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At least 163 records · Page 9

Spirochaeta americana sp. nov., a new haloalkaliphilic, obligately anaerobic spirochaete isolated from soda Mono Lake in California

A novel, obligately anaerobic, mesophilic, haloalkaliphilic spirochaete, strain ASpG1(T), was isolated from sediments of the alkaline, hypersaline Mono Lake in California, USA. Cells of the Gram-negative strain were motile and spirochaete-shaped with sizes of 0.2-0.22 x 8-18 microm. Growth of the strain was observed between 10 and 44 degrees C (optimum 37 degrees C), in 2-12% (w/v) NaCl (optimum 3% NaCl) and between pH 8 and 10.5 (optimum pH 9.5). The novel strain was strictly alkaliphilic, required high concentrations of carbonates in the medium and was capable of utilizing D-glucose, fructose, maltose, sucrose, starch and D-mannitol. End products of glucose fermentation were H2, acetate, ethanol and formate. Strain ASpG(T) was resistant to kanamycin and rifampicin, but sensitive to gentamicin, tetracycline and chloramphenicol. The G + C content of its DNA was 58.5 mol%. DNA-DNA hybridization analysis of strain ASpG1(T) with its most closely related species, Spirochaeta alkalica Z-7491(T), revealed a hybridization value of only 48.7%. On the basis of its physiological and molecular properties, strain ASpG1(T) appears to represent a novel species of the genus Spirochaeta, for which the name Spirochaeta americana is proposed (type strain ASpG1(T) =ATCC BAA-392(T) = DSM 14872(T)).

Sodium Chloride/metabolism↗

Ligand‐induced CaMKIIα hub Trp403 flip, hub domain stacking, and modulation of kinase activity

Abstract γ‐Hydroxybutyric acid (GHB) analogs are small molecules that bind competitively to a specific cavity in the oligomeric CaMKIIα hub domain. Binding affects conformation and stability of the hub domain, which may explain the neuroprotective action of some of these compounds. Here, we describe molecular details of interaction of the larger‐type GHB analog 2‐(6‐(4‐chlorophenyl)imidazo[1,2‐b]pyridazine‐2‐yl)acetic acid (PIPA). Like smaller‐type analogs, PIPA binding to the CaMKIIα hub domain promoted thermal stability. PIPA additionally modulated CaMKIIα activity under sub‐maximal CaM concentrations and ultimately led to reduced substrate phosphorylation. A high‐resolution X‐ray crystal structure of a stabilized CaMKIIα (6x mutant) hub construct revealed details of the binding mode of PIPA, which involved outward placement of tryptophan 403 (Trp403), a central residue in a flexible loop close to the upper hub cavity. Small‐angle X‐ray scattering (SAXS) solution structures and mass photometry of the CaMKIIα wild‐type hub domain in the presence of PIPA revealed a high degree of ordered self‐association (stacks of CaMKIIα hub domains). This stacking neither occurred with the smaller compound 3‐hydroxycyclopent‐1‐enecarboxylic acid (HOCPCA), nor when Trp403 was replaced with leucine (W403L). Additionally, CaMKIIα W403L hub was stabilized to a larger extent by PIPA compared to CaMKIIα hub wild type, indicating that loop flexibility is important for holoenzyme stability. Thus, we propose that ligand‐induced outward placement of Trp403 by PIPA, which promotes an unforeseen mechanism of hub domain stacking, may be involved in the observed reduction in CaMKIIα kinase activity. Altogether, this sheds new light on allosteric regulation of CaMKIIα activity via the hub domain.

