Triple Complex Research Articles

Structural basis for the synergetic neutralization of hepatitis E virus by antibody–antibody interaction

Neutralizing antibodies (nAbs) play a crucial role in virology, antibody drug development, and vaccine research. In this study, we investigated the synergistic effect of two hepatitis E virus (HEV) nAbs, 8H3, and 8C11, which have exhibited enhanced neutralizing activity in a rhesus monkey model. We presented crystal structures of 8H3 Fab alone and a triple complex of 8C11 Fab and 8H3 Fab simultaneously binding to the HEV E2s protein (8C11:E2s:8H3). Through structural analysis, we identified critical binding sites and fully elucidated the binding footprints of nAb 8H3 in the 8C11:E2s:8H3 complex using site-directed mutagenesis, pinpointing Ile 529, Glu 549, Lys 554, and Ser 566 in the E2s domain, and K66H, S67H, D88H in the 8C11 heavy chain. Interestingly, the synergetic enhancement of 8C11 to 8H3 converted to an antagonistic effect when 8C11 bound to E2s with pretreatment of 8H3, indicating a unidirectional synergistic effect associated with the sequence of antibody involvement. We demonstrated this phenomenon through structural comparisons of E2s:8C11 vs. 8C11:E2s:8H3 crystal structures and molecular dynamics simulations, found that Ile 529 played a key role in the synergistic interplay between these two nAbs. The two-antibody combination showed a more potent antibody-imposed physical disruption mechanism and enhanced coneutralization in an authentic HEV-based cell model. Our study suggests a strategy for synergistic antibody cocktail design with antibody–antibody side-by-side interaction.

Advances in plant pathogen detection: integrating recombinase polymerase amplification with CRISPR/Cas systems.

Plant pathogens are causing substantial economic losses and thus became a significant threat to global agriculture. Effective and timely detection methods are prerequisite for combating the damages caused by the plant pathogens. In the realm of plant pathogen detection, the isothermal amplification techniques, e.g., recombinase polymerase amplification (RPA) and loop-mediated isothermal amplification (LAMP), have emerged as a fast, precise, and most sensitive alternative to conventional PCR but they often comprise high rates of non-specific amplification and operational complexity. In recent advancements, clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated nuclease Cas systems, particularly Cas12, have emerged as powerful tools for highly sensitive, specific, and rapid pathogen detection. Exploiting the collateral activities of Cas12, which selectively cleaves single-stranded DNA (ssDNA), novel detection platforms have been developed. The mechanism employs the formation of a triple complex molecule comprising guide RNA, Cas12 enzyme, and the substrate target nucleotide sequence. Upon recognition of the target, Cas12 indiscriminately cleaves the DNA strand, leading to the release of fluorescence from the cleaved ssDNA reporter. Integration of isothermal amplification methods with CRISPR/Cas12 enables one-step detection assays, facilitating rapid pathogen identification within 30min at a single temperature. This integrated RPA-CRISPR/Cas12a approach eliminates the need for RNA extraction and cDNA conversion, allowing direct use of crude plant sap as a template. With an affordable fluorescence visualization system, this portable method achieves 100-fold greater sensitivity than conventional techniques. This review summarizes recent advances in RPA-CRISPR/Cas12a for detecting plant pathogens, covering primer design, field-level portability, and enhanced sensitivity.

