Role of Divalent Cations in Infections in Host-Pathogen Interaction.

With increasing numbers of patients worldwide diagnosed with diabetes mellitus, renal disease, and iatrogenic immune deficiencies, an increased understanding of the role of electrolyte interactions in mitigating pathogen virulence is necessary. The levels of divalent cations affect host susceptibility and pathogen survival in persons with relative immune insufficiency. For instance, when host cellular levels of calcium are high compared to magnesium, this relationship contributes to insulin resistance and triples the risk of clinical tuberculosis. The movement of divalent cations within intracellular spaces contributes to the host defense, causing apoptosis or autophagy of the pathogen. The control of divalent cation flow is dependent in part upon the mammalian natural resistance-associated macrophage protein (NRAMP) in the host. Survival of pathogens such as M tuberculosis within the bronchoalveolar macrophage is also dependent upon NRAMP. Pathogens evolve mutations to control the movement of calcium through external and internal channels. The host NRAMP as a metal transporter competes for divalent cations with the pathogen NRAMP in M tuberculosis (whether in latent, dormant, or active phase). This review paper summarizes mechanisms of pathogen offense and patient defense using inflow and efflux through divalent cation channels under the influence of parathyroid hormone vitamin D and calcitonin.

Synthesis and characterization of ion-induced sodium alginate/soy protein isolate microgels for the controlled release.

In this study, sodium alginate/ soy protein isolate (SPI) microgels cross-linked by various divalent cations including Cu2+, Ba2+, Ca2+, and Zn2+ were fabricated. Cryo-scanning electron microscopy observations revealed distinctive structural variations among the microgels. In the context of gastric pH conditions, the degree of shrinkage of the microgels followed the sequence of Ca2+ > Ba2+ > Cu2+ > Zn2+. Meanwhile, under intestinal pH conditions, the degree of swelling was ranked as Zn2+ > Ca2+ > Ba2+ > Cu2+. The impact of these variations was investigated through in vitro digestion studies, revealing that all microgels successfully delayed the release of ß-carotene within the stomach. Within the simulated intestinal fluid, the microgel cross-linked with Zn2+ exhibited an initial burst release, while those cross-linked with Cu2+, Ba2+, or Ca2+ displayed a sustained release pattern. This research underscores the potential of sodium alginate/SPI microgels cross-linked with different divalent cations as efficient controlled-release delivery systems.

Structural and biochemical analysis of the unique interactions of the Campylobacter jejuni CorA channel protein with divalent cations.

CorA is a Mg2+ channel that plays a key role in the homeostasis of intracellular Mg2+ in bacteria and archaea. CorA consists of a cytoplasmic domain and a transmembrane domain and generates a Mg2+ pathway by forming a pentamer in the cell membrane. CorA gating is regulated via negative feedback by Mg2+, which is accommodated by the pentamerization interface of the CorA cytoplasmic domain (CorACD). The Mg2+-binding sites of CorACD differ depending on the species, suggesting that the Mg2+-binding modes and Mg2+-mediated gating mechanisms of CorA vary across prokaryotes. To define the Mg2+-binding mechanism of CorA in the Campylobacter jejuni pathogen, we structurally and biochemically characterized C. jejuni CorACD (cjCorACD). cjCorACD adopts a three-layered α/ß/α structure as observed in other CorA orthologs. Interestingly, cjCorACD exhibited enhanced thermostability in the presence of Ca2+, Ni2+, Zn2+, or Mn2+ in addition to Mg2+, indicating that cjCorACD interacts with diverse divalent cations. This cjCorACD stabilization is mediated by divalent cation accommodation by negatively charged residues located at the bottom of the cjCorACD structure away from the pentamerization interface. Consistently, cjCorACD exists as a monomer irrespective of the presence of divalent cations. We concluded that cjCorACD binds divalent cations in a unique pentamerization-independent manner.

Functionalization of a cast NaAl/binary ZnO/SiO2 nanohybrid with amine and Schiff base ligands as an adsorbent of divalent cations in water system.

