Experimental evaluation of the processes resulting from the introduction of the transgenic microorganism Escherichia coli Z905/pPHL7 (lux+) into aquatic microcosms.

The processes resulting from the introduction of the tranagenic microorganism (TM) E. coli Z905/pPHL7 into aquatic microcosms have been modeled experimentally. It has been shown that the TM E. coli is able to adapt to a long co-existence with indigenous heterotrophic microflora in variously structured microcosms. In more complex microcosms the numerical dynamics of the introduced E. coli Z905/pPHL7 population is more stable. In the TM populations staying in the microcosms for a prolonged time, changes are recorded in the phenotypic expression of plasmid genes (ampicillin resistance and the luminescence level) and chromosome genes (morphological and physiological traits). However, in our study microcosms, the recombinant plasmid persisted in the TM cells for 6 years after the introduction, and as the population adapts to the conditions of the microcosms, the efficiency of the cloned gene expression in the cells is restored. In the microcosms with high microalgal counts (10(7) cells/ml), cells with a high threshold of sensitivity to ampicillin dominate in the population of the TM E. coli Z905/pPHL7.     

Survival and alteration of the plasmid-containing microorganism Escherichia coli Z905/pPHL7 introduced into manmade closed aquatic microcosms.

It has been demonstrated that the transgenic microorganism Escherichia coli Z905/pPHL7 (AprLux+) can exist for a long time at an elevated concentration of mineral salts. The microorganism was introduced into microcosms with sterile brackish water (salinity variable from 21 to 22 g l-1) taken from Lake Shira (Khakasia, Russia). The survival of the microorganism was estimated both by measuring the growth of the colonies on solid nutrient media and by the bioluminescence exhibited by the transgenic strain in samples from the microcosms and in the enrichment culture with the added selective factor-ampicillin (50 micrograms/ml). In the enrichment culture, the bioluminescent signal was registered through the 160-day experiment. It has been shown that in the closed microcosms with brackish water the E. coli strain becomes heterogeneous in its ampicillin resistance. The populations of the transgenic strain were mainly represented by isolates able to persist in the medium containing 50 micrograms/ml, but there were also the cells (about 10%) with the threshold of ampicillin resistance not more than 0.05 micrograms/ml. Thus, it was shown that in the microcosms with brackish water and in the absence of the selective factor the transgenic strain survives and retails the recombinant plasmid.     

Expression of lux-genes as an indicator of metabolic activity of cells in model ecosystem studies.

Quick response to different impacts and easy measurement make the luminescent systems of luminous bacteria an object convenient for application in various fields. Cloning of gene luminescence in different organisms is currently used to study both the survival of microbial cells and the effect of different factors on their metabolic activity, including the environment. A primary test-object in estimating bacteriological contamination of water bodies, Escherichia coli, can be conveniently used as an indicator of bactericidal properties of aquatic ecosystems. The application of Escherichia coli Z905/pPHL7 (lux+) as a marker microorganism can facilitate monitoring the microbiological status of closed biocenoses, including systems with higher organisms. The investigation of various parameters of microecosystems (carbon nutrition type, concentrations of inorganic ions and toxic compounds) shows that the recombinant strain E. coli Z905/pPHL7 can be effectively used as a marker.     

Population dynamics of transgenic microorganisms in the different microecosystem conditions.

The role of key environmental factors in adaptation of spore-forming and non-spore-forming transgenic microorganisms (TM) have been studied in model ecosystems. Model TM Escherichia coli Z905 (bearing plasmid genes of bacterial luminescence Ap (r) Lux+) has been found to have a higher adaptation potential than TM Bacillus subtilis 2335/105 (bearing genes of human alpha 2-interferon Km (r) Inf+), planned for employment as a living vaccine under varying environmental conditions. Effects of abiotic factors on migration of natural and recombinant plasmids between microorganisms under model ecosystem conditions has been estimated. The transgenic microorganisms with low copy number survived better under introduction conditions in the microcosms studied. This trend has been shown to be independent of the microcosm type and its complexity. Grant numbers: 99-04-96017, 25, 00-07-9011.     

