Bifunctional Promoter Research Articles

Constructing an Artificial Interface as a Bifunctional Promoter for the Li Anode and the NCM Cathode in Lithium Metal Batteries.

The bottleneck of Li metal batteries toward practical applications lies at inferior cyclability as well as Li dendrite issues. As a promising solution, an interface engineering strategy is proposed herein for the Li anode through constructing a hybrid artificial interface. It is assembled onto the Li anode using photocontrolled free radical polymerization (photo-CRP) of polyethylene glycol diacrylate-hexafluorobutyl methacrylate and hexafluorobutyl methacrylate-trifluoroethyl carbonate (PEGDA-HFMBA@HFMBA-FEMC or PH@HF layer). Among such hybrid interfaces, the interior layer of PEGDA-HFMBA exists as a protective shield with flexibility and fracture resistance, while the exterior layer of HFMBA-FEMC plays a role as a LiF reservoir to promote Li mass transfer and its even electrodeposition. In the meantime, some excess HFMBA and FEMC monomers further dissolve into the electrolyte as molecular additives, followed by in situ generation of a thin and robust LiF-rich cathode electrolyte interface (CEI). With the resulting Li anode, Li/NCM811 full cells showcase multifold cyclability amplification in comparison to cells using Bare-Li, covering durable cyclability with a capacity retention of 81.8% after 400 cycles. When the cutoff voltage is elevated to 4.5 V or the working temperature is elevated to 45 °C, the cells still maintain a stable operation for extending 300 cycles.

Unraveling promoter effect in enhancing Rh-catalyzed hydroformylation of formaldehyde

Hydroformylation is one of the most important homogenous reactions, especially formaldehyde hydroformylation is of critical importance for synthesis of ethylene glycol. Due to lack of mechanistic analysis on promoters, we have employed a combined experimental and theoretical approach to reveal the promoter effect in the Rh-catalyzed hydroformylation of formaldehyde. The base promoter was suggested to be a bifunctional promoter. The mechanism of Cl-promoted formaldehyde hydroformylation was established, which significantly different from Cl-free system, and HCHO insertion was illustrated to be the rate-determining step. The distortion/interaction-activation strain analysis and mechanism calculations of halogen types demonstrate that Cl-containing catalyst has superior promotion effects compare to F, Br, and I-containing systems. The investigation on the effect of Cl coordination number shows that one-coordinated catalyst exhibits lower energy barriers and higher reactivity.

Nucleophilic Radiofluorination Using Tri-tert-Butanol Ammonium as a Bifunctional Organocatalyst: Mechanism and Energetics.

We present a quantum chemical analysis of the 18F-fluorination of 1,3-ditosylpropane, promoted by a quaternary ammonium salt (tri-(tert-butanol)-methylammonium iodide (TBMA-I) with moderate to good radiochemical yields (RCYs), experimentally observed by Shinde et al. We obtained the mechanism of the SN2 process, focusing on the role of the –OH functional groups facilitating the reactions. We found that the counter-cation TBMA+ acts as a bifunctional promoter: the –OH groups function as a bidentate ‘anchor’ bridging the nucleophile [18F]F− and the –OTs leaving group or the third –OH. These electrostatic interactions cooperate for the formation of the transition states of a very compact configuration for facile SN2 18F-fluorination.

Michael-Addition-Initiated Chemoselective Three-Component Reaction for the Synthesis of 2-(3-Oxo-1,3-diarylpropyl)malononitrile Derivatives Using Cerium(IV) Ammonium Nitrate in Phosphorus Ionic Liquid

The aim of this research is the study of hexyltriphenylphosphonium bromide (HTPB) ionic liquid as a bifunctional promoter (solvent and catalyst) in the presence of cerium(IV) ammonium nitrate as a single electron oxidant for the Michael reaction at room temperature and solvent-free conditions. A novel, one-pot and environmentally benign the process to achieve the chemoselective of 2-(3-oxo-1,3-diarylpropyl)malononitrile derivatives instead of 2-(1,3-diarylallylidene)malononitriles using a three-component reaction involving acetophenone derivatives, various aromatic aldehydes and malononitrile was developed. This protocol is simple and provides biologically interesting products in good yields, short reaction times at ambient conditions without the need for using volatile solvents. The remarkable feature of this method is the chemoselectivity of its which makes this protocol effective and distinctive.

