Interferon‐γ suppresses Na+–H+ exchanger in cultured human endolymphatic sac epithelial cells

Adequate regulation of endolymphatic pH is essential for maintaining inner ear function. The Na⁺–H⁺ exchanger (NHE) is a major determinant of intracellular pH (pH_i), and facilitates Na⁺ and fluid absorption in various epithelia. We determined the functional and molecular expression of NHEs in cultured human endolymphatic sac (ES) epithelial cells and examined the effect of IFN‐γ on NHE function. Serial cultures of human ES epithelial cells were generated from tissue samples. The molecular expression of NHE1, ‐2, and ‐3 isoforms was determined by real‐time RT‐PCR. The functional activity of NHE isoforms was measured microfluorometrically using a pH‐sensitive fluorescent dye, 2′,7′‐bis(carbonylethyl)‐5(6)‐carboxyfluorescein (BCECF), and a NHE‐inhibitor, 3‐methylsulfonyl‐4‐piperidinobenzoyl guanidine methanesulfonate (HOE694). NHE1, ‐2, and ‐3 mRNAs were expressed in human ES epithelial cells. Functional activity of NHE1 and ‐2 was confirmed in the luminal membrane of ES epithelial cells by sequentially suppressing Na⁺‐dependent pH_i recovery from intracellular acidification using different concentrations of HOE694. Treatment with IFN‐γ (50 nM for 24 h) suppressed mRNA expression of NHE1 and ‐2. IFN‐γ also suppressed functional activity of both NHE1 and ‐2 in the luminal membrane of ES epithelial cells. This study shows that NHEs are expressed in cultured human ES epithelial cells and that treatment with IFN‐γ suppresses the expression and functional activity of NHE1 and ‐2. J. Cell. Biochem. 107: 965–972, 2009. © 2009 Wiley‐Liss, Inc.     

Orange light‐emitting Ca3Mg3(PO4)4:Sm3+ phosphors

This work reports the photoluminescence properties of Ca₃Mg₃(PO₄)₄:Sm³⁺ phosphors that were synthesized by the combustion method. The phase formation and morphology of the prepared phosphors were analysed by X‐ray diffraction studies and scanning electron microscopy. Ca₃Mg₃(PO₄)₄:Sm³⁺ phosphors give orange light emission when excited by near‐ultraviolet (NUV) and blue light. The photoluminescence characteristics of the as‐prepared phosphors were investigated and their emission spectra showed three peaks due to ⁴G_5/2 → ⁶H_5/2, ⁴G_5/2 → ⁶H_7/2 and ⁴G_5/2 → ⁶H_9/2 transitions. The mechanism responsible for the concentration quenching of luminescence was found to be an electric dipole–dipole interaction. The CIE chromaticity coordinates suggested that the prepared phosphors are potential candidates for orange light‐emitting diodes (LEDs).     

Glucose-6-phosphate dehydrogenase-dependent hydrogen peroxide production is involved in the regulation of plasma membrane H+-ATPase and Na+/H+ antiporter protein in salt-stressed callus from Carex moorcroftii

