Optimising the acid-base ratio of Mg-Al layered double oxides to enhance CO2 capture performance: the critical role of calcination conditions.

The effect of calcination conditions (ramp rate, calcination temperature and time) on the formation of Mg2Al layered double oxides (Mg2Al LDOs) as well as their CO2 capture performance, has been systematically investigated. This study explores novel insights into the intricate relationship between these calcination conditions and the resulting surface characteristics, which play a vital role in CO2 capture efficiency. Notably, it is revealed that a rapid ramp rate (100 °C min-1) significantly increases surface area and hydroxyl concentration, leading to a 69% increase in CO2 capture efficiency compared to slower ramp rate. Conversely, short calcination times (1 h) and fast ramp rates (100 °C min-1) are observed to compromise CO2 adsorption due to the presence of dehydrated LDHs. A critical acid : base ratio of 0.37, achieved from a fast ramp rate (100 °C min-1) at 400 °C for 2 h, was found as a key threshold for optimising surface properties, effectively balancing favourable hydroxyl and less favourable strong acid sites, thereby maximizing CO2 capture performance.

Molecular, biochemical and enzymatic characterization of photorespiratory 2-phosphoglycolate phosphatase (PGLP1) in rice.

Phosphoglycolate phosphatase (PGLP, EC3.1.3.18) is a key enzyme in photorespiration. However, genes encoding the rice photorespiratory PGLP have not yet been identified or characterized. Here, PGLP for photorespiration in rice was identified and its enzymatic properties were investigated. In order to define the function of PGLP homologs, rice PGLP mutants were constructed using CRISPR/Cas9, the transcriptional expressions were analyzed by RT-qPCR, and subcellular localizations were detected via rice protoplast transient expression analysis. Based on sequence alignment, proteins encoded by genes OsPGLP1, OsPGLP2, and OsPGLP3 in the rice genome were predicted to have PGLP activity. Subsequent experimentation showed that OsPGLP1 and OsPGLP3 are chloroplast proteins, while OsPGLP2 is localized in the cytoplasm. In rice leaves, levels of PGLP1 transcript were substantially higher than those of PGLP2 and PGLP3, whereas in roots, levels of PGLP2 transcript were higher than those of PGLP1 and PGLP3. There was no detectable PGLP activity in leaves of the OsPGLP1 mutant, which was non-viable in ambient air condition (400 ppm CO2 ) and high CO2 (4000 ppm) was unable to restore normal growth. In contrast, mutations of PGLP2 or PGLP3 did not result in visible phenotypes and the leaf PGLP activities were also unaffected It is suggested that PGLP1, encoded by Os04g0490800, is responsible for photorespiration. Furthermore, PGLP1 is a dimer with an apparent molecular mass of ca.65 kDa, and its Km is 272 µM, with a higher broad optimum pH (7.5 to 10.0) for PGLP activity than that in other higher plants.

Correlations of acidity-basicity of solvent treated layered double hydroxides/oxides and their CO2 capture performance.

The basicity and acidity of solvent-treated layered double hydroxide (ST-LDHs) and their layered double oxides (ST-LDOs) have been fully studied using Hammett titration, in situ FTIR, CO2-TPD and NH3-TPD. Five solvents (ethanol, acetone, isopropanol, ethyl acetate and 1-hexanol) were selected to treat [Mg0.72Al0.28(OH)2](CO3)0.14 (Mg2.5Al-CO3 LDH) and compared with traditional LDH co-precipitated from water. The Brønsted basicity strength of the ST-LDHs and ST-LDOs increased but was accompanied by a decrease in basic site density. In addition, the Lewis acidity of ST-LDOs also changes significantly, with medium strength Lewis acid sites dissapearing after solvent treatment. We found that the CO2 capture capacity of solvent treated LDOs is 50% higher than that of traditional co-precipitated LDO sample. The ethanol treated LDO exhibited the highest CO2 uptake of 1.01 mmol g-1. The observed CO2 capture performance of the ST-LDOs correlates linearly with the ratio of total acid sites to total basic sites.