Biochemistry & Molecular Biology↗

Thermodynamic, Kinetic, and Transport Contributions to Hydrogen Evolution Activity and Electrolyte-Stability Windows for Water-in-Salt Electrolytes

Concentrated water-in-salt electrolytes (WiSEs) are used in aqueous batteries and to control electrochemical reactions for fuel production. The hydrogen evolution reaction is a parasitic reaction at the negative electrode that limits cell voltage in WiSE batteries and leads to self-discharge, and affects selectivity for electrosynthesis. Mitigating and modulating these processes is hampered by a limited fundamental understanding of HER kinetics in WiSEs. Here, in this study, we quantitatively assess how thermodynamics, kinetics, and interface layers control the apparent HER activities in 20 m LiTFSI. When the LiTFSI concentration is increased from 1 to 20 m, an increase in proton activity causes a positive shift in the HER equilibrium potential of 71 mV. The exchange current density, i o , derived from the HER branch for 20 m LiTFSI in 98% purity (0.56 ± 0.05 μA/cm Pt 2 ), however, is 8 times lower than for 20 m LiTFSI in 99.95% (4.7 ± 0.2 μA/cm Pt 2 ) and 32 times lower than for 1 m LiTFSI in 98% purity (18 ± 1 μA/cm Pt 2 ), demonstrating that the WiSE's impurities and concentration are both central in significantly suppressing HER kinetics. The ability and applicability of the reported methods are extended by examining additional WiSEs formulations made of acetates and nitrates.

25 ENERGY STORAGE↗

Pineal physiology in microgravity - Relation to rat gonadal function aboard Cosmos 1887

Results are reported from an analysis of pineal glands obtained for five male rats flown aboard an orbiting satellite for their melatonin, serotonin (5-HT), 5-hydroxyindole acetic acid (5-HIA), and calcium content. Plasma 5-HT and 5-HIAA were measured. These parameters were compared to indicators of gonadal function: plasma testosterone concentration and spermatogonia development. Plasma melotonin was found to be low at the time of euthanasia and was not different among the experimental groups. Pineal calcium of flight animals was not different from ground controls. Pineal 5-HT and 5-HIAA in the flight group were significantly higher than those in ground controls. These findings suggest a possible increase in pineal 5-HT turnover in flight animals which may result in increased melatonin secretion. It is argued that the alteration of pinal 5-HT turnover and its expected effects on melatonin secretion may partially explain the lower plasma testosterone levels and 4-11 percent fewer spermatogonia cells observed in flight animals.

Holley, Daniel C.↗

Fluorescence Studies of Lysozyme Nucleation

Fluorescence is one of the most powerful tools available for the study of macromolecules. For example, fluorescence can be used to study self association through methods such as anisotropy (the rotational rate of the molecule in solution), quenching (the accessibility of a bound probe to the bulk solution), and resonance energy transfer (measurement of the distance between two species). Fluorescence can also be used to study the local environment of the probe molecules, and the changes in that environment which accompany crystal nucleation and growth. However fluorescent techniques have been very much underutilized in macromolecular growth studies. One major advantage is that the fluorescent species generally must be at low concentration, typically ca 10-5 to 10-6 M. Thus one can study a very wide range of solution conditions, ranging from very high to very low protein concentration, he latter of which are not readily accessible to scattering techniques. We have prepared a number of fluorescent derivatives of chicken egg white lysozyme (CEWL). Fluorescent probes have been attached to two different sites, ASP 101 and the N-terrninal amine, with a sought for use in different lines of study. Preliminary resonance energy transfer studies have been -carried out using pyrene acetic acid (Ex 340 mn, Em 376 nm) lysozyme as a donor and cascade blue (Ex 377 run, Em 423 nm) labeled lysozyme as an acceptor. The emission of both the pyrene and cascade blue probes was followed as a function of the salt protein concentrations. The data show an increase in cascade blue and a concomitant decrease in the pyrene fluorescence as either the salt or protein concentrations are increased, suggesting that the two species are approaching each other close enough for resonance energy transfer to occur. This data can be analyzed to measure the distance between the probe molecules and, knowing their locations on the protein molecule their distances from and orientations with respect to each other. The results of these and other studies will be discussed.