Glucose‐6‐phosphate dehydrogenase (G6PDH) is important for the activation of plant resistance to environmental stresses, and ion homeostasis is the physiological foundation for living cells. In this study, we investigated G6PDH roles in modulating ion homeostasis under salt stress in Carex moorcroftii callus. G6PDH activity increased to its maximum in 100 mM NaCl treatment and decreased with further increased NaCl concentrations. K⁺/Na⁺ ratio in 100 mM NaCl treatment did not exhibit significant difference compared with the control; however, in 300 mM NaCl treatment, it decreased. Low‐concentration NaCl (100 mM) stimulated plasma membrane (PM) H⁺‐ATPase and NADPH oxidase activities as well as Na⁺/H⁺ antiporter protein expression, whereas high‐concentration NaCl (300 mM) decreased their activity and expression. When G6PDH activity and expression were reduced by glycerol treatments, PM H⁺‐ATPase and NADPH oxidase activities, Na⁺/H⁺ antiporter protein level and K⁺/Na⁺ ratio dramatically decreased. Simultaneously, NaCl‐induced hydrogen peroxide (H₂O₂) accumulation was abolished. Exogenous application of H₂O₂ increased G6PDH, PM H⁺‐ATPase and NADPH oxidase activities, Na⁺/H⁺ antiporter protein expression and K⁺/Na⁺ ratio in the control and glycerol treatments. Diphenylene iodonium (DPI), the NADPH oxidase inhibitor, which counteracted NaCl‐induced H₂O₂ accumulation, decreased G6PDH, PM H⁺‐ATPase and NADPH oxidase activities, Na⁺/H⁺ antiporter protein level and K⁺/Na⁺ ratio. Western blot result showed that G6PDH expression was stimulated by NaCl and H₂O₂, and blocked by DPI. Taken together, G6PDH is involved in H₂O₂ accumulation under salt stress. H₂O₂, as a signal, upregulated PM H⁺‐ATPase activity and Na⁺/H⁺ antiporter protein level, which subsequently resulted in the enhanced K⁺/Na⁺ ratio. G6PDH played a central role in the process.     

Chemistry of the binary mixture of 1,10-phenanthroline and zinc perchlorate hexahydrate in methanol and gas phase. Monohydroxo species and tricoordination of zinc and water clusters

ESIMS reveals that methanol solutions of 1:1, 1:2 and 1:3 mixtures of Zn(ClO₄)₂ · 6H₂O and 1,10-phenanthroline (phen) generate [Zn(phen)(OH)]⁺, [Zn(phen)(H₂O)₄(OH)]⁺, [Zn(phen)₂(H₂O)(OH)]⁺and [Zn(phen)₂(H₂O)₄(OH)]⁺ ions in the gas phase. DFT calculations at the B3LYP/LanL2DZ level show that zinc is planar tricoordinate in [Zn(phen)(OH)]⁺ and the cis configuration is more stable than the trans one for the hexacoordinate ion [Zn(phen)₂(H₂O)(OH)]⁺. DFT calculations also show that the [Zn(phen)(H₂O)₄(OH)]⁺ and [Zn(phen)₂(H₂O)₄(OH)]⁺ ions are actually [Zn(phen)(H₂O)(OH)]⁺ · 3H₂O and [Zn(phen)₂(H₂O)(OH)]⁺ · 3H₂O containing extended motifs of H-bonded water clusters. The aqua species corresponding to the monohydroxo ions are acidic. Their acid dissociations are modeled collectively by equilibrium (see below) where other ligands around Zn are not specified. An attempt is then made to estimate K_a

     

Semi-synthetic antibiotics: Titanium(IV), zirconium(IV) and mercury(II) complexes of 6-amino penicillinic acid

A number of organometallic derivatives involving 6-amino penicillinic acid (I), of the types η⁵-R)₂M- (Cl)L^?Et₃NH⁺ (II), (η⁵-R)₂M(Cl)L (III) and R′HgL [R = cyclopentadienyl (C₅H₅), indenyl (C₉H₇), R′ = phenyl (C₆H₅), p-acetoxyphenyl (p-CH₃COOC₆H₄), o-hydroxyphenyl (o-HOC₆H₄), p-hydroxyphenyl (p-HOC₆H₄); M = Ti(IV), Zr(IV); LH = 6-amino penicillinic acid] have been synthesized and characterized. Conductance measurements indicate that while the (η⁵-R)₂M(Cl)L^?Et₃NH⁺ complexes are 1:1 electrolytes, the remaining compounds are non-electrolytes. From IR and UV spectral studies it is concluded that the penicillin moiety is bidentate. PMR and CMR studies support the stoichiometry of the complexes. Fluorescence studies have been carried out for o- and p-HOC₆H₄HgL complexes and relevant photochemical parameters have been elucidated. X-ray diffraction studies have been made for the o-HOC₆H₄HgL complex. For the C₆H₅HgL, p-CH₃COOC₆H₄HgL and p-HOC₆H₄HgL complexes, thermal studies (TG and DTA) have been carried out and kinetic parameters for thermal degradation have been enumerated. In addition, the fragmentation pattern of these complexes has been analysed on the basis of mass spectra. The C₆H₅HgL and p-CH₃COOC₆H₄HgL complexes show positive bactericidal activities.     