STRUCTURAL ANALYSIS OF THE CAUSATIVE-HOLISTIC MODEL OF HEALTH

Within this study, the necessity of a causative-holistic approach to health, viewed through philosophical and spiritual contexts, is substantiated. The author focuses on the structural analysis of the health model, based on the conducted research on the understanding of the phenomenon of health by ancient philosophers, medieval thinkers, and contemporary scientists. The article acknowledges the importance of spirituality in health and defines it as a causal factor that sets parameters for the development of physical health. The research offers a detailed description of the structure of holistic health, which includes unity, duality, and trinity, forming the foundation on the tripartite model “spirit-soul-body,” which has shaped the understanding of holistic health since medieval times and remains unchanged. Based on the metaphysical system proposed by the German mystic Jacob Böhme, the author substantiates the septenary structure of the spirit, reflecting in the septenary manifestation of the human being, and should have its analogy in the physical body and social dimension of the human being. A decenary model of health is also proven, appearing in the connection of the septenary «earthly human» with the triple metasystem complex, reflecting the spiritual aspect of holistic human. This decenary model forms a new causative-holistic view on human health and can be the basis for changing the medical paradigm, which should be based on primary spiritual factors of health for the preservation and improvement of both physical health and the health of the metasystem complex human-collective-society-planet.

Fluorometric determination of mecA gene in MRSA with a graphene-oxide based bioassay using flap endonuclease 1-assisted target recycling and Klenow fragment-triggered signal amplification

Methicillin-resistant Staphylococcus aureus (MRSA) is a multidrug-resistant bacterium that causes a wide range of illnesses, necessitating the development of new technologies for its detection. Herein, we propose a graphene oxide (GO)-based sensing platform for the detection of mecA gene in MRSA using flap endonuclease 1 (FEN1)-assisted target recycling and Klenow fragment (KF)-triggered signal amplification. Without the target, all the DNA probes were adsorbed onto GO, resulting in fluorescence quenching of the dye. Upon the addition of the target, a triple complex was formed that triggered FEN1-assisted target recycling and initiated two polymerization reactions with the assistance of KF polymerase, generating numerous dsDNA that were repelled by GO. These dsDNAs triggered fluorescence enhancement when SYBR Green I was added. Therefore, the target DNA was quantified by measuring the fluorescence at excitation and emission wavelengths of 480/526 nm. This mecA gene assay showed a good linear range from 1 to 50 nM with a lower limit of detection of 0.26 nM, and displayed good applicability to the analysis of real samples. Thus, a new method for monitoring MRSA has been developed that has great potential for early clinical diagnosis and treatment.

Cadmium Isotope Fractionation during Adsorption onto Edge Sites and Vacancies in Phyllomanganate.

Cadmium (Cd) geochemical behavior is strongly influenced by its adsorption onto natural phyllomanganates, which contain both layer edge sites and vacancies; however, Cd isotope fractionation mechanisms at these sites have not yet been addressed. In the present work, Cd isotope fractionation during adsorption onto hexagonal (containing both types of sites) and triclinic birnessite (almost only edge sites) was investigated using a combination of batch adsorption experiments, extended X-ray absorption fine structure (EXAFS) spectroscopy, surface complexation modeling, and density functional theory (DFT) calculations. Light Cd isotopes are preferentially enriched on solid surfaces, and the isotope fractionation induced by Cd2+ adsorption on edge sites (Δ114/110Cdedge-solution = -1.54 ± 0.11‰) is smaller than that on vacancies (Δ114/110Cdvacancy-solution = -0.71 ± 0.21‰), independent of surface coverage or pH. Both Cd K-edge EXAFS and DFT results indicate the formation of double corner-sharing complexes on layer edge sites and mainly triple cornering-sharing complexes on vacancies. The distortion of both complexes results in the negative isotope fractionation onto the solids, and the slightly longer first Cd-O distances and a smaller number of nearest Mn atoms around Cd at edge sites probably account for the larger fractionation magnitude compared to that of vacancies. These results provide deep insights into Cd isotope fractionation mechanisms during interactions with phyllomanganates.

Elevated RHAMM as a biomarker for predicting diabetic kidney disease in patients with type 2 diabetes.

