Divalent Metal Research Articles

Why is binding of a divalent metal cation to a structural motif containing four carboxylate residues not accompanied by a conformational change?

We earlier showed that Torpedo californica acetylcholinesterase (AChE) contains a cluster of four conserved aspartates that can strongly bind divalent cations, which we named the 4D motif. Binding of the divalent metal cations greatly increases its thermal stability. Here we systematically examined all available crystallographic structures of T. californica AChE. Two additional metal-binding sites were identified, both composed of acidic and histidine residues. Relative binding to the 4D and additional sites was studied using metadynamics simulations. It was observed that in crystal structures devoid of metal ions in the 4D site, the conformation of T. californica AChE is almost identical to that in structures in which it is occupied by a divalent metal ion. Closer examination of the 4D motif reveals that three of the four acidic residues form ion pairs with conserved basic residues surrounding them. We named this new motif the 4A/3B motif. Molecular dynamics with quantum potential simulations was used to quantify the 4D motif's binding strength compared with that of the metal-binding site in the protein fXIIIa, which consists of four aspartates, but is devoid of adjacent cationic residues. Whereas fXIIIa's 4D site, in the absence of a metal cation, expanded significantly in the simulation, that of Torpedo AChE displayed only minor periodic changes in size. Furthermore, the energy of metal ion unbinding from the two sites differs by ca. 10 kcal/mol. We identified several other proteins in the PDB that contain the 4A/3B motif, whose conformations are identical in the presence or absence of a metal ion. An animated Interactive 3D Complement (I3DC) is available in Proteopedia at https://proteopedia.org/w/Journal:Protein_Science:4.

Abstract 4138304: Macrophage ZIP8 Enhances Efferocytosis and Mitigates Atherosclerosis by Modulating cellular Manganese (Mn) Homeostasis

Introduction: ZIP8, also known as SLC39A8, is a transmembrane transporter responsible for the movement of various divalent metal ions. Multiple genome-wide association studies (GWAS) have revealed a significant association between a mutation in ZIP8 (rs13107325; Ala391Thr) and cardiovascular diseases. While its physiologic role is incompletely understood, recent studies implicate that Zn2+ influx into monocytes regulated by ZIP8 is a novel factor determining their adhesion and recruitment to atherosclerotic lesions. The potential role of ZIP8 in modulating macrophage function and thereby influencing the development of atherosclerosis warrants further investigation. Methods and Results: Using an ApoE -/- ZIP8 A393T missense mutation mouse model, we quantified atherosclerotic plaque burden and lipid profiles following induction of atherosclerosis. The presence of the ZIP8 A393T missense mutation resulted in increased necrotic regions and plaque load in the atherosclerosis mouse model, while having no impact on total cholesterol (TC) or low-density lipoprotein cholesterol (LDL-C) levels. Confocal microscopy and flow cytometry to investigate how reduced ZIP8 expression affects macrophage efferocytosis. When siRNA was used to knock down ZIP8 in vitro, macrophage efferocytosis was reduced. Subsequently, Macrophage-specific ZIP8 knockout mice (ZIP8 LysM ) demonstrated a decrease in the efferocytosis index in primary macrophage. Additionally, in vivo thymic and peritoneal efferocytosis assays showed an impaired efferocytosis phenotype in ZIP8 LysM mice. Confocal microscopy showed ZIP8 LysM mice have no effect on macrophage binding to apoptotic cells. The lack of or incompleteness of the F-ACTIN ring formed by macrophages surrounding apoptosis in the ZIP8 LysM mice, and the prolonged duration of phagocytosis ring formation suggests that the internalization process is compromised. The role of ZIP8 ion transport in the efferocytosis impairment mechanism was also investigated.The regulation of macrophage efferocytosis via the metabolism of D-galactose, glutamine, glutamate, and Zn 2+ is unaffected by ZIP8. Mn 2+ was identified as a vital regulatory factor in the efferocytosis function of ZIP8 in macrophages. Conclusions: ZIP8 influences the efferocytotic capacity of macrophages and regulates the clearance of apoptotic cells through Mn 2+ , suggesting that ZIP8 and Mn 2+ hold promise as a key therapeutic targets for the treatment of atherosclerotic cardiovascular diseases.

