Exploration of new drug delivery pathways: I. Mechanism of folate uptake in cultured human cells. II. Role of nuclear localization signal peptides in the delivery of oligonucleotide into reconstituted nuclei

The roles of the folate receptor and an anion carrier in the uptake of 5- methyltetrahydrofolate (5-MeH_4folate) were studied in cultured human (KB) cells using radioactive 5-MeH_4folate. Binding of the 5-MeH_4folate was inhibited by folic acid, but not by probenecid, an anion carrier inhibitor. The internalization of 5-MeH_4folate was inhibited by low temperature, folic acid, probenecid and methotrexate. Prolonged incubation of cells in the presence of high concentrations of probenecid appeared to inhibit endocytosis of folatereceptors as well as the anion carrier. The V_(max) and K_M values for the carrier were 8.65 ± 0.55 pmol/min/mg cell protein and 3.74 ± 0.54µM, respectively. The transport of 5-MeH4folate was competitively inhibited by folic acid, probenecid and methotrexate. The carrier dissociation constants for folic acid, probenecid and methotreate were 641 µM, 2.23 mM and 13.8 µM, respectively. Kinetic analysis suggests that 5-MeH_4folate at physiological concentration is transported through an anion carrier with the characteristics of the reduced-folate carrier after 5-MeH_4folate is endocytosed by folate receptors in KB cells. Our data with KB cells suggest that folate receptors and probenecid-sensitive carriers work in tandem to transport 5-MeH_4folate to the cytoplasm of cells, based upon the assumption that 1 mM probenecid does not interfere with the acidification of the vesicle where the folate receptors are endocytosed.

Oligodeoxynucleotides designed to hybridize to specific mRNA sequences (antisense oligonucleotides) or double stranded DNA sequences have been used to inhibit the synthesis of a number of cellular and viral proteins (Crooke, S. T. (1993) FASEB J. 7, 533-539; Carter, G. and Lemoine, N. R. (1993) Br. J. Cacer 67, 869-876; Stein, C. A. and cohen, J. S. (1988) Cancer Res. 48, 2659-2668). However, the distribution of the delivered oligonucleotides in the cell, i.e., in the cytoplasm or in the nucleus has not been clearly defined. We studied the kinetics of oligonucleotide transport into the cell nucleus using reconstituted cell nuclei as a model system. We present evidences here that oligonucleotides can freely diffuse into reconstituted nuclei. Our results are consistent with the reports by Leonetti et al. (Proc. Natl. Acad. Sci. USA, Vol. 88, pp. 2702-2706, April 1991), which were published while we were carrying this research independently. We also investigated whether a synthetic nuclear localization signal (NLS) peptide of SV40 T antigen could be used for the nuclear targeting of oligonucleotides. We synthesized a nuclear localization signal peptide-conjugated oligonucleotide to see if a nuclear localization signal peptide can enhance the uptake of oligonucleotides into reconstituted nuclei of Xenopus. Uptake of the NLS peptide-conjugated oligonucleotide was comparable to the control oligonucleotide at similar concentrations, suggesting that the NLS signal peptide does not significantly enhance the nuclear accumulation of oligonucleotides. This result is probably due to the small size of the oligonucleotide.

Development of a new modelling framework to estimate the C footprint from Basque dairy farms

19 p.

New climate scenario framework implementation in the GCAM integrated assessment model

48 p.

The new exploration for proton-implanted silicon: the conversion of a surface-region-purification-induced p-n junction into a p-i-n electrical structure approaching silicon on insulator

A surface-region-purification-induced p-n junction, a puzzle discovered at Brookhaven National Laboratory, in a silicon-on-defect-layer (SODL) material has been explored by carrying out various annealing conditions and subsequent measurements on electrical properties. The origin of the pn junction has been experimentally investigated. Furthermore, the p-n junction has been transformed into a p-i-n electrical structure by adding a high temperature annealing process to the previously used SODL procedure, making the SODL material approach silicon on insulator (SOI). The control of the initial oxygen amount in the silicon material is suggested to be critical for the experimental results.

