OXPHOS capacity is diminished and the phosphorylation system inhibited during diapause in an extremophile, embryos of Artemia franciscana.

Diapause exhibited by embryos of Artemia franciscana is accompanied by severe arrest of respiration. A large fraction of this depression is attributable to downregulation of trehalose catabolism that ultimately restricts fuel to mitochondria. This study now extends knowledge on the mechanism by revealing metabolic depression is heightened by inhibitions within mitochondria.

Target enzymes are stabilized by AfrLEA6 and a gain of α-helix coincides with protection by a group 3 LEA protein during incremental drying.

Anhydrobiotic organisms accumulate late embryogenesis abundant (LEA) proteins, a family of intrinsically disordered proteins (IDPs) reported to improve cellular tolerance to water stress. Here we show that AfrLEA6, a Group 6 LEA protein only recently discovered in animals, protects lactate dehydrogenase (LDH), citrate synthase (CS) and phosphofructokinase (PFK) against damage during desiccation. In some cases, protection is enhanced by trehalose, a naturally-occurring protective solute.

Electrochemical Disinfection in Water and Wastewater Treatment: Identifying Impacts of Water Quality and Operating Conditions on Performance.

Electrochemical disinfection-a method in which chemical oxidants are generated via redox reactions on the surface of an electrode-has attracted increased attention in recent years as an alternative to traditional chemical dosing disinfection methods. Because electrochemical disinfection does not entail the transport and storage of hazardous materials and can be scaled across centralized and distributed treatment contexts, it shows promise for use both in resource limited settings and as a supplement for aging centralized systems. In this Critical Review, we explore the significance of treatment context, oxidant selection, and operating practice on electrochemical disinfection system performance.

Transgenic expression of late embryogenesis abundant proteins improves tolerance to water stress in .

Four lines of were created that expressed transgenes encoding selected late embryogenesis abundant (LEA) proteins originally identified in embryos of the anhydrobiote The overall aim was to extend our understanding of the protective properties of LEA proteins documented with isolated cells to a desiccation-sensitive organism during exposure to drying and hyperosmotic stress. Embryos of were dried at 57% relative humidity to promote a loss of 80% tissue water and then rehydrated. Embryos that expressed AfrLEA2 or AfrLEA3m eclosed 2 days earlier than wild-type embryos or embryos expressing green fluorescent protein (Gal4GFP control).

A novel group 6 LEA protein from diapause embryos of Artemia franciscana is cytoplasmically localized.

The expression of late embryogenesis abundant (LEA) proteins is one mechanism by which anhydrobiotic organisms survive periods of severe water loss. Artemia franciscana is an animal extremophile that uniquely expresses LEA proteins concurrently from Groups 1, 3, and 6. In this study we examine the subcellular localization of AfrLEA6, a Group 6 LEA protein from embryos of A.

Technoeconomic Analysis of Brackish Water Capacitive Deionization: Navigating Tradeoffs between Performance, Lifetime, and Material Costs.

Capacitive deionization (CDI), a class of electrochemical separation technologies, has been proposed as an energy-efficient brackish water desalination method. Previous studies have focused on improving capacity and energy consumption through material (e.g.

Structural properties and cellular expression of AfrLEA6, a group 6 late embryogenesis abundant protein from embryos of Artemia franciscana.

Late embryogenesis abundant (LEA) proteins are intrinsically disordered proteins (IDPs) commonly found in anhydrobiotic organisms and are frequently correlated with desiccation tolerance. Herein we report new findings on AfrLEA6, a novel group 6 LEA protein from embryos of Artemia franciscana. Assessment of secondary structure in aqueous and dried states with circular dichroism (CD) reveals 89% random coil in the aqueous state, thus supporting classification of AfrLEA6 as an IDP.

Global Sensitivity Analysis To Characterize Operational Limits and Prioritize Performance Goals of Capacitive Deionization Technologies.

Capacitive deionization (CDI) technologies couple electronic and ionic charge storage, enabling improved thermodynamic efficiency of brackish desalination by recovering energy released during discharge. However, insight into CDI has been limited by discrete experimental observations at low desalination depths (Δ c, typically reducing influent salinity by 10 mM or less). In this study, the performance and sensitivity of three common CDI configurations [standard CDI, membrane CDI (MCDI), and flowable electrode CDI (FCDI)] were evaluated across the operational and material design landscape by varying eight common input parameters (electrode thickness, influent concentration, current density, electrode flow rate, specific capacitance, contact resistance, porosity, and fixed charge).

