Citrobacter enshiensis sp. nov., isolated from seleniferous soil.

Two Gram-stain-negative, facultative anaerobic, rod-shaped bacterial strains, S171T and S2-9, were isolated from seleniferous soil in China. Comprehensive phylogenetic analyses based on 16S rRNA genes, multilocus sequences and whole genome sequences indicated that the two strains belonged to the genus Citrobacter. The average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values of strains S171T and S2-9 with the closest relative Citrobacter koseri NCTC 10786T were 83.6-83.7% and 27.7-27.8 %, respectively, which were below the species cutoff values. The ANI and dDDH values between the two strains were 97.9% and 84.8 %, respectively. The biochemical characteristics revealed that selenite tolerance, H2S and indole production, arginine dihydrolase, ornithine decarboxylase, as well as acid production from carbon sources such as d-sorbitol and arbutin are distinctive features of the two strains. Based on these results, strain S171T and strain S2-9 represent a novel species of the genus Citrobacter, for which the name Citrobacter enshiensis sp. nov. is proposed, with strain S171T (=GDMCC 1.3637T=JCM 35851T) as the type strain. The genome size of strain S171T was 4.92 Mb with a G+C content of 52.6 mol%. The genome size of strain S2-9 was 4.89 Mb with a G+C content of 52.6 mol%.

Achromobacter seleniivolatilans sp. nov. and Buttiauxella selenatireducens sp. nov., isolated from the rhizosphere of selenium hyperaccumulator Cardamine hupingshanesis.

Two Gram-stain-negative bacterial strains, R39T and R73T, were isolated from the rhizosphere soil of the selenium hyperaccumulator Cardamine hupingshanesis in China. Strain R39T transformed selenite into elemental and volatile selenium, whereas strain R73T transformed both selenate and selenite into elemental selenium. Phylogenetic and phylogenomic analyses indicated that strain R39T belonged to the genus Achromobacter, while strain R73T belonged to the genus Buttiauxella. Strain R39T (genome size, 6.68 Mb; G+C content, 61.6 mol%) showed the closest relationship to Achromobacter marplatensis LMG 26219T and Achromobacter kerstersii LMG 3441T, with average nucleotide identity (ANI) values of 83.6 and 83.4 %, respectively. Strain R73T (genome size, 5.22 Mb; G+C content, 50.3 mol%) was most closely related to Buttiauxella ferragutiae ATCC 51602T with an ANI value of 86.4 %. Furthermore, strain A111 from the GenBank database was found to cluster with strain R73T within the genus Buttiauxella through phylogenomic analyses. The ANI and digital DNA-DNA hybridization values between strains R73T and A111 were 97.5 and 80.0% respectively, indicating that they belong to the same species. Phenotypic characteristics also differentiated strain R39T and strain R73T from their closely related species. Based on the polyphasic analyses, strain R39T and strain R73T represent novel species of the genera Achromobacter and Buttiauxella, respectively, for which the names Achromobacter seleniivolatilans sp. nov. (type strain R39T=GDMCC 1.3843T=JCM 36009T) and Buttiauxella selenatireducens sp. nov. (type strain R73T=GDMCC 1.3636T=JCM 35850T) are proposed.

Disruption of the cell division protein ftsK gene changes elemental selenium generation, selenite tolerance, and cell morphology in Rahnella aquatilis HX2.

AIMS: Some studies have indicated that the alterations in cellular morphology induced by selenite [Se(Ⅳ)] may be attributed to its inhibitory effects on cell division. However, whether the genes associated with cell division are implicated in Se(Ⅳ) metabolism remains unclear. METHODS AND RESULTS: The ftsK gene in Rahnella aquatilis HX2 was mutated with an in-frame deletion strategy. The ftsK mutation strongly reduced the tolerance to selenite [Se(Ⅳ)] and the production of red elemental selenium [Se(0)] in R. aquatilis HX2, and this effect could not be attributed solely to the inhibition of cell growth. Deleting the ftsK gene also resulted in a significant decrease in bacterial growth of R. aquatilis HX2 during both exponential and stationary phases. The deletion of ftsK inhibited cell division, resulting in the development of elongated filamentous cells. Furthermore, the loss-of-function of FtsK significantly impacted the expression of seven genes linked to cell division and Se(Ⅳ) metabolism by at least 2-fold, as unveiled by real-time quantitative PCR (RT-qPCR) under Se(Ⅳ) treatment. CONCLUSIONS: These findings suggest that FtsK is associated with Se(Ⅳ) tolerance and Se(0) generation and is a key player in coordinating bacterial growth and cell morphology in R. aquatilis HX2.