Expression of cloned genes of transgenic microorganisms introduced into man-made ecosystems.

Modeling of transgenic microorganism introduction into small man-made ecosystems can help forecast changes in expression of cloned genes under different conditions of existence. Introduction of the E. coli Z905/pPHL7 strain containing a plasmid with luminescent system genes of luminous bacteria led to changes in cell and colony morphology, reduction in metabolic activity of cells, and, as a result, a lower level of expression of cloned gene. A low concentration of nutrients has been shown to favor greatly the phenotypic change of cells of the recombinant strain. Expression of cloned genes changed due to: a lower concentration of plasmid DNA, a change in regulation of cloned genes, and a change in cells of biosynthesis of substrates needed for expression of luminescent genes. The conducted investigations can provide a basis for the use of marker transgenic microorganisms in closed ecosystems of different types. Grant numbers: 99-04-96017, 00-07-9011.     

Modelling of genetically engineered microorganisms introduction in closed artificial microcosms.

The possibility of introducing genetically engineered microorganisms (GEM) into simple biotic cycles of laboratory water microcosms was investigated. The survival of the recombinant strain Escherichia coli Z905 (Apr, Lux+) in microcosms depends on the type of model ecosystems. During the absence of algae blooming in the model ecosystem, the part of plasmid-containing cells E. coli decreased fast, and the structure of the plasmid was also modified. In conditions of algae blooming (Ankistrodesmus sp.) an almost total maintenance of plasmid-containing cells was observed in E. coli population. A mathematics model of GEM's behavior in water ecosystems with different level of complexity has been formulated. Mechanisms causing the difference in luminescent exhibition of different species are discussed, and attempts are made to forecast the GEM's behavior in water ecosystems.     

Preliminary evaluation of the landsat-4 thematic mapper data for mineral exploration

Landsat-4 Thematic Mapper (TM) data recorded over an arid terrain were analyzed to determine the applicability of using of TM data for identifying and mapping hydrothermally altered, potentially mineralized rocks. Clays, micas, and other minerals bearing the OH anion in specific crystal lattice positions have absorption bands in the 2.2-μm region (TM channel 7, TM7) and commonly lack features in the 1.6-μm region (TM5). Channel ratios TM5/TM7, TM5/TM4, and TM3/TM1 were combined into a color-ratio-composite (CRC) image and used to distinguish hydrothermally altered rocks, unaltered rocks, and vegetation. These distinctions are made possible by using the TM5 and TM7, channels which are not available in the Landsat multispectral scanner (MSS). Digital masking was used to eliminate ambiguities due to water and shadows. However, some ambiguities in identification resulted between altered volcanic rocks and unaltered sedimentary deposits that contained clays, carbonates, and gypsum, and between altered volcanic rocks and volcanic tuffs diagenetically altered to zeolites. However, compared to MSS data, TM data should greatly improve the ability to map hydrothermally altered rocks in arid terrains.     

The use of microcosms as an experimental approach to understanding terrestrial ecosystem functioning.

Since 1986, a series of microcosm experiments has been conducted at the Unit of Comparative Plant Ecology (UCPE) in an attempt to test our understanding of the principles controlling the structure and dynamics of plant communities and ecosystems. In each experiment microcosms have been seeded with a common pool of organisms, and systems have been allowed to assemble under replicated controlled conditions. Experiment variables have included mineral nutrient supply, temperature, moisture supply, soil depth, carbon dioxide concentration, mycorrhizas, rhizobia, herbivores and carnivores. Results from these experiments are presented to illustrate the value of synthesised ecosystems in ecological research.     