Converging Roles of the Metal Transporter SMF11 and the Ferric Reductase FRE1 in Iron Homeostasis of Candida albicans.

Pathogenic fungi must appropriately sense the host availability of essential metals such as Fe. In Candida albicans and other yeasts, sensing of Fe involves mitochondrial Fe-S clusters. Yeast mutants for Fe-S cluster assembly sense Fe limitation even when Fe is abundant and hyperaccumulate Fe. We observe this same disrupted Fe sensing with C. albicans mutants of SMF11, a NRAMP transporter of divalent metals. Mutants of smf11 hyperaccumulate both Mn and Fe and the elevated Mn is secondary to Fe overload. As with Fe-S biogenesis mutants, smf11∆/∆ mutants show upregulation of ferric reductases that are normally repressed under high Fe, and Fe import is activated. However, unlike Fe-S biogenesis mutants, smf11∆/∆ mutants show no defects in mitochondrial Fe-S enzymes. Intriguingly, this exact condition of disrupted Fe sensing without inhibiting Fe-S clusters occurs with C. albicans fre1∆/∆ mutants encoding a ferric reductase. Mutants of fre1 and smf11 display similar perturbations in the cell wall, in filamentation and in the ROS burst of morphogenesis, a Fe-dependent process. As with FRE1, SMF11 is important for virulence in a mouse model for disseminated candidiasis. We propose a model in which FRE1 and SMF11 operate outside the mitochondrial Fe-S pathway to donate ferrous Fe for Fe sensing.

Thermogenic Adipocytes Promote M2 Macrophage Polarization through CNNM4-Mediated Mg Secretion.

M2 macrophages promote adipose tissue thermogenesis which dissipates energy in the form of heat to combat obesity. However, the regulation of M2 macrophages by thermogenic adipocytes is unclear. Here, it is identified magnesium (Mg) as a thermogenic adipocyte-secreted factor to promote M2 macrophage polarization. Mg transporter Cyclin and CBS domain divalent metal cation transport mediator 4 (CNNM4) induced by ADRB3-PKA-CREB signaling in thermogenic adipocytes during cold exposure mediates Mg efflux and Mg in turn binds to the DFG motif in mTOR to facilitate mTORC2 activation and M2 polarization in macrophages. In obesity, downregulation of CNNM4 expression inhibits Mg secretion from thermogenic adipocytes, which leads to decreased M2 macrophage polarization and thermogenesis. As a result, CNNM4 overexpression in adipocytes or Mg supplementation in adipose tissue ameliorates obesity by promoting thermogenesis. Importantly, an Mg wire implantation (AMI) approach is introduced to achieve adipose tissue-specific long-term Mg supplement. AMI promotes M2 macrophage polarization and thermogenesis and ameliorates obesity in mice. Taken together, a reciprocal regulation of thermogenic adipocytes and M2 macrophages important for thermogenesis is identified, and AMI is offered as a promising strategy against obesity.

Metal Ion Sensing by Tetraloop-like RNA Fragment: Role of Compact Intermediates with Non-Native Metal Ion-RNA Inner-Shell Contacts.

Divalent metal ions influence the folding and function of ribonucleic acid (RNA) in the cells. The mechanism of how RNA structural elements in riboswitches sense specific metal ions is unclear. RNA interacts with ions through two distinct binding modes: direct interaction between the ion and RNA (inner-shell (IS) coordination) and indirect interaction between the ion and RNA mediated through water molecules (outer-shell (OS) coordination). To understand how RNA senses metal ions such as Mg2+ and Ca2+, we studied the folding of a small RNA segment from the Mg2+ sensing M-Box riboswitch using computer simulations. This RNA segment has the characteristics of a GNRA tetraloop motif and interestingly requires the binding of a single Mg2+ ion. The folding free energy surface of this simple tetraloop system is multidimensional, with a population of multiple intermediates where the tetraloop and cation interact through IS and OS coordination. The partially folded compact tetraloop intermediates form multiple non-native IS contacts with the metal ion. Thermal fluctuations should break these strong non-native IS contacts so that the tetraloop can fold to the native state, resulting in higher folding free energy barriers. Ca2+ undergoes rapid OS to IS transitions and vice versa due to its lower charge density than Mg2+. However, the ability of Ca2+ to stabilize the native tetraloop state is weaker, as it could not hold the loop-closing nucleotides together due to its weaker interactions with the nucleotides. These insights are critical to understanding the specific ion sensing mechanisms in riboswitches, and the predictions are amenable for verification by nuclear magnetic resonance (NMR) experiments.

