the model? Session V. Assumptions regarding future fishing scenarios and enhancement activities Session VI Where do we go from here? References Appendix 1.1 List of Participants PHASE 2 [pdf, 0.7 MB] Summary of the PICES/NPRB Workshop on Forecasting Climate Impacts on Future Production of Commercially Exploited Fish and Shellfish (October 30, 2007, Victoria, Canada) Background Workshop Agenda Forecast Feasibility Format of Information Modeling Approaches Coupled bio-physical models Stock assessment projection models Comparative approaches Similarities in Data Requests Opportunities for Coordination with Other PICES Groups and International Efforts BACKGROUND REPORTS PREPARED FOR THE PHASE 2 WORKSHOP Northern California Current (U.S.) groundfish production by Melissa Haltuch Changes in sablefish (Anoplopoma fimbria) recruitment in relation to oceanographic conditions by Michael J. Schirripa Northern California Current (British Columbia) Pacific cod (Gadus macrocephalus) production by Caihong Fu and Richard Beamish Northern California Current (British Columbia) sablefish (Anoplopoma fimbria) production by Richard Beamish Northern California Current (British Columbia) pink (Oncorhynchus gorbuscha) and chum (O. keta) salmon production by Richard Beamish Northern California Current (British Columbia) ocean shrimp (Pandalus jordani) production by Caihong Fu Alaska salmon production by Anne Hollowed U.S. walleye pollock (Theragra chalcogramma) production in the eastern Bering Sea and Gulf of Alaska by Kevin Bailey and Anne Hollowed U.S. groundfish production in the eastern Bering Sea by Tom Wilderbuer U.S. crab production in the eastern Bering Sea by Gordon H. Kruse Forecasting Japanese commercially exploited species by Shin-ichi Ito, Kazuaki Tadokoro and Yasuhiro Yamanka Russian fish production in the Japan/East Sea by Yury Zuenko, Vladimir Nuzhdin and Natalia Dolganova Chum salmon (Oncorhynchus keta) production in Korea by Sukyung Kang, Suam Kim and Hyunju Seo Jack mackerel (Trachurus japonicus) production in Korea by Jae Bong Lee and Chang-Ik Zhang Chub mackerel (Scomber japonicus) production in Korea by Jae Bong Lee, Sukyung Kang, Suam Kim, Chang-Ik Zhang and Jin Yeong Kim References Appendix 2.1 List of Participants PHASE 3 [pdf, < 0.1 MB] Summary of the PICES Workshop on Linking Global Climate Model Output to (a) Trends in Commercial Species Productivity and (b) Changes in Broader Biological Communities in the World’s Oceans (May 18, 2008, Gijón, Spain) Appendix 3.1 List of Participants Appendix 3.2 Workshop Agenda (Document contains 101 pages)

Reproduction, maturation, and seed production of cultured species: Proceedings of the Twelfth U.S.-Japan Meeting on Aquaculture, Baton Rouge, Louisiana, October 25-29, 1983

The United States and Japanese counterpart panels on aquaculture were formed in 1969 under the United States-Japan Cooperative Program in Natural Resources (UJNR). The panels currently include specialists drawn from the federal departments most concerned with aquaculture. Charged with exploring and developing bilateral cooperation, the panels have focused their efforts on exchanging information related to aquaculture which could be of benefit to both countries. The UJNR was begun during the Third Cabinet-Level Meeting of the Joint United States-Japan Committee on Trade and Economic Affairs in January 1964. In addition to aquaculture, current subjects in the program include desalination of seawater, toxic microorganisms, air pollution, energy, forage crops, national park management, mycoplasmosis, wind and seismic effects, protein resources, forestry, and several joint panels and committees in marine resources research, development, and utilization. Accomplishments include: Increased communication and cooperation among technical specialists; exchanges of information, data, and research findings; annual meetings of the panels, a policy-coordinative body; administrative staff meetings; exchanges of equipment, materials, and samples; several major technical conferences; and beneficial effects on international relations. (PDF file contains 79 pages.)

Investment prospects in the production of the common carp (Cyprinus carpio) fingerlings in freshwater ponds

Since the introduction of Common carp Cyprinus carpio in Oyo State (Nigeria) from Israel in 1964, various local breeding methods have been employed in carp rearing to improve the survival rate at all stages of development during breeding. The physico-chemical parameters of the ponds which were simultaneously investigated for carp rearing in this study includes temperature (t), dissolved oxygen (DO) and hydrogen-ion concentration (pH). However, high rates of water displacement in the breeding ponds were unfavourable to the development of zooplankton which play important role in the food web of C. carpio. The survival rates of 15.88-69.50% and 19.60-33.83% obtained for the egg-hatchling and hatchling-fingerling stages respectively were encouraging. A breeding performance of this magnitude was found to be viable, breaking even in the fourth year. However, an increase in size of this trial project would be more profitable and increase fingerling supply as well as provide employment opportunities. This study thereby provides some baseline information on some local techniques and progress in the propagation of C. carpio and scope for further improvement

Photographic database development of Nigerian fisheries: a complementary source of information documentation

The paper traces the history of the different documentation media used for information dissemination. Such early media are clay tablets, papyrus, and vellum or parchment codex. The invention of printing however revolutionized the information industry, enabling the production of books in multiple copies. Photography came into documentation mainly to preserve rare materials and those that easily deteriorate. This paper reports the efforts of National Institute for Freshwater Fisheries Research (NIFFR) and Kainji Lake Fisheries Promotion Project (KLFPPP), Nigeria, to develop an Object Oriented Database (OOD) using photographs. The photographs are stored in digitized form on commercial computers, using the program ACDSee 32 for classification, description and retrieval. Specifically the paper focuses on photographs in fisheries as visual communication and expression. Presently, the database contains photo documents about the following aspects of Kainji Lake fisheries: fishing gears and crafts, fish preservation methods

Effects of El Niño events on energy demand and egg production of rockfish (Scorpaenidae: Sebastes): a bioenergetics approach

Fish bioenergetics models estimate relationships between energy budgets and environmental and physiological variables. This study presents a generic rockfish (Sebastes) bioenergetics model and estimates energy consumption by northern California blue rockf ish (S. mystinus) under average (baseline) and El Niño conditions. Compared to males, female S. mystinus required more energy because they were larger and had greater reproductive costs. When El Niño conditions (warmer temperatures; lower growth, condition, and fecundity) were experienced every 3−7 years, energy consumption decreased on an individual and a per-recruit basis in relation to baseline conditions, but the decrease was minor (<4% at the individual scale, <7% at the per-recruit scale) compared to decreases in female egg production (12−19% at the individual scale, 15−23% at the per-recruit scale). When mortality in per-recruit models was increased by adding fishing, energy consumption in El Niño models grew more similar to that seen in the baseline model. However, egg production decreased significantly — an effect exacerbated by the frequency of El Niño events. Sensitivity analyses showed that energy consumption estimates were most sensitive to respiration parameters, energy density, and female fecundity, and that estimated consumption increased as parameter uncertainty increased. This model provides a means of understanding rockfish trophic ecology in the context of community structure and environmental change by synthesizing metabolic, demographic, and environmental information. Future research should focus on acquiring such information so that models like the bioenergetics model can be used to estimate the effect of climate change, community shifts, and different harvesting strategies on rockfish energy demands.

Preliminary report on the first production of the crab stages of alimasag, Portunus pelagicus (Linnaeus)

An experiment was undertaken studying the early life history of Portunus pelagicus in order to obtain information of relevance to rearing techniques for crabs.