辐射条件下微藻基因表达内参基因的选择

作为空间辐射的一种主要成分,紫外辐射可广泛引起陆地植物与水生生物细胞及其组份的破坏.荧光定量PCR技术广泛应用于各类胁迫环境下,研究目的基因的转录水平.在荧光定量PCR中选用合适的内参基因,能够更加准确地校正和标准化目的基因转录水平.本实验研究了辐射条件下水生生物莱茵衣藻6个传统内参基因18SrRNA,GAPDH,β-actin,β-tubulin,EF1-α和UBC基因表达的稳定性.经GeNorm软件研究分析,在辐射条件下,莱茵衣藻18SrRNA基因表达最不稳定,而选用β-actin和GAPDH作为双内参,可以得到更精确的实验结果.      

Early evaluation of thematic mapper data for coastal process studies

Two sets of TM data taken over the ocean off the coast of the Southeastern U.S. Bight were studied for the applicability of TM data to marine environments. First, the results of applying TM and TMS data to determine chlorophyll concentration in the ocean are presented. Chlorophyll quantification in the range of 0.5 to 2.0 mg/M³ was achieved by taking the ratio of TM band-1/band-2. Second, the results of applying TM band-6 data to monitor sea surface temperature are described. A comparison of TM data with AVHRR data shows TM readings coincide with AVHRR data within a scatter of 0.5°C in most of areas studied. Lastly, the results of a technique to map the water depths of coral reefs in the Great Bahama Bank are demonstrated. Depths from 0 to 20 meters, were delineated using TM band-1. The classification accuracy and origins of anomalous depth points are discussed.     

一种被动更新LBTS的HLA时间管理方案

LBTS(Lower Bound Time Stamp)查询是影响时间管理性能的重要因素.为提高高层体系结构(HLA,High Level Architecture)中时间管理的性能,提出了一种减小最小时间戳下限LBTS传递次数的优化方法,将主动查询LBTS变成当LBTS改变时才通知相应的受控成员.应用可靠组播服务,降低了时间管理的通讯代价,将算法应用在具有自主知识产权的AST-RTI(Advanced Simulation Technology-Run Time Infrastructure)中,应用系统证明能有效提高RTI的实时性.最后针对RTI时间管理机制在仿真应用中带来的问题以及解决方法进行了讨论.      

Mutagenic effects of heavy ions in bacteria.

Various mutagenic effects by heavy ions were studied in bacteria, irradiated at accelerators in Dubna, Prague, Berkeley or Darmstadt. Endpoints investigated are histidine reversion (B. subtilis, S. typhimurium), azide resistance (B. subtilis), mutation in the lactose operon (E. coli), SOS chromotest (E. coli) and lambda-prophage induction (E. coli). It was found that the cross sections of the different endpoints show a similar dependence on energy. For light ions (Z < or = 4) the cross section decreases with increasing energy. For ions of Z = 10, it is nearly independent of energy. For heavier ions (Z > or = 26) it increases with energy up to a maximum or saturation. The increment becomes steeper with increasing Z. This dependence on energy suggests a "mutagenic belt" inside the track that is restricted to an area where the density of departed energy is low enough not to kill the cell, but high enough to induce mutations.     

Physical assessment of coastal vulnerability under enhanced land subsidence in Semarang,Indonesia, using multi-sensor satellite data

Research has been conducted in Semarang, Indonesia, to assess coastal vulnerability under enhanced land subsidence using multi-sensor satellite data, including the Advanced Land Observing Satellite (ALOS) Phased Array type L-band SAR (PALSAR), Landsat TM, IKONOS, and TOPEX/Poseidon. A coastal vulnerability index (CVI) was constructed to estimate the level of vulnerability of a coastline approximately 48.68?km in length using seven physical variables, namely, land subsidence, relative sea level change, coastal geomorphology, coastal slope, shoreline change, mean tidal range, and significant wave height. A comparison was also performed between a CVI calculated using seven parameters and a CVI using six parameters, the latter of which excludes the land subsidence parameter, to determine the effects of land subsidence during the coastal vulnerability assessment. This study showed that the accuracy of coastal vulnerability was increased 40% by adding the land subsidence factor (i.e., CVI 6 parameters?=?53%, CVI 7 parameters?=?93%). Moreover, Kappa coefficient indicated very good agreement (0.90) for CVI 7 parameters and fair agreement (0.3) for CVI 6 parameters. The results indicate that the area of very high vulnerability increased by 7% when land subsidence was added. Hence, using the CVI calculation including land subsidence parameters, the very high vulnerability area is determined to be 20% of the total coastline or 9.7?km of the total 48.7?km of coastline. This study proved that land subsidence has significant influence on coastal vulnerability in Semarang.     