Diabetic kidney disease (DKD) poses a significant challenge globally as a complication of diabetes. Hyaluronan (HA), a critical non-sulfated glycosaminoglycan in the extracellular matrix, plays a pivotal role in the progression of DKD. This study assesses the predictive significance of HA's corresponding receptor, RHAMM (receptor for HA-mediated motility), in DKD pathogenesis in type 2 diabetes (T2DM) patients. Enzyme-linked immunosorbent assays were utilized to measure plasma and urine levels of HA, CD44 and RHAMM in 99 diabetic patients. Immunohistochemistry staining was employed to examine HA deposition, CD44 and RHAMM expressions from 18 biopsy-proven DKD patients. Spearman correlation analysis, linear regression and receiver operating characteristic (ROC) analysis were conducted to establish associations between plasma HA, CD44 and RHAMM levels, and clinical parameters in DKD patients with T2DM. Elevated plasma and urine HA, CD44 and RHAMM levels were notably observed in the severe renal dysfunction group. Plasma RHAMM exhibited positive correlations with HA (r=0.616, P<.001) and CD44 (r=0.220, P<.001), and a negative correlation with estimated glomerular filtration rate (eGFR) (r=-0.618, P<.001). After adjusting for other potential predictors, plasma RHAMM emerged as an independent predictor of declining eGFR (β=-0.160, P<.05). Increased HA, CD44 and RHAMM levels in kidney biopsies of DKD patients were closely associated with heightened kidney injury. The ROC curve analysis highlighted an area under the curve (AUC) of 0.876 for plasma RHAMM, indicating superior diagnostic efficacy compared to CD44 in predicting DKD pathogenesis. The combined AUC of 0.968 for plasma RHAMM, HA and CD44 also suggested even greater diagnostic potential for DKD pathogenesis. These findings provide initial evidence that elevated RHAMM levels predict DKD pathogenesis in T2DM patients. The formation of a triple complex involving HA, CD44 and RHAMM on the cell surface shows promise as a targetable biomarker for early intervention to mitigate severe renal dysfunctions.

Harnessing the Power of a Novel Triple Chelate Complex in Fermented Probiotic Dairy Products: A Promising Solution for Combating Iron Deficiency Anemia.

This study discovered and examined novel triple chelate complexes involving iron, ascorbic acid, and essential amino acids (AsA-Fe-AmA triple chelate complexes) for the first time. The mechanism of complex formation was studied using FTIR spectroscopy and quantum chemical modeling. The produced complexes were shown to be suitable for fortifying food items with a pH of 3-7 that have not been exposed to heat treatment at temperatures over 75 °C for more than 15 min. Thus, it can be said that the concentration for milk fortification should be 0.005 mol/L or less. In vivo experiments in rats models revealed that the synthesized complexes increased serum iron levels after a single application to reference values within 24 h of oral administration. The iron level increased by 14.0 mmol/L at 2 mL dose of the complex. This fact makes it possible to consider the use of developed complexes and developed fermented dairy products for the prevention of iron deficiency and iron deficiency anemia. Research on the effect of discovered compounds on the physicochemical and organoleptic qualities of milk was conducted. Furthermore, iron ascorbate threoninate, iron ascorbate methioninate, iron ascorbate lysinate, and iron ascorbate tryptophanate all had a beneficial effect on Lacticaseibacillus rhamnosus at concentrations as low as 0.0005 mol/L, which is significant for milk fermentation. A study of fermented milk products revealed that the most effective AsA-Fe-AmA triple chelate complex is iron ascorbate lysinate, which might be further investigated as a viable molecule for dietary fortification in iron deficiency anemia. It was found that fortified fermented milk products had a titratable acidity of 67 ± 1°T, pH of 4.38 ± 0.05, and a viscosity of 2018 ± 142 Pa·s.

Stimulus-Responsive Four-Stranded DNA Nanoring Assembly to Host Multiple Nanosilver Clusters for Cooperatively Enhanced Fluorescence Biosensing.