Site‐dependent biological activity of valinomycin analogs bearing derivatizable hydroxyl sites

Valinomycin (VLM, 1) is a K⁺ ionophore cyclodepsipeptide capable of depolarizing mitochondria and inducing apoptosis to several mammalian cell types, including a number of tumor cell lines. With the aim of creating VLM‐based ligand‐targeted anticancer drugs that may selectively convey VLM to pathological cells, we have previously introduced derivatizable hydroxyl handles into the VLM structure, allowing to access a three‐entity library of monohydroxyl VLMs (HyVLMs) bearing the OH group at the isopropyl side chain of a d ‐Hyi, d ‐Val, or l ‐Val residue (analogs 2–4, respectively). Herein, the levels of bioactivity retained by the conjugable HyVLMs have been assessed on the basis of their ability to alter the functionality of isolated rat‐liver mitochondria. Experiments run with HyVLMs in the range 1–10 nM and in 20 or 125 mM KCl medium show that the hydroxyl group reduces the potency of HyVLMs relative to VLM to an extent that depends upon the molecular site involved in the hydroxylation. On the other hand, estimation of the stability constants of complexes (in methanol at 25 °C) of each analog with Na⁺, K⁺, and Cs⁺ reveals that HyVLMs nicely retain the VLM binding features, except for a moderate increase in the stability of Na⁺ complexes. These findings, along with pertinent structural considerations, suggest that the incorporation of OH into the VLM structure might actually have altered its K⁺ transporting ability across mitochondrial membranes. Besides facing new aspects of VLM structure–activity relationship, these studies set the basis for the rational design of ligand‐HyVLMs conjugates through derivatization of hanging OH group. Copyright © 2013 European Peptide Society and John Wiley & Sons, Ltd.     

Studies on the aerobic utilization of synthesis gas (syngas) by wild type and recombinant strains of Ralstonia eutropha H16

The biotechnical platform strain Ralstonia eutropha H16 was genetically engineered to express a cox subcluster of the carboxydotrophic Oligotropha carboxidovoransOM5, including (i) the structural genes coxM, ‐S and ‐L, coding for an aerobic carbon monoxide dehydrogenase (CODH) and (ii) the genes coxD, ‐E, ‐F and ‐G, essential for the maturation of CODH. The cox_Oc genes expressed under control of the CO₂‐inducible promoter P_L enabled R. eutropha to oxidize CO to CO₂ for the use as carbon source, as demonstrated by ¹³CO experiments, but the recombinant strains remained dependent on H₂ as external energy supply. Therefore, a synthetic metabolism, which could be described as ‘carboxyhydrogenotrophic’, was established in R. eutropha. With this extension of the bacterium's substrate range, growth in CO‐, H₂‐ and CO₂‐containing artificial synthesis gas atmosphere was enhanced, and poly(3‐hydroxybutyrate) synthesis was increased by more than 20%.     

Esr Spectra of Radicals of Single-Stranded and Double-Stranded DNA in Aqueous Solution. Implications for Oh-Induced Strand Breakage