Casting method was used to synthesize a novel sodium alginate nanohybrid functionalized with aminated ZnO/SiO2 Schiff base for adsorption of nickel (Ni2+) and copper (Cu2+) divalent cations in single and binary water systems. The cast Schiff base nanohybrids were investigated using FESEM, XRD, BET, FTIR, TGA, and XPS analyses. The influence of unfunctionalized binary ZnO/SiO2 nano oxides and aminated Schiff base ligands formed by the reaction between salicylaldehyde and O-phenylenediamine on the adsorption of Ni2+ and Cu2+ cations was evaluated. The results confirmed that the aminated Schiff base ligands led to a higher adsorption ability of the cast nanohybrids containing interaction of divalent cations with nitrogen and oxygen atoms, as well as carboxyl and hydroxyl groups. The adsorption kinetics and isotherm for both cations followed a double-exponential model and the Redlich-Peterson model, respectively. The maximum monolayer capacity was found to be 249.8 mg/g for Cu2+ cation and 96.4 mg/g for Ni2+ cation. Thermodynamic analysis revealed an endothermic and spontaneous adsorption process with an increase in entropy. Furthermore, the synthesized Schiff base adsorbent could be easily reused over five times. The simultaneous adsorption in binary system exhibited a higher adsorption selectivity of the cast Schiff base nanohybrid for Cu2+ cation compared to Ni2+ cation. It was found that the removal percentages of Cu2+ and Ni2+ from industrial electroplating wastewater were 91.3 and 64.5%, respectively. Lastly, cost analysis of the synthesized nanohybrid was investigated.

Decoding the divalent cation effect on sulfidation of zero-valent iron: Phase evolution and FeSx assembly.

The decontamination ability of sulfidated zero-valent iron (S-ZVI) can be enhanced by the effective assembly of iron sulfides (FeSx) on neglected heterogeneous surfaces by liquid-phase precipitation. However, S-ZVI preparation with the usual pickling is detrimental to orderly interfacial assembly and leads to an imbalance between electron transfer optimization and electron storage. In this work, S-ZVI was prepared in solutions containing trace divalent cation, and it removed Cr(VI) up to 323.25 times higher than ZVI. This result is achieved by surface sites protonation of divalent cations regulating the phase evolution on the ZVI surface and inducing FeSx chemical assembly. Regulation of divalent cation and S(-II) content further promotes FeSx targeted assembly and reduces electron storage consumption as much as possible. The barrier for FeSx assembly is found to lie at the ZVI interface rather than in the deposition between FeSx. Chemical assembly at heterogeneous interfaces is a prerequisite for the ordered assembly of FeSx. In addition, S-ZVI prepared in simulated groundwater showed extensive preparation pH and universality for remediation scenarios. These findings provide new insights into the development of in-situ sulfidation mechanisms with particular implications for S-ZVI applied to soil and groundwater remediation by the regulation of heterogeneous interfacial assembly.

Klebsiella pneumoniae DedA family proteins have redundant roles in divalent cation homeostasis and resistance to phagocytosis.

The DedA superfamily is a highly conserved family of membrane proteins. Deletion of Escherichia coli yqjA and yghB, encoding related DedA family proteins, results in sensitivity to elevated temperature, antibiotics, and alkaline pH. The human pathogen Klebsiella pneumoniae possesses genes encoding DedA family proteins with >90% amino acid identity to E. coli YqjA and YghB. We hypothesized that the deletion of K. pneumoniae yqjA and yghB will impact its physiology and may reduce its virulence. The K. pneumoniae ΔyqjA ΔyghB mutant (strain VT101) displayed a growth defect at 42°C and alkaline pH sensitivity, not unlike its E. coli counterpart. However, VT101 retained mostly wild-type resistance to antibiotics. We found VT101 was sensitive to the chelating agent EDTA, the anionic detergent SDS, and agents capable of alkalizing the bacterial cytoplasm such as bicarbonate or chloroquine. We could restore growth at alkaline pH and at elevated temperature by addition of 0.5-2 mM Ca2+ or Mg2+ to the culture media. VT101 displayed a slower uptake of calcium, which was dependent upon calcium channel activity. VT201, with similar deletions as VT101 but derived from a virulent K. pneumoniae strain, was highly susceptible to phagocytosis by alveolar macrophages and displayed a defect in the production of capsule. These findings suggest divalent cation homeostasis and virulence are interlinked by common functions of the DedA family.IMPORTANCEKlebsiella pneumoniae is a dangerous human pathogen. The DedA protein family is found in all bacteria and is a membrane transporter often required for virulence and antibiotic resistance. K. pneumoniae possesses homologs of E. coli YqjA and YghB, with 60% amino acid identity and redundant functions, which we have previously shown to be required for tolerance to biocides and alkaline pH. A K. pneumoniae strain lacking yqjA and yghB was found to be sensitive to alkaline pH, elevated temperature, and EDTA/SDS and displayed a defect in calcium uptake. Sensitivity to these conditions was reversed by addition of calcium or magnesium to the growth medium. Introduction of ΔyqjA and ΔyghB mutations into virulent K. pneumoniae resulted in the loss of capsule, increased phagocytosis by macrophages, and a partial loss of virulence. These results show that targeting the Klebsiella DedA family results in impaired divalent cation transport and, in turn, loss of virulence.