Pusey, Marc L.↗

Chemokinetic motility responses of the cyanobacterium oscillatoria terebriformis

Oscillatoria terebriformis, a gliding, filamentous, thermophilic cyanobacterium, exhibited an inhibition of gliding motility upon exposure to fructose. The observed response was transient, and the duration of nonmotility was directly proportional to the concentration of fructose. Upon resumption of motility, the rate of motility was also inversely proportional to the concentration of fructose. Sulfide caused a similar response. The effect of sulfide was specific and not due to either anoxia or negative redox potential. Exposure to glucose, acetate, lactate, or mat interstitial water did not elicit any motility response.

Richardson, Laurie L.↗

Response of Two Legumes to Two Ultraviolet-B Radiation Regimes

Depletion of the stratospheric ozone layer has been directly linked to increased levels of UV radiation at the earth's surface. The purpose of this study was to evaluate the responses of soybean (Glycine max) and alfalfa (Medicago sativa) to increased UV-B radiation (280-320 nm). Soybean and alfalfa were grown successively in a growth chamber that provided UV-B intensities 45% above nominal summer field levels. Mylar-D (UVB opaque) and mono-acetate (UV-B transparent) films were used to establish the two UV-B treatments. Soybean grown under increased UV showed 21% smaller internodal lengths and higher concentrations of UV-B absorbing pigments (i.e. flavonoids) compared to plants grown under no UV. Significant results for alfalfa included 22% greater leaf flavonoid concentration under increased UV, 14% greater leaf chlorophyll concentration under no UV, and 32% greater above-ground biomass with no UV. These leguminous species possess mechanisms that protect against UV-B damage as indicated by increases in foliar concentrations of UV-B absorbing compounds. Alfalfa appears to be more sensitive to UV-B damage than soybean. Remote sensing of chlorophyll fluorescence may offer a means of monitoring UV-induced plant stress and damage.

Levy, Daniel L.↗

Modeling of photodegradation in solar cell modules of substrate and superstrate design made with ethylene-vinyl acetate as pottant material

The rates of hydrogen abstraction by peroxy radicals were proven to be too slow for significant oxidation of the alkane substrate to be important. The numerical procedure, independent of our particular data base was verified by reproducing concentration time profiles for a model reaction set describing the cesium flare system in the upper atmosphere. Simulation was identical to that given in the literature. Experimental verification of the data base is to be attempted by weatherometry studies in the coming year. Work on the new diagnostic techniques was completed. The adapted automated viscometer was demonstrated to be an efficient and reliable tool for routine measurements of viscosity (molecular weight) changes in solid samples after batch solutions have been made up. The laser photolysis GC method for monitoring extremely low levels of oxidation in polymers proved to be impractical because the yields of carbon monoxide were too low for quantification. Much progress was made with the computer model. The reaction matrix was completely revised, resulting in a new scheme of 31 reactions and time, lifetimes in excess of ten years. The results to date lead us to some tentative observations.

Somersall, A. C.↗

Thermophysical properties of lysozyme (protein) solutions

Thermophysical properties of protein solutions composed of the lysozyme crystals with a 0.1 M sodium acetate and 5 percent NaCl solution as the buffer (pH = 4.0) are determined. The properties being measured include specific heat, thermal conductivity, dynamic viscosity, and surface tension. The protein concentrations are varied. Thermal diffusivity is calculated using the measured results. The purpose of the research is to measure thermophysical properties of lysozyme solutions which would serve as the data bank for controlling and modeling the crystal growth process on earth as well as in space.

Liu, Jiaching↗

Development of a sparging technique for volatile emissions from potato (Solanum tuberosum)

Accumulation of volatile emissions from plants grown in tightly closed growth chambers may have allelopathic or phytotoxic properties. Whole air analysis of a closed chamber includes both biotic and abiotic volatile emissions. A method for characterization and quantification of biogenic emissions solely from plantlets was developed to investigate this complex mixture of volatile organic compounds. Volatile organic compounds from potato (Solanum tuberosum L. cv. Norland) were isolated, separated and identified using an in-line configuration consisting of a purge and trap concentrator with sparging vessels coupled to a GC/MS system. Analyses identified plant volatile compounds: transcaryophyllene, alpha-humulene, thiobismethane, hexanal, cis-3-hexen-1-ol, and cis-3-hexenyl acetate.