Exploring the ability of four-stranded DNA nanorings (fsDNRs) to host multiple nanosilver clusters (NAgCs) for cooperatively amplifiable fluorescence biosensing to a specific initiator (tI*) is fascinating. By designing three DNA single strands and three analogous stem-loop hairpins, we developed a functional fsDNR through sequential cross-opening and overlapped hybridization. Note that a substrate strand (SS) was programmed with six modules: two severed splits (sT and sT') of NAgCs template, two sequestered segments by a middle unpaired spacer, and a partition for tI*-recognizable displacement, while sT and sT' were also tethered in two ends of three hairpins. At first, a triple dsDNA complex with stimulus-responsiveness was formed to guide the specific binding to tI*, while the exposed toehold of the SS activated the forward cascade hybridization of three hairpins, until the ring closure in the tailored self-assembly pathway for forming the fsDNR. The resulting four duplexes forced each pair of sT/sT' to be merged as the parent template in four nicks, guiding the preferential synthesis of four clusters in the shared fsDNR, thereby cooperatively amplifying the green fluorescence signal for sensitive assay of tI*. Meanwhile, the topological conformation of fsDNR can be stabilized by the as-formed cluster adducts to rivet the pair of two splits in the nicks. Benefitting from the self-enhanced effect of multiple emitters, this label-free fluorescent sensing strategy features simplicity, rapidity, and high on-off contrast, without involving complicated nucleic acid amplifiers.

Dairy product enriched with triple manganese complex

Micronutrient deficiency is a serious problem in Russia. In this article, we developed a complex of the essential microelement manganese with ascorbic acid and L-isoleucine. The properties of the resulting compound were studied by X-ray phase analysis, IR spectroscopy, and computer quantum chemical modeling. It was established that the complex has an amorphous structure. As a result of the simulation, the optimal interaction model was determined: through the carboxyl group and α-amino group of L-isoleucine, and through the C2 and C3 atoms of ascorbic acid ( E = -2264.757 kcal/mol). Quantum chemical modeling data were confirmed by IR spectroscopy. The stability of the complex was studied under various technological parameters (pH, temperature and exposure time). To do this, a multifactorial experiment was carried out with three input parameters and three levels of variation. It has been shown that the stability of the ternary complex of the essential microelement manganese with ascorbic acid and the essential amino acid L-isoleucine is significantly influenced by all parameters (pH, temperature and exposure time). It was found that with increasing pH, temperature and exposure time, ∆ Eh increases, which indicates a loss of stability of the complex and its destruction. The parameters at which the stability of the samples is observed correspond to the lowest values of ∆ Eh: pH = 3–7, t = 25–60°, τ = 5–15. At the next stage, milk was enriched with the resulting compound at the rate of 30% of the daily dose of manganese consumption and studied the physicochemical parameters of milk depending on the concentration of the developed complex. It was found that for the enrichment of manganese ascorbate isoleucinate it is necessary to use concentrations of 0.005 mol/l or less, since the obtained indicators correspond to the quality standards established in the Russian Federation. According to the organoleptic assessment, the values of the indicators (smell and taste) of milk enriched with manganese ascorbate isoleucinate exceed the values of the indicators of the control sample (JSC MKS, Stavropol, Russia) and the sample enriched with the inorganic form of iron — manganese sulfate.

An innovative form of the essential trace element copper for fortification of dairy products

