Effect of Aβ1-40 on membrane permeability and intracellular free Ca2+ of nerve cells

The effects of soluble and fibrillar Aβ1-40 on membrane permeability and intracellular free Ca2+ of nerve cells were investigated by the laser confocal microscopy. Results indicate that: i) Effects of soluble and fibrillar Aβ1-40 on cell membrane permeability are both concentration-de- pendent. Soluble Aβ1-40 increases membrane permeability only at concentration of 3 ?mol/L, while the toxic effect of fibrillar Aβ1-40 is much stronger, its evident effect begins from 1 ?mol/L. When its concentration rose to 3 ?mol/L, not only the membrane permeability increased, but also the nuclear membrane broke seriously. ii) Both soluble and fibrillar Aβ1-40 at high concentrations increased the intracellular free Ca2+, and the increased amplitudes are concentration-dependent. However, the fibrillar one induces the increase of intracellular Ca2+ much quicker and synchronously. These results indicate that some correlation exists between the neurotoxicity of high concentration soluble and fibrillar Aβ1-40 and the change of physico-chemical properties and intracellular Ca ion imbalance.     

Two forms of the membrane-bound state of the first C2 domain (C2A) of synaptotagminⅠand calcium-triggered membrane insertion

The synaptic vesicle protein synaptotagmin I(syt I) is a vesicle transmembrane protein present in synaptic vesicles, which has been proposed as the Ca^2 sensor that regulates secretion. The C2A domain is the membrane proximal part of its cytoplasmic domain. The interaction between C2A and lipid bilayer has been considered to be essential for triggering neurotransmitter release. In the present work, the measurements of membrane surface tension and surface concentration showed that the C2A domain of syt I exhibited two membrane-bound states: the surface adsorption state and the membrane insertion state. The surface absorption state formed in a Ca2~-independent manner with lower affinity, while the membrane insertion state formed with high affinity was only found in the presence of Ca^2 . Both the Ca^2 -independent and Ca^2 -dependent syt I membrane interactions required anionic phospholipids, such as phosphatidylserine (PS). When expressed into rat pheo-chromocytoma (PC12) cells and human embryonic kidney (HEK-293) cells, as demonstrated by immunofluorescence staining and subcellular fractionation, most of the C2A was found at the plasma membrane, even when the cells weredepleted of Ca^2 by incubation with EGTA. These resultssuggested a new molecular mechanism of syt I as a Ca^2 sensor in membrane fusion. Ca^2 -independent surface adsorption might attach syt I to the release site during the docking or priming step. When intracellular Ca^2 increased,syt I triggered the neurotransmitter release following the Ca^2 -dependent penetration into the target membrane.     

Influence of green tea polyphenols on mitochondrial permeability transition pore and Ca^2＋ transport

The authors investigated the influence of green tea polyphenols(GPSs) on the liver mitochondria permeability transition pore(PTP) opening through mitochondria sweling and change of mitochcodrin membrane potetial .The data showed that GPSs had obvious protective effect on the Ca^2 -induced PTP opening in a dose-dependent manner detected by mitochondria swelling.The results were obtained by meating the change of mitochondria membrane potential through Rh 123.Further experiments were conducted to examine the detailed influence of GTPs on Ca^2 import and export of mitochondria.The results showed that GTPs had remarkably inhibitory effect on the Ca^2 -induced Ca^2 import in mitochondria.and they could accelerate Ca^2 -release from from mitochondria.Our data provide an altemate interpretation of the potent protective function of GPSs on cell against apoptosis.     

Physiological responses of osteoblasts to cyclic stretching and the change of intracellular calcium concentration

The development of bone tissues is regulated by mechanical stimulation.Cyclic stretching was applied to the osteoblasts that were delivered from rat calvarie,The results showed that stretching at 500με increased the cell proliferation while loading at 1000με and 1500με inhabited cell growth ,Loading also increased the adhesive force between cells and substrate as well as spreading areas of osteobalsts.Furthermore,the fluorescence probe Fluo-3/AM was used to investigate the effect of stretching stimulation on the intracellular calcium concentration of osteoblasts.The intracellular calcium concentration of osteoblasts that were stretched at 500 με for 5 min was 92.9% higher than the control ,After being treated with the panax ontoginseng saponins,the streteched osteoblasts still expressed 28.6% higher intracellular calcium concentration than that of the control ,which proved that both the influx of extracellualr calcium and the release of intracellular calcium store were involved in the increase of intracellular calcium concentration when osteoblasts responded to the cyclic stretching And the influx of extracellular calcium through transmembrance channel played a main role.     

