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At least 145 records · Page 8

Molecular Pathogenesis and Immune Evasion of Vesicular Stomatitis New Jersey Virus Inferred from Genes Expression Changes in Infected Porcine Macrophages

The molecular mechanisms associated with the pathogenesis of vesicular stomatitis virus (VSV) in livestock remain poorly understood. Several studies have highlighted the relevant role of macrophages in controlling the systemic dissemination of VSV during infection in different animal models, including mice, cattle, and pigs. To gain more insight into the molecular mechanisms used by VSV to impair the immune response in macrophages, we used microarrays to determine the transcriptomic changes produced by VSV infection in primary cultures of porcine macrophages. The results indicated that VSV infection induced the massive expression of multiple anorexic, pyrogenic, proinflammatory, and immunosuppressive genes. Overall, the interferon (IFN) response appeared to be suppressed, leading to the absence of stimulation of interferon-stimulated genes (ISG). Interestingly, VSV infection promoted the expression of several genes known to downregulate the expression of IFNβ. This represents an alternate mechanism for VSV control of the IFN response, beyond the recognized mechanisms mediated by the matrix protein. Although there was no significant differential gene expression in macrophages infected with a highly virulent epidemic strain compared to a less virulent endemic strain, the endemic strain consistently induced higher expression of all upregulated cytokines and chemokines. Collectively, this study provides novel insights into VSV molecular pathogenesis and immune evasion that warrant further investigation.

59 BASIC BIOLOGICAL SCIENCES↗

Variable Inhibition of Nitrous Oxide Reduction in Denitrifying Bacteria by Different Forms of Methanobactin

Aerobic methanotrophic activity is highly dependent on copper availability, and methanotrophs have developed multiple strategies to collect copper. Specifically, when copper is limiting (ambient concentrations < 1 μM), some methanotrophs produce and secret a small modified peptide (< 1,300 Da) termed methanobactin (MB) that binds copper with high affinity. As MB is secreted into the environment, other microbes that require copper for their metabolism may be inhibited as MB may make copper unavailable; e.g., inhibition of denitrifiers as complete conversion nitrate to dinitrogen involves multiple enzymes, some of which are copper-dependent. Of key concern is inhibition of the copper-dependent nitrous oxide reductase (NosZ), the only known enzyme capable of converting nitrous oxide (N 2 O) to dinitrogen. Herein, we show that different forms of MB differentially affect copper uptake and N2O reduction by Pseudomonas stutzeri strain DCP-Ps1 (that expresses clade I NosZ) and Dechloromonas aromatica strain RCB (that expresses clade II NosZ). Specifically, in the presence of MB from Methylocystis sp. strain SB2 (SB2-MB), copper uptake and nosZ expression were more significantly reduced than in the presence of MB from Methylosinus trichosporium OB3b (OB3b-MB). Further, N2O accumulation increased more significantly for both P. stutzeri strain DCP-Ps1 and D. aromatica strain RCB in the presence of SB2-MB versus OB3b-MB. These data illustrate that copper competition between methanotrophs and denitrifying bacteria can be significant and that the extent of such competition is dependent on the form of MB that methanotrophs produce.

54 ENVIRONMENTAL SCIENCES↗

A second-order modeling study of confined swirling flow

A numerical study of a confined strong swirling flow is presented. Computations are performed using a differential second-moment closure. The effect of inlet dissipation rate on calculated mean and turbulence fields is investigated. Two pressure-strain models with various model constants are examined to demonstrate their influences on the predicted results. Finally, comparison of the differential second-moment calculation is made with algebraic second-moment results to determine a better suited model for complex swirling flows.

Nikjooy, M.↗

Measurements of fuselage skin strains and displacements near a longitudinal lap joint in a pressurized aircraft

Strains and displacements in a small area near a longitudinal lap joint in the fuselage skin of a B737 aircraft were measured during a pressurization cycle to a differential pressure of 6.2 psi while the aircraft was on the ground. It was found that hoop strains were higher than longitudinal strains at each location; membrane strains in the unreinforced skin were higher than in the joint; membrane strains in the hoop direction, as well as radial displacements, tended to be highest at the mid-bay location between skin reinforcements; significant bending in the hoop direction occurred in the joint and in the skin near the joint, and the bending was unsymmetrically distributed about the stringer at the middle of the joint; and radial displacements were unsymmetrically distributed across the lap joint. The interpretation of the strain gage data for locations on the bonded and riveted lap joint assumed that the joint did not contain disbonded areas.