Enhancing capacitive deionization performance with charged structural polysaccharide electrode binders.

Capacitive deionization (CDI) performance, as measured by salt adsorption capacity (SAC) and energy normalized adsorption of salt (ENAS), is frequently limited by anion repulsion at the positive electrode. In this work, we investigate the ability to prevent co-ion repulsion by increasing complementary fixed charged within the electrode macropores by binding composite CDI electrodes with the ionically charged structural polysaccharides chitosan and carboxymethyl cellulose. When employing asymmetrically charged electrode binders, co-ion repulsion was prevented, resulting in SAC and ENAS values that were three times greater than composite electrodes bound with polyvinylidene fluoride (PVDF) and similar to CDI electrodes composed of chemically modified carbon.

Challenges during diapause and anhydrobiosis: Mitochondrial bioenergetics and desiccation tolerance.

In preparation for the onset of environmental challenges like overwintering, food limitation, anoxia, or water stress, many invertebrates and certain killifish enter diapause. Diapause is a developmentally-programed dormancy characterized by suppression of development and metabolism. For embryos of Artemia franciscana (brine shrimp), the metabolic arrest is profound.

Characterizing the Impacts of Deposition Techniques on the Performance of MnO Cathodes for Sodium Electrosorption in Hybrid Capacitive Deionization.

Capacitive deionization (CDI) is currently limited by poor ion-selectivity and low salt adsorption capacity of porous carbon electrodes. To enhance selectivity and capacity via sodium insertion reactions, carbon aerogel electrodes were modified by depositing amorphous manganese dioxide layers via cyclic voltammetry (CV) and electroless deposition (ED). MnO-coated electrodes were evaluated in a hybrid capacitive deionization system to understand the relationship between oxide coating morphology, electrode capacitance, and sodium removal efficacy.

Cryopreservation of lipid bilayers by LEA proteins from Artemia franciscana and trehalose.

The capacity of Late Embryogenesis Abundant (LEA) proteins and trehalose to protect liposomes against freezing-induced damage was examined by measuring the leakage of 5(6)-carboxyfluorescein (CF). Liposomes were prepared to simulate the lipid compositions of the inner leaflet of the plasma membrane, outer mitochondrial membrane (OMM), and inner mitochondrial membrane (IMM). Two recombinant LEA proteins belonging to Group 3 (AfrLEA2 and AfrLEA3m) were expressed and purified from embryos of Artemia franciscana.

Mechanisms of animal diapause: recent developments from nematodes, crustaceans, insects, and fish.

Life cycle delays are beneficial for opportunistic species encountering suboptimal environments. Many animals display a programmed arrest of development (diapause) at some stage(s) of their development, and the diapause state may or may not be associated with some degree of metabolic depression. In this review, we will evaluate current advancements in our understanding of the mechanisms responsible for the remarkable phenotype, as well as environmental cues that signal entry and termination of the state.

Phage-based extraction of polyhydroxybutyrate (PHB) produced from synthetic crude glycerol.

Polyhydroxybutyrate (PHB), a biodegradable plastic, is an attractive alternative to traditional petrochemical-derived plastics. However, its production is expensive due to high feedstock and extraction costs. As bacteriophages are natural predators to bacteria and specific to their hosts, bacteriophages offer a new and unique means to release PHB from bacteria via cell lysis.

Liposomes with diverse compositions are protected during desiccation by LEA proteins from Artemia franciscana and trehalose.

Intracellular accumulation of Late Embryogenesis Abundant (LEA) proteins and the disaccharide trehalose is associated with cellular desiccation tolerance in a number of animal species. Two LEA proteins from anhydrobiotic embryos of the brine shrimp Artemia franciscana were tested for the ability to protect liposomes of various compositions against desiccation-induced damage in the presence and absence of trehalose. Damage was assessed by carboxyfluorescein leakage after drying and rehydration.

Developmental changes in hypoxic exposure and responses to anoxia in Drosophila melanogaster.

Holometabolous insects undergo dramatic morphological and physiological changes during ontogeny. In particular, the larvae of many holometabolous insects are specialized to feed in soil, water or dung, inside plant structures, or inside other organisms as parasites where they may commonly experience hypoxia or anoxia. In contrast, holometabolous adults usually are winged and live with access to air.

Physiological strategies during animal diapause: lessons from brine shrimp and annual killifish.