Functional expression of the Vibrio parahaemolyticus Na+/galactose (vSGLT) cotransporter in Xenopus laevis oocytes.

We have successfully expressed a bacterial cotransporter in a functional form in the Xenopus laevis oocyte expression system. The goals were to compare the kinetics and selectivity of the cotransporter expressed in oocytes with those obtained in bacteria and in proteoliposomes, and to determine if it is possible to measure the electrical properties of the bacterial cotransporter expressed in oocytes. The Vibrio parahaemolyticus Na+/galactose cotransporter (vSGLT) expressed in oocytes has functional properties that are similar to those expressed in bacteria and those of the purified cotransporter reconstituted into liposomes. vSGLT is a Na+-dependent transporter that is saturable with Na+ (K(0.5)=17 mM) and D-galactose (K(0.5)=237 microM) and is sensitive to both D-fucose and phlorizin. In addition, vSGLT in oocytes shows a sugar specificity in the order of D-galactose >D-fucose > D-glucose, distinguishing it from the animal members of the Na+/glucose cotransporter family. The level of transport by vSGLT in oocytes is lower overall (V(max) approximately 10 pmol/oocyte/hour) compared to other plant and animal cotransporters (V(max) approximately 1000 pmol/oocyte/hour). The low level of expression does not permit us to carry out electrophysiological studies of the bacterial cotransporter. This study shows the potential and unique advantages of utilizing a eukaryotic oocyte expression system to study bacterial cotransporters.

Inhibition of gap junction hemichannels by chloride channel blockers.

Electrophysiological methods were used to assess the effect of chloride-channel blockers on the macroscopic and microscopic currents of mouse connexin50 (Cx50) and rat connexin46 (Cx46) hemichannels expressed in Xenopus laevis oocytes. Oocytes were voltage-clamped at -50 mV and hemichannel currents (ICx50 or ICx46) were activated by lowering the extracellular Ca2+ concentration ([Ca2+]o) from 5 mM to 10 microM. Ion-replacement experiments suggested that ICx50 is carried primarily (>95%) by monovalent cations (PK : PNa : PCl = 1.0 : 0.74 : 0.05). ICx50 was inhibited by 18beta-glycyrrhetinic acid (apparent Ki, 2 microM), gadolinium (3 microM), flufenamic acid (3 microM), niflumic acid (11 microM), NPPB (15 microM), diphenyl-2-carboxylate (26 microM), and octanol (177 microM). With the exception of octanol, niflumic acid, and diphenyl-2-carboxylate, the above agents also inhibited ICx46. Anthracene-9-carboxylate, furosemide, DIDS, SITS, IAA-94, and tamoxifen had no inhibitory effect on either ICx50 or ICx46. The kinetics of ICx50 inhibition were not altered at widely different [Ca2+]o (10-500 microM), suggesting that drug-hemichannel interaction does not involve the Ca2+ binding site. In excised membrane patches, application of flufenamic acid or octanol to the extracellular surface of Cx50 hemichannels reduced single channel-open probability without altering the single-channel conductance, but application to the cytoplasmic surface had no effect on the channels. We conclude that some chloride-channel blockers inhibit lens-connexin hemichannels by acting on a site accessible only from the extracellular space, and that drug-hemichannel interaction involves a high-affinity site other than the Ca2+ binding site.

The structure and functions of human lysophosphatidic acid acyltransferases.

