Role of yceA-cybB-yceB operon in oxidative stress tolerance, swimming motility and antibiotic susceptibility of Stenotrophomonas maltophilia.

BACKGROUND: Stenotrophomonas maltophilia is ubiquitous in the environment and is an important MDR opportunistic pathogen. Oxidative stress is an inevitable challenge to an aerobic bacterium. Accordingly, S. maltophilia has many capabilities to face variable oxidative stress. Some of the oxidative stress alleviation systems cross-protect bacteria from antibiotics. In our recent RNA-sequencing transcriptome analysis, we documented the increased expression of a three-gene cluster, yceA-cybB-yceB, in the presence of hydrogen peroxide (H2O2). The YceI-like, cytochrome b561 and YceI-like proteins encoded by yceA, cybB and yceB are located in the cytoplasm, inner membrane and periplasm, respectively. OBJECTIVES: To characterize the role of the yceA-cybB-yceB operon of S. maltophilia in oxidative stress tolerance, swimming motility and antibiotic susceptibility. METHODS: The presence of the yceA-cybB-yceB operon was verified by RT-PCR. The functions of this operon were revealed by in-frame deletion mutant construction and complementation assay. Expression of the yceA-cybB-yceB operon was assessed by quantitative RT-PCR. RESULTS: The yceA, cybB and yceB genes form an operon. Loss of function of the yceA-cybB-yceB operon compromised menadione tolerance, enhanced swimming motility and increased susceptibility to fluoroquinolone and ß-lactam antibiotics. The expression of the yceA-cybB-yceB operon was up-regulated by oxidative stress, such as H2O2 and superoxide, and not impacted by antibiotics, such as fluoroquinolone and ß-lactams. CONCLUSIONS: The evidence strongly supports the view that the physiological function of the yceA-cybB-yceB operon is to alleviate oxidative stress. The operon provides an additional example that oxidative stress alleviation systems can cross-protect S. maltophilia from antibiotics.

Hyperspectral Tensor Completion Using Low-Rank Modeling and Convex Functional Analysis.

Hyperspectral tensor completion (HTC) for remote sensing, critical for advancing space exploration and other satellite imaging technologies, has drawn considerable attention from recent machine learning community. Hyperspectral image (HSI) contains a wide range of narrowly spaced spectral bands hence forming unique electrical magnetic signatures for distinct materials, and thus plays an irreplaceable role in remote material identification. Nevertheless, remotely acquired HSIs are of low data purity and quite often incompletely observed or corrupted during transmission. Therefore, completing the 3-D hyperspectral tensor, involving two spatial dimensions and one spectral dimension, is a crucial signal processing task for facilitating the subsequent applications. Benchmark HTC methods rely on either supervised learning or nonconvex optimization. As reported in recent machine learning literature, John ellipsoid (JE) in functional analysis is a fundamental topology for effective hyperspectral analysis. We therefore attempt to adopt this key topology in this work, but this induces a dilemma that the computation of JE requires the complete information of the entire HSI tensor that is, however, unavailable under the HTC problem setting. We resolve the dilemma, decouple HTC into convex subproblems ensuring computational efficiency, and show state-of-the-art HTC performances of our algorithm. We also demonstrate that our method has improved the subsequent land cover classification accuracy on the recovered hyperspectral tensor.

Implication of the σE Regulon Members OmpO and σN in the ΔompA299-356-Mediated Decrease of Oxidative Stress Tolerance in Stenotrophomonas maltophilia.