Berdis, Elizabeth↗

Spirochaeta Americana sp. Nov., A New Haloalkaliphilic, Obligately Anaerobic Spirochaete Isolated from Soda Mona Lake in California

A novel, obligately anaerobic, mesophilic, haloalkaliphilic spirochaete, strain ASpG1(sup T), was isolated from sediments of the alkaline, hypersaline Mono Lake in California, USA. Cells of the Gram-negative strain were motile and spirochaete-shaped with sizes of 0.2-0.22 x 8-18 microns. Growth of the strain was observed between 10 and 44 C (optimum 37 C), in 2-12% (w/v) NaCl (optimum 3 % NaCl) and between pH 8 and 10.5 (optimum pH 9.5). The novel strain was strictly alkaliphilic, required high concentrations of carbonates in the medium and was capable of utilizing D-glucose, fructose, maltose, sucrose, starch and D-mannitol. End products of glucose fermentation were H2, acetate, ethanol and formate. Strain ASpG1(sup T) was resistant to kanamycin and rifampicin, but sensitive to gentamicin, tetracycline and chloramphenicol. The G + C content of its DNA was 58.5 mol%. DNA-DNA hybridization analysis of strain ASpG1(sup T) with its most closely related species, Spirochaeta alkalica Z-7491(sup T) revealed a hybridization value of only 48.7 %. On the basis of its physiological and molecular properties, strain ASpG1(sup T) appears to represent a novel species of the genus Spirochaeta, for which the name Spirochaeta americana is proposed (type strain ASpG1(sup T) = ATCC 13AA-392(sup T) = DSM 14872(sup T)).

Hoover, Richard B.↗

Minimizing Oxygen Permeability in Chitin/Cellulose Nanomaterial Coatings by Tuning Chitin Deacetylation

The demand for packaging materials with low gas permeabilities is increasing, but commonly used petroleum-derived single-use plastics are not renewable, biodegradable, or easy to recycle. Nanomaterials composed of chitin and cellulose, which are abundant in nature, have high crystallinities and low oxygen permeabilities (OP), providing a viable alternative. In this work, we explore how deacetylation conditions of crab shell chitin can be used to tune the charge and size of resulting chitin nanowhiskers (ChNWs) and the resulting OP of layered film structures. Three deacetylation factors, the concentration of sodium hydroxide (%NaOH), temperature (T), and reaction time, were explored using a three-factor three-level Taguchi design with an orthogonal array. Furthermore, the resulting ChNW suspensions were sequentially spray-coated with suspensions of cellulose nanocrystals (CNCs) onto cellulose acetate (CA) films to form a multilayer structure. We show that ChNWs prepared under more aggressive deacetylation conditions inside the original orthogonal array (higher %NaOH, T, and time) had shorter lengths but the surface charge was significantly influenced only by more aggressive deacetylation outside of the original design conditions. With only ~10% decrease in the ultimate tensile strength and no significant loss in failure strain, the ChNW-CNC coating resulted in ~20% decrease in WVTR in comparison to uncoated CA films. The optimization of process conditions resulted in CA-ChNW-CNC films with a 91-99% decrease in OP (132-16.7 cm 3 ∙μm/m 2 /day/kPa versus 1553 cm3∙μm/m2/day/kPa for uncoated CA).

