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 Ca^2 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-dependent. Soluble Aβ1-40 increases membrane permeability only at concentration of 3μmol/L, while the toxic effect of fibrillar Aβ1-40 is much stronger, its evident effect begins from 1μmol/L. When its concentration rose to 3μmol/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 Ca^2 , and the increased amplitudes are concentration-dependent. However, the fibrillar one induces the increase of intracellular Ca^2 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.     

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.     

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.     

Glucose-induced Ca2+ signals in rat pancreatic β cells

Using microfluorometry to assay intracellular Ca2+ , the influences of varied factors on glucose induced Ca22+ signals, such as glucose-induced initial decline phase (GIDP), Ca2+ oscillation, and Ca2+ release from internal stores, were investigated in single rat pancreatic β cells. Glucose was able to evoke GIDP even at non-stimulus concentration (5 mol/L), which is insufficient to induce Ca2+ spikes. GIDP was dependent on neither membrane depo larization nor extraeellular Ca2+ . However, GIDP was inhibited by thapsigargin, indicating a dependence on Ca2+ up take by Ca22+ stores. The glucose-induced calcium oscillation was inhibited when external Ca2+ was removed. However, thapsigargin could not block the Ca2+ oscillation. These results suggest that maintenance of Ca22+ oscillation requires ex tracellular Ca2+ but not Ca2+ stores. Glucose was able to evoke Ca2+ signals even in the absence of external Ca2+ . The glucose-induced Ca2+ release from intracellular Ca2+ stores was blocked by TTX. However, TTX had no effect on high K--induced Ca2+ store release, suggesting that membrane depolarization can directly release Ca2+ from some internal Ca2+ stores in β cells.     

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.     

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.     

Icariin suppresses bone resorption activity of rabbit osteoclasts in vitro

The effect of icariin on the bone resorption activity of rabbit osteoclasts is assessed in vitro. Osteoclasts were isolated from Japanese white rabbits and cultured on plates with a sterilized bone slice in each well. After treatment with icariin at various concentrations, the bone resorption activity of osteoclasts was evaluated by examining pit areas, superoxide anion (·O2-) generation, size and number of actin rings and intracellular calcium concentration [Ca2 ]i. As revealed by these data, icariin elicited continuous decline of [Ca2 ]i, making actin ring constricted and ·O2- generation decreased. These events resulted in smaller and fewer pits which indicate suppressed bone resorption activity of rabbit osteoclasts by icariin.     

Effect of Membrane Thickness on Permeability Coefficient of Microcap-sule

The effect of membrane thickness of mieroeapsule, measured by optical microseope duplex focusing, on its permeability coefflcient is discussed. The results reveal that the membrane thickness is of great imiportance on the permeabillty coefficient with changes of formaldehyde dosage, complex coacervation times, core dosage, agitation rate and emulsification time.     

Cinobufacini-induced HeLa cell apoptosis enhanced by curcumin

When used in combination with certain chemotherapies, curcumin has been shown to increase apoptosis in several cancer cell lines. Here, we report the combined effects of curcumin and cinobufacini on human cervical carcinoma cells. The aim of this study was to examine whether curcumin could enhance apoptosis induced by cinobufacini. 3-(4,5-Dimethylthiazol-2-y1)-2,5- diphenytetrazolium bromide (MTT) assays revealed that the growth and proliferation of HeLa cells could be inhibited by 75% after a combined treatment of 25 μg/mL cinobufacini and 8 μg/mL curcumin. The combined treatment is 3 times more effective than treatment with 25 μg/mL cinobufacini alone. Annexin V-FITC/PI staining, morphological changes and immunofluorescence verified a significant enhancement in cinobufacini-induced apoptosis when cells were also exposed to curcumin. The data showed that the proportion of early apoptotic cells significantly increased from 15.43% in cells treated only with 25 μg/mL cinobufacini to 49.2% in cells treated with 25 μg/mL cinobufacini and 8 μg/mL curcumin. Moreover, compared with treatment of only 25 μg/mL cinobufacini, ROS production increased 1.7-fold, the intracellular free Ca 2+ concentration increased 1.5-fold, and the mitochon- drial membrane potential decreased by 20% in the combined treatment. Simultaneously, the atomic force microscopy (AFM) re- sults suggest that cells treated with a combination of cinobufacini and curcumin varied significantly in shape and ultrastructure. Collapsed cells with leaking cytoplasm, blebbing pores and emerging apoptotic bodies were prevalent. The nanoparticle size increased from 70 nm when the cells were treated with 25 μg/mL cinobufacini to 190 nm when the cells were treated with 25 μg/mL cinobufacini and 8 μg/mL curcumin. The size increase resulted in the cell membrane becoming considerably rough. These results can improve our understanding of combination treatments. Specifically, the combination of cinobufacini and curcumin may potentially find use as a novel cervical carcinoma treatment. Additionally, AFM is a powerful tool that can be used to explore cellular morphologies and ultrastructures.     

