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In our previous study, pyrrolnitrin produced in Pseudomonas chlororaphis G05 plays more critical role in suppression of mycelial growth of some fungal pathogens that cause plant diseases in agriculture. Although some regulators for pyrrolnitrin biosynthesis were identified, the pyrrolnitrin regulation pathway was not fully constructed. During our screening novel regulator candidates, we obtained a white conjugant G05W02 while transposon mutagenesis was carried out between a fusion mutant G05ΔphzΔprn::lacZ and E. coli S17-1 (pUT/mini-Tn5Kan). By cloning and sequencing of the transposon-flanking DNA fragment, we found that a vfr gene in the conjugant G05W02 was disrupted with mini-Tn5Kan. In one other previous study on P. fluorescens, however, it was reported that the deletion of the vfr caused increased production of pyrrolnitrin and other antifungal metabolites. To confirm its regulatory function, we constructed the vfr-knockout mutant G05Δvfr and G05ΔphzΔprn::lacZΔvfr. By quantifying β-galactosidase activities, we found that deletion of the vfr decreased the prn operon expression dramatically. Meanwhile, by quantifying pyrrolnitrin production in the mutant G05Δvfr, we found that deficiency of the Vfr caused decreased pyrrolnitrin production. However, production of phenazine-1-carboxylic acid was same to that in the wild-type strain G05. Taken together, Vfr is required for pyrrolnitrin but not for phenazine-1-carboxylic acid biosynthesis in P. chlororaphis G05.

Background: Neuroblastoma (NBL) is an extracranial malignant tumor in children deriving from the neural crest in the sympathetic nervous system. Although various immunotherapy interventions have made significant breakthroughs in many adult cancers, the efficacy of these immunotherapies was still limited in NBL. NBL has low immunogenicity which results in a lack of tumor-infiltrating T lymphocytes in the tumor microenvironment (TME). Moreover, tumor cells can wield many immune evasion strategies both in the TME and systemically to impede lymphocyte infiltration and activation. All these factors hamper the anti-tumor effects of CD8+ T cells during immunotherapy and the levels of infiltrating CD8+ T cells correlate with therapy response. Materials and methods: In this study, we utilized multidimensional bioinformatic methods to establish a risk model based on CD8+ T cells -related genes (CD8+ TRGs). Results: We obtained 33 CD8+ TRGs with well-predictive ability for prognosis in both GSE49711 and E-MTAB-8248 cohorts. Then, 12 CD8+ TRGs including HK2, RP2, HPSE, ELL2, GFI1, SLC22A16, FCGR3A, CTSS, SH2D1A, RBP5, ATF5, and ADAM9 were finally identified for risk model construction and validation. This model revealed a stable performance in prognostic prediction of the overall survival (OS) and event-free survival (EFS) in patients with NBL. Additionally, our research indicated that the immune and stromal scores, immune-related pathways, immune cell infiltration, the expression of major histocompatibility complex (MHC) and immune checkpoint molecules, immunotherapy response, and drug susceptibility revealed significant differences between high and low-risk groups. Conclusions: According to our analyses, the constructed CD8+ TRGs-based risk model may be promising for the clinical prediction of anti-tumor therapy responses and prognoses in NBL. [ABSTRACT FROM AUTHOR]

Copy number variations have been found in patients with neural tube abnormalities. In this study, we performed genome-wide screening using high-resolution array-based comparative genomic hybridization in three children with tethered spinal cord syndrome and two healthy parents. Of eight copy number variations, four were non-polymorphic. These non-polymorphic copy number variations were associated with Angelman and Prader-Willi syndromes, and microcephaly. Gene function enrichment analysis revealed that COX8C, a gene associated with metabolic disorders of the nervous system, was located in the copy number variation region of Patient 1. Our results indicate that array-based comparative genomic hybridization can be used to diagnose tethered spinal cord syndrome. Our results may help determine the pathogenesis of tethered spinal cord syndrome and prevent occurrence of this disease.

This present study aimed to clarify the effect of contact scale and surface topography of substrates with different roughnesses on the actual contact area, tangential stiffness, and tangential deformation of the substrate at micro- and macro-scales via finite element method (FEM) simulations, as well as the final tribological performances of MoS2 coatings by experiments. The MoS2 coatings were deposited on stainless steel (SS) substrates with different roughnesses, and the settings in the simulation models were based on the roughness of the SS substrates. The predicted tribological behavior of the simulation results was confirmed by the morphological and compositional analysis of the wear track using scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDS), 3D profilometer, and Raman spectroscopy. The results showed that the substrate with a surface roughness of Ra 600 nm (R600), coated by MoS2 nanosheets, exhibited excellent tribological properties at both micro- and macro-scales. At the micro-scale, the lubrication lifetime of R600 was as long as 930 cycles, while the substrates with surface roughnesses of Ra 60 nm (R60) and Ra 6 nm (R6) had a lubrication lifetime of 290 cycles and 47 cycles, respectively. At the macro-scale, the lifetime of the substrate R600 was 9509 cycles, which was nearly six times longer than the 1616 cycles of substrate R60. For the rough surface of substrate, the surface grooves could not only effectively preserve the lubricant but also continuously release them, ensuring that the lubricants with low shear strength were always present in the contact interface. It was further verified that the high surface roughness of the substrate reduced friction and wear by reducing the actual contact area and enhancing the tangential stiffness of asperities, thereby prolonging the lubrication lifetime. The wear mechanisms were discussed in terms of the morphology and chemical composition of the wear tracks. [ABSTRACT FROM AUTHOR]

The structure and morphology of Cu films deposited by DC magnetron sputtering on silicon and stainless-steel substrates at different deposition temperatures of −140 °C, −95 °C, −55 °C, 25 °C (RT), 50 °C, and 200 °C were investigated by X-ray diffraction (XRD), scanning electron microscopy (SEM), and atomic force microscopy (AFM). It was found that all Cu films presented strong orientation of the (111) and (200) peaks. The Cu films deposited at low temperatures (lower than −55 °C) showed the bilayer structures, in which the upper layer appeared to be a loose and porous structure and the lower layer near the substrate had a fine and dense structure that consisted of small grains. In addition, the Cu films deposited at low temperatures could be observed a large roughness. The roughness tended to decline and then increase with the rising of deposition temperature. The ball-on-disc reciprocating sliding tribometer was employed to evaluate the tribological behaviors of the Cu films at current-carrying levels of 0 A, 0.5 A, and 1.0 A. The results revealed that the Cu films deposited at low temperatures exhibited outstanding current-carrying friction performance and low electrical contact resistance (ECR), peeling only at 0.5 A and 1.0 A. Nevertheless, the Cu films deposited at the relatively high temperature exhibited oxidative wear caused by electric arc ablation at 0.5 A and 1.0 A. Additionally, the wear mechanism was discussed in terms of the structure and morphology of the wear track and formation of the tribo-film. [ABSTRACT FROM AUTHOR]