Electrochemical Aptamer-Based Biosensors for Measurements in Undiluted Human Saliva.

Although blood remains a gold standard diagnostic fluid for most health exams, it involves an unpleasant and relatively invasive sampling procedure (finger pricking or venous draw). Saliva contains many relevant and useful biomarkers for diagnostic purposes, and its collection, in contrast, is noninvasive and can be obtained with minimal effort. Current saliva analyses are, however, achieved using chromatography or lateral flow assays, which, despite their high accuracy and sensitivity, can demand expensive laboratory-based instruments operated by trained personnel or offer only semiquantitative results. In response, we investigated electrochemical aptamer-based (E-AB) biosensors, a reagentless sensing platform, to allow for continuous and real-time measurements directly in undiluted, unstimulated human whole saliva. As a proof-of-concept study, we developed E-AB biosensors capable of detecting low-molecular-weight analytes (glucose and adenosine monophosphate (AMP)). To our knowledge, we report the first E-AB sensor for glucose, an approach that is inherently independent of its chemical reactivity in contrast to home glucometers. For these three sensors, we evaluated their figures of merits, stability, and reusability over short- and long-term exposure directly in saliva. In doing so, we found that E-AB sensors allow rapid and convenient molecular measurements in whole saliva with unprecedented sensitivities in the pico- to nanomolar regime and could be regenerated and reused up to 7 days when washed and stored in phosphate-buffered saline at room temperature. We envision that salivary molecular measurements using E-AB sensors are a promising alternative to invasive techniques and can be used for improved point-of-care clinical diagnosis and at-home measurements.

Collagen IV of basement membranes: IV. Adaptive mechanism of collagen IV scaffold assembly in Drosophila.

Collagen IV is an essential structural protein in all metazoans. It provides a scaffold for the assembly of basement membranes, a specialized form of extracellular matrix, which anchors and signals cells and provides microscale tensile strength. Defective scaffolds cause basement membrane destabilization and tissue dysfunction. Scaffolds are composed of α-chains that coassemble into triple-helical protomers of distinct chain compositions, which in turn oligomerize into supramolecular scaffolds. Chloride ions mediate the oligomerization via NC1 trimeric domains, forming an NC1 hexamer at the protomer-protomer interface. The chloride concentration-"chloride pressure"-on the outside of cells is a primordial innovation that drives the assembly and dynamic stabilization of collagen IV scaffolds. However, a Cl-independent mechanism is operative in Ctenophora, Ecdysozoa, and Rotifera, which suggests evolutionary adaptations to environmental or tissue conditions. An understanding of these exceptions, such as the example of Drosophila, could shed light on the fundamentals of how NC1 trimers direct the oligomerization of protomers into scaffolds. Here, we investigated the NC1 assembly of Drosophila. We solved the crystal structure of the NC1 hexamer, determined the chain composition of protomers, and found that Drosophila adapted an evolutionarily unique mechanism of scaffold assembly that requires divalent cations. By studying the Drosophila case we highlighted the mechanistic role of chloride pressure for maintaining functionality of the NC1 domain in humans. Moreover, we discovered that the NC1 trimers encode information for homing protomers to distant tissue locations, providing clues for the development of protein replacement therapy for collagen IV genetic diseases.