Establishment of a stable transfection method in Babesia microti and identification of a novel bidirectional promoter of Babesia microti

Babesia microti, an emerging human pathogen, is primarily transmitted through a bite of an infected tick and blood transfusions in human. Stable transfection technique has been reported in many protozoan parasites over the past few years. However, in vivo transient and stable transfection method has not been established for Babesia microti. Here, for the first time, we present a method of transient as well as stable transfection of the Babesia microti (B. microti) in the in vivo conditions. We have identified a novel promoter of B. microti. We also demonstrated that Plasmodium berghei DHFR promoter is recognized and functional in B. microti. We show that BM-CTQ41297 promoter control the expression of two genes, which are present on either side and thus represents a bi-functional promoter in B. microti. The predicted promoter activity values using Promoter 2.0 program is higher for BM- CTQ41297 promoter than strong promoters such as β-actin, ef-1β, and many other promoters. Furthermore, we discovered a non-essential locus for the genetic manipulation of the parasite, allowing us to stably integrate foreign genes; GFP, mCherry, into the B. microti. The transfection using an electroporation method and genetic manipulation of B. microti is now achievable and it is possible to obtain transfected viable parasites under in vivo growing conditions. The growth curve analysis of transfected and WT B. microti are similar indicating no defects in the transgenic parasites. This study will enable other researchers in understanding the B. microti biology, host modulation and diverse parasite developmental stages using reverse genetics and holds great potential to identify novel drug targets and vaccine development.

NiO as a Bifunctional Promoter for RuO2 toward Superior Overall Water Splitting.

Conventional development of nanomaterials for efficient electrocatalysis is largely based on performance-oriented trial-and-error/iterative approaches, while a rational design approach at the atomic/molecular level is yet to be found. Here, inspired by a fundamental understanding of the mechanism for both oxygen and hydrogen evolution half reactions (OER/HER), a unique strategy is presented to engineer RuO2 for superior alkaline water electrolysis through coupling with NiO as an efficient bifunctional promoter. Benefitting from desired potential-induced interfacial synergies, NiO-derived NiOOH improves the oxygen binding energy of RuO2 for enhanced OER, and NiO also promotes water dissociation for enhanced HER on RuO2 -derived Ru. The resulting hybrid material exhibits remarkable bifunctional activities, affording 2.6 times higher OER activity than that of RuO2 and an HER activity comparable to Pt/C. As a result, the simple system requires only 1.5 V to deliver 10 mA cm-2 for overall alkaline water splitting, outperforming the benchmark PtC/NF||IrO2 /NF couple with high mass loading. Comprehensive electrochemical investigation reveals the unique and critical role of NiO on the optimized RuO2 /NiO interface for synergistically enhanced activities, which may be extended to broader (electro)catalytic systems.

Active Platinum Nanoparticles as a Bifunctional Promoter for Lithium−Sulfur Batteries

AbstractAs a widely used electrocatalyst in fuel cells, metallic platinum has achieved great success in terms of kinetic promotion. Still plagued by a familiar problem, owing to the sluggish kinetics of irreversible Li2S2/Li2S species deposition, lithium−sulfur batteries with high theoretical energy are far from being realized. Inspired by the similarity between oxygen and sulfur, being in the same group, commercially available Pt/C was investigated here to explore the possible effect on the sulfur cathode. The electrochemical tests show that the electrode with 2 wt% Pt added delivers a 61 % enhancement in the discharging specific capacity compared to the control electrode after 200 cycles at 0.5 C, along with an average coulombic efficiency of 98.3 %. It is proven that these beneficial results can be attributed to the favorable capability of polysulfide immobilization and effective inhibition of self‐discharging behavior. The ex situ XPS analysis and the increased exchange current density further reveal that the involved platinum nanoparticles could not only function as a static chemical absorber for polysulfide species, but, more importantly, to promote the redox kinetic effectively as an electrochemical catalyst. The positive effect of utilizing commercial Pt/C materials directly on sulfur cathodes is of great significance in developing sulfur cathodes for Li/S batteries.