In situ photolysis at 20^oC (argon plasma light source, $, $ 200 mm) of oxygen-free solutions containing 2mM H₂0₂ and heat-denatured, single-stranded (sS)DNA from calf-thymus resulted in the ESR spectra of the 6-hydroxy-5,6-dihydro-thymin-5-yl {1} and 5-methyleneuracil {3} radicals linked to the sugar-phosphate backbone. They were generated by reaction of OH radicals with DNA. By comparison of the decay characteristics of the ESR signals with rate constants from pulse-conductivity measurements [E. Bothe, G.A. Qureshi and D. Schulte-Frohlinde, Z. Naturforsch. 38c 1030, (1983)] the thymine-derived radicals {1} and {3} can be excluded as precursors of the fast, dominating component of strand breakage of ssDNA. In the absence of H₂0₂ from native, doubie-stranded (ds)DNA an ESR signal was obtained (singlet, g ～ 2.004, $1/2 ～ 0.8 mT) which was assigned to the deprotonated guanine radical cation, {G'(-H)} of a DNA subunit. It is assumed that by the UV irradiation the guanine radical cation, {G⁺}, is generated, either by monophotonic photoionisation or by electron transfer to pyrimidine bases. By rapid transfer of the bridging proton from {G⁺} to the hydrogen bonded cytosine {G'(-H)} is formed. When photolysis of dsDNA was carried out in the presence of H₂0₂, reaction of photolytically generated OH resulted in peroxyl radicals and purine radicals. The oxygen for formation of the peroxyl radicals is probably produced by reaction of {G' (-H)} with H₂0₂. Photolysis of N₂0-saturated solutions containing dsDNA or ssDNA provided another possibility of generation of OH radicals. Under those conditions the OH-induced radicals {1} and {3} were obtained not only from ssDNA but also from dsDNA.     

Understanding Li‐Ion Dynamics in Lithium Hydroxychloride (Li2OHCl) Solid State Electrolyte via Addressing the Role of Protons

Low‐melting‐point solid‐state electrolytes (SSE) are critically important for low‐cost manufacturing of all‐solid‐state batteries. Lithium hydroxychloride (Li₂OHCl) is a promising material within the SSE domain due to its low melting point (mp < 300 °C), cheap ingredients (Li, H, O, and Cl), and rapid synthesis. Another unique feature of this compound is the presence of Li vacancies and rotating hydroxyl groups which promote Li‐ion diffusion, yet the role of the protons in the ion transport remains poorly understood. To examine lithium and proton dynamics, a set of solid‐state NMR experiments are conducted, such as magic‐angle spinning ⁷Li NMR, static ⁷Li and ¹H NMR, and spin‐lattice T₁(⁷Li)/T₁(¹H) relaxation experiments. It is determined that only Li⁺ contributes to long‐range ion transport, while H⁺ dynamics is constrained to an incomplete isotropic rotation of the OH group. The results uncover detailed mechanistic understanding of the ion transport in Li₂OHCl. It is shown that two distinct phases of ionic motions appear at low and elevated temperatures, and that the rotation of the OH group controls Li⁺ and H⁺ dynamics in both phases. The model based on the NMR experiments is fully consistent with crystallographic information, ionic conductivity measurements, and Born–Oppenheimer molecular dynamic simulations.     

Expression of OCTN2 and OCTN3 in the apical membrane of rat renal cortex and medulla

Immunological assays and transport measurements in apical membrane vesicles revealed that the apical membrane of rat kidney cortex and medulla presents OCTN2 and OCTN3 proteins and transports L ‐[³H]‐carnitine in a Na⁺‐dependent and ‐independent manner. OCTN2 mediates the Na⁺/L ‐carnitine transport activity measured in medulla because (i) the transport showed the same characteristics as the cortical Na⁺/L ‐carnitine transporter and (ii) the medulla expressed OCTN2 mRNA and protein. The Na⁺‐independent L ‐carnitine transport activity appears to be mediated by both OCTN2 and OCTN3 since: (i) Na⁺‐independent L ‐carnitine uptake was inhibited by both, anti‐OCTN2 and anti‐OCTN3 antibodies, (ii) kinetics studies revealed the involvement of a high‐ and a low‐affinity transport systems, and (iii) Western and immunohistochemistry studies revealed that OCTN3 protein is located at the apical membrane of the kidney epithelia. The Na⁺‐independent L ‐carnitine uptake exhibited trans‐stimulation by intravesicular L ‐carnitine or betaine. This trans‐stimulation was inhibited by anti‐OCTN3 antibody, but not by anti‐OCTN2 antibody, indicating that OCTN3 can function as an L ‐carnitine/organic compound exchanger. This is the first report showing a functional apical OCTN2 in the renal medulla and a functional apical OCTN3 in both renal cortex and medulla. J. Cell. Physiol. 223: 451–459, 2010. © 2010 Wiley‐Liss, Inc.     