The ASTROCULTURE(TM) flight experiment series, validating technologies for growing plants in space.

A flight experiment, ASTROCULTURE(TM)-1 (ASC-1), to evaluate the operational characteristics and hardware performance of a porous tube nutrient delivery system (PTNDS) was flown on STS-50 as part of the U.S. Microgravity Laboratory-1 mission, 25 June to 9 July, 1992. This experiment is the first in a series of planned ASTROCULTURE(TM) flights to validate the performance of subsystems required to grow plants in microgravity environments. Results indicated that the PTNDS was capable of supplying water and nutrients to plants in microgravity and that its performance was similar in microgravity to that in 1g on Earth. The data demonstrated that water transfer rates through a rooting matrix are a function of pore size of the tubes, the degree of negative pressure on the 'supply' fluid, and the pressure differential between the 'supply' and 'recovery' fluid loops. A slightly greater transfer rate was seen in microgravity than in 1g, but differences were likely related to the presence of hydrostatic pressure effects at 1g. Thus, this system can be used to support plant growth in microgravity or in partial gravity as on a lunar or Mars base. Additional subsystems to be evaluated in the ASTROCULTURE(TM) flight series of experiments include lighting, humidity control and condensate recovery, temperature control, nutrient composition control, CO2 and O2 control, and gaseous contaminant control.     

X-ray induced alterations in the differentiation and mineralization potential of murine preosteoblastic cells

To evaluate the effects of ionizing radiation (IR) on murine preosteoblastic cell differentiation, we directed OCT-1 cells to the osteoblastic lineage by treatment with a combination of β-glycerophosphate (β-GP), ascorbic acid (AA), and dexamethasone (Dex). In vitro mineralization was evaluated based on histochemical staining and quantification of the hydroxyapatite content of the extracellular bone matrix. Expression of mRNA encoding Runx2, transforming growth factor β1 (TGF-β1), osteocalcin (OCN), and p21^CDKN1A was analyzed. Exposure to IR reduced the growth rate and diminished cell survival of OCT-1 cells under standard conditions. Notably, calcium content analysis revealed that deposition of mineralized matrix increased significantly under osteogenic conditions after X-ray exposure in a time-dependent manner. In this study, higher radiation doses exert significant overall effects on TGF-β1, OCN, and p21^CDKN1A gene expression, suggesting that gene expression following X-ray treatment is affected in a dose-dependent manner. Additionally, we verified that Runx2 was suppressed within 24 h after irradiation at 2 and 4 Gy. Although further studies are required to verify the molecular mechanism, our observations strongly suggest that treatment with IR markedly alters the differentiation and mineralization process of preosteoblastic cells.     

Temporal regulation of global gene expression and cellular morphology in Xenopus kidney cells in response to clinorotation.