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.     

Frequency dependence of magnetization and giant magneto impedance effect of amorphous wires

The frequency dependence of magnetization process and giant magneto impedance (GMI) effect of Co-rich melt-extracted amorphous wires was studied by Kerr effect and impedance analyzer, respectively. It is demonstrated that the transverse Kerr intensity and the corresponding GMI response increase with increasing frequency, which contributes to the upgraded skin effect. However, the skin depth has a slothful trend with frequency when it is up to the megahertz range, which gives rise to the transformation of magnetization. The process is much more sensitive to the direct current magnetic field and the sensitive change of the circular permeability, and GMI response is observed as its consequence. This proves that the evolution of circumferential magnetization and the corresponding permeability with the direct current magnetic field is the essence of GMI response, and a much more sensitive magnetization promises a better GMI response.     

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.     

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.     

Synthesis of Zeolites by Alkaline Activation of Fly Ash

In terms of mineral transformation, and chemical composition of acid-soluble component as a function of reaction time, the effect of alkaline solution on zeolite-like fly ash was studied by employing fly ash and NaOH solution as starting materials. When fly ash and 1-10 mol/L NaOH solution were processed at 100 ℃ for 24 h with 1:10 W/S ratio in a relatively closed system, powder XRD patterns of resulting products indicated the formation of various zeolites. Zeolite P crystallized early at low alkaline concentration, which was replaced then by zeolites X and A. At high concentration, hydroxy sodalite was the only new phase. Quartz, in fly ash and NaOH solution system, gradually dissolved, and mullite, however, remained stable. It was concluded that, with Al/Si and Na/Si finally reaching equilibrium in molar ratio, composition of starting mixtures affects the crystallization of zeolite from fly ash.     

Heterotrimeric G protein participated in modulation of cytoplasmic calcium concentration in pollen cells

Cytoplasmic free calcium concentration([Ca2+]c) in pollen cells of Lilium daviddi is measured with confocal laser scanning microscopy to investigate the effect of heterotrimeric G protein (G protein) on [Ca2+]c and the possible signal transduction pathway of G protein triggering cellular calcium signal. After application, cholera toxin (CTX), an agonist of G protein, triggers a transient increase of [Ca2+]c in pollen cells, and evokes a spatial-temporal characteristic calcium dynamics; while pertussis toxin (PTX), a G protein antagonist, leads to the decrease of [Ca2+]c. Both L-type Ca2+ channel blocker verapamil and inhibitor of IP3 receptor heparin inhibit CTX-induced [Ca2+]c increase. The results show that G protein may play a role in the modulation of [Ca2+]c through enhancing the extracellular Ca2+ influx and releasing of Ca2+ from intracellular stores.     

Effects of ytterbium on outward K<Superscript>+</Superscript> currents in NIH3T3 cell

Using the whole cell patch-clamp technique, we observed the outward K+ currents and studied for the first time the effects of Yb3+ on the currents and kinetics of activation and inactivation in non-excitable NIH3T3 cell. Our results show that the outward K+ currents were promoted with increasing concentration of Ca2+ in pipette solution and saturated at the concentration of 100 μmol/L Ca2+. Yb3+ in bath solution inhibited the currents in a concentration-dependent manner. At the concentration of 1 μmol/L, Yb...     

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.     

Effect of ytterbium on sodium current and its kinetics in rat hippocampal neurons

This study addressed the effects of Yb3＋ on voltage-gated sodium currents in rat hippocampal neurons using the whole-cell patch-clamp technique. Voltage-clamp recordings in single neurons were filtered and stored in a computer. Yb3＋ increased the amplitude of sodium currents in a concentration-dependent and voltage-dependent man- ner. The 50 % enhancement concentration of Yb3＋ on sodium currents was about 8.97 μmol/L, which was dif- ferent from the inhibitory effects of Yb3＋ on potassium current. The analysis on the activation and inactivation kinetics of Na＋ current showed that 100 μmol/L Yb3＋ did not change the process of activation and inactivation. In addition, the times reaching the peak of current （t） and inactivated time constant （τ） were voltage dependent. 100 μmol/L Yb3＋ significantly prolonged the time to peak at -70 and -80 mV. The effect disappeared at the positive direction of -70 mV. Furthermore, Yb3＋ decreased r val- ues to more positive values than -80 mV. In total, Yb3＋ did not change the process of activation, but impelled inacti- vated process. Yb3＋ mainly increased the Na＋ current through changing its conductance. It might be one of the mechanisms that Yb3＋ affected the hippocampal neurons.