Microelementosis is a disease associated with a deficiency of certain vital microelements. Today, there are ways to combat the lack of essential micronutrients. In this article, the mechanochemical synthesis of a triple copper-containing complex with ascorbic acid and L-isoleucine, an essential amino acid, was carried out. Examination of the sample by X-ray diffractometry showed that the sample has a trigonal crystal structure with space group P31c. As a result of computer quantum chemical modeling, the values of the difference in total energy, chemical hardness, energy of the highest occupied molecular orbital and the lowest unoccupied molecular orbital were determined. We determined the optimal option for the interaction of copper with vitamin C and L-isoleucine, where the binding occurs through hydroxyl groups attached to the C2 and C3 atoms of ascorbic acid, and through the carboxyl group and α-amino group of isoleucine. The sample was examined by IR spectroscopy and confirmed by computer modeling data. The stability of the resulting complex was studied depending on the technological parameters — pH, temperature and exposure time. Using the Greco-Latin square method, a matrix for planning a multifactorial experiment was compiled. An analysis of the dependence of the change in optical density (∆ D) on pH, temperature and exposure time showed that the pH of the medium and the temperature of the solution have a significant effect on ∆ D: an increase in the pH of the medium and the temperature of the solution leads to an increase in ∆ D. Exposure time does not have a significant effect to change the optical density value (∆ D). The parameters at which sample stability is observed correspond to the lowest values of ∆ D: pH = 3–8, t = 25–70, τ = 5–15. The effect of the concentration of a copper-containing complex on the physicochemical parameters of milk was studied. It has been established that the optimal concentration of copper ascorbate isoleucinate for fortifying dairy products is 0.005 mol/l or less. Next, an organoleptic assessment of the indicators of milk enriched with copper ascorbate isoleucinate was carried out. The analysis of the results showed that the organoleptic parameters (smell and taste) of milk enriched with ascorbate-isoleucinate of copper are 0.1 points higher than those of milk enriched with the inorganic form of the essential trace element copper and 0.3 points lower than that of the control milk sample.

Triple alignment-enhanced complex question answering over knowledge bases

Program induction is a crucial paradigm for complex question answering over knowledge bases. In the existing learning framework, the predicted program is required to strictly align (word-by-word) with a gold program, which could cause over penalization for minor deviations. Meanwhile, due to the existence of synonyms, program induction question answering often fails to retrieve answer from the knowledge base because individual function argument cannot perfectly replicate the target argument. To address above misalignment problems, we propose a triple alignment-enhanced complex question answering (TACQA) method by incorporating global token alignment, function alignment, and argument alignment. First, apart from the classical global token alignment, the predicted functions are extracted and aligned separately with the gold functions, enabling efficient learning of implicit structural information related to the query framework of program from input questions. Second, an argument alignment is introduced to correct the ambiguous function arguments, which enhances the disambiguation processing efficiency of multi-argument by optimizing candidate pool construction and similarity calculation. The experiments on KQA Pro show that our method consistently outperforms the SOTA methods, demonstrating the effectiveness of triple alignment processing mechanism for simultaneously addressing function misalignment and argument ambiguity in program induction and further improving the model performance.

Triple Inverse Sandwich versus End-On Diazenido: Bonding Motifs across a Series of Rhenium-Lanthanide and -Actinide Complexes.

While synthesizing a series of rhenium-lanthanide triple inverse sandwich complexes, we unexpectedly uncovered evidence for rare examples of end-on lanthanide dinitrogen coordination for certain heavy lanthanide elements as well as for uranium. We begin our report with the synthesis and characterization of a series of trirhenium triple inverse sandwich complexes with the early lanthanides, Ln[(μ-η5:η5-Cp)Re(BDI)]3(THF) (1-Ln, Ln = La, Ce, Pr, Nd, Sm; Cp = cyclopentadienide, BDI = N,N'-bis(2,6-diisopropylphenyl)-3,5-dimethyl-β-diketiminate). However, as we moved across the lanthanide series, we ran into an unexpected result for gadolinium in which we structurally characterized two products for gadolinium, namely, 1-Gd (analogous to 1-Ln) and a diazenido dirhenium double inverse sandwich complex Gd[(μ-η1:η1-N2)Re(η5-Cp)(BDI)][(μ-η5:η5-Cp)Re(BDI)]2(THF)2 (2-Gd). Evidence for analogues of 2-Gd was spectroscopically observed for other heavy lanthanides (2-Ln, Ln = Tb, Dy, Er), and, in the case of 2-Er, structurally authenticated. These complexes represent the first observed examples of heterobimetallic end-on lanthanide dinitrogen coordination. Density functional theory (DFT) calculations were utilized to probe relevant bonding interactions and reveal energetic differences between both the experimental and putative 1-Ln and 2-Ln complexes. We also present additional examples of novel end-on heterobimetallic lanthanide and actinide diazenido moieties in the erbium-rhenium complex (η8-COT)Er[(μ-η1:η1-N2)Re(η5-Cp)(BDI)](THF)(Et2O) (3-Er) and uranium-rhenium complex [Na(2.2.2-cryptand)][(η5-C5H4SiMe3)3U(μ-η1:η1-N2)Re(η5-Cp)(BDI)] (4-U). Finally, we expand the scope of rhenium inverse sandwich coordination by synthesizing divalent double inverse sandwich complex Yb[(μ-η5:η5-Cp)Re(BDI)]2(THF)2 (5-Yb), as well as base-free, homoleptic rhenium-rare earth triple inverse sandwich complex Y[(μ-η5:η5-Cp)Re(BDI)]3 (6-Y).