Phillips, Edward P.↗

Three-Dimensional Postbuckling Analysis of Curved Beams

Presented here is a method of solving highly flexible curved beam undergoing huge static or quasi-static deformations. A geometrically exact beam theory based on the use of Jaumann stresses and strains and exact coordinate transformation is presented in terms of 17 first-order ordinary differential equations, and a multiple shooting method is used to solve the corresponding nonlinear two-point boundary value problems. The geometrically exact beam theory accounts far large rotations, large displacements, initial curvatures, extensionality, and transverse shear strains. Four examples are used to demonstrate this method, including a rotating clamped-free beam under the influence of gravity and centrifugal forces, an L-frame subjected to an in-plane tip load, a circular arch subjected to a concentrated load, and a clamped-hinged helical spring subjected to an axial displacement. Results show that the combination of the multiple shooting method and the geometrically exact beam theory works very well. Moreover, the obtained numerically exact solutions can be used to verify the accuracy of nonlinear finite element codes for nonlinear analysis of complex structures.

Pai, P. Frank↗

Genomic and morphological characterization of Knufia obscura isolated from the Mars 2020 spacecraft assembly facility

Members of the family Trichomeriaceae, belonging to the Chaetothyriales order and the Ascomycota phylum, are known for their capability to inhabit hostile environments characterized by extreme temperatures, oligotrophic conditions, drought, or presence of toxic compounds. The genus Knufia encompasses many polyextremophilic species. In this report, the genomic and morphological features of the strain FJI-L2-BK-P2 presented, which was isolated from the Mars 2020 mission spacecraft assembly facility located at the Jet Propulsion Laboratory in Pasadena, California. The identification is based on sequence alignment for marker genes, multi-locus sequence analysis, and whole genome sequence phylogeny. The morphological features were studied using a diverse range of microscopic techniques (bright field, phase contrast, differential interference contrast and scanning electron microscopy). The phylogenetic marker genes of the strain FJI-L2-BK-P2 exhibited highest similarities with type strain of Knufia obscura (CBS 148926 T ) that was isolated from the gas tank of a car in Italy. To validate the species identity, whole genomes of both strains (FJI-L2-BK-P2 and CBS 148926 T ) were sequenced, annotated, and strain FJI-L2-BK-P2 was confirmed as K. obscura. The morphological analysis and description of the genomic characteristics of K. obscura FJI-L2-BK-P2 may contribute to refining the taxonomy of Knufia species. Key morphological features are reported in this K. obscura strain, resembling microsclerotia and chlamydospore-like propagules. These features known to be characteristic features in black fungi which could potentially facilitate their adaptation to harsh environments.

59 BASIC BIOLOGICAL SCIENCES↗

Metalloproteomics Reveals Multi-Level Stress Response in Escherichia coli When Exposed to Arsenite

The arsRBC operon encodes a three-protein arsenic resistance system. ArsR regulates the transcription of the operon, while ArsB and ArsC are involved in exporting trivalent arsenic and reducing pentavalent arsenic, respectively. Previous research into Agrobacterium tumefaciens 5A has demonstrated that ArsR has regulatory control over a wide range of metal-related proteins and metabolic pathways. We hypothesized that ArsR has broad regulatory control in other Gram-negative bacteria and set out to test this. Here, we use differential proteomics to investigate changes caused by the presence of the arsR gene in human microbiome-relevant Escherichia coli during arsenite (AsIII) exposure. We show that ArsR has broad-ranging impacts such as the expression of TCA cycle enzymes during AsIII stress. Additionally, we found that the Isc [Fe-S] cluster and molybdenum cofactor assembly proteins are upregulated regardless of the presence of ArsR under these same conditions. An important finding from this differential proteomics analysis was the identification of response mechanisms that were strain-, ArsR-, and arsenic-specific, providing new clarity to this complex regulon. Given the widespread occurrence of the arsRBC operon, these findings should have broad applicability across microbial genera, including sensitive environments such as the human gastrointestinal tract.

Biochemistry & Molecular Biology↗

On the oscillation of the laterally heterogeneous earth, 1

The perturbative effects, as cause by lateral inhomogeneities in the earth structure and by Coriolis force, contaminate the originally toroidal and spheroidal earth's oscillations, making them of mixed type. For this reason, in order to make the computation of the perturbations more uniform and homogeneous, it was suggested that the earth's free oscillations be expanded into a series in terms of generalized harmonics familiar from the theory of angular momentum in quantum mechanics. Making use of Gibbsian symbolism and of some operators from the theory of angular momentum, explicit expressions were deduced for the perturbative terms in the differential equation of the earth's free oscillations. Decomposition of the strain tensor in terms of canonical vectors was also obtained.

Musen, P.↗

X-Ray-Scattering Measurements Of Strain In PEEK

Internal stress relieved by heating above glass-transition temperature. Report describes wide-angle x-ray scattering and differential scanning calorimetry of specimens of poly(etheretherketone) having undergone various thermal treatments. Wide-angle x-ray scattering particularly useful in determining distances between atoms, crystallinity, and related microstructurally generated phenomena, as thermal expansion and strain. Calorimetric measurements aid interpretation of scattering measurements by enabling correlation with thermal effects.