Characterization of promoter activity of the ethylene receptor gene OgERS1 from Oncidesa in transgenic Arabidopsis

Physiological changes associated with senescence of flowers and abscission of floral parts in Oncidesa (formerly Oncidium) cv. Gower Ramsey are caused by a plant hormone ethylene which is produced by pollinia cap dislodgment during postharvest handling and transportation. The ethylene receptor gene OgERS1 of Oncidesa has been previously cloned and characterized. To analyze promoter activity of OgERS1, transgenic Arabidopsis thaliana plants were generated to express the s-glucuronidase (GUS) reporter gene under the control of 5’-upstream sequence of OgERS1 from Oncidesa. The expression pattern of the OgERS1 promoter at the cellular level was investigated by analysis of GUS activity. This promoter can activate gene expression in both actively dividing young tissues and abscission-related aging tissues. Expression of GUS was detected in the shoot meristem uniquely in 10 to 30 d-old-plants and was found in flower buds, axillary buds, flower stems, and abscission layers during later development. In 2- to 3-week-old transgenic Arabidopsis, exogenous ethylene, glucose, lactose, and maltose enhanced promoter activity implying that crosstalk between sugar and an ethylene receptor may exist. However, indole-3-acetic acid, benzylaminopurine, abscisic acid, heat, wounding, salinity, drought, and flooding slightly suppressed promoter activity. These results demonstrate that the promoter of OgERS1 was developmentally and environmentally regulated, and imply a potential for application of this bi-functional promoter to increase branching or enhanced dwarfing.

Mechanistic Study of the Palladium–Phenanthroline Catalyzed Carbonylation of Nitroarenes and Amines: Palladium–Carbonyl Intermediates and Bifunctional Effects

Palladium-phenanthroline complexes catalyze both the nitroarene carbonylation reaction and the amine oxidative carbonylation reaction to give, depending on the conditions, carbamates and ureas. There is evidence that the key step in both processes is the amine carbonylation. Here, we show that when the reaction is run in methanol key intermediate compounds have the general formula [Pd(RPhen)(COOMe)(2)] (1) (RPhen = 1,10-phenanthroline or one of its substituted derivatives). The kinetics of the reaction of 1 with toluidine in the presence of a carboxylic or phosphorus acid is first-order with respect to complex, acid, and toluidine. A CO atmosphere is also required for the reaction to proceed. Acid dimerization was shown not to be influential under the concentration conditions examined, but reaction between the acid and toluidine is not negligible and a correction has to be applied. Diphenylphosphinic acid is more effective than any carboxylic acid in promoting this reaction, as also observed under catalytic conditions. A series of equilibria and an irreversible acid-assisted proton transfer explain the observed data. Formation of an adduct between complexes of the kind 1 and CO was spectroscopically observed when RPhen = 2,9-Me(2)Phen. Several analogous complexes were also spectroscopically characterized and the X-ray structure of [Pd(2,9-Me(2)Phen)Cl(2)(CO)] was solved. This shows an asymmetric coordination of the nitrogen ligand. Kinetic measurements were also conducted under catalytic conditions. An Eyring plot shows that the effect of the acidic promoter is to decrease the DeltaS(double dagger) value, whereas no positive effect is observed on DeltaH(double dagger). A temperature-dependent correction for the reaction between the acid and aniline and phenanthroline present under the reaction conditions has to be applied. Comparison of the results obtained under stoichiometric and catalytic conditions strongly supports the view that 1 is involved even in the latter and that the acid is acting as a bifunctional promoter.