Alcohol-induced Golgiphagy is triggered by the downregulation of Golgi GTPase RAB3D

ABSTRACT The development of alcohol-associated liver disease (ALD) is associated with disorganized Golgi apparatus and accelerated phagophore formation. While Golgi membranes may contribute to phagophores, association between Golgi alterations and macroautophagy/autophagy remains unclear. GOLGA4/p230 (golgin A4), a dimeric Golgi matrix protein, participates in phagophore formation, but the underlying mechanism is elusive. Our prior research identified ethanol (EtOH)-induced Golgi scattering, disrupting intra-Golgi trafficking and depleting RAB3D GTPase from the trans-Golgi. Employing various techniques, we analyzed diverse cellular and animal models representing chronic and chronic/binge alcohol consumption. In trans-Golgi of non-treated hepatocytes, we found a triple complex formed between RAB3D, GOLGA4, and MYH10/NMIIB (myosin, heavy polypeptide 10, non-muscle). However, EtOH-induced RAB3D downregulation led to MYH10 segregation from the Golgi, accompanied by Golgi fragmentation and tethering of the MYH10 isoform, MYH9/NMIIA, to dispersed Golgi membranes. EtOH-activated autophagic flux is evident through increased WIPI2 recruitment to the Golgi, phagophore formation, enhanced LC3B lipidation, and reduced SQSTM1/p62. Although GOLGA4 dimerization and intra-Golgi localization are unaffected, loss of RAB3D leads to an extension of the cytoplasmic N terminal domain of GOLGA4, forming GOLGA4-positive phagophores. Autophagy inhibition by hydroxychloroquine (HCQ) prevents alcohol-mediated Golgi disorganization, restores distribution of ASGR (asialoglycoprotein receptor), and mitigates COL (collagen) deposition and steatosis. In contrast to short-term exposure to HCQ, extended co-treatment with both EtOH and HCQ results in the depletion of LC3B protein via proteasomal degradation. Thus, (a) RAB3D deficiency and GOLGA4 conformational changes are pivotal in MYH9-driven, EtOH-mediated Golgiphagy, and (b) HCQ treatment holds promise as a therapeutic approach for alcohol-induced liver injury. Abbreviation: ACTB: actin, beta; ALD: alcohol-associated liver disease; ASGR: asialoglycoprotein receptor; AV: autophagic vacuoles; EM: electron microscopy; ER: endoplasmic reticulum; EtOH: ethanol; HCQ: hydroxychloroquine; IP: immunoprecipitation; KD: knockdown; KO: knockout; MYH10/NMIIB: myosin, heavy polypeptide 10, non-muscle; MYH9/NMIIA: myosin, heavy polypeptide 9, non-muscle; PLA: proximity ligation assay; ORO: Oil Red O staining; PM: plasma membrane; TGN: trans-Golgi network; SIM: structured illumination super-resolution microscopy.

