Secreted MbovP0145 Promotes IL-8 Expression through Its Interactive β-Actin and MAPK Activation and Contributes to Neutrophil Migration

Mycoplasma bovis (M. bovis) is an important pathogen of cattle responsible for huge economic losses in the dairy and beef industries worldwide. The proteins secreted by M. bovis are mainly related to its adhesion, invasion, virulence, and intracellular survival and play a role in mycoplasma–host interactions. In our previous study, we found MbovP0145, a secreted protein present in the M. bovis secretome, but little is known about its function. In this study, we assessed the inflammatory characteristics and underlined mechanism of this inflammation of recombinant MbovP0145 (rMbovP0145). For this, bovine lung epithelial cells (EBL) were stimulated by rMbovP0145 to see the IL-8 production in a time- and dose-dependent manner. We observed that rMbovP0145 increased the production of IL-8 via ERK1/2 and P38 pathway activation. Further, the effect of the M. bovis ΔMbov_0145 mutant and its complementary strain on IL-8 mRNA expression was also confirmed. A pulldown assay of the GST-tagged MbovP0145 protein with mass spectrometry demonstrated that β-actin could specifically interact with rMbovP0145 to mediate the IL-8 signaling. As knockdown of β-actin expression with RNA interference in EBL cells decreased the mRNA expression of IL-8 and the phosphorylated ERK1/2 and P38 proteins, whereas disrupted actin polymerization by cytochalasin D led to a significantly higher IL-8 expression and MAPK phosphorylation in rMbovP0145-stimulated cells. Compared to M. bovis HB0801 and its complementary strain, the culture supernatant of EBL cells infected with the M. bovis ΔMbov_0145 mutant induced less neutrophil migration to the lower chamber in a transwell system. In conclusion, MbovP0145 promoted IL-8 expression by interacting with β-actin through activation of the MAPK pathway, thus contributing to neutrophil migration.

sRNA STnc150 is involved in virulence regulation of Salmonella Typhimurium by targeting fimA mRNA

ABSTRACT Small RNAs (sRNAs) are essential virulent regulators in Salmonella typhimurium (STM). To explore the role of sRNA STnc150 in regulating STM virulence, we constructed a STnc150 deletion strain (ΔSTnc150) and its complementary strain (ΔSTnc150/C). Then, we compared their characteristics to their original parent strain experimentally, identified the target genes of STnc150 and determined the expression levels of target genes. The results showed that the ΔSTnc150 strain exhibited delayed biofilm formation, enhanced adhesion to macrophages, significantly reduced LD50, increased liver and spleen viral loads and more vital pathological damaging ability than its parent and complementary strains. Further, bioinformatics combined with the bacterial dual plasmid reporter system confirmed that the bases 72–88 of STnc150 locating at the secondary stem-loop structure of the STnc150 are complementary with the bases 1–19 in the 5′-terminal of fimA mRNA of the type 1 fimbriae subunit. Western blot analysis showed that fimA protein level was increased in STnc150 strain compared with its parent and complementary strains. Together, this study suggested that STnc150 can down-regulate STM fimA expression at the translation level, which provided insights into the regulatory mechanisms of sRNAs in virulence of STM.

The Fungi-specific histone Acetyltransferase Rtt109 mediates morphogenesis, Aflatoxin synthesis and pathogenicity in Aspergillus flavus by acetylating H3K9

Aspergillus flavus is a common saprophytic filamentous fungus that produces the highly toxic natural compound aflatoxin during its growth process. Synthesis of the aflatoxins, which can contaminate food crops causing huge losses to the agricultural economy, is often regulated by epigenetic modification, such as the histone acetyltransferase. In this study, we used Aspergillus flavus as an experimental model to construct the acetyltransferase gene rtt109 knockout strain (△rtt109) and its complementary strain (△rtt109·com) by homologous recombination. The growth of △rtt109 was significantly suppressed compared to the wild type (WT) strain and the △rtt109·com strain. The sclerotium of △rtt109 grew smaller, and the amount of sclerotia generated by △rtt109 was significantly reduced. The number of conidiums of △rtt109 was significantly reduced, especially on the yeast extract sucrose (YES) solid medium. The amount of aflatoxins synthesized by △rtt109 in the PDB liquid medium was significantly decreased We also found that the △rtt109 strain was extremely sensitive to DNA damage stress. Through the maize seed infection experiment, we found that the growth of △rtt109 on the surface of affected corn was largely reduced, and the amount of aerial mycelium decreased significantly, which was consistent with the results on the artificial medium. We further found that H3K9 was the acetylated target of Rtt109 in A. flavus. In conclusion, Rtt109 participated in the growth, conidium formation, sclerotia generation, aflatoxin synthesis, environmental stress response, regulation of infection of A. flavus. The results from this study of rtt109 showed data for acetylation in the regulation of life processes and provided a new thought regarding the prevention and control of A. flavus hazards.