Role of Lipids and Divalent Cations in Membrane Fusion Mediated by the Heptad Repeat Domain 1 of Mitofusin.

Mitochondria are highly dynamic organelles that constantly undergo fusion and fission events to maintain their shape, distribution and cellular function. Mitofusin 1 and 2 proteins are two dynamin-like GTPases involved in the fusion of outer mitochondrial membranes (OMM). Mitofusins are anchored to the OMM through their transmembrane domain and possess two heptad repeat domains (HR1 and HR2) in addition to their N-terminal GTPase domain. The HR1 domain was found to induce fusion via its amphipathic helix, which interacts with the lipid bilayer structure. The lipid composition of mitochondrial membranes can also impact fusion. However, the precise mode of action of lipids in mitochondrial fusion is not fully understood. In this study, we examined the role of the mitochondrial lipids phosphatidylethanolamine (PE), cardiolipin (CL) and phosphatidic acid (PA) in membrane fusion induced by the HR1 domain, both in the presence and absence of divalent cations (Ca2+ or Mg2+). Our results showed that PE, as well as PA in the presence of Ca2+, effectively stimulated HR1-mediated fusion, while CL had a slight inhibitory effect. By considering the biophysical properties of these lipids in the absence or presence of divalent cations, we inferred that the interplay between divalent cations and specific cone-shaped lipids creates regions with packing defects in the membrane, which provides a favorable environment for the amphipathic helix of HR1 to bind to the membrane and initiate fusion.

Amyloid-ß Tetramers and Divalent Cations at the Membrane/Water Interface: Simple Models Support a Functional Role.

Charge polarization at the membrane interface is a fundamental process in biology. Despite the lower concentration compared to the abundant monovalent ions, the relative abundance of divalent cations (Ca2+, Mg2+, Zn2+, Fe2+, Cu2+) in particular spaces, such as the neuron synapse, raised many questions on the possible effects of free multivalent ions and of the required protection of membranes by the eventual defects caused by the free forms of the cations. In this work, we first applied a recent realistic model of divalent cations to a well-investigated model of a polar lipid bilayer, di-myristoyl phosphatidyl choline (DMPC). The full atomistic model allows a fairly good description of changes in the hydration of charged and polar groups upon the association of cations to lipid atoms. The lipid-bound configurations were analyzed in detail. In parallel, amyloid-ß 1-42 (Aß42) peptides assembled into tetramers were modeled at the surface of the same bilayer. Two of the protein tetramers' models were loaded with four Cu2+ ions, the latter bound as in DMPC-free Aß42 oligomers. The two Cu-bound models differ in the binding topology: one with each Cu ion binding each of the monomers in the tetramer; one with pairs of Cu ions linking two monomers into dimers, forming tetramers as dimers of dimers. The models here described provide hints on the possible role of Cu ions in synaptic plasticity and of Aß42 oligomers in storing the same ions away from lipids. The release of structurally disordered peptides in the synapse can be a mechanism to recover ion homeostasis and lipid membranes from changes in the divalent cation concentration.

Direct Effects of Toxic Divalent Cations on Contractile Proteins with Implications for the Heart: Unraveling Mechanisms of Dysfunction.

The binding of calcium and magnesium ions to proteins is crucial for regulating heart contraction. However, other divalent cations, including xenobiotics, can accumulate in the myocardium and enter cardiomyocytes, where they can bind to proteins. In this article, we summarized the impact of these cations on myosin ATPase activity and EF-hand proteins, with special attention given to toxic cations. Optimal binding to EF-hand proteins occurs at an ionic radius close to that of Mg2+ and Ca2+. In skeletal Troponin C, Cd2+, Sr2+, Pb2+, Mn2+, Co2+, Ni2+, Ba2+, Mg2+, Zn2+, and trivalent lanthanides can substitute for Ca2+. As myosin ATPase is not a specific MgATPase, Ca2+, Fe2+, Mn2+, Ni2+, and Sr2+ could support myosin ATPase activity. On the other hand, Zn2+ and Cu2 significantly inhibit ATPase activity. The affinity to various divalent cations depends on certain proteins or their isoforms and can alter with amino acid substitution and post-translational modification. Cardiac EF-hand proteins and the myosin ATP-binding pocket are potential molecular targets for toxic cations, which could significantly alter the mechanical characteristics of the heart muscle at the molecular level.