Effects of Cu2+ and Ph on osteoclastic bone resorption in vitro

The effects of Cu2+ and pH on osteoclastic bone resorption lacunae are studied by culturing Japanese white rabbit osteoclasts on bone slices. The number and surface area of bone resorption lacunae are measured by photomicrography and image analysis. Concentration of calcium ion in the supernatant is measured by atomic absorption spectrometry to evaluate the activity of osteoclasts. The morphology of the lacunae is observed under a scanning electron microscope. The results indicate that Cu2+ at concentration of 1.00×10-6 mol/L and 1.00×10-7 mol/L inhibits osteoclastic activity and causes a dose-dependent reduction in the number and surface area of the lacunae. While the number of lanunae is increased and osteoclastic bone resorbing function is significantly improved at Cu2+ concentration of 1.00×10-8 mol/L. It is suggested that the effect of Cu2+ on osteoclastic bone resorption depends on Cu2+ concentration. pH had no significant effect on osteoclastic function in the near neutral range.     

Recent progress in the study of intracellular toxicity of amyloid P peptide in Alzheimer's disease

Amyloidβ(Aβ) deposition is one of the major pathological markers of Alzheimer's disease (AD). Extracellular Afi toxi-city has been studied for a long time in AD research field. However, controversial data show that extracellular Aβload does not correlate with the dementia levels of AD patients and extracellular Aβonly induces significant cell death at non-physiological high concentrations. With the evolvement of Afi hypothesis, considerable attention has been devoted to the study of intracellular Aβtoxicity recently. Intracellular Aβinduces dramatic cell loss in AD transgenic models and in human primary neurons (at pM concentrations) through p53, Bax and caspase-6 pathways. Here, we review the generation, toxicity and possible pathways of intracellular Aβtoxicity, and discuss the implication and current knowledge of intracellular Aβin neuronal cell loss in neurodegenerative diseases.     

Ca^2＋ is involved in muscarine-acetylcholine-receptor-mediated acetylcholine signal transduction in guard cells of Vicia faba L.

Acetyicholine (ACh) is an important neuro-chemical transmitter in animals; it also exists in plants and plays a significant role in various kinds of physiological functions in plants. ACh has been known to induce the stomatal opening. By monitoring the changes of cytusolic Ca^2 with fluorescent probe Fiuo-3 AM under the confocal microscopy, we found that exogenous ACh increased cytosolic Ca^2 concentration of guard cells of Vicia faba L. Muscarlne, an agonist of muscarine acetyicholine receptor (mAChR), could do so as well. In contrast, atropine, the antagonist of mAChR abolished the ability of ACh to increase Ca^2 in guard cells. This mechanism is similar to mAChR in animals. When EGTA was used to chelate Ca^2 or ruthenium red to block Ca^2 released from vacuole respectively, the results showed that the increased cytosolic Ca^2 mainly come from intracellular Ca^2 store. The evidence supports that Ca^2 is involved in guard-cell response to ACh and that Ca^2 sigual is coupled to mAChRs in ACh signal transduction in guard cells.     

Preparation of pH-responsive ceramic composite membranes by grafting acrylic acid onto α-alumina membranes

A pH-responsive ceramic composite membrane was prepared by chemical graft polymerization of acrylic acid （AA） onto the KH-570 modified （x-alumina membrane. The influence of monomer concentration on the gating characteristics of the pH-responsive membrane was investigated. The FT-IR spectrum, contact angle and water filtration rate of the membrane were measured. The monomer concentration was found to have a remarkable effect on the pH-response coefficient and the water filtration rate. In addition, the grafted membrane exhibited fast and reversible response to the pH change in the external solution.     