Diapause is a programmed state of developmental arrest that typically occurs as part of the natural developmental progression of organisms that inhabit seasonal environments. The brine shrimp Artemia franciscana and annual killifish Austrofundulus limnaeus share strikingly similar life histories that include embryonic diapause as a means to synchronize the growth and reproduction phases of their life history to favorable environmental conditions. In both species, respiration rate is severely depressed during diapause and thus alterations in mitochondrial physiology are a key component of the suite of characters associated with cessation of development.

Biodegradation of 1,4-dioxane: effects of enzyme inducers and trichloroethylene.

1,4-Dioxane is a groundwater contaminant and probable human carcinogen. In this study, two well-studied degradative bacteria Mycobacterium vaccae JOB5 and Rhodococcus jostii RHA1 were examined for their 1,4-dioxane degradation ability in the presence and absence of its co-contaminant, trichloroethylene (TCE), under different oxygenase-expression conditions. These two strains were precultured with R2A broth (complex nutrient medium) before supplementation with propane or 1-butanol to induce the expression of different oxygenases.

Molecular approaches for improving desiccation tolerance: insights from the brine shrimp Artemia franciscana.

We have evaluated the endogenous expression and molecular properties of selected Group 3 LEA proteins from Artemia franciscana , and the capacity of selected Groups 1 and 3 proteins transfected into various desiccation-sensitive cell lines to improve tolerance to drying. Organisms inhabiting both aquatic and terrestrial ecosystems frequently are confronted with the problem of water loss for multiple reasons--exposure to hypersalinity, evaporative water loss, and restriction of intracellular water due to freezing of extracellular fluids. Seasonal desiccation can become severe and lead to the production of tolerant propagules and entry into the state of anhydrobiosis at various stages of the life cycle.

A Raman microspectroscopy study of water and trehalose in spin-dried cells.

Long-term storage of desiccated nucleated mammalian cells at ambient temperature may be accomplished in a stable glassy state, which can be achieved by removal of water from the biological sample in the presence of glass-forming agents including trehalose. The stability of the glass may be compromised due to a nonuniform distribution of residual water and trehalose within and around the desiccated cells. Thus, quantification of water and trehalose contents at the single-cell level is critical for predicting the glass formation and stability for dry storage.

Intracellular localization of group 3 LEA proteins in embryos of Artemia franciscana.

Late embryogenesis abundant (LEA) proteins are accumulated by anhydrobiotic organisms in response to desiccation and improve survivorship during water stress. In this study we provide the first direct evidence for the subcellular localizations of AfrLEA2 and AfrLEA3m (and its subforms) in anhydrobiotic embryos of Artemia franciscana. Immunohistochemistry shows AfrLEA2 to reside in the cytoplasm and nucleus, and the four AfrLEA3m proteins to be localized to the mitochondrion.

Group 3 late embryogenesis abundant proteins from embryos of Artemia franciscana: structural properties and protective abilities during desiccation.

Group 3 late embryogenesis abundant (LEA) proteins are highly hydrophilic, and their expression is associated with desiccation tolerance in both plants and animals. Here we show that two LEA proteins from embryos of Artemia franciscana, AfrLEA2 and AfrLEA3m, are intrinsically disordered in solution but upon desiccation gain secondary structure, as measured by circular dichroism. Trifluoroethanol and sodium dodecyl sulfate are both shown to induce α-helical structure in AfrLEA2 and AfrLEA3m.

Quantification of cellular protein expression and molecular features of group 3 LEA proteins from embryos of Artemia franciscana.

Late embryogenesis abundant (LEA) proteins are highly hydrophilic, low complexity proteins whose expression has been correlated with desiccation tolerance in anhydrobiotic organisms. Here, we report the identification of three new mitochondrial LEA proteins in anhydrobiotic embryos of Artemia franciscana, AfrLEA3m_47, AfrLEA3m_43, and AfrLEA3m_29. These new isoforms are recognized by antibody raised against recombinant AfrLEA3m, the original mitochondrial-targeted LEA protein previously reported from these embryos; mass spectrometry confirms all four proteins share sequence similarity.

Metabolic downregulation and inhibition of carbohydrate catabolism during diapause in embryos of Artemia franciscana.

Diapause embryos were collected from ovigerous females of Artemia franciscana at the Great Salt Lake, Utah, and were synchronized to within 4 h of release. Respiration rate for these freshly released embryos across a subsequent 26-d time course showed a rapid decrease during the first several days followed thereafter by a much slower decline. The overall metabolic depression was estimated to be greater than 99%.