Strain gradient theory based on a new framework of non-local model

A new framework of non-local model for the strain energy density is proposed in this paper. The global strain energy density of the representative volume element is treated as a non-local variable and can be obtained through a special integral of the local strain energy density. The local strain energy density is assumed to be dependent on both the strain and the rotation-gradient. As a result of the non-local model, a new strain gradient theory is derived directly, in which the first and second strain gradients, as well as the triadic and tetradic stress, are introduced in the context of work conjugate. For power law hardening materials, size effects in thin metallic wire torsion and ultra-thin cantilever beam bend are investigated. It is found that the result predicted by the theoretical model is well consistent with the experimental data for the thin wire torsion. On the other hand, the calculation result for the micro-cantilever beam bend clearly shows the size effect.

济阳-昌潍坳陷火成岩油气藏体系形成机理和分布规律研究

Jiyang & Changwei depressions are two neighboring depressions in Bahai Bay Basin, the famous oil rich basin in East China. The exploration activities in the past 40 years has proved that, within the basins, there exists not only plentiful sandstone hydrocarbon reservoirs (conventional), but also abundant special reservoirs as igneous rock, mudstone and conglomerate ones which have been knowing as the unconventional in the past, and with the prospecting activity is getting more and more detailed, the unconventional reservoirs are also getting more and more important for further resources, among which, the igneous lithological reservoir be of significance as a new research and exploration area. The purpose of this paper is, with the historical researches and data as base, the System Theory, Practice Theory and Modern Comprehensive Petroleum Geology Theory as guide, the theoretical and practice break through as the goal, and the existing problems in the past as the break through direction, to explore and establish a valid reservoir formation and distribution models for igneous strata in the profile of the eastern faulted basins. After investigating the distribution of the igneous rocks and review the history of the igneous rocks reservoirs in basins, the author focused on the following issues and correspondingly the following progresses have been made: 1.Come to a new basin evolution and structure model named "Combined-Basin-bodies Model" for Jiyang even Eastern faulted basins based on the study on the origin and evolution of Jiyang & Changwei basins, depending on this model, every faulted basin in the Bo-hai Bay Basin is consisted of three Basin-Bodies including the Lower (Mesozoic), Middle (Early Tertiary) and the Upper (Late Tertiary) Bodies, each evolved in different geo-stress setting and with different basin trend, shape and igneous-sedimentary buildings system, and from this one to next one, the basin experienced a kind of process named "shape changing" and "Style changing". 2. Supposed a serious of new realizations as follows (1) There were "multi-level magma sources" including Upper mantel and the Lower, Middle and even the Upper Shell magma Chambers in the historical Magma Processes in the basins; (2) There were "multi-magma accessing or pass" from the first level (Mantel faults) to the second, third and fourth levels (that is the different levels of fault in the basin sediment strata) worked in the geo-historical and magma processes; (3) Three tectonic magma cycles and more periods have been recognized those are matched with the "Basin -body-Model" and (4)The geo-historical magma processes were non-homogeneous in time and space scale and so the magma rocks distributed in "zones" or "belts". 3. The study of magma process's effect on basin petroleum conditions have been made and the following new conclusions were reached: (1) the eruptive rocks were tend to be matched with the "caped source rock", and the magma process were favorable to the maturing of the source rocks. (2) The magma process were fruitful to the accumulation of the non-hydrocarbon reservoirs however a over magma process may damage the grade of resource rock; (3) Eruptive activity provided a fruitful environment for the formation of such new reservoir rocks as "co-eruptive turbidity sandstones" and "thermal water carbonate rocks" and the intrusive process can lead to the origin of "metamorphism rock reservoir"; (4) even if the intrusive process may cause the cap rock broken, the late Tertiary intrusive rocks may indeed provide the lateral seal and act as the cap rock locally even regionally. All above progresses are valuable for reconstructing the magma-sedimentary process history and enriching the theory system of modem petroleum geology. 