Multi-mode stable, high peak power Nd:YAG laser device for laser cleaning.

High average power and peak power solid-state lasers are of great interest in the field of laser cleaning. In this research, a high peak power laser with over 400 W average power using a multi-mode stable resonator in a diode-pumped Nd:YAG master oscillator power amplifier has been demonstrated. A maximum peak power over 1.08 MW was achieved at a repetition frequency of 5 kHz. Delivery fiber with a 400 µm core diameter was utilized for flexible laser transmission. A prototype of the laser cleaning system was developed and inspiring application effects for paint and mold cleaning were achieved.

Uptake of Selenite by Rahnella aquatilis HX2 Involves the Aquaporin AqpZ and Na+/H+ Antiporter NhaA.

Microbial transformation of selenite [Se(IV)] to elemental selenium nanoparticles (SeNPs) is known to be an important process for removing toxic soluble selenium (Se) oxyanions and recovery of Se from the environment as valuable nanoparticles. However, the mechanism of selenite uptake by microorganisms, the first step through which Se exerts its cellular function, remains not well studied. In this study, the effects of selenite concentration, time, pH, metabolic inhibitors, and anionic analogues on selenite uptake in Rahnella aquatilis HX2 were investigated. Selenite uptake by R. aquatilis HX2 was concentration- and time-dependent, and its transport activity was significantly dependent on pH. In addition, selenite uptake in R. aquatilis HX2 was significantly inhibited by the aquaporin inhibitor AgNO3 and sulfite (SO32-), and partially inhibited by carbonyl cyanide m-chlorophenyl hydrazone (CCCP) and 2,4-dinitrophenol (2,4-DNP) treatments. Three mutants with in-frame deletions of aqpZ, glpF, and nhaA genes were constructed. The transport assay showed that the water channel protein AqpZ, and not GlpF, was a key channel of selenite uptake by R. aquatilis HX2, and sulfite and selenite had a common uptake pathway. In addition, the Na+/H+ antiporter NhaA is also involved in selenite uptake in R. aquatilis HX2.

Multifactorial interaction of selenium, iron, xylose, and glycine on cordycepin metabolism in Cordyceps militaris.

Cordycepin, a nucleoside analog, is the main antioxidative and antimicrobial substance in Cordyceps militaris. To improve the metabolism of cordycepin, carbon sources, nitrogen sources, trace elements, and precursors were studied by single factor, Plackett-Burman, and central composite designs in C. militaris mycelial fermentation. Under the regulation of the multifactorial interactions of selenite, ferrous chloride, xylose, and glycine, cordycepin production was increased by 5.2-fold compared with the control. The gene expression of hexokinase, ATP phosphoribosyltransferase, adenylosuccinate synthetase, and cns1-3 in the glycolysis, pentose phosphate, and adenosine synthesis pathways were increased by 3.2-7.5 times due to multifactorial interactions, while the gene expression of histidine biosynthesis trifunctional protein and histidinol-phosphate aminotransferase in histidine synthesis pathway were decreased by 23.4%-56.2%. Increasing with cordycepin production, glucose uptake was accelerated, mycelia growth was inhibited, and the cell wall was damaged. Selenomethionine (SeMet), selenocysteine (SeCys), and selenium nanoparticles (SeNPs) were the major Se species in C. militaris mycelia. This study provides a new insight for promoting cordycepin production by regulating glycolysis, pentose phosphate, and histidine metabolism. KEY POINTS: • Cordycepin production in the CCDmax group was 5.2-fold than that of the control. • Glucose uptake of the CCDmax group was accelerated and cell wall was damaged. • The metabolic flux was concentrated to the cordycepin synthesis pathway.

Xanthones from securidaca inappendiculata antagonizes the anti-rheumatic effect of methotrexate by inhibiting reduced folate carrier 1.