Cebe, Peggy↗

Alternate routes to acetate tolerance lead to varied isoprenol production from mixed carbon sources in Pseudomonas putida

ABSTRACT Lignocellulose is a renewable resource for the production of a diverse array of platform chemicals, including the biofuel isoprenol. Although this carbon stream provides a rich source of sugars, other organic compounds, such as acetate, can be used by microbial hosts. Here, we examined the growth and isoprenol production in a Pseudomonas putida strain pre-tolerized (“PT”) background where its native isoprenol catabolism pathway is deleted, using glucose and acetate as carbon sources. We found that PT displays impaired growth in minimal medium containing acetate and often fails to grow in glucose-acetate medium. Using a mutant recovery-based approach, we generated tolerized strains that overcame these limitations, achieving fast growth and isoprenol production in the mixed carbon feed. Changes in the glucose and acetate assimilation routes, including an upregulation in PP_0154 (SpcC, succinyl-CoA:acetate CoA-transferase) and differential expression of the gluconate assimilation pathways, were key for higher isoprenol titers in the tolerized strains, whereas a different set of mechanisms were likely enabling tolerance phenotypes in media containing acetate. Among these, a coproporphyrinogen-III oxidase (HemN) was upregulated across all tolerized strains and in one isolate required for acetate tolerance. Utilizing a defined glucose and acetate mixture ratio reflective of lignocellulosic feedstocks for isoprenol production in P. putida allowed us to obtain insights into the dynamics and challenges unique to dual carbon source utilization that are obscured when studied separately. Together, this enabled the development of a P. putida bioconversion chassis able to use a more complex carbon stream to produce isoprenol. IMPORTANCE Acetate is a relatively abundant component of many lignocellulosic carbon streams and has the potential to be used together with sugars, especially in microbes with versatile catabolism such as P. putida . However, the use of mixed carbon streams necessitates additional optimization. Furthermore, the use of P. putida for the production of the biofuel target, isoprenol, requires the use of engineered strains that have additional growth and production constraints when cultivated in acetate and glucose mixtures. In this study, we generate acetate-tolerant P. putida strains that overcome these challenges and examine their ability to produce isoprenol. We show that acetate tolerance and isoprenol production, although independent phenotypes, can both be optimized in a given P. putida strain. Using proteomics and whole genome sequencing, we examine the molecular basis of both phenotypes and show that tolerance to acetate can occur via alternate routes and result in different impacts on isoprenol production.

de Siqueira, Guilherme M. V. (ORCID:00000002364563↗

The Exact Solution for Linear Thermoelastic Axisymmetric Deformations of Generally Laminated Circular Cylindrical Shells

A detailed exact solution is presented for laminated-composite circular cylinders with general wall construction and that undergo axisymmetric deformations. The overall solution is formulated in a general, systematic way and is based on the solution of a single fourth-order, nonhomogeneous ordinary differential equation with constant coefficients in which the radial displacement is the dependent variable. Moreover, the effects of general anisotropy are included and positive-definiteness of the strain energy is used to define uniquely the form of the basis functions spanning the solution space of the ordinary differential equation. Loading conditions are considered that include axisymmetric edge loads, surface tractions, and temperature fields. Likewise, all possible axisymmetric boundary conditions are considered. Results are presented for five examples that demonstrate a wide range of behavior for specially orthotropic and fully anisotropic cylinders.

Nemeth, Michael P.↗

Bioinspired aligned magnetic features in aerogels for humidity sensing

Many natural plant tissues, such as pinecones, utilize structures to transduce changes in humidity into movement or strain. These often employ layered structures with aligned features in order to create differential stress across the material as different portions swell at different rates. Here, by replicating these types of structures in synthetic materials we can create structures that respond to humidity in a similar manner. Taking these aligned features as inspiration, silica aerogels with tailored humidity reactive properties were created. This behavior was achieved by creating aligned features in these aerogels using a Helmholtz coil to produce a uniform magnetic field that aligned ferrofluid droplets into chains and needle like features. These magnetite-based structures were shown to retain their superparamagnetic properties, allowing them to easily be removed from the aerogels, leaving a large number of aligned channels. Aerogels with the removed needles experienced significantly different strains than both unmodified aerogels and aerogels with the aligned features still in place. Aerogels with these tunable aligned features could be used in humidity sensors that can transduce changes in humidity into strain and could be used in applications such as volatile organic compound capture.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Shear-strain contours from moire interferometry

The development of whole-field contour maps of shear strains gamma (xy), derived from displacement fields obtained by moire interferometry with 2400 lines/mm, is described. The use of mechanical differentiation to obtain cross-derivatives of displacements and the use of graphical additive moire to sum the cross-derivatives are explained. Quantitative analysis in the small-strain domain is possible because of the high sensitivity of moire interferometry. The applicability of this technique is shown by the testing of a short epoxy beam under three-point bending.