The Role of Type IV Collagen and Basement Membranes in Cancer Progression and Metastasis

The basement membrane acts as a barrier separating the epithelium from the surrounding stroma. While the structural role of the basement membrane in defining tissue architecture and compartmentalization has been appreciated for decades, its dynamic role in the regulation of cell behavior and recruitment of signaling pathways is only now emerging.1 The basement membrane further defines the tumor microenvironment and provides significant host-derived regulatory signals during progression of tumor growth and metastasis. The major component of the basement membrane is known to be type IV collagen.1 Nevertheless, a precise role for type IV collagen in its assembled suprastructural form versus its unassembled/degraded form is primarily unknown. In this issue of The American Journal of Pathology, Ikeda and colleagues2 demonstrate the effect of promoter hypermethylation on the expression levels of COL4A5/COL4A6 collagen genes. They suggest that such hypermethylation leads to diminished levels of α5(IV) and α6(IV) collagen in colorectal cancer tissue and colon cancer cells. The normal production and assembly of the basement membrane is disrupted in this malignant process. This data is consistent with evidence emerging from recent studies examining different cancer tissues, including prostate cancer, using antibodies generated in the Sado Ninomiya laboratory.3–8 However, a lack of free access to these antibodies to all investigators has limited independent evaluation. In vitro studies demonstrate that the expression of α5(IV) and α6(IV) collagen is down-regulated in association with hypermethylation of the bidirectional promoter of COL4A5/COL4A6 genes in colon cancer cell lines.2 These results suggest that normal assembly and organization of the basement membrane is disrupted in cancer progression.2 In an earlier study, Tanaka and colleagues9 analyzed the expression of the α1(IV) to α6(IV) collagen chain assembly by immunohistochemical analysis in colorectal cancer tissue, revealing normal expression of α1(IV)/α2(IV) and α5(IV)/α6(IV) chains but lost expression of α5(IV)/α6(IV) chains upon cancer invasion. These authors suggested that loss of α5(IV)/α6(IV) expression in the invasive cancer may be attributable to remodeling of basement membrane assembly, which in turn may be regulated by gene expression. The present article by Ikeda and colleagues2 now further demonstrates that such loss may be at the gene expression level.

Characterization and Use of Catabolite-Repressed Promoters from Gluconate Genes in Corynebacterium glutamicum

The genes involved in gluconate catabolism (gntP and gntK) in Corynebacterium glutamicum are scattered in the chromosome, and no regulatory genes are apparently associated with them, in contrast with the organization of the gnt operon in Escherichia coli and Bacillus subtilis. In C. glutamicum, gntP and gntK are essential genes when gluconate is the only carbon and energy source. Both genes contain upstream regulatory regions consisting of a typical promoter and a hypothetical cyclic AMP (cAMP) receptor protein (CRP) binding region but lack the expected consensus operator region for binding of the GntR repressor protein. Expression analysis by Northern blotting showed monocistronic transcripts for both genes. The expression of gntP and gntK is not induced by gluconate, and the gnt genes are subject to catabolite repression by sugars, such as glucose, fructose, and sucrose, as was detected by quantitative reverse transcription-PCR (qRT-PCR). Specific analysis of the DNA promoter sequences (PgntK and PgntP) was performed using bifunctional promoter probe vectors containing mel (involved in melanin production) or egfp2 (encoding a green fluorescent protein derivative) as the reporter gene. Using this approach, we obtained results parallel to those from qRT-PCR. An applied example of in vivo gene expression modulation of the divIVA gene in C. glutamicum is shown, corroborating the possible use of the gnt promoters to control gene expression. glxR (which encodes GlxR, the hypothetical CRP protein) was subcloned from the C. glutamicum chromosomal DNA and overexpressed in corynebacteria; we found that the level of gnt expression was slightly decreased compared to that of the control strains. The purified GlxR protein was used in gel shift mobility assays, and a specific interaction of GlxR with sequences present on PgntP and PgntK fragments was detected only in the presence of cAMP.