Purification and characterization of phenoloxidase from the hemolymph of healthy and diseased Antheraea assamensis Helfer (Lepidoptera: Saturniidae): Effects of certain biological components and chemical agents on enzyme activity

In the current study, a dimeric phenoloxidase (PO) from the hemolymph of healthy and diseased (pebrine infected) larvae of Antheraea assamensis Helfer was extracted and purified. The protein was subjected to purification using Sephacryl S‐100 and CM Sepharose chromatography. The enzyme comprised of two subunits of ~76.8 and 76 kDa that showed PO activity in 6 mM l ‐3,4‐dihydroxyphenylalanine (L ‐DOPA) and 8 mM catechol but not in hydroquinone. Optimum temperature for PO activity was 30°C in l ‐DOPA and 37°C in catechol. Optimum pH ranged from 6.8 to 7.0 in L ‐DOPA and 7.0–7.2 in catechol. Specific activity of the purified PO from healthy larvae was 53.9 µM/min per mg of protein per ml in L ‐DOPA and 50.77 µM/min per mg of protein per ml in catechol. Specific activity of PO from diseased larvae was 30.0 µM/min per mg of protein per ml in L ‐DOPA and 28.55 µM/min per mg of protein per ml in catechol. Purification fold was 3.27–4.21 for healthy and 2.38–2.56 for diseased fractions. The enzyme showed the Michaelis constant (K_m) of 2.46–2.85 mM for healthy and diseased fractions in L ‐DOPA. In catechol K_m of 9.23–17.71 mM was observed. Peptidoglycan was the best activator of purified PO from both healthy and diseased fractions. Interactions between controls and activators appeared statistically significant (F = 767.5; df = 3; P < 0.0001). Na⁺, K⁺, and Cu²⁺ increased, whereas Ca²⁺, Zn²⁺, Mg²⁺, and Co²⁺ decreased PO activity. The overall interactions appeared highly significant (F = 217.0; df = 27; P < 0.0001). Kojic acid, dithiothreitol, thiourea, phenylthiourea, carbendazim, N‐bromosuccinimide, N,N,N′,N′‐tetraacetic acid, and diethyldithiocarbamate inhibited PO activity.     

A theoretical investigation of the interactions between hydroxyl-functionalized ionic liquid and water/methanol/dimethyl sulfoxide

Density functional calculations have been used to investigate the interactions of 1-(2-hydroxyethyl)-3-methylimidazolium ([C₂OHmim]⁺)-based ionic liquids (hydroxyl ILs) with water (H₂O), methanol (CH₃OH), and dimethyl sulfoxide (DMSO). It was found that the cosolvent molecules interact with the anion and cation of each ionic liquid through different atoms, i.e., H and O atoms, respectively. The interactions between the cosolvent molecules and 1-ethyl-3-methylimizolium ([C₂mim]⁺)-based ionic liquids (nonhydroxyl ILs) were also studied for comparison. In the cosolvent–[nonhydroxyl ILs] systems, a furcated H-bond was formed between the O atom of the cosolvent molecule and the C2-H and C6-H, while there were always H-bonds involving the OH group of the cation in the cosolvent–[hydroxyl ILs] systems. Introducing an OH group on the ethyl side of the imidazolium ring may change the order of solubility of the molecular liquids.     

Interactions of tertiary phosphines with p-benzoquinones; X-ray structures of [HO(CH2)3]3PC6H2(O)(OH)(MeO) and Ph3PC6H3(O)(OH)