Geochemical Decoupling of Iron and Zinc during Transformation of Zn-Bearing Ferrihydrite in Reducing Sediments.

The transformation of the mineral ferrihydrite in reducing environments, and its impact on the mobility of incorporated trace metals, has been investigated in model laboratory studies, but studies using complex soil or sediment matrices are lacking. Here, we studied the transformation of zinc (Zn)-bearing ferrihydrite labeled with 57Fe and mixed with natural sediments, incubated in reducing conditions for up to six months. We tracked the evolution of Fe and Zn speciation with 57Fe Mössbauer spectroscopy and with bulk and micro-X-ray absorption spectroscopy. We show that Fe was readily reduced and incorporated into a poorly crystalline mixed-valence Fe(II)-Fe(III) phase resembling green rust. In parallel, Zn was released in the surrounding porewater and scavenged by precipitation with available ligands, particularly as zinc sulfide (ZnS) or Zn-carbonates. Early in the mineral transformation process, the chemical behavior of Fe was decoupled from Zn, suppressing the impact of Zn on the rates and products of the ferrihydrite transformation. Our results underline the discrepancy between model experiments and complex field-like conditions and highlight the importance of sediment and soil geochemistry and ligand competition on the fate of divalent metal contaminants in the environment.

Modulatory effects of inorganic mercury (Hg (II)) and lead (Pb (II)) on immune responses of Pekin ducklings (Anas platyrhynchos domesticus) upon a viral-like immune challenge

Trace metal contamination is ubiquitous around the world and may affect the health of wildlife. Divalent trace metals, including ions of mercury (Hg) and lead (Pb), have been shown to be immunotoxic to avian species. However, little is known about the immunomodulatory effects of trace metal exposure on viral infections, especially in young birds, who may be more sensitive. Therefore, the objective of the current study is to provide more insights in the causality between trace metal exposure and the effects of exposure on the immune responses in young waterfowls. Pekin duckling was used as an animal model to investigate the effects of inorganic divalent Hg (II) and Pb (II) on avian immune responses upon a viral-like challenge with double-stranded RNA. Our results indicate that Hg (II) altered the immune gene expression 24 h post-challenge, as reflected by induction of pro-inflammatory genes IL-8, iNOS, TLR3 and TLR7, and a significant decrease of microRNA-155. Ducklings exposed to Pb (II) showed lower levels of natural antibodies, reduced white blood cell counts and lower heterophil proportions 24 h post-challenge. Although inorganic divalent Hg (II) and Pb (II) showed specific differential effects on the immune response of Pekin ducklings, the overall adverse immunomodulatory outcomes in both cases point to inflammation, impaired B-cell function, and weaker immunocompetence.

One-pot synthesis of graphene nanosheets‑nickel cobalt LDHs nanocomposite for electrocatalysis of oxygen evolution reaction