Permeation of vanadium（Ⅲ, Ⅳ, Ⅴ）-dipicolinate complexes across MDCK cell monolayer and comparison with Caco-2 cells

The permeation and eytotoxicity of three insulin-mimetic vanadium（Ⅲ, Ⅳ, Ⅴ）-dipicolinate complexes were studied using the MDCK cell monolayer in comparison with the Caeo-2 cells. On MDCK cell monolayer, the apparent permeation coefficients （Papp） were estimated to be （7.5±1.0）×10^-6, （1.0±0.2）×10^-6, （1.7±0.4）× 10^-6cm/s for V（Ⅴ）, V（Ⅵ）, and V（Ⅲ）-dipie complexes, respectively. The permeability of V（Ⅴ）-dipie complexes is much better than the others, which is in agreement with its better hypoglycemie effect in animal tests. On Caeo-2 cell monolayer, Papp were found to be in the range of 1-3×10^-6 ends and not to be affected by excessive amounts of dipieolinate ligand. By contrast, the permeability in the AP→BL direction across the MDCK monolayer increased greatly in the presence of free ligands, suggesting existence of active transport mechanism of vanadium complex anions on the MDCK cells. The eytotoxieity of the three complexes was found similar and the IC50 were measured in the range of 0.6-0.9 mmol/L for MDCK cells and 1.6--2 mmol/L for Caco-2 cells. The cytotoxicity of three vanadium complexes was conceivably in consistence with their permeability, suggesting that the toxicity, permeation and cellular metabolism of vanadium complexes are closely related.     

The Feasibility of ASP Flooding with Low Alkali Concentration in Heterogeneous Reservoirs

In this paper, as far as the ASP flooding in the pilot area of Daqing oilfield is concerned, the effect of apparent viscosity of ASP solution on enhanced oil recovery have been studied by comparison of experiments on artificial heterogeneous cores with on natural cores.The results showed that the decrease in the concentration of alkali could increase tile apparent viscosity of ASP solutions, namely, the apparent viscosity increased and the IFF couldn‘t get ultra low, and the oil recovery efficiency by ASP solution with a reasonably lower concentration of alkali was not less than that with a higher concentration of alkali and ultra low IFF. The experiments with models having impermeable interlayers between adjacent layers, and by means of “co-injection and separate production” showed that the ASP solution with a higher viscosity and a lower alkali concentration was more beneficial to starting-up medium and low permeability layers and enhancing total oil recovery. Therefore, to heterogeneous reservoir, as factors of affecting oil recovery, the apparent viscosity may be more important than the ultralow IFF. Furthermore, on the premise that the concentration of surfactant and the concentration of polymer keep constant, ASP flooding with low alkali concentration not only enlarges the selectable range of surfactants and decreases the cost of ASP flooding, but also solves the problem of alkali scales. Finally, a dynamic absorption experiment was performed with a 0.5 meter long linked-up natural core to determine alkali wastage in formation. The result being accord with the application case in the pilot confirmed that it was not necessary to add the alkali too much in ASP flooding from a new direction.     

Detection of immunoglobulin E using an aptamer based dot-blot assay

A novel aptamer based dot-blot assay for the detection of immunoglobulin E (IgE) was developed. A biotinylated aptamer was employed as the bio-recognition element to specifically interact with the target protein immobilized onto a nitrocellulose membrane substrate. Avidin conjugated horseradish peroxidase was introduced onto the membrane through the biotin-avidin system to catalyze the hydrogen peroxide mediated oxidation of 3,3’,5,5’-tetramethylbenzidine, thereby producing the blue-colored insoluble product. The intensity of the dots increased as the concentration of IgE increased. The spot intensity was quantified using a simple portable instrument. A linear response relationship between the spot intensity and the concentration of IgE over the range of 50 nmol/L to 1 μmol/L was obtained. The detection limit for IgE using the aptamer-based assay was 2.89 nmol/L. This assay was found to discriminate IgE from non-target proteins such as thrombin, bovine serum albumin and immunoglobulin G.     