Phosphonium zwitterions of a known type were obtained in high yield via a 1:1 reaction of p-benzoquinone or methoxy-p-benzoquinone with the tertiary phosphines R₃P [R = (CH₂)₃OH, Ph, Et, Me] and Ph₂MeP, in acetone or benzene at room temperature. In all cases, attack of the P-atom occurs at a C-atom rather than at an O-atom. The products were characterized to various degrees by elemental analysis, ³¹P{¹H}, ¹H and ¹³C NMR spectroscopies, and mass spectrometry, and two of the zwitterions, the new [HO(CH₂)₃]₃P⁺C₆H₂(O⁻)(OH)(MeO) and the known Ph₃P⁺C₆H₃(O⁻)(OH), were structurally characterized by X-ray analysis. The PEt₃ reaction also produces small amounts of the ‘dimeric’, μ-oxo co-product Et₃P⁺C₆H₂(O⁻)(OH)-O-C₆H₃(O⁻)P⁺Et₃ that is tentatively characterized by 1D- and 2D-NMR data. 2,5-Di-tert-butyl- and 2,3,5,6-tetramethyl-p-benzoquinone do not react with [HO(CH₂)₃]₃P under the conditions noted above. Heating D₂O solutions of the water-soluble zwitterions R₃P⁺C₆H₃(O⁻)(OH) [R = (CH₂)₃OH, Et] at 90 °C for 72 h leads to complete H/D exchange of the H-atom in the position ortho to the phosphonium center.     

A diphenyldiselenide derivative induces autophagy via JNK in HTB‐54 lung cancer cells

Symmetric aromatic diselenides are potential anticancer agents with strong cytotoxic activity. In this study, the in vitro anticancer activities of a novel series of diarylseleno derivatives from the diphenyldiselenide (DPDS) scaffold were evaluated. Most of the compounds exhibited high efficacy for inducing cytotoxicity against different human cancer cell lines. DPDS 2 , the compound with the lowest mean GI₅₀ value, induced both caspase‐dependent apoptosis and arrest at the G₀/G₁ phase in acute lymphoblastic leucemia CCRF‐CEM cells. Consistent with this, PARP cleavage; enhanced caspase‐2, ‐3, ‐8 and ‐9 activity; reduced CDK4 expression and increased levels of p53 were detected in these cells upon DPDS 2 treatment. Mutated p53 expressed in CCRF‐CEM cells retains its transactivating activity. Therefore, increased levels of p21^CIP1 and BAX proteins were also detected. On the other hand, DPDS 6 , the compound with the highest selectivity index for cancer cells, resulted in G₂/M cell cycle arrest and caspase‐independent cell death in p53 deficient HTB‐54 lung cancer cells. Autophagy inhibitors 3‐methyladenine, wortmannin and chloroquine inhibited DPDS 6 ‐induced cell death. Consistent with autophagy, increased LC3‐II and decreased SQSTM1/p62 levels were detected in HTB‐54 cells in response to DPDS 6 . Induction of JNK phosphorylation and a reduction in phospho‐p38 MAPK were also detected. Moreover, the JNK inhibitor SP600125‐protected HTB‐54 cells from DPDS 6 ‐induced cell death indicating that JNK activation is involved in DPDS 6 ‐induced autophagy. These results highlight the anticancer effects of these derivatives and warrant future studies examining their clinical potential.     

Novel Electrochemical Method for the Characterization of the Degree of Chirality in Chiral Polyaniline

A novel method to indicate the degree of chirality in polyaniline (PANI) was developed. The ( d ‐camphorsulfonic acid)‐ and (HCl)‐PANI‐based electrodes exhibited significantly different electrochemical performances in d ‐ and l ‐Alanine (Ala) aqueous solution, respectively, which can be used for the characterization the optical activity of chiral PANI. Cyclic voltammogram, tafel, and open circuit potential of PANI‐based electrodes were measured within d ‐ and l ‐Ala electrolyte solution, respectively. The open circuit potentials under different reacting conditions were analyzed by Doblhofer model formula, in which [C⁺]_poly1/[C⁺]_poly2 was used as a parameter to characterize the degree of chirality in chiral PANI. The results showed that [C⁺]_poly1/[C⁺]_poly2 can be increased with increasing concentrations of (1S)‐(+)‐ and (1R)‐(?)‐10‐camphorsulfonic acid. In addition, we detected that appropriate response time and lower temperature are necessary to improve the degree of chirality. Chirality 25:39‐42, 2013.© 2012 Wiley Periodicals, Inc.     