4. A systematic classification system has been provided and the dominating factors for the origin and distribution of igneous rock reservoirs have been worked out based on the systematic case studies, which are as follows: (1) The classification is given based on multi-factors as the origin type, litho-phase, type of reservoir pore, reservoir ability etc., (2) Each type of reservoir was characterized in a detailed way; (3) There are 7 factors dominated the intrusive reservoir's characteristics including depth of intrusion, litho-facies of surrounding rocks, thickness of intrusive rock, intrusive facies, frequency and size of the working faults, shape and tectonic deformation of rock, erosion strength of the rock and the time of the intrusion ect., in the contrast, 4 factors are for eruptive rocks as volcanic facies, frequency and size of the working faults, strength of erosion and the thermal water processing. 5. Several new concept including "reservoir litho-facies", "composite-volcanic facies" and "reservoir system" ect. Were suggested, based on which the following models were established: (1) A seven reservoir belts model for a intrusive unit profile and further more, (2) a three layers cubic model consisted of three layer as "metamorphic roe layer", "marginal layer" and "the core"; (3) A five zones vertical reservoir sequence model consisted of five litho-facies named A, B, C, D and E for a original lava unit and furthermore three models respectively for a erosion, subsidence and faulted lava unit; (4) A composite volcanic face model for a lava cone or a composite cone that is consisted of three facies as "crater and nearby face", "middle slope" and "far slope", among which, the middle slope face is the most potential reservoir area and producible for oil & gas. 6. The concept of "igneous reservoir" was redefined as the igneous, and then a new concept of "igneous reservoir system" was supposed which means the reservoir system consisted of igneous and associated non-igneous reservoirs, with non-hydrocarbon reservoir included. 7. The origin and distribution of igneous reservoir system were probed and generalized for the exploration applications, and origin models of the main reservoir sub-systems have been established including those of igneous, related non-igneous and non-hydrocarbon. For intrusive rocks, two reservoir formation models have been suggested, one is called "Original or Primary Model", and the another one is "Secondary Model"; Similarly, the eruptive rock reservoirs were divided in three types including "Highly Produced", "Moderately Produced" and "Lowly Produced" and accordingly their formation models were given off; the related non-igneous reservoir system was considered combination of eight reservoirs, among which some ones like the Above Anticline Trap are highly produced; Also, the non-hydrocarbon. Trap system including five kinds of traps was discussed. 8. The concept models for four reservoir systems were suggested, which include the intrusive system consisted of 7 kinds of traps, the land eruptive system with 6 traps, the under water eruptive system including 6 kinds of traps and the non-hydrocarbon system combined by 5 kinds of traps. In this part, the techniques for exploration of igneous reservoir system were also generalized and probed, and based on which and the geological progresses of this paper, the potential resources and distributions of every reservoir system was evaluated and about 186 millions of reserves and eight most potential non-hydrocarbon areas were predicted and outlined. The author believe that the igneous reservoir system is a very important exploration area and its study is only in its early stage, the framework of this paper should be filled with more detailed studies, and only along way, the exploration of igneous reservoir system can go into it's really effective stage.