Lysophosphatidic acid (LPA) and phosphatidic acid (PA) are two phospholipids involved in signal transduction and in lipid biosynthesis in cells. LPA acyltransferase (LPAAT), also known as 1-acyl sn-glycerol-3-phosphate acyltransferase (1-AGPAT) (EC 2.3.1.51), catalyzes the conversion of LPA to PA. Two human isoforms of LPAAT, designated as LPAAT-alpha (AGPAT1) and LPAAT-beta (AGPAT2), have been extensively characterized. These two proteins contain extensive sequence similarities to microbial, plant and animal LPAAT sequences. LPAAT-alpha mRNA is uniformly expressed throughout most tissues with the highest level found in skeletal muscle; whereas LPAAT-beta is differentially expressed, with the highest level found in heart and liver, and negligible level in brain and placenta. The LPAAT-alpha gene is located on chromosome 6p21.3, an area within the class III region of the major hiscompatibility complex (MHC) and the LPAAT-beta gene is mapped to chromosome 9q34.3. Enhanced transcription of LPAAT-beta is suggested for neoplasm of the female genital tract. Additionally, ectopic LPAAT expression in certain cytokine-responsive cell lines can effect amplification of cellular signaling processes, such as those leading to enhancement of synthesis of tumor necrosis factor-alpha and interleukin-6 from cells following stimulation with interleukin-1beta; this suggests that the LPAAT genes represent candidates for affecting the development of certain cancers or inflammation-associated diseases.

Hiding bits in bell states.

We present a scheme for hiding bits in Bell states that is secure even when the sharers, Alice and Bob, are allowed to carry out local quantum operations and classical communication. We prove that the information that Alice and Bob can gain about a hidden bit is exponentially small in n, the number of qubits in each share, and can be made arbitrarily small for hiding multiple bits. We indicate an alternative efficient low-entanglement method for preparing the shared quantum states. We discuss how our scheme can be implemented using present-day quantum optics.

Variations in phytoremediation performance with diesel-contaminated soil.

The potential for phytoremediation of high concentrations of petroleum hydrocarbons is poorly understood. This study examines variations in phytoremediation performance for a soil contaminated with diesel at 6400 mg TPH kg-1 dry mixture. Experiments on diesel-contaminated soil were conducted in cups using 200 g of soil, and in columns using 4,000 g. Root development and TPH levels were measured in both experiments. In addition, CO2 soil gas concentrations were measured in the column experiments. The results show that ryegrass enhanced the loss of TPH over controls, and that this benefit only became evident after full root establishment. A comparison of the two experiments shows that rooting intensity (mg root kg-1 soil) is the key factor leading to higher TPH loss rates in the smaller containers. No clear difference in TPH loss occurred at 100 and 260 mm depths. Soil gas CO2 did not correlate well with TPH loss rates. The research concludes that an understanding of root development is crucial to evaluating the potential for ryegrass phytoremediation.

Effects of medication, behavioral, and combined treatments on parents' and children's attributions for the behavior of children with attention-deficit hyperactivity disorder.

Seventy-four mothers and 41 fathers and their 6 to 13 year old sons with attention-deficit hyperactivity disorder (ADHD) watched videos of child ADHD symptoms, compliance, and noncompliance. Participants were told either that the child was receiving medication, behavioral treatment, a combination of the two, or was not receiving treatment and were asked to rate the cause of the behavior. Parents attributed less control but greater stability to positive child behaviors when the child was receiving medication. However, for negative behaviors, medication increased attributions of control but diminished stability. With behavior management. compliance was seen as more external and stable and noncompliance as more controllable but less stable. For all treatments, boys reported increased control over ADHD symptoms and noncompliance. The implications of these treatment-related attributions for parenting and children's self-perceptions are discussed.

Losses of diesel oil by volatilisation and effects of diesel oil on seed germination and seedling growth.

For phytoremediation to be effective, seeds must germinate and subsequently grow, or seedlings must become established, in contaminated soil. In this study, the effect of diesel oil on the viability of seeds of white clover and ryegrass together with long term abiotic diesel oil loss were investigated. Losses of diesel by volatilisation were found to be as high as 58% over 360 days suggesting that volatilisation (abiotic loss) may be a significant method of contaminant removal that may have been previously underestimated or overlooked in short term studies. White clover and ryegrass seeds were able to germinate in the presence of volatile diesel components and following 24 weeks immersion in diesel oil, which suggested that properties of their seed coats prevented diesel oil causing damage to the seeds.

Urea transport by cotransporters.