Conformational investigations on three large dinuclear triple helicates by single crystal X-ray diffraction

Three new dinuclear triple helicates were synthesised using a ditopic semi-rigid pyridylylimine ligand L, separated by a diphenoxy-biphenol spacer providing considerable length to the backbone. L and the new large dinuclear triple helicate complexes [Fe2L3](BF4)4 (1), [Ni2L3](BF4)4 (2) and [Zn2L3](BF4)4 (3) have been characterised in solution and solid state. Single crystal X-ray diffraction was used to investigate overall complex ion shape as the coordination sphere was modulated by metal ion selection. Small differences in complex shape were seen to arise due to subtle distortions in coordination sphere environments. This study sheds light on how the length and twist of dinuclear triple helicates can be tuned by selection of coordinating metal ion.

Normal and malaligned talonavicular fusion alters cadaveric foot pressure and kinematics.

Talonavicular (TN) fusion is a common treatment for TN arthritis or deformity correction. There is incongruous evidence regarding remaining motion at the talocalcaneal and calcaneocuboid joints after TN fusion. Additionally, the effects of a malaligned TN fusion are not well understood and alignment of the fusion may be important for overall foot integrity. This project assessed the kinematic and kinetic effects of neutral and malaligned TN fusions. Ten cadaveric feet were tested on a gait simulator in four conditions: unfused, fused in neutral, fused in varus, and fused in valgus. The fusions were simulated with external fixation hardware. An eight-camera motion analysis system and a 10-segment foot model generated kinematic data, and a pressure mat captured pressure data. Simulated TN fusion was achieved in eight feet. From unfused to fused-neutral, range of motion (ROM) was not eliminated in the adjacent joints, but the positions of the joints changed significantly throughout stance phase. Furthermore, the ROM increased at the tibiotalar joint. Plantar pressure and center of pressure shifted laterally with neutral fusion. The malalignments marginally affected the ROM but changed joint positions throughout stance phase. Pressure patterns were shifted laterally in varus malalignment and medially in valgus malalignment. The residual motion and the altered kinematics at the joints in the triple joint complex after TN fusion may subsequently increase the incidence of arthritis. Clinical significance: This study quantifies the effects of talonavicular fusion and malalignment on the other joints of the triple joint complex.

The impact of stoichiometry on the initial steps of crystal formation: Stability and lifetime of charged triple-ion complexes.

Minerals form in natural systems from solutions with varying ratios of their lattice ions, yet non-stoichiometric conditions have generally been overlooked in investigations of new formation (nucleation) of ionic crystals. Here, we investigated the influence of cation:anion ratio in the solution on the initial steps of nucleation by studying positively and negatively charged triple ion complexes and subsequent particle size evolution. Our model systems are carbonates and sulfates of calcium and barium, as it was recently shown that solution stoichiometry affects the timing and rate of their nucleation. Molecular dynamics (MD) simulations and dynamic light scattering (DLS) flow experiments show that nucleation correlates with the stability and lifetime of the initial complexes, which were significantly impacted by the cation:anion stoichiometry and ion type. Specifically, was found to have higher association constants and its lifetime was twofold longer than . Similar trends were observed for and . Contrastingly, for , was found to have lower association constants and its lifetime was shorter than . These trends in stability and lifetime follow the same asymmetrical behaviour as observed experimentally for particle formation using techniques like DLS. This suggests a causal relationship between the stability and lifetime of the initial charged complexes and the nucleation under non-stoichiometric conditions.

RBM10 regulates the tumorigenic potential of human cancer cells by modulating PPM1B and YBX1 activities

RNA binding protein RBM10 participates in various RNA metabolism, and its decreased expression or loss of function by mutation has been identified in many human cancers. However, how its dysregulation contributes to human cancer pathogenesis remains to be determined. Here, we found that RBM10 expression was decreased in breast tumors, and breast cancer patients with low RBM10 expression presented poorer survival rates. RBM10 depletion in breast cancer cells significantly promotes the cellular proliferation and migration. We further demonstrated that RBM10 forms a triple complex with YBX1 and phosphatase 1B (PPM1B), in which PPM1B serves as the phosphatase of YBX1. RBM10 knock-down markedly attenuated association between YBX1 and PPM1B, leading to elevated levels of YBX1 phosphorylation and its nuclear translocation. Furthermore, cancer cells with RBM10 depletion had a significantly accelerated tumor growth in nude mice. Importantly, these enhanced tumorigenic phenotypes can be reversed by overexpression of PPM1B. Our findings provide the mechanistic bases for functional loss of RBM10 in promoting tumorigenicity, and are potentially useful in the development of combined therapeutic strategies for cancer patients with defective RBM10.