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Spirochaeta americana sp. nov.: A New Haloalkaliphilic, Obligately Anaerobic Spirochete Isolated from Soda Mono Lake, California

A novel obligately anaerobic, mesophilic, haloalkaliphilic spirochete, strain ASpG1, was isolated from sediments of the alkaline, hypersaline Mono Lake in California, U.S.A. The gram-negative cells are motile and spirochete-shaped with sizes of 0.22 x 10-15 micron. Growth was observed over the temperature range of 10 C to 44 C (optimum 37 C), NaCl concentration range of greater than 1 - 12 % (wt/vol) (optimum 3%), and pH range 7.5 - 10.5 (optimum pH 9.5). The novel isolate is strictly alkaliphilic, requires high concentrations of carbonate in the medium, and is capable of utilizing D-glucose, fructose, maltose, sucrose, starch, and D-mannitol. Main end products of glucose fermentation are: H2, acetate, ethanol, and formate. Strain AspG1 is resistant to kanamycin, but sensitive to chloramphenicol, gentamycin and tetracycline. The G+C content of its DNA is 58.5 mol%. On the basis of its physiological and molecular properties, the isolate appears to be a novel species among the genus Spirochaeta; and the name Spirochaeta americana sp. nov., is proposed for the taxon (type strain ASpG1(sup T) = ATCC BAA_392(sup T) = DSMZ 14872(sup T)).

Hoover, Richard B.↗

Protein solubilities determined by a rapid technique and modification of that technique to a micro-method

A simple, rapid method for determination of protein solubilities has been developed which is based upon maximization of the free solution volume to be brought into equilibrium. The tetragonal lysozome solubility diagram has been determined from pH 4.0 to 5.2 (0.1 M sodium acetate), 2-7 percent NaCl, 3-25 C, and portions of the orthorhombic solubility diagram using this technique. Both tetragonal and orthorhombic solubilities were found to increase smoothly with decreasing salt concentration and increasing temperature; no retrograde solubilities were observed. Using column volumes of 75, 300, and 900 microliters, identical tetragonal lysozyme solubility diagrams were obtained. Chymotrypsinogen solubilities have also been determined using this apparatus, being retrograde over the temperature range tested. It is noted that the primary limiting factor in reducing the crystalline volume is the minimum solution sample size needed to accurately quantitate the protein.

Cacioppo, Elizabeth↗

Spirochaeta Americana Sp. Nov., A new Haloalkaliphilic, Obligately Anaerobic Spirochete Isolated from Soda Mono Lake in California

A novel obligately anaerobic, mesophilic, haloalkaliphilic spirochete, strain ASpG1(sup T), was isolated from sediments of the alkaline, hypersaline Mono Lake in California, U.S.A. The Gram-negative cells are motile and spirochete-shaped with sizes of 0.2 - 0.22 X 8-15 microns. Growth was observed over the following ranges: temperature 10 C to 44 C; optimum +37 C; NaCl concentration 2 - 12 % (w/v); optimum NaCl3 % and pH 8 - 10.5; optimum pH 9.5. The novel isolate is strictly alkaliphilic, requires high concentrations of carbonate in the medium, and is capable of utilizing D-glucose, fructose, maltose, sucrose, starch, and D-mannitol. The main end products of glucose fermentation are: H2, acetate, ethanol, and formate. Strain ASpG(sup T) is resistant to kanamycin, and rifampin, but sensitive to chloramphenicol, gentamycin and tetracycline. The G+C content of its DNA is 58.5 mol%, genome size is 2.98 x l0(exp 9) Daltons, Tm of the genomic DNA is 68 +/- 2 C, and DNA-DNA hybridization with the most closely related species, Spirocheta alkalica Strain Z-7491(sup T), exhibited 48.7% homology. On the basis of its physiological and molecular properties, the isolate appears to be a novel species of the genus Spirochaeta; and the name Spirochaeta americana sp. nov., is proposed for the taxon (type strain ASpG1(sup T) = ATCC BAA-392(sup T) = DSMZ 14872(sup T)).

