In shrimp, small GTPases in the Ras superfamily can regulate hemolytic phagocytosis against WSSV infection. However, the ADP ribosylation factors (Arfs), also belonging to the regulatory GTP-binding ...proteins and playing a central role in membrane trafficking, have not been reported yet in shrimp and their relationship with WSSV infection is completely unknown to date. Here, a novel class II Arf (designated as MjArf4) was cloned and characterized from the shrimp
Marsupenaeus japonicus. Like other Arfs, MjArf4 contains an N-terminal myristoylated site, a p loop, switch regions, as well as an interswitch region. In High Five cells, when MjArf4 was in its GDP-bound form, it dispersed into the whole cell, whereas in the GTP-bound form it promoted formation of a punctuate Golgi-like structure, indicating that the MjArf4 distribution was dependent on its GDP/GTP binding. After challenge with WSSV, the mRNA level of MjArf4 was up-regulated significantly as WSSV propagated. Thus, a member of the Arf family was characterized for the first time in shrimp and found to be involved in WSSV infection.
•Enhanced progesterone inhibition of IL-1β-driven COX-2 expression.•Reduced NFκB nuclear translocation and DNA binding.•Increased progesterone-induced gene expression.•Increased progesterone receptor ...levels.•Reduced the interaction between progesterone receptor and the repressor, NCoR.
Cyclic AMP (cAMP) has been shown to promote progesterone and glucocorticoid action in a variety of cellular settings. In this study, we have used human myometrial cells to investigate whether cAMP potentiates the ability of progesterone to repress IL-1β-driven COX-2 expression. We found that forskolin enhanced progesterone-repression of IL-1β-driven COX-2 expression in association with delayed IL-1β-induced nuclear phospho-p65 entry and reduced NF-κB binding to the COX-2 promoter. Further, forskolin enhanced the progesterone-induced expression of FKBP5 and 11βHSD1, progesterone-driven activity of a progesterone response element (PRE) and progesterone receptor (PR)-B binding to a transfected PRE. In addition, forskolin treatment increased PR-B levels and reduced the PR-A:PR-B ratio while acutely decreasing the association between PR and nuclear receptor co-repressor (NCoR) and reducing NCoR levels after 6h. These findings are of importance in situations where enhancing progesterone activity is desirable, for example in the management of endometrial cancer, the promotion of endometrial receptivity or the maintenance of myometrial quiescence during pregnancy.
High throughput sequencing has previously identified differentially expressed genes (DEGs) and enriched signalling networks in human myometrium for term (≥37 weeks) gestation labour, when defined as ...a singular state of activity at comparison to the non-labouring state. However, transcriptome changes that occur during transition from early to established labour (defined as ≤3 and >3 cm cervical dilatation, respectively) and potentially altered by fetal membrane rupture (ROM), when adapting from onset to completion of childbirth, remained to be defined. In the present study, we assessed whether differences for these two clinically observable factors of labour are associated with different myometrial transcriptome profiles. Analysis of our tissue ('bulk') RNA-seq data (NCBI Gene Expression Omnibus: GSE80172) with classification of labour into four groups, each compared to the same non-labour group, identified more DEGs for early than established labour; ROM was the strongest up-regulator of DEGs. We propose that lower DEGs frequency for early labour and/or ROM negative myometrium was attributed to bulk RNA-seq limitations associated with tissue heterogeneity, as well as the possibility that processes other than gene transcription are of more importance at labour onset. Integrative analysis with future data from additional samples, which have at least equivalent refined clinical classification for labour status, and alternative omics approaches will help to explain what truly contributes to transcriptomic changes that are critical for labour onset. Lastly, we identified five DEGs common to all labour groupings; two of which (AREG and PER3) were validated by qPCR and not differentially expressed in placenta and choriodecidua.
Bleaching of red beds, a type of hydrocarbon-induced alteration, is generally attributed to redox reactions between ferric iron minerals and hydrocarbon-bearing solutions. Herein, we report sandstone ...bleaching occurs interbedded with the coal- and dark mudstone-bearing strata at shallow depths below two unconformity surfaces separating sandstone formations of Triassic-Jurassic age in the Ordos Basin, China. Field observations, petrography, and geochemistry suggest that uplift events controlled the formation of red beds via supergene alteration and bleaching via hydrocarbon circulation. The color of sandstones below the unconformities grade from red, yellow, and white colors at shallow depths (few meters to tens of meters) to dark yellow, gray-green and gray colors at deeper depths. Organic matter (carbonaceous plant debris) and pyrite in the unaltered sandstone gave rise to the gray color. The red/yellow sandstones are characterized by the presence of extensive iron oxide/hydroxide grain coatings, exhibit intense dissolution and extensive kaolinization of detrital feldspar and biotite and lithics and are mainly composed of detrital quartz. The white, bleached sandstone presents similar petrographic characteristics as the unbleached sandstone except for a lack of iron oxide/hydroxide cements. δ18OVSMOW (9.8‰ to 15.8‰) and δDVSMOW (−103‰ to −119‰) values of kaolinite, and chemical indices of alteration of sandstones indicate a weathering origin for the kaolinite and the dissolution of labile minerals in the red and yellow sandstones. The original color of the bleached sandstone was gray during very early diagenesis and shifted to red/yellow due to the oxidation of pyrite and ferromagnesian silicate minerals (e.g., biotite) into hematite or goethite cements by the meteoric water circulation during regional uplift following the deposition of each formation. Supergene alteration associated with unconformities also created significant secondary porosity, and allowed later hydrocarbons to flow along the unconformities. The lithological properties of the weathered rocks below unconformities are highly heterogeneous both vertically and laterally and have a significant influence on fluid flow. This study provides direct evidence for hydrocarbon migration along unconformities and improves understanding of fluid-rock interaction in subsurface reservoirs.