Post, D.↗

Influence of Space Flight Factors on the Genetic Properties of Streptomyces Lividans 66 (PIJ702)

Gram-positive Streptomyces bacteria display genetic instability in response to external factors. Strain S. lividans 66 harbors the multicopy plasmid pIJ702 with selective and differential marker genes for antibiotic thiostrepton resistance and melanin production. Culture plates of modified ISP agar medium with and without thiostrepton were flown on Foton-M2. Suboptimal flight temperatures, which were simulated for asynchronous ground controls, resulted in slow growth and failure to differentiate and sporulate. Flight samples and asynchronous controls showed a high frequency of failing to express plasmid markers compared to laboratory controls. This was associated with loss of plasmid DNA and likely resulted from suboptimal temperatures for flight cultures and controls. Neither restriction fragment length polymorphism, nor polymerase chain reaction amplification coupled with denaturing gradient gel electrophoresis, revealed differences between pIJ702 DNA from flight vs. control clones. Mutations of the plasmid marker genes resulting from specific spaceflight factors, e.g., microgravity and radiation, were not detected.

Tabakov, V. Yu.↗

Measurement of high temperature strain by the laser-speckle strain gauge

By using the laser-speckle strain gauge, the strain of metal at the temperature lower than 250 C is measured. The principle of the gauge is to measure the expansion or contraction of the fine structures of surface by detecting the resultant speckle displacement in an optoelectronic way, whereby the effect of rigid-body motion is automatically cancelled out with the aid of a differential detection system. A transportable apparatus was built and a comparison experiment performed with a resistance strain gauge at room temperature. It has a strain sensitivity of .00002, a gauge length smaller than 1 mm, and no upper limit in a range of strain measurement. In the measurement of high-temperature strain it is free from the need for a dummy gauge and insensitive to an electric drift effect. As examples of strain measurement at high-temperature, thermal expansion and contraction of a top of a soldering iron are measured. The interval of the measurement can be made at shortest 1.6 sec. and the change in the strain is clearly followed until the ultimate stationary temperature is reached.

Yamaguchi, I.↗

Thermomechanical conversion in metals: dislocation plasticity model evaluation of the Taylor-Quinney coefficient

Using a partitioned-energy thermodynamic framework which assigns energy to that of atomic configurational stored energy of cold work and kinetic-vibrational, in this study we derive an important constraint on the Taylor-Quinney coefficient, which quantifies the fraction of plastic work that is converted into heat during plastic deformation. Associated with the two energy contributions are two separate temperatures – the ordinary temperature for the thermal energy and the effective temperature for the configurational energy. We show that the Taylor-Quinney coefficient is a function of the thermodynamically defined effective temperature that measures the atomic configurational disorder in the material. Finite-element analysis of recently published experiments on the aluminum alloy 6016-T4 [1], using the thermodynamic dislocation theory (TDT), shows good agreement between theory and experiment for both stress-strain behavior and temporal evolution of the temperature. The simulations include both conductive and convective thermal energy loss during the experiments, and significant thermal gradients exist within the simulation results. Computed values of the differential Taylor-Quinney coefficient are also presented and suggest a value which differs between materials and increases with increasing strain.

36 MATERIALS SCIENCE↗

Reused cultivation water from a self-inhibiting alga does not inhibit other algae but alters their microbiomes

Economical production of algal commodities (food, feed, and fuels) requires reusing cultivation water to reduce operating costs. While some algae strains show growth inhibition in reused water, other strains appear un-affected. Reusing water to grow different strains (i.e., crop rotation) could potentially improve overall biomass production compared to water reuse with the same strain by reducing accumulation of strain-specific waste products, limiting carryover of population-specific pathogens such as viruses, and introducing bacteria that degrade residual organic matter. Here, batch culture experiments tested the effects of reused cultivation water from the self-inhibiting diatom Staurosira sp. C323 on the diatom Navicula sp. SFP and green alga Chlorella sp. D046, as well as their microbiomes. Navicula sp. and Chlorella sp. grew well in Staurosira sp. reused water, indicating that previously observed self-inhibition was likely specific to this Staurosira strain. While algal mi-crobiomes were not significantly different between controls and reused water treatments, specific bacteria taxa were differentially abundant in reused treatments, suggesting that some taxa responded to compounds re-maining in the reused water. Finally, results suggest that algae growth responses in reused water may depend largely on strain-specific factors, and that crop rotation can support biomass production.

09 BIOMASS FUELS↗