Here, we report changes gene expression and morphology of the renal epithelial cell line, A6, which was derived from Xenopus laevis adult kidney that had been induced by long-term culturing with a three-dimensional clinostat. An oligo microarray analysis on the A6 cells showed that mRNA levels for 52 out of 8091 genes were significantly altered in response to clinorotation. On day 5, there was no dramatic change in expression level, but by day 8 and day 10, either upregulation or downregulation of gene expression became evident. By day 15, the expression levels of 18 out of 52 genes had returned to the original levels, while the remaining 34 genes maintained the altered levels of expression. Quantitative analyses of gene expression by real-time PCR confirmed that changes in the mRNA levels of selected genes were found only under clinorotation and not under hypergravity (7 g) or ground control. Morphological changes including loss of dome-like structures and disorganization of both E-cadherin adherence junctions and cortical actin were also observed after 10 days of culturing with clinorotation. These results revealed that the expression of selected genes was altered specifically in A6 cells cultured under clinorotation.     

Computer modeling of the biotic cycle formation in a closed ecological system.

The process of biotic turnover in a closed ecological system (CES) with an external energy flow was analyzed by mathematical modeling of the biotic cycle formation. The formation of hierarchical structure in model CESs is governed by energy criteria. Energy flow through the ecosystem increases when a predator is introduced into a "producer-reducer" system at steady state. Analysis of the model shows that under certain conditions the presence of the primary predator with its high mineralization ability accelerates the biotic turnover measured by primary production. We, therefore, conclude that for every system it is possible to find a suitable predator able to provide the system with a higher biotic turnover rate and energy consumption. Grant numbers: 99-04-96017/2000.     

Spectral transformation of ASTER and Landsat TM bands for lithological mapping of Soghan ophiolite complex,south Iran

Spectral transformation methods, including correlation coefficient (CC) and Optimum Index Factor (OIF), band ratio (BR) and principal component analysis (PCA) were applied to ASTER and Landsat TM bands for lithological mapping of Soghan ophiolitic complex in south of Iran. The results indicated that the methods used evidently showed superior outputs for detecting lithological units in ophiolitic complexes. CC and OIF methods were used to establish enhanced Red–Green–Blue (RGB) color combination bands for discriminating lithological units. A specialized band ratio (4/1, 4/5, 4/7 in RGB) was developed using ASTER bands to differentiate lithological units in ophiolitic complexes. The band ratio effectively detected serpentinite dunite as host rock of chromite ore deposits from surrounding lithological units in the study area. Principal component images derived from first three bands of ASTER and Landsat TM produced well results for lithological mapping applications. ASTER bands contain improved spectral characteristics and higher spatial resolution for detecting serpentinite dunite in ophiolitic complexes. The developed approach used in this study offers great potential for lithological mapping using ASTER and Landsat TM bands, which contributes in economic geology for prospecting chromite ore deposits associated with ophiolitic complexes.     

Preliminary evaluation of soil moisture probe for use with Arcillite.

There is a need for reliable methods of measuring the level and distribution of water in the solid substrates that are used for growing plants in space. In a microgravity environment, water distribution is governed generally by capillary forces. Arcillite is the solid substrate used in the ASTROCULTURE (TM) system which was developed for growing plants in space. The goal of this study is to evaluate the applicability of heat pulse moisture sensors for measuring moisture levels in Arcillite. The ASTROCULTURE system uses suction as a means of controlling the moisture level in Arcillite, but the spatial distribution of the moisture is left unknown. Studies of the moisture content in a cell experiment were conducted to calibrate a heat pulse moisture sensor and then the sensor was used in a suction experiment to verify moisture content and distribution. Results of the studies demonstrate that head pulse moisture sensors can be used to monitor moisture content and distribution within the root module of the ASTROCULTURE system.     

垂直电流对赤道电射流中双流不稳定性的影响

本文导出了在由中性风引起的垂直电流的条件下, 赤道电射流中双流不稳定性的相速度, 增长率和饱和水平的修正公式.在通常电射流条件下, 密度为2×10-7A/m2的垂直电流对不稳定性有一定的影响.当驱动电场接近不稳定性的临界值时, 上述垂直电流有着决定性的影响.本文结果说明反向电射流期间十分强的半周日潮汐风引起的垂直电流可能是使“H”型不稳定性饱和水平十分高的原因之一.