Outer membrane protein A (OmpA) is the most abundant porin in bacterial outer membranes. KJΔOmpA299-356, an ompA C-terminal in-frame deletion mutant of Stenotrophomonas maltophilia KJ, exhibits pleiotropic defects, including decreased tolerance to menadione (MD)-mediated oxidative stress. Here, we elucidated the underlying mechanism of the decreased MD tolerance mediated by ΔompA299-356. The transcriptomes of wild-type S. maltophilia and the KJΔOmpA299-356 mutant strain were compared, focusing on 27 genes known to be associated with oxidative stress alleviation; however, no significant differences were identified. OmpO was the most downregulated gene in KJΔOmpA299-356. KJΔOmpA299-356 complementation with the chromosomally integrated ompO gene restored MD tolerance to the wild-type level, indicating the role of OmpO in MD tolerance. To further clarify the possible regulatory circuit involved in ompA defects and ompO downregulation, σ factor expression levels were examined based on the transcriptome results. The expression levels of three σ factors were significantly different (downregulated levels of rpoN and upregulated levels of rpoP and rpoE) in KJΔOmpA299-356. Next, the involvement of the three σ factors in the ΔompA299-356-mediated decrease in MD tolerance was evaluated using mutant strains and complementation assays. rpoN downregulation and rpoE upregulation contributed to the ΔompA299-356-mediated decrease in MD tolerance. OmpA C-terminal domain loss induced an envelope stress response. Activated σE decreased rpoN and ompO expression levels, in turn decreasing swimming motility and oxidative stress tolerance. Finally, we revealed both the ΔompA299-356-rpoE-ompO regulatory circuit and rpoE-rpoN cross regulation. IMPORTANCE The cell envelope is a morphological hallmark of Gram-negative bacteria. It consists of an inner membrane, a peptidoglycan layer, and an outer membrane. OmpA, an outer membrane protein, is characterized by an N-terminal ß-barrel domain that is embedded in the outer membrane and a C-terminal globular domain that is suspended in the periplasmic space and connected to the peptidoglycan layer. OmpA is crucial for the maintenance of envelope integrity. Stress resulting from the destruction of envelope integrity is sensed by extracytoplasmic function (ECF) σ factors, which induce responses to various stressors. In this study, we revealed that loss of the OmpA-peptidoglycan (PG) interaction causes peptidoglycan and envelope stress while simultaneously upregulating σP and σE expression levels. The outcomes of σP and σE activation are different and are linked to ß-lactam and oxidative stress tolerance, respectively. These findings establish that outer membrane proteins (OMPs) play a critical role in envelope integrity and stress tolerance.

Comparison between orthogonal subspace projection and background subtraction techniques applied to remote-sensing data.

The basic measurement equation r = B + alphad + n is solved for alpha (the weight or abundance of the spectral target vector d) by two methods: (a) by subtracting the stochastic spectral background vector B from the spectral measurement's vector r (subtraction solution) and (b) by orthogonal subspace projection (OSP) of the measurements to a subspace orthogonal to B (the OSP solution). The different geometry of the two solutions and in particular the geometry of the noise vector n is explored. The angular distribution of the noise angle between B and n is the key factor for determining and predicting which solution is better. When the noise-angle distribution is uniform, the subtraction solution is always superior regardless of the orientation of the spectral target vector d. When the noise is more concentrated in the direction orthogonal to B, the OSP solution becomes better (as expected). Simulations and one-dimensional hyperspectral measurements of vapor concentration in the presence of background radiation and noise are given to illustrate these two solutions.

Detection, identification, and estimation of biological aerosols and vapors with a Fourier-transform infrared spectrometer.

Two experiments were conducted with a Fourier-transform infrared (FTIR) spectrometer. The purpose of the first experiment was to detect and identify Bacillus subtilis subsp. niger (BG) bioaerosol spores and kaolin dust in an open-air release for which the thermal contrast between the aerosol temperature and background brightness temperature is small. The second experiment estimated the concentration of a small amount of triethyl phosphate (TEP) vapor in a closed chamber in which an external blackbody radiation source was used and where the thermal contrast was large. The deduced BG (TEP) extinction spectrum (identification) showed an excellent match to the library BG (TEP) extinction spectrum. Analysis of the time sequence of the measurements coincided well with the presence (detection) of the BG during the measurements, and the estimated concentration of time-dependent TEP vapor was excellent. The data were analyzed with hyperspectral detection, identification, and estimation algorithms. The algorithms were based on radiative transfer theory and statistical signal-processing methods. A subspace orthogonal projection operator was used to statistically subtract the large thermal background contribution to the measurements, and a robust maximum-likelihood solution was used to deduce the target (aerosol or vapor cloud) spectrum and estimate its mass-column concentration. A Gaussian-mixture probability model for the deduced mass-column concentration was computed with an expectation-maximization algorithm to produce the detection threshold, the probability of detection, and the probability of false alarm. The results of this study are encouraging, as they suggest for the first time to the authors' knowledge the feasibility of detecting biological aerosols with passive FTIR sensors.