Measurement of the free concentrations of alkyl phenols and bisphenol A to determine their biodegradation kinetics by activated sludge

A negligible depletion-solid phase microextraction (nd-SPME) method is developed to measure free concentrations of two alkyl phenols (4-t-octylphenol, nonylphenol) and bisphenol A in activated sludge, with which aerobic biodegradation kinetics of three pollutants was determined. For the degradation of octylphenol, nonylphenol and bisphenol A, the apparent rate constant based on total concentration is 0.02, 0.01 and 0.03 h-1, respectively; with the half life being 34.9, 54.3 and 22.3 h correspondingly. Meanwhile, real rate constants based on free concentration are 0.74, 0.60 and 1.03 h-1, respectively; with the half life being 0.9, 1.2 and 0.7 h accordingly. Differences between two constants revealed that desorption of targeted pollutants from activated sludge should be the rate-limiting step. Compared to the rapid biodegradation of alkyl phenols and bisphenol A in free state, the desorption of pollutants from the sludge is relatively slow, which made the apparent biodegradation rate constants much lower.     

Importance of NPA motifs in the expression and function of water channel aquaporin-1

The asparagine-proline-alanine sequences (NPA motifs) are highly conserved in aquaporin water channel family. Crystallographic studies of AQP1 structure demonstrated that the two NPA motifs are in the narrow central constriction of the channel, serving to bind water molecules for selective and effi-cient water passage. To investigate the importance of the two NPA motifs in the structure, function and biogenesis of aquaporin water channels, we generated AQP1 mutations with NPA1 deletion, NPA2 de-letion and NPA1,2 double deletion. The coding sequences of the three mutated cDNAs were subcloned into the mammalian expression vector pcDNA3.1 to form expression plasmids. We established stably transfected CHO cell lines expressing these AQP1 mutants. Immunofluorescence indicated that all the three mutated AQP1 proteins are expressed normally on the plasma membrane of stably transfected CHO cells, suggesting that deletion of NPA motifs does not influence the expression and intracellular processing of AQP1. Functional analysis demonstrated that NPA1 or NPA2 deletion reduced AQP1 water permeability by 49.6% and 46.7%, respectively, while NPA1,2 double deletion had little effect on AQP1 water permeability. These results provide evidence that NPA motifs are important for water per-meation but not essential for the expression, intracellular processing and the basic structure of AQP1 water channel.     

Effect of the free surface on the earthquake energy: A case study of reverse faulting

Based on the representation theorem of seismic energy radiation, in this study, we have quantitatively investigated the effect of free surface on the radiation energy distribution due to a coupling interaction between free surface and near surface finite fault for the reverse earthquake faulting. Corresponding to the finite faulting, a 2-D pseudostatic-reverse-fault-dislocation solution has been used in the calculation of the work done by the seismic response against free surface. The results indicate that, due to a strong coupling interaction between the free surface and near surface fault, the total radiated seismic energy ER is much larger than that radiated only from the fault itself （EF）, especially for the shallow reverse faulting. In convention, EF is commonly used in the estimation of earthquake energy radiation. However, when the fault depth H, the distance between the free surface and top of fault location, increases, the effect of the coupling interaction between the fault and free surface decreases gradually. Therefore, the total radiated energy ER approaches to the EF when the depth H is about 2 times the fault length L The current study could provide us a partial explanation of the apparent stress discrepancy observed at the far field and near field in the recent large earthquake. Moreover, the current study also has a significant implication of how to quantitatively describe the near fault strong ground motion and associated seismic hazard from the earthquake source energy point of view.     

A STUDY ON THE SELECTED FACTORS AFFECTING CONCRETE PERMEABILITY

The effect of w/c ratio,incorporation of silica fume and maximum aggregate sizes on permeabilityof concrete were studied.The test results both from chloride permeability and water absorptionshowed that the permeability of concrete increased as the w/c increases whilst a limitedamount of replacement of cement with silica fume the permeability decreased significantly.Themaximum aggregate size in the range of 8 to 25 mm seems also affect permeability but with a muchless significant level. A linear relationship between chloride permeability and capillary absorptionwas found.     

Recent progress in the study of intracellular toxicity of amyloid β peptide in Alzheimer's disease

Amyloid β (Aβ) deposition is one of the major pathological markers of Alzheimer's disease (AD). Extracellular Aβ toxicity has been studied for a long time in AD research field. However, controversial data show that extracellular Aβ load does not correlate with the dementia levels of AD patients and extracellular Aβ only induces significant cell death at non-physiological high concentrations. With the evolvement of Aβ hypothesis, considerable attention has been devoted to the study of intracellular Aβ toxicity recently. Intracellular Aβ induces dramatic cell loss in AD transgenic models and in human primary neurons (at pM concentrations) through p53, Bax and caspase-6 pathways. Here, we review the generation, toxicity and possible pathways of intracellular Aβ toxicity, and discuss the implication and current knowledge of intracellular Aβ in neuronal cell loss in neurodegenerative diseases.     