Oxygen evolution reaction (OER) is known as a bottleneck for the overall water-splitting process due to its slow kinetics. In this regard, constructing a high-performance, cost-effective electrode material may act as a game changer. Here, a 2D/2D nanocomposite has prepared out of graphene nanosheets and nickel‑cobalt layered double hydroxide (NiCo LDH) by a simple solvothermal method. The 2D flower-like structures of NiCo LDH attached to graphene nanosheets have observed in field-emission scanning (FE-SEM) and transmission (TEM) electron microscopy images. The X-ray diffraction (XRD) patterns have revealed the formation of Ni(OH)2 and Co(OH)2 lattices along with hexagonal graphene crystals. X-ray energy dispersive spectroscopy (EDS) has noted the 2.2:1 ratio of nickel to cobalt in the graphene/NiCo LDH nanocomposite. X-ray elemental maps have shown the uniform distribution of elements in the sample, and the corresponding functional groups have observed in the Fourier-transform infrared (FTIR) and Raman spectra. X-ray photoelectron spectroscopy (XPS) has determined the coexistence of divalent and trivalent metals in the nanocomposite. The N2 adsorption-desorption study has shown signs of slit-shaped ion-accessible micro and mesopores with high specific surface area for the nanocomposite. It has benefited the ion diffusion process during the exposure of the modified electrode to the electrolyte. The graphene/NiCo LDH nanocomposite has provided the onset potential of 1.56 V, overpotential of 338 mV at 10 mA cm−2, Tafel slope of 69 mV dec−1, charge-transfer resistance (Rct) of 27 Ω, double layer capacitance (Cdl) of 24 μF, electrochemically surface area (ECSA) of 0.6 cm2, and roughness factor (RF) of 20. The electrode has maintained 98.3 % of its initial signal after 10 h continuous measurement at OER potential. This electrocatalytic activity refers to the sheet-like morphology, effective hydroxide ion transfer through interlayer spaces, enhanced conductivity, and high chemical stability achieved by a constructive synergy between graphene nanosheets and NiCo LDH.

Biochanin A inhibits excitotoxicity-triggered ferroptosis in hippocampal neurons

Excitatory neurotransmitter-induced neuronal ferroptosis has been implicated in multiple neurodegenerative diseases such as Alzheimer’s disease and Parkinson’s disease. Although there are several reports pertaining to the pharmacological activities of biochanin A, the effects of this isoflavone on excitotoxicity-triggered neuronal ferroptosis remain unclear. In this study, we demonstrate that biochanin A inhibits ferroptosis of mouse hippocampal neurons induced by glutamate or the glutamate analog, kainic acid. Biochanin A significantly inhibited accumulation of intracellular iron and lipid peroxidation in glutamate- or kainic acid-treated mouse hippocampal neurons. Furthermore, biochanin A regulated the level of glutathione peroxidase 4, a master regulator of ferroptosis, by modulating its autophagy-dependent degradation. We observed that biochanin A reduced the glutamate-induced accumulation of intracellular iron by regulating expression of iron metabolism-related proteins including ferroportin-1, divalent metal transferase 1, and transferrin receptor 1. Taken together, these results indicate that biochanin A effectively inhibits hippocampal neuronal death triggered by glutamate or kainic acid. Our study is the first to report that biochanin A has therapeutic potential for the treatment of diseases associated with hippocampal neuronal death, particularly ferroptosis induced by excitatory neurotransmitter.

Influence mechanism of divalent metals in the laminates of M2+Al-layered double hydroxides on peroxymonosulfate activation reaction pathway: Radical vs nonradical

Radical and nonradical reaction pathways have been commonly validated in peroxymonosulfate (PMS) based advanced oxidation progresses (AOPs) using layered double hydroxides (LDHs) as catalysts, but achieving efficient and selective degradation of organic pollutants remains challenging due to the unclear regulatory mechanism of laminate M2+ in PMS activation. Here, the d-orbital electrons configuration of M2+ was found to influence the degree of overlap between M−OH and O of PMS, which results in the selective dissociation of PMS to form reactive oxygen species (ROS) or achieve electron transfer pathway. Further analysis revealed that Co(II, 3d7) and Ni(II, 3d8) with filled 3d orbital electrons can serve as electron donors for PMS, leading to the cleavage of O–O bond and the efficient generation of radical-based ROS. In contrast, Mg(II, 3d0) with unfilled 3d orbital electrons and Zn(II, 3d10) with full filled 3d orbital electrons tend to act as electron shuttles, and the outermost orbitals substantially overlap with the O of PMS to form metastable complexes. Consequently, MgAl-LDH interacts with PMS to selectively degrade quinolones as electron donor through electron transfer pathway, and its performance was not inhibited by common anions and organic matter in actual water. The above results provide a basis for optimizing the design of LDH-based catalysts according to the type of laminate M2+ to achieve high selectivity and efficiency of PMS-AOPs in water purification applications.