东营凹陷下第三系岩性油气藏形成条件与预测

The Dongying depression, located in the northern part of the jiyang Sag in the Buohaiwan Basin, comprises one of the major oil-producing bases of the Shengli oil-field. The prediction and exploration of subtle or litho1ogical oil traps in the oil-field has become the major confronted target. This is also one of the frontier study areas in the highly-explored oil-bearing basins in East China and abroad. Based on the integrated analysis of the geological, seismic and logging data and the theories of sequence stratigraphy, tectono-stratigraphy and petroleum system, the paper has attempted to document the characteristics of the sequence stratigraphic and structural frameworks of the low Tertiary, the syndepositional faults and their control on deposition, and then to investigate the forming conditions and distribution of the tithological oil traps in the depression. The study has set up a set of analysis methods, which can be used to effectively analysis the sequence stratigraphy of inland basins and predict the distribution of sandstone reservoirs in the basins. The major achievements of the study are as follows: 1. The low Tertiary can be divided into 4 second-order sequences and 13 third-order sequences, and the systems tracts in the third-order sequences have been also identified based on the examination and correction of well logging data and seismic profiles. At the same time, the parasequences and their stacking pattern in the deltaic systems of the third member of the Shahejie Formation have been recognized in the key study area. It has been documented that the genetic relation of different order sequences to tectonic, climatic and sediment supply changes. The study suggested that the formation of the second-order sequences was related to multiple rifting, while the activity of the syndepositional faults controlled the stacking pattern of parasequences of the axial deltaic system in the depression. 2. A number of depositional facies have been recognized in the low Tertiary on the basis of seismic facies and well logging analysis. They include alluvial fan, fan delta or braided delta, axial delta, lowstand fan, lacustrine and gravity flow deposits. The lacustrine lowstand fan deposits are firstly recognized in the depression, and their facies architecture and distribution have been investigated. The study has shown that the lowstand fan deposits are the important sandstone reservoirs as lithological oil traps in the depression. 3. The mapping of depositional systems within sequences has revealed the time and special distrbution of depositional systems developed in the basin. It is pointed out that major elastic systems comprise the northern marginal depositional systems consisting of alluvial fan, fan delta and offshore lowstand fan deposits, the southern gentle slope elastic deposits composed of shallow lacustrine, braided delta and lowstand fan deposits and the axial deltaic systems including those from eastern and western ends of the depression. 4. The genetic relationship between the syndepositional faults and the distribution of sandstones has been studied in the paper, upper on the analysis of structural framework and syndepositional fault systems in the depression. The concept of structural slope-break has been firstly introduced into the study and the role of syndepositional faults controlling the development of sequence architecture and distribution of sandstones along the hinged and faulted margins have been widely investigated. It is suggested that structural styles of the structural slope-break controlled the distribution of lowstand fan deposits and formed a favorable zone for the formation of lithological or structure-lithological oil traps in the basin. 5. The paper has made a deep investigation into the forming condition and processes of the lithological traps in the depression, based the analysis of composition of reservoir, seal and resource rocks. It is pointed out that there were two major oil pool-forming periods, namely the end of the Dongying and Guangtao periods, and the later one is the most important. 6. The study has finally predicted a number of favorable targets for exploration of lithologieal traps in the depression. Most of them have been drilled and made great succeed with new discovered thousands tons of raw oil reserves.

丘陵油田三间房组储层三维地质模型和砂体预测

This paper is belonging to Chinese Petrochemical Corporation's science and technology project. Although it is difficult, it has important theoretical and practical value. The study was aimed to reveal inhomogeneity of two kinds of reservoirs of fan-shaped delta and braided river by using new theories, new methods and new technology about 3-D model building and reservoir knowledge repository throughout the world, and to build reservoir knowledge repository and 3-D geological model which would predict the type of sand body forming reason and distribution rule in order to improve exploration result in Qiuling oil fields. Multi-discipline theories such as petroleum structure geology, reservoir geology, petroleum geology, sequence geology, logging geology, geomathematics and so on are used as guide. The information of geology, seism, logging and production test is combined. Outcrop area and overlap area are combined. By making full use of computer, stable structure, reservoir geometric shape, spatial distribution and inhomogeneity of bed of interest are investigated, described and characterized. Petroleum pool 3-D static geological model of reservoir knowledge repository was built. Sand body distribution was predicted. It has guided oil development, lowed the investment and improved development benefits. Several results are achieved as follows: (1) Strata framework of Sanjianfang group in Qiuling oil field has been established. (2) Geometric shape, spatial distribution and evolve rule of two different forming reason's reservoir of fan-shaped delta and braided river of Sanjianfang group in Qiuling oil field are discussed. (3) The two kinds of reservoirs have lower pore and permeability and very strong inhomogeneity. (4) Reservoir knowledge repository of two different forming reasons has been built of Sanjianfang group, which includes 5 geological knowledge sublibrary. (5) 3-D geological model of two kinds of forming reason's reservoirs has been built. (6) That same sequence instruction a simulation and probability field were used to predict sand body of Sanjianfang group was put forward. Coincidence rate is high after production test. It shows this method has great popularity value. (7) A set of theories, methods and technologies of knowledge repository of two kinds of reservoir of braided river and fan-shaped delta and 3-D geological model building were finished. (8) A set of theories, methods and technologies of investigating, describing, characterizing and predicting two kinds of oil pool were developed. It gets noticeable economic benefit after exploration. Theory and method about extrusion basin are developed.