Effects of 1,25(OH)2D3 and vitamin D receptor on peripheral CD4+/CD8+ double‐positive T lymphocytes in a mouse model of systemic lupus erythematosus

This study aims to explore effects of 1,25(OH)₂D₃ and vitamin D receptor (VDR) on peripheral CD4⁺/CD8⁺ double‐positive (DP) T lymphocytes in systemic lupus erythematosus (SLE). MRL‐LPr/LPr mice with SLE (n = 20) and normal MRL mice (n = 20) were assigned into the control group (normal mice, without feeding with 1,25(OH)₂D₃), the 1,25(OH)₂D₃ group (SLE mice, feeding with 1,25(OH)₂D₃), the VDR‐knock‐in + 1,25(OH)₂D₃ group (SLE mice, VDR‐knock‐in, feeding with 1,25(OH)₂D₃) and the VDR‐knockout group (normal mice, VDR‐knockout, without feeding with 1,25(OH)₂D₃) (n = 10 per group). Levels of T lymphocytes were measured by flow cytometry. The mRNA and proteins expressions of inflammatory factors were measured by qRT‐PCR and ELISA. Extracellular signal‐regulated kinase‐1/2 (ERK1/2) expression was measured by Western blotting. Compared with normal mice, SLE mice showed reduced levels of CD4⁺, CD4⁺/CD8⁺ ratio, and DP lymphocytes. The levels of SLE‐related indicators all increased significantly, followed with severe skin ulcers and urinary system infection. With the increase in time, skin ulcers and urinary system infection were significantly improved, levels of CD4⁺, CD4⁺/CD8⁺ ratio, and DP lymphocytes increased, and levels of SLE‐related indicators all decreased in the 1,25(OH)₂D₃ group. There were no significant changes in bioindicators in the control and the VDR‐knock‐in + 1,25(OH)₂D₃ groups. The symptoms of SLE gradually occurred in the VDR‐knockout group. This study demonstrates that VDR and 1,25(OH)₂D₃ could elevate CD4⁺/CD8⁺ DP T lymphocytes and reduce expressions of inflammatory factors, thus inhibiting the development and progression of SLE.     

Mechanistic differences in the uptake of salicylic acid glucose conjugates by vacuolar membrane‐enriched vesicles isolated from Arabidopsis thaliana

Salicylic acid (SA) is a plant hormone involved in a number of physiological responses including both local and systemic resistance of plants to pathogens. In Arabidopsis, SA is glucosylated to form either SA 2‐O‐β‐d ‐glucose (SAG) or SA glucose ester (SGE). In this study, we show that SAG accumulates in the vacuole of Arabidopsis, while the majority of SGE was located outside the vacuole. The uptake of SAG by vacuolar membrane‐enriched vesicles isolated from Arabidopsis was stimulated by the addition of MgATP and was inhibited by both vanadate (ABC transporter inhibitor) and bafilomycin A₁ (vacuolar H⁺‐ATPase inhibitor), suggesting that SAG uptake involves both an ABC transporter and H⁺‐antiporter. Despite its absence in the vacuole, we observed the MgATP‐dependent uptake of SGE by Arabidopsis vacuolar membrane‐enriched vesicles. SGE uptake was not inhibited by vanadate but was inhibited by bafilomycin A₁ and gramicidin D providing evidence that uptake was dependent on an H⁺‐antiporter. The uptake of both SAG and SGE was also inhibited by quercetin and verapamil (two known inhibitors of multidrug efflux pumps) and salicin and arbutin. MgATP‐dependent SAG and SGE uptake exhibited Michaelis–Menten‐type saturation kinetics. The vacuolar enriched‐membrane vesicles had a 46‐fold greater affinity and a 10‐fold greater transport activity with SGE than with SAG. We propose that in Arabidopsis, SAG is transported into the vacuole to serve as a long‐term storage form of SA while SGE, although also transported into the vacuole, is easily hydrolyzed to release the active hormone which can then be remobilized to other cellular locations.     

