A green procedure for the determination of bioavailable forms of aluminum in natural waters by electrothermal atomic absorption spectroscopy combined with microextraction technique.

Aluminum is a prevalent element in nature, but bioavailable forms of aluminum are toxic to plants, animals, and humans. The present study is dedicated to the development of an ecologically friendly, fast, simple, reliable, sensitive, and accurate improved procedure for the determination of subtrace concentrations of bioavailable forms of aluminum in natural waters. The procedure includes the separation and pre-concentration of bioavailable forms of aluminum using vortex-assisted liquid-liquid microextraction (VALLME) of ionic associates with salicylaldehyde 4-picolinhydrazone (SAPH) and sodium dodecyl sulfate (DDSNa) by isoamylacetate (200 μl) and direct electrothermal atomic absorption spectroscopy (ET AAS).

The dynamic hypoosmotic response of relies on the mechanosensitive channel MscS.

adapts to osmotic down-shifts by releasing metabolites through two mechanosensitive (MS) channels, low-threshold MscS and high-threshold MscL. To investigate each channel's contribution to the osmotic response, we generated , , and double mutants in O395. We characterized their tension-dependent activation in patch-clamp, and the millisecond-scale osmolyte release kinetics using a stopped-flow light scattering technique.

Polymer-extracted structure of the mechanosensitive channel MscS reveals the role of protein-lipid interactions in the gating cycle.

Membrane protein structure determination is not only technically challenging but is further complicated by the removal or displacement of lipids, which can result in non-native conformations or a strong preference for certain states at the exclusion of others. This is especially applicable to mechanosensitive channels (MSC's) that evolved to gate in response to subtle changes in membrane tension transmitted through the lipid bilayer. MscS, a model bacterial system, is an ancestral member of the large family of MSCs found across all phyla of walled organisms.

On the lipid dependence of bacterial mechanosensitive channel gating in situ.

For bacterial mechanosensitive channels acting as turgor-adjusting osmolyte release valves, membrane tension is the primary stimulus driving opening transitions. Because tension is transmitted through the surrounding lipid bilayer, it is possible that the presence or absence of different lipid species may influence the function of these channels. In this work, we characterize the lipid dependence of chromosome-encoded MscS and MscL in E.

MscS inactivation and recovery are slow voltage-dependent processes sensitive to interactions with lipids.

Mechanosensitive channel MscS, the major bacterial osmolyte release valve, shows a characteristic adaptive behavior. With a sharp onset of activating tension the channel population readily opens, but under prolonged action of moderate tension it inactivates. The inactivated state is non-conductive and tension insensitive, which suggests that the gate becomes uncoupled from the lipid-facing domains.

Tight hydrophobic core and flexible helices yield MscL with a high tension gating threshold and a membrane area mechanical strain buffer.

The mechanosensitive (MS) channel of large conductance, MscL, is the high-tension threshold osmolyte release valve that limits turgor pressure in bacterial cells in the event of drastic hypoosmotic shock. Despite MscL from (TbMscL) being the first structurally characterized MS channel, its protective mechanism of activation at nearly-lytic tensions has not been fully understood. Here, we describe atomistic simulations of expansion and opening of wild-type (WT) TbMscL in comparison with five of its gain-of-function (GOF) mutants.

Dissipation during the Gating Cycle of the Bacterial Mechanosensitive Ion Channel Approaches the Landauer Limit.

The Landauer principle sets a thermodynamic bound of kBT ln 2 on the energetic cost of erasing each bit of information. It holds for any memory device, regardless of its physical implementation. It was recently shown that carefully built artificial devices can attain this bound.

MscS inactivation and recovery are slow voltage-dependent processes sensitive to interactions with lipids.

Mechanosensitive channel MscS, the major bacterial osmolyte release valve, shows a characteristic adaptive behavior. With a sharp onset of activating tension, the channel population readily opens, but under prolonged action of moderate near-threshold tension, it inactivates. The inactivated state is non-conductive and tension-insensitive, which suggests that the gate gets uncoupled from the lipid-facing domains.

The dynamic hypoosmotic response of Vibrio cholerae relies on the mechanosensitive channel MscS.

Like other intestinal bacteria, the facultative pathogen Vibrio cholerae adapts to a wide range of osmotic environments. Under drastic osmotic down-shifts, Vibrio avoids mechanical rupture by rapidly releasing excessive metabolites through mechanosensitive (MS) channels that belong to two major types, low-threshold MscS and high-threshold MscL. To investigate each channel individual contribution to V.