The rabbit Na+-glucose cotransporter (rbSGLT1) was expressed in Xenopus laevis oocytes and urea transport in rbSGLT1 and non-injected (control) oocytes was studied using [14C]urea as a tracer. The level of rbSGLT1 expression in these batches of oocytes was monitored by measuring the uptake of alpha-methyl-D-[14C]glucopyranoside ([14C]alphaMDG). In rbSGLT1-expressing oocytes, there was a 4-fold increase in urea transport in the absence of sugar relative to that in control oocytes. Urea uptake was not Na+ dependent and was linear with both time of incubation (5-120 min) and increasing urea concentration (50 microM to 100 mM) in the bathing medium. rbSGLT1 urea uptake was blocked by the rbSGLT1-specific inhibitor phlorizin (Ki 1 microM) in 100 mM NaCl buffer, but was not affected in 100 mM choline chloride buffer. Phloretin inhibited rbSGLT1 urea uptake with a low affinity (Ki > 1 mM) in the presence and absence of Na+. The uptake of 55 m[mu]M urea through rbSGLT1 was not blocked by 100 mM urea analogues including thiourea, 1,3-dimethyl urea, 1,1-dimethyl urea and acetamide. The activation energies (Ea) of urea transport for control and rbSGLT1-expressing oocytes were 14+/-3 and 6+/-1 kcal mol(-1), respectively. The low Ea for urea transport through rbSGLT1 is comparable to the Ea of passive water transport through rbSGLT1. Urea transport through rbSGLT1 was further increased when the cotransporter was activated by the addition of sugar to the external medium. The rate of sugar-dependent urea uptake was directly proportional to the rate of Na+-glucose-H2O cotransport such that the amount of urea transport was approximately proportional to the molar concentration ratio of urea to H2O (55 microM/55 M). The low affinity Na+-glucose (pSGLT3), the Na+-iodide (rNIS) and the Na+-(Cl-)-GABA (hGAT1) cotransporters expressed in oocytes demonstrated similar urea transport properties. These observations suggest that cotransporters behave as urea channels in the absence of substrates. Furthermore, under substrate-transporting conditions, the same cotransporters serve as urea cotransporters. This could account for urea transport in cells that appear not to have urea uniporters or channels.

Purification and functional reconstitution of a truncated human Na(+)/glucose cotransporter (SGLT1) expressed in E. coli.

A truncated human Na(+)/glucose cotransporter (C(5), residues 407-664) was expressed and purified from Escherichia coli using a GST fusion vector and glutathione affinity chromatography. The truncated transporter (C(5)) was cleaved from GST-C(5) by Factor Xa proteolysis and purified by gel filtration chromatography. Up to 1 mg of purified GST-C(5) was obtained from 1 l bacterial culture. Reconstitution of both GST-C(5) and C(5) proteins into lipid vesicles resulted in 2.5-fold higher initial uptake rates of [(3)H]D-glucose into C(5)-proteoliposomes than into liposomes. Transport was stereospecific, saturable, and inhibited by phloretin. These properties are similar to those obtained for C(5) in Xenopus laevis oocytes, and provide additional evidence that the five C-terminal transmembrane helices in SGLT1 form the sugar translocation pathway.

Molecular cloning of two alternatively spliced forms of human phosphatidic acid phosphatase cDNAs that are differentially expressed in normal and tumor cells.