Recent progress in the study of intracellular toxicity of amyloid β peptide in Alzheimer''s disease

Amyloid β (Aβ) deposition is one of the major pathological markers of Alzheimer's disease (AD). Extracellular Aβ toxicity has been studied for a long time in AD research field. However, controversial data show that extracellular Aβ load does not correlate with the dementia levels of AD patients and extracellular Aβ only induces significant cell death at non-physiological high concentrations. With the evolvement of Aβ hypothesis, considerable attention has been devoted to the study of intracellular Aβ toxicity recently. Intracellular Aβ induces dramatic cell loss in AD transgenic models and in human primary neurons (at pM concentrations) through p53, Bax and caspase-6 pathways. Here, we review the generation, toxicity and possible pathways of intracellular Aβ toxicity, and discuss the implication and current knowledge of intracellular Aβ in neuronal cell loss in neurodegenerative diseases.     

Effect of Thrombin on the Apoptosis of Hippocampal Neurons in vitro

Hippocampal neurons were treated by thrombin and thrombin receptor activating peptides (TRAP). Cell survival rate was decreased in a dose-dependent manner by MTT assay. The numbers of apoptotic cell and apoptotic rate of hippocampal neurons treated by different concentrations of thrombin were increased in a dose-dependent manner by terminal deoxynucleotidyl transferase (TdT) mediated dUTP-biotin nick end-labeling (TUNEL) method and Flow Cytometry. When the concentration of thrombin is 40 U/mL, TUNEL positive cells and apoptotic rate of hippocampal neurons reached peak value, were 27. 3 4.0 and (29.333 4.633 ) %, respectively. Immunocytochemistry assay show that Bcl-2 protein expression was down- regulated and Bax protein expression was up-regulated with the concentration of thrombin increased. TRAP can mimic the effect of thrombin to induce apoptosis on hippocampal neurons. These data demonstrated that thrombin induced hippocampal neuron apoptosis in a dose-dependent manner through activating protease-activated protein-1 (PAR-1). The change in expression of Bcl-2 and Bax was related with the effect of high concentration thrombin induced apoptosis on hippocampal neurons.     

Characteristic parameters and operational curves of γ-Al2O3 nanofiltration membranes

The γ-Al2O3 nanofiltration membranes were manufactured by the sol-gel technique. The result of N2 adsorption and desorption test indicates that the characteristic parameters of the membranes: BJH desorption average pore diameter, BJH desorption cumulative volume of pores and BET surface area are about 3.9 nm, 0.33 cm^3/g and 245 m^3/g respectively, and the pore size distribution is very narrow.The operational curves of γ-Al2O3 nanofiltration membranes of the Ca^2 retention rate vs the trans-membrane pressure,feed concentration of solution treated and pH of solution treated were studied for the first time. It is found that the retention rate for Ca2~ increases with the transmembrane pressure increasing and decreases with the feed concentration of CaCl2 solution increasing. The retention of Ca^2 is very rmuch dependent on the pH of the solution. Minima Ca^2 retention rate is found at the isoelectric point (pH=7.5).     

The induction of Sinorhizobium meliloti C4-dicarboxylate transport system （Dct） is regulated by oxygen concentration

The Sinorhizobium meliloti C4-dicarboxylate transport （Dct） system is essential for symbiotic nitrogen fixation. The dctA gene, encoding the C4-dicarboxylate permease, is expressed in both free living and symbiotic cells. But in free living cells expression of dctD and dctB is absolutely required for the expression of dctA. In this study, in order to investigate the effect of oxygen concentration on the induction of Dct system, E. coli DH5a strain which carries the plasmid-encoded dctABD operon was used in tube assays. It was found that the specific induction of Dct system oc- curred only at a certain depth under the surface of M63-0.6% agar media, suggesting that Dct system could respond to oxygen concentration during succinate-induced expression. Furthermore, when measured at different oxygen concentrations, the highest expression level was observed at oxygen concentration of 2%. Thus, we predict that in addition to dicarboxylates, the induction of Dct system may also regulated by oxygen concentration.