Alteration of the -35 and -10 sequences and deletion the upstream sequence of the -35 region of the promoter A1 of the phage T7 in dsDNA confirm the contribution of non-specific interactions with E. coli RNA polymerase to the transcription initiation process.

Transcription initiation is a multi-step process, in which the RNA polymerase holoenzyme binds to the specific promoter sequences to form a closed complex, which, through intermediate stages, isomerizes into an open complex capable of initiating the productive phase of transcription. The aim of this work was to determine the contribution of the -10 and -35 regions of the promoter, as well as the role of non-specific interactions, in the binding of RNA polymerase and the formation of an active initiation complex capable of transcription. Therefore, fragments of promoter DNA, derived from the strong promoter A1 of the phage T7, containing completely and partially altered elements -35 and -10, and devoid of an upstream region, were constructed using genetic engineering methods. Functional analyses of modified promoter fragments were carried out, checking their ability to form binary complexes with Escherichia coli RNA polymerase (RNAP) and the efficiency of converting binary complexes into triple complexes characteristic of the productive phase of transcription. The obtained results suggest that, in relation to the A1 promoter of the T7 phage, the most important role of the -35 region is carrying the open complex through the next phases of transcription initiation. The weakening of specific impacts within the region -35 is a reason for the defect associated with the transformation of the open complex, formed by a DNA fragment containing the completely altered -35 region, into elongation and the impairment of RNA synthesis. This leads to breaking contacts with the RNA polymerase holoenzyme, and destabilization and disintegration of the complex in the initial phase of productive transcription. This confirms the hypothesis of the so-called stressed intermediate state associated with the stage of transition from the open complex to the elongation complex. The experiments carried out in this work confirm also that the process of promoter localization and recognition, as well as the formation of binary complexes, is sequential in nature, and that the region located upstream of the -35 hexamer, and the hexamer itself, plays here an additive role.

A dysprosium(III)-based triple helical-like complex as a turn-on/off fluorescence sensor for Al(III) and 4,5-dimethyl-2-nitroaniline.

A novel triple helical-like complex [Dy2K2L3(NO3)2]·3DMF (1) based on a designed Schiff base N'1,N'3-bis((E)-3-ethoxy-2-hydroxybenzylidene)-malonohydrazide (H4L) was synthesized with good chemical and thermal stabilities. Single-crystal X-ray structural analysis showed that 1 presents a tetranuclear triple helical-like structure via the coordination mode of Dy : K : L with 2 : 2 : 3 stoichiometry. Fluorescence measurements showed that 1@EtOH has excellent fluorescence turn-on/off response ability for aluminium ions and 4,5-dimethyl-2-nitroaniline (DMNA) with outstanding selectivity, sensitivity, and anti-interference ability. The calculated limit of detection (LOD) values for 1@EtOH to Al3+ and DMNA were found to be 0.53 and 3.33 μM, respectively. Density functional theory (DFT) calculation showed that the fluorescence response mechanism can be explained by the photoinduced electron transfer (PET) mechanism; meanwhile, the inner filter effect (IFE) of DMNA can also affect the emission of 1@EtOH.

A triple hydrogen-bond dominated fluorescent probe for monitoring the melting transition of polyurethane

Accurately characterizing the phase transition temperatures of polyurethanes (PU) is indispensable in controlling over the material processing and usage conditions yet challenging. Herein, a DDA-AAD-type triple hydrogen-bonded luminescent complex was...