Bifunctional promoter of type IV collagen COL4A5 and COL4A6 genes regulates the expression of α5 and α6 chains in a distinct cell-specific fashion

Type IV collagen is present ubiquitously in basement membranes. A bifunctional promoter regulates the expression of the alpha1/alpha2 genes, and the alpha3/alpha4 and the alpha5/alpha6 genes are also considered to be regulated by putative bifunctional promoters. Unlike the other type IV collagen chains, the alpha5(IV) and alpha6(IV) chains do not always co-localize and are present in distinct basement membranes. To address such dichotomy in the alpha5(IV) and alpha6(IV) gene regulation, we cloned a mouse genomic DNA fragment containing the promoter region between the two transcription start sites of these genes and we then placed this putative promoter sequence between the chloramphenicol acetyltransferase and Luciferase reporter genes, so that these genes would be transcribed in opposite directions in this unique construct. Glomerular endothelial cells and mesangial cells generate the kidney glomerular basement membrane, which always contains the alpha5(IV) chain but not the alpha6(IV) chain. In contrast, the basement membranes of Bowman's capsule and distal tubuli (produced by the tubular epithelial cells) contain the alpha6(IV) chain. We demonstrate that, in response to TGF-beta (transforming growth factor beta), epidermal growth factor, vascular endothelial growth factor and platelet-derived growth factor, expression from the alpha5(IV) gene is significantly enhanced in the glomerular endothelial cells and mesangial cells, but not expression from the alpha6(IV) gene. In contrast, the expression from the alpha6(IV) gene, and not that from the alpha5(IV) gene, was significantly enhanced in response to growth factors in the tubular epithelial cells. Our results demonstrate that the proximal bifunctional promoter regulates the expression of the alpha5(IV) and alpha6(IV) genes in a cell-specific manner and offers the first demonstration of the promoter plasticity in growth factor regulation of type IV collagen genes in different tissues of the body.

Direct epoxidation of propylene with molecular oxygen over Ag–MoO 3/ZrO 2 catalyst

Direct gas-phase epoxidation of propylene to propylene oxide (PO) by molecular oxygen was studied over the MoO 3-modified supported-silver catalyst. The effect of promoter and support on the performance of the Ag–MoO 3 catalyst and their role were investigated by XPS, XRD, SEM, BET surface area, NH 3-TPD, CO 2-TPD techniques and so on. As the promoter, MoO 3 plays a great role of electron- and structure-type bi-functional promoter. Of all the supports studied, ZrO 2 is the most suitable one for the Ag–MoO 3 catalyst. The presence of support can regulate the size of the Ag–MoO 3 particles and the pore radii of catalyst, and decrease the loading of MoO 3 from ∼50 wt.% in the unsupported Ag–MoO 3 catalyst to ∼4 wt.% in the Ag–MoO 3/ZrO 2 catalyst. The properly weak Lewis acidic sites, the larger size of the Ag–MoO 3 particles and pore channel on the Ag–MoO 3/ZrO 2 catalyst are beneficial to improve epoxidation performance of the catalyst. The effect of the reaction temperature and space velocity on the catalytic epoxidation of propylene was also investigated. The low temperature or high space velocity can decrease the deep oxidation of propylene oxide to improve the selectivity to PO. Over the 20%Ag–4%MoO 3/ZrO 2 catalyst at 400 °C, 0.1 MPa and space velocity of 7500 h −1, the selectivity to PO of 60.3% was achieved with the O 2 conversion of 4.8%; under the space velocity of 12 000 h −1, the selectivity to PO was 71.5% with 2.5% O 2 conversion.

Specific binding of nuclear proteins to a bifunctional promoter element upstream of the H1/AC box of the testis-specific histone H1t gene.

The testis-specific histone H1t gene is transcribed exclusively in primary spermatocytes during spermatogenesis. Studies with transgenic mice show that 141 base pairs (bp) of the H1t proximal promoter accompanied with 800 bp of downstream sequence are sufficient for tissue-specific transcription. Nuclear proteins from testis and pachytene spermatocytes produce footprints spanning the region covering the repressor element (RE) from 100 to 125 nucleotides upstream of the H1t transcriptional initiation site. Only testis nuclear proteins bind to the 5'-end of the element and produce a unique, low-mobility complex in electrophoretic mobility shift assays. This testis complex is distinct from the complex formed by a repressor protein derived from several cell lines that binds to the 3'-end of the element. The testis complex band is formed when using nuclear proteins from primary spermatocytes, where the H1t gene is transcribed, and band intensity drops 70%-80% when using nuclear proteins from early spermatids, where H1t gene transcription ceases. Protein-DNA cross-linking experiments using testis nuclear proteins produce electrophoretic bands of 59, 52, and 50 kDa on SDS/PAGE gels.