Zn2+ is Essential for Ca2+ Oscillations in Mouse Eggs.

Changes in the intracellular concentration of free calcium (Ca2+) underpin egg activation and initiation of development in animals and plants. In mammals, the Ca2+ release is periodical, known as Ca2+ oscillations, and mediated by the type 1 inositol 1,4,5-trisphosphate receptor (IP3R1). Another divalent cation, zinc (Zn2+), increases exponentially during oocyte maturation and is vital for meiotic transitions, arrests, and polyspermy prevention. It is unknown if these pivotal cations interplay during fertilization. Here, using mouse eggs, we showed that basal concentrations of labile Zn2+ are indispensable for sperm-initiated Ca2+ oscillations because Zn2+-deficient conditions induced by cell-permeable chelators abrogated Ca2+ responses evoked by fertilization and other physiological and pharmacological agonists. We also found that chemically- or genetically generated eggs with lower levels of labile Zn2+ displayed reduced IP3R1 sensitivity and diminished ER Ca2+ leak despite the stable content of the stores and IP3R1 mass. Resupplying Zn2+ restarted Ca2+ oscillations, but excessive Zn2+ prevented and terminated them, hindering IP3R1 responsiveness. The findings suggest that a window of Zn2+ concentrations is required for Ca2+ responses and IP3R1 function in eggs, ensuring optimal response to fertilization and egg activation.

Divalent Cation Signaling in Clostridium perfringens Spore Germination.

Spore germination plays an essential role in the pathogenesis of Clostridium perfringens-associated food poisoning. Germination is initiated when bacterial spores sense various stimuli, including chemicals and enzymes. A previous study showed that dipicolinic acid (DPA) chelated with calcium (Ca-DPA) significantly stimulated spore germination in C. perfringens. However, whether Ca2+ or DPA alone can induce germination is unknown. Therefore, we aimed to evaluate the possible roles of Ca2+ and other divalent cations present in the spore core, such as Mn2+ and Mg2+, in C. perfringens spore germination. Our study demonstrated that (i) Ca-DPA, but not DPA alone, induced C. perfringens spore germination, suggesting that Ca2+ might play a signaling role; (ii) all tested calcium salts induced spore germination, indicating that Ca2+ is critical for germination; (iii) the spore-specific divalent cations Mn2+ and Mg2+, but not Zn2+, induced spore germination, suggesting that spore core-specific divalent cations are involved in C. perfringens spore germination; and (iv) endogenous Ca2+ and Mg2+ are not required for induction of C. perfringens spore germination, whereas exogenous and partly endogenous Mn2+ are required. Collectively, our results suggest that exogenous spore core-specific divalent cation signals are more important than endogenous signals for the induction of spore germination.

Novel Chimeric Amino Acid-Fatty Alcohol Ester Amphiphiles Self-Assemble into Stable Primitive Membranes in Diverse Geological Settings.

Primitive cells are believed to have been self-assembled vesicular structures with minimal metabolic components, that were capable of self-maintenance and self-propagation in early Earth geological settings. The coevolution and self-assembly of biomolecules, such as amphiphiles, peptides, and nucleic acids, or their precursors, were essential for protocell emergence. Here, we present a novel class of amphiphiles-amino acid-fatty alcohol esters-that self-assemble into stable primitive membrane compartments under a wide range of geochemical conditions. Glycine n-octyl ester (GOE) and isoleucine n-octyl ester (IOE), the condensation ester products of glycine or isoleucine with octanol (OcOH), are expected to form at a mild temperature by wet-dry cycles. The GOE forms micelles in acidic aqueous solutions (pH 2-7) and vesicles at intermediate pH (pH 7.3-8.2). When mixed with cosurfactants (octanoic acid [OcA]; OcOH, or decanol) in different mole fractions [XCosurfactant = 0.1-0.5], the vesicle stability range expands significantly to span the extremely acidic to mildly alkaline (pH 2-8) and extremely alkaline (pH 10-11) regions. Only a small mole fraction of cosurfactant [XCosurfactant = 0.1] is needed to make stable vesicular structures. Notably, these GOE-based vesicles are also stable in the presence of high concentrations of divalent cations, even at low pHs and in simulated Hadean seawater composition (without sulfate). To better understand the self-assembly behavior of GOE-based systems, we devised complementary molecular dynamics computer simulations for a series of mixed GOE/OcA systems under simulated acidic pHs. The resulting calculated critical packing parameter values and self-assembly behavior were consistent with our experimental findings. The IOE is expected to show similar self-assembly behavior. Thus, amino acid-fatty alcohol esters, a novel chimeric amphiphile class composed of an amino acid head group and a fatty alcohol tail, may have aided in building protocell membranes, which were stable in a wide variety of geochemical circumstances and were conducive to supporting replication and self-maintenance. The present work contributes to our body of work supporting our hypothesis for synergism and coevolution of (proto)biomolecules on early Earth.