BACKGROUND: The first-line anti-rheumatic drug methotrexate (MTX) is used in the combination. Because of the unpredictable adverse reactions, optimization of relevant regimens is necessary and meaningful. This study aimed to study the possible interaction between Securidaca inappendiculate Hassk. Derived xanthones and MTX. METHODS: We established adjuvant-induced arthritis (AIA) model, which was treated with MTX and MTX + xanthone-rich fraction (XRF). The clinical efficacy was evaluated by histopathological examination, and LC-MS was used to monitor the blood concentration of MTX. Western blotting and immunohistochemistry were used to detect protein expression. In vitro, we assessed the activity of related transporters by cellular uptake assay based on HEK-293T cells. RESULTS: Compared with MTX-treated rats, inflammation in the immunized rats in the MTX + XRF group was obvious, indicating that XRF antagonized the anti-rheumatic effect of MTX. Meanwhile, XRF reduced liver and kidney injuries caused by MTX in addition to MTX. Results from immunohistochemical and nappendiculat assays suggested that XRF may reduce uptake of MTX by down-regulating reduced folate carrier 1 (RFC1). CONCLUSION: This study indicated that XRF could reduce the plasma concentration of MTX by inhibiting the expression of RFC1, antagonize the therapeutic effect of MTX on AIA rats, and reduce its oral bioavailability. The combination of S. inappendiculate and MTX should be further optimized to achieve the goal of increasing efficiency and reducing toxicity.

Adaptively Optimized Gas Analysis Model with Deep Learning for Near-Infrared Methane Sensors.

Noise significantly limits the accuracy and stability of retrieving gas concentration with the traditional direct absorption spectroscopy (DAS). Here, we developed an adaptively optimized gas analysis model (AOGAM) composed of a neural sequence filter (NSF) and a neural concentration retriever (NCR) based on deep learning algorithms for extraction of methane absorption information from the noisy transmission spectra and obtaining the corresponding concentrations from the denoised spectra. The model was trained on two data sets, including a computationally generated one and the experimental one. We have applied this model for retrieving methane concentration from its transmission spectra in the near-infrared (NIR) region. The NSF was implemented through an encoder-decoder structure enhanced by the attention mechanism, improving robustness under noisy conditions. Further, the NCR was employed based on a combination of a principal component analysis (PCA) layer, which focuses the algorithm on the most significant spectral components, and a fully connected layer for solving the nonlinear inversion problem of the determination of methane concentration from the denoised spectra without manual computation. Evaluation results show that the proposed NSF outperforms widely used digital filters as well as the state-of-the-art filtering algorithms, improving the signal-to-noise ratio by 7.3 dB, and the concentrations determined with the NCR are more accurate than those determined with the traditional DAS method. With the AOGAM enhancement, the optimized methane sensor features precision and stability in real-time measurements and achieves the minimum detectable column density of 1.40 ppm·m (1σ). The promising results of the present study demonstrate that the combination of deep learning and absorption spectroscopy provides a more effective, accurate, and stable solution for a gas monitoring system.

The Effect of α7nAChR Signaling on T Cells and Macrophages and Their Clinical Implication in the Treatment of Rheumatic Diseases.

Rheumatoid arthritis (RA) is one of the most common autoimmune disease and until now, the etiology and pathogenesis of RA is not fully understood, although dysregulation of immune cells is one of the leading cause of RA-related pathological changes. Based on current understanding, the priority of anti-rheumatic treatments is to restore immune homeostasis. There are several anti-rheumatic drugs with immunomodulatory effects available nowadays, but most of them have obvious safety or efficacy shortcomings. Therefore, the development of novel anti-rheumatic drugs is still in urgently needed. Cholinergic anti-inflammatory pathway (CAP) has been identified as an important aspect of the so-called neuro-immune regulation feedback, and the interaction between acetylcholine and alpha 7 nicotinic acetylcholine receptor (α7nAChR) serves as the foundation for this signaling. Consistent to its immunomodulatory functions, α7nAChR is extensively expressed by immune cells. Accordingly, CAP activation greatly affects the differentiation and function of α7nAChR-expressing immune cells. As a result, targeting α7nAChR will bring profound therapeutic impacts on the treatment of inflammatory diseases like RA. RA is widely recognized as a CD4+ T cells-driven disease. As a major component of innate immunity, macrophages also significantly contribute to RA-related immune abnormalities. Theoretically, manipulation of CAP in immune cells is a feasible way to treat RA. In this review, we summarized the roles of different T cells and macrophages subsets in the occurrence and progression of RA, and highlighted the immune consequences of CAP activation in these cells under RA circumstances. The in-depth discussion is supposed to inspire the development of novel cell-specific CAP-targeting anti-rheumatic regimens.