Mechanosensitive channel MscS is critical for termination of the bacterial hypoosmotic permeability response.

Free-living microorganisms are subjected to drastic changes in osmolarity. To avoid lysis under sudden osmotic down-shock, bacteria quickly expel small metabolites through the tension-activated channels MscL, MscS, and MscK. We examined five chromosomal knockout strains, ∆mscL, ∆mscS, a double knockout ∆mscS ∆mscK, and a triple knockout ∆mscL ∆mscS ∆mscK, in comparison to the wild-type parental strain.

Mechanosensitive channel MscS is critical for termination of the bacterial hypoosmotic permeability response.

Unlabelled: Free-living microorganisms are subjected to drastic changes in osmolarity. To avoid lysis under sudden osmotic down-shock, bacteria quickly expel small metabolites through the tension-activated channels MscL, MscS, and MscK. We examined five chromosomal knockout strains, Δ , Δ , a double knockout Δ Δ , and a triple knockout Δ Δ Δ in comparison to the wild-type parental strain.

A novel mechanosensitive channel controls osmoregulation, differentiation, and infectivity in .

The causative agent of Chagas disease undergoes drastic morphological and biochemical modifications as it passes between hosts and transitions from extracellular to intracellular stages. The osmotic and mechanical aspects of these cellular transformations are not understood. Here we identify and characterize a novel mechanosensitive channel in (TcMscS) belonging to the superfamily of small-conductance mechanosensitive channels (MscS).

A skin-inspired soft material with directional mechanosensation.

Lessons about artificial sensor design may be taken from evolutionarily perfected physiological systems. Mechanosensory cells in human skin are exquisitely sensitive to gentle touch and enable us to distinguish objects of different stiffnesses and textures. These cells are embedded in soft epidermal layers of gel-like consistency.

Partitioning of Seven Different Classes of Antibiotics into LPS Monolayers Supports Three Different Permeation Mechanisms through the Outer Bacterial Membrane.

The outer membrane (OM) of Gram-negative (G-) bacteria presents a barrier for many classes of antibacterial agents. Lipopolysaccharide (LPS), present in the outer leaflet of the OM, is stabilized by divalent cations and is considered to be the major impediment for antibacterial agent permeation. However, the actual affinities of major antibiotic classes toward LPS have not yet been determined.

Mechanical Activation of MscL Revealed by a Locally Distributed Tension Molecular Dynamics Approach.

Membrane tension perceived by mechanosensitive (MS) proteins mediates cellular responses to mechanical stimuli and osmotic stresses, and it also guides multiple biological functions including cardiovascular control and development. In bacteria, MS channels function as tension-activated pores limiting excessive turgor pressure, with MS channel of large conductance (MscL) acting as an emergency release valve preventing cell lysis. Previous attempts to simulate gating transitions in MscL by either directly applying steering forces to the protein or by increasing the whole-system tension were not fully successful and often disrupted the integrity of the system.

Application of microextraction techniques for indirect spectrophotometric determination of fluorides in river waters.

The present study is dedicated to development of improved method for determination of trace amounts of fluorides in natural waters which is based on the interaction of fluorides with ion associate (IA) of Al(III), salicylic aldehyde acylhydrazones (benzhydrazone (SABH) and 4-picolinhydrazone (SAPH)) and polymethine dye Astra Phloxine FF (AP). Comparison of analytical forms [Al(SABH)]⋅AP and [Al(SAPH)]⋅AP showed that the analytical system Al(III)-SAPH-AP is more effective, namely, a higher level of preconcentration of the analytical form is ensured by and extraction equilibrium is achieved faster. Based on the study, we propose a new, fast, simple, reliable, sensitive, and accurate method of the indirect UV-Vis-spectrophotometric determination of fluorides grounded on the interaction of fluorides with IA of Al(III), SAPH and AP with the utilization of vortex-assisted liquid-liquid microextraction (VALLME).

Biologically Active Substance Content in Edible Plants of Zakarpattia and Their Elemental Composition Model.

Consumption of edible plants satisfies a significant part of human body needs in macro- and micronutrients while biologically active substances contain strong antioxidant properties and reduce the risk of a number of diseases. Balanced nutrition and design of personalized diets and treatment rely on the data on the content of macro- and micronutrients and biologically active substances. We determined polyphenol and anthocyanin content in 22 species of local edible plants using modified spectrophotometric method with Folin-Ciocalteu reagent as well as chemical elements' content in a mixture of edible plants from 13 regions using standard procedures.