Phosphatidic acid (PA) and diacylglycerol (DG) are lipids involved in signal transduction and in structural membrane-lipid biosynthesis in cells. Phosphatidic acid phosphatase (PAP) catalyzes the conversion of PA to DG. This enzyme exists in at least two isoforms, one of which (PAP1) is presumed to be cytosolic and membrane associated and the other (PAP2) to be an integral membrane protein. Homology search of the GenBank database using a murine sequence probe enabled the cloning of several putative human isoenzymes. Two isoforms, presumed to be alternative splice variants from a single gene, designated as PAP2-alpha1 and PAP2-alpha2, have been cloned and expressed. The PAP2-alpha1 and PAP2-alpha2 have a 84% and a 72% overall match, respectively, with the published mouse PAP amino acid sequence. The area of alternative exon usage was confined to the coding region at amino acids 20 to 70. Ectopic expression of PAP2-alpha1 and PAP2-alpha2 cDNAs in ECV304 endothelial cells led to a 6- to 8-fold and a 2-fold increase in PAP activity, respectively, in cell-free extracts using an in vitro assay that measured the conversion of [14C]PA to [14C]DG. The increase in PAP activity in PAP2-alpha-transfected cells correlated with a >50% decrease in the steady-state PA level. Northern analysis showed that PAP2-alpha mRNA expression was suppressed in several tumor tissues, notably those derived from the lower alimentary tract. Subsequent analysis of colon tumor tissue derived from four donors confirmed lower expression of PAP2-alpha than in matching normal colon tissue. Considering these data and previous demonstrations that certain transformed cell lines have lower PAP activity, we suggest that human PAP cDNAs may be candidates for gene therapy for certain tumors.

Cloning and expression of two human lysophosphatidic acid acyltransferase cDNAs that enhance cytokine-induced signaling responses in cells.

Lysophosphatidic acid (LPA) and phosphatidic acid (PA) are two phospholipids involved in signal transduction and in lipid biosynthesis in cells. LPA acyltransferase (LPAAT), also known as 1-acyl sn-glycerol-3-phosphate acetyltransferase (EC 2.3.1.51), catalyzes the conversion of LPA to PA. In this study, we describe the isolation and characterization of two human cDNAs that encode proteins possessing LPAAT activities. These two proteins, designated here as LPAAT-alpha and LPAAT-beta, contain extensive sequence sequence similarities to microbial or plant LPAAT sequences. LPAAT-alpha mRNA was detected in all tissues with highest expression in skeletal muscle whereas LPAAT-beta was expressed predominantly in heart and liver tissues. Expression of these two cDNAs in an Escherichia coli strain with a mutated LPAAT gene (plsC) complements its growth defect and shifts the equilibrium of cellular lipid content from LPA to PA and other lipids. Overexpression of these two cDNAs in mammalian cells leads to increased LPAAT activity in cell-free extracts using an in vitro assay that measures the conversion of fluorescently labeled LPA to PA. This increase in LPAAT activity correlates with enhancement of transcription and synthesis of tumor necrosis factor-alpha and interleukin-6 from cells upon stimulation with interleukin-1beta, suggesting LPAAT overexpression may amplify cellular signaling responses from cytokines.

Isolation and expression of an isoform of human CDP-diacylglycerol synthase cDNA.

Phosphatidic acid (PA) is a phospholipid involved in signal transduction and in glycerolipid biosynthesis. CDP-diacylglycerol synthase (CDS) or CTP:phosphatidate cytidylyltransferase (EC 2.7.7.41) catalyzes the conversion of PA to CDP-diacylglycerol (CDP-DAG), an important precursor for the synthesis of phosphatidylinositol, phosphatidylglycerol, and cardiolipin. We describe in this study the isolation and characterization of a human cDNA clone that encodes amino acid sequences homologous to Escherichia coli, yeast, and Drosophila CDS sequences. Expression of this human cDNA under the control of a GAL1 promoter in a null cds1 mutant yeast strain complements its growth defect and produces CDS activity when induced with galactose. Transfection of this cDNA into mammalian cells leads to increased CDS activity in cell-free extracts using an in vitro assay that measures the conversion of [alpha-32P]CTP to [32P]CDP-DAG. This increase in CDS activity also leads to increased secretion of tumor necrosis factor-alpha and interleukin-6 from endothelial ECV304 cells upon stimulation with interleukin-1beta, suggesting that CDS overexpression may amplify cellular signaling responses from cytokines.

Endogenous natural killer enhancing factor-B increases cellular resistance to oxidative stresses.