Study of spinel LiTi2O4 superconductors via near-infrared reflection experiments.

We present an optical study on high-quality and single-phase LiTi2O4 (LTO) superconductor thin films grown on MgAl2O4 substrates by pulsed laser deposition. The near infrared (NIR) reflectivity is measured for samples with (001) and (111) lattice orientations. The temperature-induced metal-superconductor transition can be observed, and the superconducting transition temperature can be measured for both samples. We find that the NIR reflection experiment can reflect rightly the basic features of LTO superconductor thin films. Furthermore, the results obtained from this simple optical measurement suggest that the photo-induced electronic localization effect can be present in LTO thin films in a metallic state. Such information cannot be obtained directly from conventional transport and magneto-transport measurements. These interesting and important findings demonstrate that the NIR reflection experiment is a powerful optical technique for contactless characterizations and investigations of superconductor materials.

High power broadband LiF:F(2)(-) color center laser.

A high power LiF:F(2)(-) color center laser is demonstrated with broadband emission. The excitation source is a quasi-continuous wave diode side-pumped acousto-optically Q-switched Nd:YAG laser. Under an incident 1064-nm laser power of 25.4 W, the highest output power of up to 4.7 W is obtained with a macro pulse repetition rate of 400 Hz and a micro pulse repetition rate of 50 kHz. The broadband emission is centered at 1142 nm with a bandwidth of 13 nm.

Single-frequency CaWO4 Raman amplifier at 1178 nm.

Single-frequency crystalline Raman amplifications at 1178 nm were demonstrated. The seeding laser was generated from a single-frequency continuous wave fiber Raman amplifier. Three stages of Raman amplifications from CaWO4 single crystals were realized with a pulsed 1064 nm Nd:YAG laser as the pumping source. The final output pulse energy at 1178 nm was 26.7 mJ, and the pulse width was 2.9 ns, corresponding to a peak power of 5.2 MW. The overall Raman amplification ratio was up to 4.6×10(6). The linewidth was less than 500 MHz.

Four-wavelength laser based on intracavity BaWO4 Raman conversions of a dual-wavelength Q-switched Nd:YLF laser.

By using diode-end-pumped acousto-optically Q-switched intracavity Raman laser configurations, we demonstrate a four-wavelength laser emitting at 1047.0, 1053.0, 1159.4 and 1166.8 nm. Two Nd:YLiF4 crystals are employed to generate 1047.0-nm and 1053.0-nm laser radiations. These two lasers are then frequency converted by a BaWO4 Raman crystal to generate 1159.4-nm and 1166.8-nm first-Stokes waves. With pulse synchronization realized, we obtain the maximum output powers of 427, 418, 423 and 332 mW for 1047.0-nm, 1053.0-nm, 1159.4-nm and 1166.8-nm lasers, respectively. The total optical-to-optical conversion efficiency is 15.1%.

Synchronized dual tunable wavelength Q-switched Nd:Glass laser.

A flash-lamp pumped electro-optically Q-switched dual tunable wavelength Nd:Glass laser is demonstrated. An Nd:Glass rod was selected as the laser medium and a DKDP crystal was employed as the Q-switch. A cubic polarizer was placed in the cavity to divide the randomly polarized radiation into two orthogonally polarized beams. The two beams were tuned by two Littman-gratings and output independently. For Q-switched operation, the tuning range was 1050.1 - 1061.6 nm and 1050.5 - 1060.2 nm for the horizontal and vertical polarizations, respectively. When the horizontally polarized output was fixed at 1059.1 nm, the synchronized vertically polarized one can be tuned from 1051.5 to 1057.5 nm.

Nitrate reductase involves in selenite reduction in Rahnella aquatilis HX2 and the characterization and anticancer activity of the biogenic selenium nanoparticles.