Human liver-derived cells stably modified for regulated proinsulin secretion function as bioimplants in vivo.

Insulin deficiency is currently treated with pharmacological insulin secretagogues, insulin injections or islet transplants. Secondary failure of pharmacological agents is common; insulin injections often fail to achieve euglycemic control; and islet transplants are rare. Non-beta cells capable of regulated insulin secretion in vivo could be a functional cure for diabetes. Hepatocytes are good candidates, being naturally glucose-responsive, protein-secreting cells, while the liver is positioned to receive direct nutrient signals that regulate insulin production. Human liver-derived Chang cells were modified with a plasmid construct in which a bifunctional promoter comprising carbohydrate response elements and the human metallothionein IIA promoter controlled human proinsulin cDNA expression. Secretory responses of stable cell clones were characterized in vitro and in vivo by proinsulin radioimmunoassay. Transfected Chang cells secreted 5-8 pmol proinsulin/10(6) cells per 24 h in continuous passage for at least a year in response to 5-25 mM glucose and 10-90 microM zinc in vitro. Glucose and zinc synergistically increased proinsulin production by up to 30-fold. Non-glucose secretagogues were also active. Glucose transporter 2 (GLUT2) and glucokinase cDNA co-transfection enhanced glucose responsiveness. Intraperitoneally implanted Chang cells secreted proinsulin in scid and Balb/c mice. Serum proinsulin levels were further increased 1.3-fold (p<0.05) after glucose and 1.4- to 1.6-fold (p<0.005) after zinc administration in vivo. These results are the first to demonstrate stable proinsulin production in a human liver-derived cell line with activity in vitro and in vivo and provide a basis for engineering hepatocytes as in vivo bioimplants for future diabetes treatment.

Bifunctional promoter mediates suppressing effect of pathogen infection on UV-induced responses

Plants are continuously challenged by multiple stresses in the natural environment. The responses mounted by the plant to fight a given type of stress can influence its ability to cope with other environmental perturbations. An intriguing example of interacting responses is the cross-talk between signaling cascades activated by UV radiation and plant consumers (pathogens and herbivores). Recent work on a variety of scales, from molecular to ecological, has revealed unexpected interdependencies between these responses. Ecological studies have shown that UV can affect plant resistance to attack by insect herbivores and pathogens, with the direction of the effect varying, in an apparently idiosyncratic manner, from system to system (i.e. depending on the combination of plant and consumer species). At the molecular level, interactions between UV and pathogen- or wound-induced signaling elements have also been shown. However, the nuts and bolts of these signaling connections, and their ecological and evolutionary implications, are still unclear.

Stress‐activated expression of a Streptomyces pristinaespiralis multidrug resistance gene (ptr) in various Streptomyces spp. and Escherichia coli

A promoter which controls expression of the pristinamycin multidrug resistance gene (ptr) in Streptomyces pristinaspiralis could be induced by physiological stresses in both Streptomyces spp. and Escherichia coli. In S. pristinaspiralis, the ptr promoter (Pptr) was induced by pristinamycin I (PI) or pristinamycin II (PII). Streptomyces lividans was adopted as a convenient heterologous host for studies of Pptr regulation since it has no known pristinamycin biosynthetic genes. Two key regulatory features were documented in these studies: many (19 of 70) antibiotics and chemicals with no common targets or structural features induced the Pptr; induction with PI was most efficient during a transition phase when antibiotic biosynthetic genes are switched on. In Streptomyces coelicolor, Pptr activity was similarly inducible by PI and not dependent on sigma factors HrdA, HrdC, or HrdD. In E. coli, Pptr cloned in the bifunctional promoter probe vector pIJ2839 was functional and activated upon entry into stationary phase in the absence of exogenous inducer. Finally, gel-retardation studies demonstrated a Pptr-binding protein in S. lividans (where its activity was PI-inducible), S. coelicolor and S. pristinaespiralis. The fact that this activity was not detected in E. coli suggested the existence of another regulatory system perhaps also present in Streptomyces.