Microstructural Evaluation and Linkage to the Engineering Properties of Metal-Ion-Contaminated Clay.

The rapid progress of urbanization and industrialization has led to the accumulation of large amounts of metal ions in the environment. These metal ions are adsorbed onto the negatively charged surfaces of clay particles, altering the total surface charge, double-layer thickness, and chemical bonds between the particles, which in turn affects the interactions between them. This causes changes in the microstructure, such as particle rearrangement and pore morphology adjustments, ultimately altering the mechanical behavior of the soil and reducing its stability. This study explores the effects of four common metal ions, including monovalent alkali metal ions (Na+, K+) and divalent heavy metal ions (Pb2+, Zn2+), with a focus on how ion valence and concentration impact the soil's microstructure and mechanical properties. Microstructural tests show that metal ion incorporation reduces particle size, increases clay content, and transforms the structure from layered to honeycomb-like. Small pores decrease while large pores dominate, reducing the specific surface area and pore volume, while the average pore size increases. Although cation exchange capacity decreases, cation adsorption density per unit surface area increases. Monovalent ions primarily disperse the soil structure, while divalent ions induce coagulation. Macro-mechanical tests reveal that metal ion contamination reduces porosity under loading, with compressibility rises as the ion concentration increases. Soils contaminated with alkali metal ions shows higher compression coefficients at all loads, while heavy metal ions cause higher compression under lower loads. Shear strength, the internal friction angle, and cohesion in metal-ion-contaminated clay decrease compared to uncontaminated field-state clay, with greater declines at higher ion concentrations. The Micropore Morphology Index and hydro-pore structural parameter effectively characterize both micro- and macrostructural properties, establishing a quantitative relationship between HPSP and the engineering properties of metal-ion-contaminated clay.

Adsorption behavior of zirconium metal–organic frameworks in multicomponent metal-ion solutions

Abstract In this study, the adsorptive behavior of zirconium metal–organic frameworks (Zr-MOFs) was investigated for the adsorption of alkali and alkaline earth metal ions from multi-element aqueous solutions. Zr-MOFs exhibit high water stability, notable surface area, and a significant pore volume, all of which contribute to their enhanced adsorption capacity. Three Zr-MOFs, namely bare UiO-66, amine-functionalized NH2-UiO-66, and carboxyl-functionalized UiO-66-(COOH)2, were tested for the adsorption of alkali and alkaline earth metal ions. Among the 3 MOFs that were tested, only the carboxyl group-functionalized Zr-MOF showed significant adsorption capacity toward divalent metal ions. Further, a thorough investigation was conducted to understand how the pH, initial concentration of the solution, and Zr-MOF dosage impact the adsorption properties of UiO-66-(COOH)2. At the natural pH (6.5) of the solution, UiO-66-(COOH)2 exhibited a superior adsorption capacity toward Sr2+ (15.3 mg g−1) and Ca2+ (7.9 mg g−1), which was attributable to the stronger electrostatic attraction of these ions relative to monovalent ions. The kinetic study results indicated that the preferred mechanism of adsorption was chemisorption for divalent metal ions. Additionally, the adsorption behavior of UiO-66-(COOH)2 for 24 elements was evaluated and the MOF showed significant adsorption of Sr2+ and Ca2+ alongside other divalent and trivalent metal ions. The experimental findings of the present study suggest that carboxylic-functionalized Zr-MOF holds significant potential toward the preparation of suitable sorbents for the extraction of higher-valence metal ions.

Identification of a divalent metal transporter required for cellular iron metabolism in malaria parasites