Activation of Na+ /H+ exchange on rat preadipocyte plasma membrane and its role in cell proliferation and differentiation

Anin vitro cultured rat perirenal preadipocyte (PA) was established as a model system to investigate the role of the intracellular pH (pHi) and of the Na⁺ /H⁺ exchanger during PA proliferation and differentiation. pH sensitive probe, 2′,7′-bis-(2-carboxyethyl)-5(6)-carboxyfluorescein(BCECF), was employed to measure the pHi of PA and to determine the Na⁺/H⁺ exchange activity. The results showed that there was Na⁺/H⁺ exchange activity in the plasma membrane of PA, FCS stimulated DNA synthesis measured by³H-TdR incorporation, and the activation of Na⁺ /H⁺ exchanger resulted in pHi increase (nearly 0.2 pH unit) within 2 min. Ethyl-isopropyl-amiloride (EIPA), a specific Na⁺/H⁺ exchange inhibitor, inhibited Na⁺/H⁺ exchange activity and DNA synthesis. In the absence of serum insulin did not stimulate DNA synthesis but did induce PA differentiation characterized by the appearance of adiposome in the cell and the enhancement of glyeerol-3-phosphate dehydrogenase (G₃PDHase) activity. Meantime, insulin was also found to stimulate Na⁺/H⁺ exchange activity and pHi increase. EIPA inhibited Na⁺/H⁺ exchanger activation induced by insulin and also partially inhibited the enhancement of G₃PDHase activity. These results demonstrated that the activation of Na⁺ /H⁺ exchange and the resulting pHi increase are the early events related to both proliferation and differentiation of PA.     

Photoluminescence properties of a new orange–red‐emitting Sm3+–La3SbO7 phosphor

The antimonate compound La₃SbO₇ has high chemical stability, lattice stiffness and thermal stability. Orange–red‐emitting antimonate‐based phosphors La₃SbO₇:xSm³⁺ (x = 0.02, 0.05, 0.08, 0.10, 0.15, 0.20 and 0.25) were synthesized. The phase structure and photoluminescence properties of these phosphors were investigated. The emission spectrum obtained on excitation at 407 nm contained exclusively the characteristic emissions of Sm³⁺ at 568, 608, 654 and 716 nm, which correspond to the transitions from ⁴G_5/2 to ⁶H_5/2, ⁶H_7/2, ⁶H_9/2 and ⁶H_11/2 of Sm³⁺, respectively. The strongest emission was located at 608 nm due to the ⁴G_5/2→⁶H_7/2 transition of Sm³⁺, generating bright orange–red light. The critical quenching concentration of Sm³⁺ in La₃SbO₇:Sm³⁺ phosphor was determined as 10% and the energy transfer between Sm³⁺ was found to be through an exchange interaction. The International Commission on Illumination chromaticity coordinates of the La₃SbO₇:0.10Sm³⁺ phosphors are located in the orange–red region. The La₃SbO₇:Sm³⁺ phosphors may be potentially used as red phosphors for white light‐emitting diodes. Copyright © 2015 John Wiley & Sons, Ltd.     

Sulfur and nitrogen co‐doped graphene quantum dot‐assisted chemiluminescence for sensitive detection of tryptophan and mercury (II)

A simple one‐step thermal treatment to prepare strong fluorescent sulfur and nitrogen co‐doped graphene quantum dots (SN‐GQD) using citric acid and l ‐cysteine as precursors was developed. The ultra‐weak chemiluminescence (CL) from the reaction of hydrogen peroxide (H₂O₂) and periodate (IO₄^?) was significantly enhanced by SN‐GQD in acidic medium. The enhanced CL was induced by excited‐state SN‐GQD (SN‐GQD*), which was produced from the transfer energy of (O₂)₂* and ¹O₂ to SN‐GQD and recombination of oxidant‐injected holes and electrons in SN‐GQD. In the presence of tryptophan (Trp), the CL intensity of the SN‐GQD–H₂O₂–KIO₄ system was greatly diminished. This finding was used to design a novel method for determination of Trp in the linear range 0.6–20.0 μM, with a limit of detection (LOD) of 58.0 nM. Furthermore, Hg²⁺ was detectable in the range 0.1–9.0 μM with a LOD of 64.0 nM, based on its marked enhancement of the SN‐GQD–H₂O₂–KIO₄ CL system. The proposed method was successfully applied to detect Trp in milk and human plasma samples and Hg²⁺ in drinking water samples, with recoveries in the range 95.7–107.0%.