On the objectivity of the nonlinear along-fibre shear stress–strain relationship for unidirectionally fibre-reinforced composites

The nonlinear along-fibre shear stress–strain relationship for unidirectionally fibre-reinforced composites has been investigated in this paper aiming at its applications in general 3D stress conditions in a consistent manner. So far, such relationship has only been addressed in plane stress conditions. In this paper, it has been shown that its straightforward generalisation to 3D stress states lacks objectivity, which is a basic requirement for all theoretical studies of physical problems. A new formulation is proposed based on the stress invariants and the complementary strain energy guided by the rational theoretical framework of nonlinear elasticity. A consistent and objective stress–strain relationship has been obtained and verified through an example of application to a torsion problem.

An Energy-Based Framework for Nonlinear Kinetostatic Modeling of Compliant Mechanisms Utilizing Beam Flexures

Although energy-based methods have advantages over the Newtonian methods for kinetostatic modeling, the geometric nonlinearities inherent in deflections of compliant mechanisms preclude most of the energy-based theorems. Castigliano's first theorem and the Crotti-Engesser theorem, which don't require the problem being solved to be linear, are selected to construct the energy-based kinetostatic modeling framework for compliant mechanisms in this work. Utilization of these two theorems requires explicitly formulating the strain energy in terms of deflections and the complementary strain energy in terms of loads, which are derived based on the beam constraint model. The kinetostatic modeling of two compliant mechanisms are provided to demonstrate the effectiveness of the explicit formulations in this framework derived from Castigliano's first theorem and the Crotti-Engesser theorem.

Optimal Plastic Analysis and Design of Pile Foundations Under Reliable Conditions

In this research, in order to evaluate the plastic limit load and also plastic design parameters of the long pile foundations subjected to horizontal loads, shakedown method is applied. In carrying out shakedown analysis and design methods, large plastic deformations and residual displacements could develop in the pile foundation which might lead to the failure of the structure. For this reason, complementary strain energy of residual forces proposed as a limit condition to control the plastic deformation of the pile structure. Furthermore, considering the uncertainties (strength, manufacturing, geometry) the limit conditions on the complementary strain energy of residual forces are assumed randomly and the reliability condition was formed by the use of the strict reliability index. The influence of the limit conditions on the plastic limit load and design parameters of the long pile in cohesionless soil subjected to lateral load were investigated and limit curves for shakedown load factors are presented. The numerical results show that the probabilistic given limit conditions on the complementary strain energy of residual forces have significant influence on the load bearing limit and the design parameters of pile foundations. The formulations of the reliability based problems lead to mathematical programming which were carried out by the use of non-linear algorithm.

Extension of Castigliano’s method for isotropic beams

In the present contribution Castigliano’s theorem is extended to find more accurate results for the deflection curves of beam-type structures. The notion extension in the context of the second Castigliano’s theorem means that all stress components are included for the computation of the complementary strain energy, and not only the dominant axial stress and the shear stress. The derivation shows that the partial derivative of the complementary strain energy with respect to a scalar dummy parameter is equal to the displacement field multiplied by the normalized traction vector caused by the dummy load distribution. Knowing the Airy stress function of an isotropic beam as a function of the bending moment, the normal force, the shear force and the axial and vertical load distributions, higher-order formulae for the deflection curves and the cross section rotation are obtained. The analytical results for statically determinate and indeterminate beams for various load cases are validated by analytical and finite element results. Furthermore, the results of the extended Castigliano theory (ECT) are compared to Bernoulli–Euler and Timoshenko results, which are special cases of ECT, if only the energies caused by the bending moment and the shear force are considered. It is shown that lower-order terms for the vertical deflection exist that yield more accurate results than the Timoshenko theory. Additionally, it is shown that a distributed load is responsible for shrinking or elongation in the axial direction.