Natural killer-enhancing factor (NKEF) was identified and cloned on the basis of its ability to increase NK cytotoxicity. Two genes, NKEF-A and -B, encode NKEF proteins and sequence analysis presented suggests that each belongs to a highly conserved family of antioxidants. To examine the antioxidant potential of NKEF, we transfected the coding region of NKEF-B cDNA into the human endothelial cell line ECV304. The stable transfectant, B/1, was found to overexpress NKEF-B gene transcript and protein. We subjected B/1 to oxidative stress by either culturing them with glucose oxidase (GO), which continuously generates hydrogen peroxide, or by direct addition of hydrogen peroxide. We found that B/1 cells were more resistant than control cell lines. Resistance to hydrogen peroxide was originally thought to be mediated mainly by catalase and the glutathione cycle. Therefore, we used inhibitors to block the two pathways and found that B/1 cells were more resistant to oxidative stress than control cells when we used inhibitors to preblock either pathway. We also examined the cellular inflammatory responses to oxidized low-density lipoprotein (LDL) and bacterial lipopolysaccharide (LPS) by measuring monocyte adhesion to endothelial cells in vitro and found that B/1 cells were resistant to such responses. Lastly, we found that B/1 cells were more resistant to a novel chemotherapeutic agent CT-2584, which appears to kill tumor cells by stimulating production of reactive oxygen intermediates in mitochondria. These results demonstrate that the NKEF-B is an antioxidant that protects cells from oxidative stress, chemotherapy agents, and inflammation-induced monocyte adhesion. Furthermore, its expression may mediate cellular responses to proinflammatory molecules.

Influence of BA and sucrose on the competence and determination of pepper (Capsicum annuum L. var. Sweet Banana) hypocotyl cultures during shoot formation.

The simultaneous presence of 6-benzyladenine (BA) and sucrose in a Murashige and Skoog medium (SIM) during the initial stages of shoot initiation have been found to be obligatory for high-frequency shoot formation in the Capsicum annuum L. var. 'Sweet Banana' upper hypocotyl explants. The explants are determined for shoot formation following a minimum of 8 days of culture on SIM. Deprivation of exogenous sucrose from day 6 to day 20 of culture had no effect on the shoot forming response of the explants. BA and sucrose appear to act independently on different aspects of the competence of explants to respond to SIM during shoot initiation.

CT-2576, an inhibitor of phospholipid signaling, suppresses constitutive and induced expression of human immunodeficiency virus.

Viruses such as human immunodeficiency virus (HIV) require cellular activation for expression. Cellular activation in lymphoid cells is associated with augmented accumulation of certain phosphatidic acid (PA) species derived from the hydrolysis of glycan phosphatidylinositol (GPI). This suggests that activation of a phospholipid pathway may play a role in initiation of viral replication. To test this hypothesis, we examined the effect of tat gene expression on the production of cellular PA species, as the Tat protein is essential for HIV expression and has been implicated in activating the expression of multiple host cellular genes. Expression of tat increased the expression of PA. We then tested whether synthetic inhibitors of PA metabolism would inhibit activation of the HIV long terminal repeat by Tat and tumor necrosis factor alpha (TNF-alpha). CT-2576 suppressed both PA generation induced by Tat and HIV long terminal repeat-directed gene expression in response to Tat or TNF-alpha at a posttranscriptional step. CT-2576 also inhibited constitutive as well as TNF-alpha- and interleukin 6-induced expression of HIV p24 antigen in chronically infected U1 cells and in peripheral blood lymphocytes acutely infected with a clinical isolate of HIV. Pharmacological inhibition of synthesis of selected PA species may therefore provide a therapeutic approach to suppression of HIV replication.

Epstein-Barr virus-derived vectors for transient and stable expression of recombinant proteins.

The transient expression level of foreign proteins in primate cells can be enhanced by incorporating the replication elements derived from Epstein-Barr virus (EBV). Specifically, we have constructed expression plasmids with the replication origin region (OriP) from EBV and an adenovirus-transformed cell line that expresses the EBV nuclear antigens-1 (EBNA-1). As EBV vectors can replicate as episomes in the nuclei, such vectors can have a stable transfection efficiency as high as 25% and provide a straightforward way of obtaining large amounts of recombinant proteins transiently or stably.