Plasmodium falciparum malaria parasites invade and multiply inside red blood cells (RBCs), the most iron-rich compartment in humans. Like all cells, P. falciparum requires nutritional iron to support essential metabolic pathways, but the critical mechanisms of iron acquisition and trafficking during RBC infection have remained obscure. Parasites internalize and liberate massive amounts of heme during large-scale digestion of RBC hemoglobin within an acidic food vacuole (FV) but lack a heme oxygenase to release porphyrin-bound iron. Although most FV heme is sequestered into inert hemozoin crystals, prior studies indicate that trace heme escapes biomineralization and is susceptible to nonenzymatic degradation within the oxidizing FV environment to release labile iron. Parasites retain a homolog of divalent metal transporter 1 (DMT1), a known mammalian iron transporter, but its role in P. falciparum iron acquisition has not been tested. Our phylogenetic studies indicate that P. falciparum DMT1 (PfDMT1) retains conserved molecular features critical for metal transport. We localized this protein to the FV membrane and defined its orientation in an export-competent topology. Conditional knockdown of PfDMT1 expression is lethal to parasites, which display broad cellular defects in iron-dependent functions, including impaired apicoplast biogenesis and mitochondrial polarization. Parasites are selectively rescued from partial PfDMT1 knockdown by supplementation with exogenous iron, but not other metals. These results support a cellular paradigm whereby PfDMT1 is the molecular gatekeeper to essential iron acquisition by blood-stage malaria parasites and suggest that therapeutic targeting of PfDMT1 may be a potent antimalarial strategy.

EDTA-treated fibrillated cellulose bio-adsorbents from okra plant biomass and their performance on toxic divalent metal ions attenuation from aqueous solution

EDTA-treated fibrillated cellulose bio-adsorbents from okra plant biomass and their performance on toxic divalent metal ions attenuation from aqueous solution

Computational Exploration of the Impact of Low‐Spin and High‐Spin Ground State on the Chelating Ability of Dimethylglyoxime Ligand on Dihalo Transition Metal: A QTAIM, EDA, and CDA Analysis

ABSTRACTWe have explored the chelation of dimethylglyoxime ligand to divalent (ndx: x = 6, 7, 8) transition metal (TM) cations in two media (gas phase and water) at the B3LYP//LANL2DZ/6–311+G (d,p) and B3LYP/def2‐TZVP level at lower multiplicity and higher multiplicity states. Majority of the 18 optimized halide (chloride and bromide) complexes prefer square planar configuration. The correlations discerned between the experimental structural data and their estimated counterparts demonstrate a good credibility for complexes at lower multiplicity state. The basis set superposition errors (BSSEs) estimated is very small which reflects the fact that the choice of different basis sets (B3LYP//LANL2DZ/6–311+G (d,p)) introduces a slight bias in the calculation of energies. The ADMP (atom‐centered density matrix propagation) simulations in water on chloride complexes indicate the irreversible nature of these M—N dissociation in trajectory simulation process. This fact explains our exclusive focus on the examination of the [glyoxime ligand]…[MX2] interactions. In addition, the solvation of (3d and 4d) transition metal chloride complexes causes a sensitive augmentation of the metal ion affinity (MIA) with an average of 0.29 and 0.24 kcal/mol. In both multiplicity states, the topological parameters have illustrated that the M—N and M—X bonds are typical metal–ligand in both media. The average ΔEorb/ΔEsteric ratio equal to 0.45 and 0.11 in gas phase and water, respectively, reveals the predominance of the contributions from non‐covalent bonding interactions (NCI) compared to those of covalent bonding. But, the maximal value equal to 6.760 is obtained for bromide rhodium complex in water. NBO analysis in both media highlights the fact that a more pronounced ionic character is observed for the majority of the chloride complexes at both spin multiplicity states because of their higher retained charges on the metal atom. For [dimethylglyoxime]…[MX2] interaction (X = Cl and Br), the charge decomposition analysis demonstrates that the lowest value of the d/b ratio is found for the chloride platinum complex at lower multiplicity state in water. This is a proof of its strong relativistic effects.

Divalent transition metal complexes with mixed of β-enaminone and N,O-donor ligands: synthesis, characterization and biological assessment