Effect of divalent cations on electrochemiluminescence of metal-organic frameworks in bioassay.

The structural characteristics of electrochemiluminescent (ECL) microreticula enabled flexible designs for probing specific molecules. However, bioanalysts paid little attention to the impact of concomitant electrolytic carriers on ECL responsiveness of these grids. Our previous finding confirmed the collisional quenching of ECL radiative secondary building units from polarized Br- and I-. To further address this concern, herein typical cationic commonplaces including Na+, K+, Ca2+, … in buffer plus regular transition metals - their influences upon the ECL performance of a well-defined zinc porphyrin-organic framework (ZnPOF) were inspected in a one-by-one manner. Except for Na+/K+, a dozen of divalent metal chlorides exerted an adverse effect in the form of Stern-Volmer quenching on the ECL brightness, which was illuminated to be cation channeling in open voids of ZnPOFs and bonding with O2-reactive sites as exemplified by the model Ca2+ via systematic compositional investigation. Following this principle, a simplistic Ca2+-sensitive sensor was developed for quantitative evaluation of health-care calcium supplements with high precision. Above all, this work highlighted the non-negligible interference from those Mn + requisites to the susceptible MOF-based ECL, which should be paid extra attention in bioassays and mechanistic analyses.

The influence of pH and monovalent/divalent cations on the structural and physicochemical properties of myofibrillar protein from silver carp.

The present study aimed to investigate effects of pH and monovalent (Na+ and K+)/divalent (Ca2+ and Mg2+) cations on the structural and physicochemical properties of myofibrillar protein (MP) from silver carp. MP treated with divalent cation had lesser change for the structure than that treated with monovalent cation. Ca2+-ATPase activity of MP treated with monovalent cation was increased firstly and then decreased, while that treated with divalent cation was decreased with increasing ionic strength. Surface hydrophobicity and Z-average of MP treated with divalent cations was lower than that with monovalent cations, while they decreased and then increased with the pH shifting from 3.0 to 9.0. Zeta potential of MP was increased and then decreased with increasing the pH but decreased and then increased with increasing ionic strength. In general, the pH and monovalent/divalent cations could cause various hydrophobic and electrostatic interactions, resulting in changes of the physicochemical properties of MP.

Formation of Organic Fouling during Membrane Desalination: The Effect of Divalent Cations and the Use of an Online Visual Monitoring Method.

Reverse osmosis (RO) is the most popular technology for brackish, seawater and wastewater desalination. An important drawback of RO is membrane fouling, which reduces filtration effectiveness and increase the cost of produced water. This study addresses two important topics of membrane fouling: (i) the impact of different divalent ions on the formation of organic fouling and (ii) online monitoring and prediction of fouling formation. In the absence of divalent ions, dissolved organic matter had little effect on fouling formation, even at 3.5 mgC/L, in the upper range of groundwater concentration. Calcium, strontium and iron enhanced (organic) fouling formation, whereas barium had negligible effect. However, while iron affected fouling throughout the entire tested range (0-0.5 mg/L), calcium and strontium enhanced organic fouling only at high concentrations: more than 140 mg/L and 10 mg/L for calcium and strontium, respectively. An online system was developed for monitoring the formation of organic fouling, consisting of (i) an ex-situ RO cell with a transparent cover, (ii) a video camera continually monitoring the surface of the membrane and (iii) an algorithm which automatically identified changes in the color of the membrane caused by fouling, using a specially designed membrane spacer with colored reference dots. Changes in the color of the membrane surface were normalized to the reference colors, to eliminate all non-fouling related interference. The system was used to record and analyze changes in membrane color during numerous filtration tests. The data was successfully correlated to changes in specific flux (and subsequently to fouling formation rate) and can be applied to monitor and predict the formation of membrane fouling during desalination.