BACKGROUND: Biogenic selenium nanoparticles (SeNPs) show numerous advantages including their high stability, low toxicity, and high bioactivity. While metabolism of SeNPs remains not well studied and need more investigation to reveal the process. PURPOSE: The objective of the study was to investigate the relationship between nitrate reductase and selenite reduction in Rahnella aquatilis HX2, characterize the properties of HX2 produced SeNPs, and explore their potential applications, particularly their anticancer activity. PROCEDURES: Selenium species were measured by high-performance liquid chromatography coupled to inductively coupled plasma - Mass spectrometry (HPLC-ICP-MS). Transcription level of nitrate reductase was determined by Real-time quantitative PCR. Morphology, particle size, crystal structure and surface chemistry of SeNPs were determined by electron microscopy, dynamic light scattering method, Raman scattering, X-ray photoelectron spectroscopy, respectively. Anti cancer cell activity was measured by CCK-8 assay. MAIN FINDINGS: SeNP production in R. aquatilis HX2 was correlated with the cell growth. The products of selenite reduction in HX2 detected by HPLC-ICP-MS included SeNPs, selenocysteine (SeCys), Se-Methylselenocysteine (MeSeCys), and 7 unknown compounds. Nitrate addition experiments suggested the involvement of nitrate reductase in selenite reduction in HX2. Both the cellular membrane and cytoplasm of HX2 exhibited selenite-reducing ability, indicating that membrane-associated nitrate reductase was not the sole selenite reductase in HX2. Characterization of the biogenic SeNPs revealed a spherical morphology and amorphous structure of them. Surface chemistry analysis implicated the binding of extracellular polymeric substances to the biogenic SeNPs, and the presence of Se0, Se2-, and electron-rich Se atoms on the surface of SeNPs. Finally, the IC50 values of the biogenic SeNPs were 36.49 µM for HepG2 and 3.70 µM for HeLa cells. CONCLUSIONS: The study first revealed that the nitrate reductase is involving in selenite reduction in R. aquatilis HX2. The biogenic SeNPs coordinated with organic substances in the surface. And SeNPs produced by R. aquatilis HX2 showed excellent anticancer activities on HepG2 and HeLa cells.

Identification and Functional Characterization of a Surfactant-like Protein Region in Flagellin FliC for Stabilizing Selenium Nanoparticles and Enhancing Bioavailability.

Biogenic selenium nanoparticles (SeNPs) are the most favorable Se form for nutritional supplementation due to their high stability, low toxicity, and high activity. However, the interaction between the surface-binding proteins and their stable biogenic SeNPs, as well as their impact on the stability and bioavailability of SeNPs, remains to be understood. In vitro stabilization experiments revealed an amino acid segment (F(235-386)) in Rahnella aquatilis' flagellin FliC, with surfactant-like properties, stabilizing SeNPs under harsh conditions. FliC and F(235-386) were employed as stabilizers to synthesize SeNPs (FliC@SeNPs and F(235-386)@SeNPs), and surface chemistry analysis revealed coordination reactions between the proteins and Se atoms on the surface of SeNPs. Both FliC and F(235-386) enhanced SeNPs uptake in wheat seedlings but reduced it in bacteria and yeast. This study highlights FliC's core function in stabilizing SeNPs and enhancing their bioavailability, paving the way for agricultural and nutritional applications.

Amorphous structure and crystal stability determine the bioavailability of selenium nanoparticles.

Microorganisms play a critical role in the biogeochemical cycling of selenium, often reducing selenite/selenate to elemental selenium nanoparticles (SeNPs). These SeNPs typically exist in an amorphous structure but can transform into a trigonal allotrope. However, the crystal structural transition process and its impact on selenium bioavailability have not been well studied. To shed light on this, we prepared chemosynthetic and biogenic SeNPs and investigated the stability of their crystal structure. We found that biogenic SeNPs exhibited a highly stable amorphous structure in various conditions, such as lyophilization, washing, and laser irradiation, whereas chemosynthetic SeNPs transformed into a trigonal structure in the same conditions. Additionally, a core-shell structure was observed in biogenic SeNPs after electron beam irradiation. Further analysis revealed that biogenic SeNPs showed a coordination reaction between Se atoms and surface binding biomacromolecules, indicating that the outer layer of Se-biomacromolecules complex prevented the SeNPs from crystallizing. We also investigated the effects of SeNPs crystal structures on the bioavailability in bacteria, yeast, and plants, finding that the amorphous structure of SeNPs determined Se bioavailability.