This study extensively describes the synthesizing and characterizing of new β-enaminone complexes derived from dimedone and amines coordinated with transition metals Fe(II), Co(II), Ni(II), and Cu(II), also two ligand's metformin (Met) and 8-hydroxyquinoline (8-hq). These were determined via FT-IR, 1H-NMR, electron and mass spectroscopy, molar electrical conductivity, thermal analysis (TGA, DTA and DSC) and characterized by metal content determination (%), magnetic susceptibility and elemental analysis (C.H.N.) and SEM technique. The measurements indicate that the complexes have six coordination numbers, where the β-enaminone is coordinated as tetradentate ligand through the two nitrogen atoms and the two oxygen atoms beside metformin is coordinated through the two nitrogen atoms and 8-hydroxyquinoline is coordinated by oxygen and nitrogen atoms as bidentate ligands. The conductance suggests that all complexes have electrolytic behavior, being conductive in a ratio of (1:2). SEM results revealed the presence of nanostructures on sample surfaces. The biological activity of the prepared ligands and complexes are studied in different concentrations for four types of bacteria Staphylococcus aureus has (+G) and Escherichia coli, Pseudomonas aeruginosa, Klebsiella have (-G), well as, the influence of the fungus. The ligands have disparate effectiveness while complexes have high effectiveness inhibiting the bacteria and fungus. KEY WORDS: Mixed ligand systems, Dimedone, β-Enaminone ligands, Transition metals, Biological application Bull. Chem. Soc. Ethiop. 2025, 39(1), 65-78. DOI: https://dx.doi.org/10.4314/bcse.v39i1.5

Effect of "Xingnao Kaiqiao" needling on expression of ferroptosis-related factors in rats with cerebral ischemia-reperfusion injury

To observe the effect of "Xingnao Kaiqiao" needling on the expression of ferroptosis-related proteins in neurons of rats with cerebral ischemia-reperfusion injury (CIRI), so as to explore its mechanism underlying improvement of CIRI. Male Wistar rats were randomly divided into sham operation, model, acupuncture and deferoxamine (DFO) groups, with 18 rats in each group. The CIRI model was established by occlusion of the middle cerebral artery. In the acupuncture group, "Xingnao Kaiqiao" needling was applied to "Shuigou" (GV26), "Neiguan" (PC6) and "Sanyinjiao"(SP6) for 20 min with electroacupuncture (2 Hz/15 Hz, 1 mA) at PC6 and SP6, twice daily for continuous 3 days. Rats of the DFO group received intraperitoneal injection of iron chelator DFO (0.1 g/kg, once daily). The severity of neurological impairment (neurological deficit score, 0-5 points, the lower the score, the severer is the neurological impairment) was evaluated by using Zausinger 6-poins scaling method. The cerebral infarct volume was measured after 2, 3, 5-triphenyltetrazolium chloride (TTC) staining, and the histopathological changes of the ischemic brain tissue were observed after H.E. staining. The mitochondrial structure of the hippocampal neurons on the ischemic side of the brain was observed by using transmission electron microscope. The levels of iron deposition rate (%) in the ischemic penumbra of the brain tissue and hippocampus were observed after Prussian blue staining, and the reactive oxygen species (ROS) content of the cerebral ischemic penumbra was assayed using flow cytometry, and the content of glutathione (GSH) content in the ischemic penumbra was detected by using microplate method. The real-time quantitative PCR was used to detect the expression of glutathione peroxidase 4 (GPX4), divalent metal transporter 1 (DMT1), transferrin (TF), transferrin receptor 1 (TFR1), and ferroportin 1 (FPN1) mRNAs in the ischemic penumbra, and the Western blot was used to detect the expression of GPX4, DMT1, TF, TFR1, FPN1, and ferritin (FER) proteins in the ischemic penumbra. Compared with the sham operation group, the neurological deficit score, GSH content, expression of GPX4 and FPN1 mRNAs and proteins were significantly decreased (P<0.01), while the percentage of cerebral infarct volume, iron deposit rates of the cerebral ischemic penumbra and hippocampus, ROS content, and the expression levels of DMT1, TF, and TFR1 mRNAs and proteins and FER protein were considerably increased (P<0.01) in the model group. In comparison with the model group, the decrease of neurological deficit score, GSH content, expression of GPX4 and FPN1 mRNAs and proteins, and the increase of the percentage of cerebral infarct volume, iron deposit rates of the cerebral ischemic penumbra and hippocampus, ROS content, and the expression levels of DMT1, TF, and TFR1 mRNAs and proteins and FER protein were all reversed in both DFO and acupuncture groups (P<0.01, P<0.05). The effects of acupuncture were significantly superior to those of DFO in lowering the levels of cerebral cortical and hippocampal iron deposit rates, ROS content and in elevating the expression of GPX4 mRNA and protein (P<0.01, P<0.05). H.E. staining showed large necrotic cells, disordered arrangement of cells in the cerebral cortex and hippocampus, with hyperchromic nuclei, vacuole-like changes, widening of cellular space, and cell swelling in the model group, which was relatively milder in the cell damage in both acupuncture and DFO groups. In addition, the ultrastructure of cells in the hippocampus showed irregular cellular nuclear morphology, atrophy of some mitochondria in the cytoplasm, partial mitochondrial membrane rupture and edema, and loosening of the ridge structure in the model group, which was milder in the mitochondrial impairment (including reduced number of mitochondria, broken mitochondrial membrane and reduced ridge structure in fewer cells) in the acupuncture group. The "Xingnao Kaiqiao" needling intervention has a neuroprotective effect in CIRI rats, which may be related to its functions in regulating ferroptosis-related targets and iron metabolism in cerebral ischemic penumbra, reducing oxidative stress injury, and suppressing neuronal ferroptosis.