Hoover, Richard B.↗

Protection of 2-(3-Thienyl)ethanol with 3-Thienylacetic Acid and Hard Cross- Linked Conducting Films by Electropolymerization of the Ester

The ester (compound 1) of 2-(3-thienyl)ethanol (T-etOH) with 3-thienylacetic acid was synthesized as a monomer whose two thiophene groups could be electropolymerized independently, becoming members of different polymer chains in a highly-crosslinked highly-insoluble polymer. Indeed, 1 was electropolymerized successfully alone and together with 3-methylthiophene (3MeT). Films of poly(1) are hard (3H, as opposed to less than 6B for poly(3MeT)), and the close proximity of the polymeric strands creates pi-stacking interactions. The behavior of 1 suggests that by: (a) limiting the potential used for the oxidation of monomeric esters of T-etOH at the foot of their oxidation waves (less than 1.8 V vs. Ag/AgCl); and, (b) compensating for the decrease in the electrogenerated radical concentration by increasing the monomer concentration, practically all esters of T-etOH should be electropolymerizable. This was confirmed by durable film formation from the archetypical ester of T-etOH, the 2-(3-thienyl)ethyl acetate (T-etOAc), whose homo-electropolymerization is reported for the first time.

Leventis, Nicholas↗

Pioneering a Biobased UAS

With the exponential growth of interest in unmanned aerial vehicles (UAVs) and their vast array of applications in both space exploration and terrestrial uses such as the delivery of medicine and monitoring the environment, the 2014 Stanford-Brown-Spelman iGEM team is pioneering the development of a fully biological UAV for scientific and humanitarian missions. The prospect of a biologically-produced UAV presents numerous advantages over the current manufacturing paradigm. First, a foundational architecture built by cells allows for construction or repair in locations where it would be difficult to bring traditional tools of production. Second, a major limitation of current research with UAVs is the size and high power consumption of analytical instruments, which require bulky electrical components and large fuselages to support their weight. By moving these functions into cells with biosensing capabilities - for example, a series of cells engineered to report GFP, green fluorescent protein, when conditions exceed a certain threshold concentration of a compound of interest, enabling their detection post-flight - these problems of scale can be avoided. To this end, we are working to engineer cells to synthesize cellulose acetate as a novel bioplastic, characterize biological methods of waterproofing the material, and program this material's systemic biodegradation. In addition, we aim to use an "amberless" system to prevent horizontal gene transfer from live cells on the material to microorganisms in the flight environment. So far, we have: successfully transformed Gluconacetobacter hansenii, a cellulose-producing bacterium, with a series of promoters to test transformation efficiency before adding the acetylation genes; isolated protein bands present in the wasp nest material; transformed the cellulose-degrading genes into Escherichia coli; and we have confirmed that the amberless construct prevents protein expression in wild-type cells. In addition, as part of our human outreach project, we have been in touch with leaders in the fields of UAVs, synthetic biology, and earth sciences, and it is clear that biodegradable UAVs could have a significant impact on the industry.

Escherichia↗

Towards a Biosynthetic UAV

We are currently working on a series of projects towards the construction of a fully biological unmanned aerial vehicle (UAV) for use in scientific and humanitarian missions. The prospect of a biologically-produced UAV presents numerous advantages over the current manufacturing paradigm. First, a foundational architecture built by cells allows for construction or repair in locations where it would be difficult to bring traditional tools of production. Second, a major limitation of current research with UAVs is the size and high power consumption of analytical instruments, which require bulky electrical components and large fuselages to support their weight. By moving these functions into cells with biosensing capabilities - for example, a series of cells engineered to report GFP, green fluorescent protein, when conditions exceed a certain threshold concentration of a compound of interest, enabling their detection post-flight - these problems of scale can be avoided. To this end, we are working to engineer cells to synthesize cellulose acetate as a novel bioplastic, characterize biological methods of waterproofing the material, and program this material's systemic biodegradation. In addition, we aim to use an "amberless" system to prevent horizontal gene transfer from live cells on the material to microorganisms in the flight environment.

Biological↗