Role of Inhibiting Inflammation of LC3-Associated Phagocytosis in Dry Eye Disease.

PURPOSE: To confirm the expression and investigate the role of LC3-associated phagocytosis (LAP) in dry eye disease (DED). METHODS: The DED model of mice was established by scopolamine subcutaneous injection in a low-humidity environment chamber. Tear secretion test and corneal fluorescein sodium staining were used to evaluate the severity of DED. Expression levels of Rubicon, microtubule-associated protein light chain 3-II (LC3-II), Beclin-1 and autophagy-related gene-7 (Atg-7) in corneas of mice with DED were tested by western blot. Cell Counting Kit-8 (CCK-8) assay was used to detect the effects of different concentrations of hypertonic solutions on the proliferation activity of human corneal epithelial cells (HCECs). The expression levels of Dectin-1, IL-6 and IL-1ß in HCECs after stimulation with different concentrations of hypertonic solutions were tested. The expressions of Rubicon, LC3-II, Beclin-1 and ATG-7 in HCECs were detected by reverse transcription polymerase chain reaction (RT-PCR). After being pretreated with 10 µM si-Rubicon, the severity of the disease was documented by corneal fluorescein sodium staining. And the expression levels of IL-6 and IL-1ß were also tested by RT-PCR. RESULTS: Compared with the normal control group, the corneal fluorescein sodium staining scores and tear secretion were significantly reduced. Rubicon, LC3-II, Beclin-1 and ATG-7 were significantly elevated. CCK-8 showed that the 400 and 450 mOsM hypertonic solutions did not affect the proliferation activity of HCECs. The expression of Dectin-1, IL-1ß and IL-6 were elevated after stimulation with 450 mOsM solution. LC3-II, Rubicon, ATG-7 and Beclin-1 increased after stimulation with 450 mOsM hyperosmolar solution in HCECs. Corneal fluorescein staining showed that si-Rubicon increased the severity of DED in mice. Moreover, the mRNA expressions of inflammatory factors IL-1ß and IL-6 in the cornea of mice were significantly increased. CONCLUSION: DED increased the expression of proteins associated with LAP. LAP could play an anti-inflammatory effect in DED.

Acquisition of phase-shift fiber grating spectra with 23.5 femtometer spectral resolution using DFB-LD.

A novel method based on distributed-feedback laser diode (DFB-LD) continuous wavelength-scanning for acquiring precise spectra of phase-shift fiber gratings is presented. Compared to the traditional method, the spectral resolution retrieved by this method is only limited by the optical line-width of the light source, which can reach up to the order of femtometer and is much higher than that of high-resolution optical spectrum analyzer (generally on the order of picometer). In addition, a Signal-to-Noise Ratio (SNR) advantage can be provided owing to a much higher spectral density of DFB-LD than amplified spontaneous emission (ASE) source. Precise spectra of three phase-shift fiber grating samples have been obtained at a resolution of 23.5 femtometer.

Measurement and analysis of water vapor inside optical components for optical fiber H2O sensing system.

Water vapor existing inside internal end-face gaps of optical components of an optical fiber H2O sensing system makes it possible to influence the measurement accuracy and stability. The influence principle has been briefly analyzed based on the structure of three main optical components: a distributed feedback laser diode (DFB-LD), a collimator, and a photoelectric diode (PD). With application of a differential technique, the influence of water vapor inside the DFB-LD can be removed. With reasonable recombination of the collimator and the PD in a dual-beam detection system, the influence of water vapor inside the collimator and the PD's end-face gaps has been suppressed from more than 1.57×10(-3) to as low as -2.175×10(-5) in absorbance. After H2O isolation processing water vapor inside the end-face gaps of the DFB-LD, the collimator, and the PD can be utilized as a reference to design a simple but feasible H2O sensor. As a result, good linearity with an R2 of 0.9964 has been realized in a concentration range of 39-2110 ppm during an application test, and a long-term test of the designed H2O sensor against the S8000 with a difference of 10 ppm has been achieved.