Remote chirality transfer in low-dimensional hybrid metal halide semiconductors.

In hybrid metal halide perovskites, chiroptical properties typically arise from structural symmetry breaking by incorporating a chiral A-site organic cation within the structure, which may limit the compositional space. Here we demonstrate highly efficient remote chirality transfer where chirality is imposed on an otherwise achiral hybrid metal halide semiconductor by a proximal chiral molecule that is not interspersed as part of the structure yet leads to large circular dichroism dissymmetry factors (gCD) of up to 10-2. Density functional theory calculations reveal that the transfer of stereochemical information from the chiral proximal molecule to the inorganic framework is mediated by selective interaction with divalent metal cations. Anchoring of the chiral molecule induces a centro-asymmetric distortion, which is discernible up to four inorganic layers into the metal halide lattice. This concept is broadly applicable to low-dimensional hybrid metal halides with various dimensionalities (1D and 2D) allowing independent control of the composition and degree of chirality.

Catalytic cleave of an RNA substrate that bypasses the reorganization of its secondary structure during substrate recognition by a trans-acting VS ribozyme

Varkud satellite ribozyme (VS ribozyme) is a class of catalytic RNA with self-cleavage activity. The wild-type VS ribozyme has structural modularity with a relatively large catalytic module (H2–H6 elements) and a small substrate module (H1 element). The two modules can be dissected physically, and the substrate H1 RNA is recognized and then cleaved by the rest of the parent ribozyme serving as catalytic RNA. We characterized the catalytic properties of a bimolecular VS ribozyme developed and employed for an in-droplet evolution experiment of the VS ribozyme. We examined the effects of polyamines and several divalent metal ions. The results obtained in this study would be useful for the optimization of laboratory evolution of the VS ribozyme.

Structural and theoretical insights into the influence of thiocyanate ligands on divalent metal complexes with terpyridine derivatives

Four complexes with terpyridine derivative, thiocyanate anion, and selected d-electron metals were studied. The obtained complexes of the formulation [M(ftpy)(SCN)2(CH3OH)]•DMF (M = Mn (1), Co (2), Ni (3)) and [Cu(ftpy)(NCS)2]•H2O (4), where ftpy = 4′‐(furan-2-yl)-2,2′:6′,2′′‐terpyridine, were characterized using FTIR and UV–Vis spectroscopies, magnetic studies, elemental analysis, and single crystal X-ray crystallography. All compounds crystallize in a triclinic crystal system with space group P1¯ and 1–3 are isomorphous and isostructural. The effect of the modification of the terpyridine ligand and the presence of the thiocyanate ligand on the structure has been verified. Interesting dependence was found in the structure of compound 4, where the two thiocyanate ligands are differently coordinated, one via N and the other via S, and the formation of complex dimers connected through hydrogen bonds is observed. The π-stacking interactions and the resulting solid-state architectures of compounds 1–4 were inspected through a combination of DFT calculations, QTAIM, and NCIPlot analyses, and the stability of the complexes was studied by UV–Vis spectroscopy in various solvents.