Manipulating polydispersity of lens ß-crystallins using divalent cations demonstrates evidence of calcium regulation.

Crystallins comprise the protein-rich tissue of the eye lens. Of the three most common vertebrate subtypes, ß-crystallins exhibit the widest degree of polydispersity due to their complex multimerization properties in situ. While polydispersity enables precise packing densities across the concentration gradient of the lens for vision, it is unclear why there is such a high degree of structural complexity within the ß-crystallin subtype and what the role of this feature is in the lens. To investigate this, we first characterized ß-crystallin polydispersity and then established a method to dynamically disrupt it in a process that is dependent on isoform composition and the presence of divalent cationic salts (CaCl2 or MgCl2). We used size-exclusion chromatography together with dynamic light scattering and mass spectrometry to show how high concentrations of divalent cations dissociate ß-crystallin oligomers, reduce polydispersity, and shift the overall protein surface charge-properties that can be reversed when salts are removed. While the direct, physiological relevance of these divalent cations in the lens is still under investigation, our results support that specific isoforms of ß-crystallin modulate polydispersity through multiple chemical equilibria and that this native state is disrupted by cation binding. This dynamic process may be essential to facilitating the molecular packing and optical function of the lens.

Response of anaerobic digestion of waste activated sludge to types of alkalis: Contribution identification of metal ions.

Alkaline pretreatment is one promising strategy for promoting anaerobic digestion of waste activated sludge (WAS). This study selected three types of alkalis with monovalent (NaOH and KOH), divalent (Ca(OH)2 and Mg(OH)2), and trivalent (Fe(OH)3 and Al(OH)3) cations to reveal the roles of metal ions on short chain fatty acids (SCFAs) production. The enhanced production potentials of SCFAs were reduced by order of alkalis with monovalent, divalent, and trivalent cations. Na+, K+, Ca2+, and Mg2+ did no contributions on SCFAs production, while Fe3+ and Al3+ performed better than control, especially the latter. The mechanism analysis proved that Na+, K+, Ca2+, and Mg2+ did no significant effects on solubilization, hydrolysis, acidification and methanogenesis stages, while the first three stages were improved by Fe3+ and Al3+ and the methanogenesis stage was inhibited. The findings may provide some new insights when using alkalis or residual metal ions to improve anaerobic digestion of WAS.

Complex effects of Mg-biomaterials on the osteoblast cell machinery: A proteomic study.

The cell-biomaterial interface is highly complex; thousands of molecules and many processes participate in its formation. Growing demand for improved biomaterials has highlighted the need to understand the structure and functions of this interface. Proteomic methods offer a viable alternative to the traditional in vitro techniques for analyzing such systems. Magnesium is a promoter of cell adhesion and osteogenesis. Here, we used the LC-MS/MS to compare the protein expression profiles of human osteoblasts (HOb) exposed to sol-gel coatings without (MT) and with Mg (MT1.5Mg) for 1, 3, and 7 days. PANTHER, DAVID, and IPA databases were employed for protein identification and data analysis. Confocal microscopy and gene expression analysis were used for further characterization. Exposure to MT1.5Mg increased the HOb cell area and the expression of SP7, RUNX2, IBP3, COL3A1, MXRA8, and FBN1 genes. Proteomic analysis showed that MT1.5Mg affected the early osteoblast maturation (PI3/AKT, mTOR, ERK/MAPK), insulin metabolism, cell adhesion (integrin, FAK, actin cytoskeleton regulation) and oxidative stress pathways. Thus, the effects of Mg on cell adhesion and osteogenesis are rather complex, affecting several pathways rather than single processes. Our analysis also confirms the potential of proteomics in biomaterial characterization, showing a good correlation with in vitro results.