Recovery of Equilibrium Free Energy from Nonequilibrium Thermodynamics with Mechanosensitive Ion Channels in E. coli.

In situ measurements of the free energy difference between the open and closed states of ion channels are challenging due to hysteresis effects and inactivation. Exploiting recent developments in statistical physics, we present a general formalism to extract the free energy difference ΔF between the closed and open states of mechanosensitive ion channels from nonequilibrium work distributions associated with the opening and closing of the channels (gating) in response to ramp stimulation protocols recorded in native patches. We show that the work distributions obtained from the gating of MscS channels in E.

Mechanism of Catalysis by l-Asparaginase.

Two bacterial type II l-asparaginases, from and , have played a critical role for more than 40 years as therapeutic agents against juvenile leukemias and lymphomas. Despite a long history of successful pharmacological applications and the apparent simplicity of the catalytic reaction, controversies still exist regarding major steps of the mechanism. In this report, we provide a detailed description of the reaction catalyzed by type II l-asparaginase (EcAII).

Differential Interactions of Piscidins with Phospholipids and Lipopolysaccharides at Membrane Interfaces.

Piscidins 1 and 3 (P1 and P3) are potent antimicrobial peptides isolated from striped bass. Their mechanism of action involves formation of amphipathic α-helices on contact with phospholipids and destabilization of the microbial cytoplasmic membrane. The peptides are active against both Gram-positive and Gram-negative bacteria, suggesting easy passage across the outer membrane.

Screening of the microelements composition of drinking well water of Transcarpathian region, Ukraine.

Background: For Transcarpathian region, with a pronounced landscape diversity of territories and significant areas of nature reserves, well water is an important source of drinking water. Screening of the microelement composition of drinking well water in Transcarpathia has not been carried out before. The microelement composition of such well water can be considered as the natural (baseline) indicator of quality.

The host-defense peptide piscidin P1 reorganizes lipid domains in membranes and decreases activation energies in mechanosensitive ion channels.

The host-defense peptide (HDP) piscidin 1 (P1), isolated from the mast cells of striped bass, has potent activities against bacteria, viruses, fungi, and cancer cells and can also modulate the activity of membrane receptors. Given its broad pharmacological potential, here we used several approaches to better understand its interactions with multicomponent bilayers representing models of bacterial (phosphatidylethanolamine (PE)/phosphatidylglycerol) and mammalian (phosphatidylcholine/cholesterol (PC/Chol)) membranes. Using solid-state NMR, we solved the structure of P1 bound to PC/Chol and compared it with that of P3, a less potent homolog.

Glutaminase Activity of L-Asparaginase Contributes to Durable Preclinical Activity against Acute Lymphoblastic Leukemia.

We and others have reported that the anticancer activity of L-asparaginase (ASNase) against asparagine synthetase (ASNS)-positive cell types requires ASNase glutaminase activity, whereas anticancer activity against ASNS-negative cell types does not. Here, we attempted to disentangle the relationship between asparagine metabolism, glutamine metabolism, and downstream pathways that modulate cell viability by testing the hypothesis that ASNase anticancer activity is based on asparagine depletion rather than glutamine depletion per se. We tested ASNase wild-type (ASNase) and its glutaminase-deficient Q59L mutant (ASNase) and found that ASNase glutaminase activity contributed to durable anticancer activity against xenografts of the ASNS-negative Sup-B15 leukemia cell line in NOD/SCID gamma mice, whereas asparaginase activity alone yielded a mere growth delay.

Isothermal Titration Calorimetry of Be with Phosphatidylserine Models Guides All-Atom Force-Field Development for Lipid-Ion Interactions.

Beryllium has multiple industrial applications but exposure to its dust during manufacturing is associated with developing chronic inflammation in lungs known as berylliosis. Besides binding to specific alleles of MHC-II, Be was recently found to compete with Ca for binding sites on phosphatidylserine-containing membranes and inhibit recognition of this lipid by phagocytes. Computational studies of possible molecular targets for this small toxic dication are impeded by the absence of a reliable force field.

The voltage-dependence of MscL has dipolar and dielectric contributions and is governed by local intramembrane electric field.

Channels without canonical voltage sensors can be modulated by voltage acting on other domains. Here we show that besides protein dipoles, pore hydration can be affected by electric fields. In patches, both WT MscL and its V23T mutant show a decrease in the tension midpoint with hyperpolarization.