•Bleaching is restricted to shallow depths below two unconformities.•Uplift events controlled the formation of red beds via oxidation of iron-bearing minerals.•Secondary porosity generation allowed fluid flow along the unconformities.
Progesterone is widely used to prolong gestation in women at risk of preterm labour (PTL), and acts at least in part via the inhibition of inflammatory cytokine-induced prostaglandin synthesis. This ...study investigates the mechanisms responsible for this inhibition in human myometrial cells. We used reporter constructs to demonstrate that interleukin 1beta (IL-1beta) inhibits progesterone driven PRE activation via p65 activation and that IL-1beta reduced progesterone driven gene expression (FKBP5). Conversely, we found that the activity of a p65-driven NFkappaB reporter construct was reduced by overexpression of progesterone receptor B (PRB) alone and that this was enhanced by the addition of MPA and that both MPA and progesterone suppressed IL-1beta-driven cyclo-oxygenase-2 (COX-2) expression. We found that over-expressed Halo-tagged PRB, but not PRA, bound to p65 and that in IL-1beta-treated cells, with no overexpression of either PR or p65, activated p65 bound to PR. However, we found that the ability of MPA to repress IL-1beta-driven COX-2 expression was not enhanced by overexpression of either PRB or PRA and that although the combined PR and GR antagonist Ru486 blocked the effects of progesterone and MPA, the specific PR antagonist, Org31710, did not, suggesting that progesterone and MPA act via GR and not PR. Knockdown using siRNA confirmed that both MPA and progesterone acted via GR and not PR or AR to repress IL-1beta-driven COX-2 expression. We conclude that progesterone acts via GR to repress IL-1beta-driven COX-2 activation and that although the interaction between p65 and PRB may be involved in the repression of progesterone driven gene expression it does not seem to be responsible for progesterone repression of IL-1beta-induced COX-2 expression.
The shrimp translationally controlled tumor protein (TCTP) gene was previously found up-regulated in response to viral infection. In order to further understand the transcriptional regulation of
TCTP
...gene expression under viral challenge, the 5′-flanking promoter region of the
TCTP
gene in shrimp
Marsupenaeus japonicus
(
Mj-TCTP
) was cloned by using genomic walking method. The sequence shows that this region contains many important potential binding sites for transcription factors, such as activator protein1 (AP1), cAMP response element binding protein (CREB), Ets like transcription factor-1 (Elk-1), suggesting its expression is highly regulated. The promoter activity of this 5′-flanking region was confirmed by fusing to the enhanced green fluorescence protein (EGFP) gene and testing in
Trichoplusia ni
High Five cell line. Along with the introns and exons cloned in this study, the
Mj-TCTP
DNA sequence was reported here as the first
TCTP
gene with complete genomic organization in crustaceans.
Progesterone administration reduces the risk of preterm labour in high-risk women with singleton pregnancies but has no effect in women with a multiple pregnancy. However, it should be noted that it ...is not clear why progesterone is effective in the singleton pregnancies and in addition stretch-induced preterm labour is an attractive but not fully proven explanation. The data from my studies showed that progesterone did not inhibit stretch-induced MAPK activation or gene expression possibly explaining why progesterone is ineffective in the prevention of preterm labour in multiple pregnancies. Although stretch did reduce PR expression in a NF-κB-dependent manner, this was not sufficient to inhibit progesterone action, suggesting that it is not responsible for the functional progesterone withdrawal observed with the onset of human labour. Progesterone is thought to reduce the risk of preterm labour by inhibiting inflammatory cytokine-induced increases in prostaglandin synthesis. Initially, I demonstrated that IL-1β inhibited progesterone-driven PRE activation via p65. Conversely, p65-driven NF-κB reporter construct activity was reduced by overexpression of PR-B and this was enhanced by the addition of MPA. Both MPA and progesterone repression of IL-1β-driven COX-2 expression was lost by knockdown of GR. Subsequently, a series of in vitro studies suggested that progesterone acted via progesterone-induced and GR-mediated MKP-1 activation to repress IL-1β-driven COX-2 and that although the interaction between p65 and PR-B might be involved in the repression of progesterone-driven gene expression, it did not seem to be responsible for progesterone repression of IL-1β-induced COX-2 expression. Finally, the cDNA microarray analysis showed that the PR and GR regulated distinct gene networks and cellular functions in the absence or presence of ligand. However, the ability of progesterone to modulate gene expression can be mediated via both PR and GR. These data broaden our view of progesterone action and suggest alternative roles for PR and GR in human parturition.
Progesterone administration reduces the risk of preterm labour in high-risk women with singleton pregnancies but has no effect in women with a multiple pregnancy. However, it should be noted that it ...is not clear why progesterone is effective in the singleton pregnancies and in addition stretch-induced preterm labour is an attractive but not fully proven explanation. The data from my studies showed that progesterone did not inhibit stretch-induced MAPK activation or gene expression possibly explaining why progesterone is ineffective in the prevention of preterm labour in multiple pregnancies. Although stretch did reduce PR expression in a NF-κB-dependent manner, this was not sufficient to inhibit progesterone action, suggesting that it is not responsible for the functional progesterone withdrawal observed with the onset of human labour. Progesterone is thought to reduce the risk of preterm labour by inhibiting inflammatory cytokine-induced increases in prostaglandin synthesis. Initially, I demonstrated that IL-1β inhibited progesterone-driven PRE activation via p65. Conversely, p65-driven NF-κB reporter construct activity was reduced by overexpression of PR-B and this was enhanced by the addition of MPA. Both MPA and progesterone repression of IL-1β-driven COX-2 expression was lost by knockdown of GR. Subsequently, a series of in vitro studies suggested that progesterone acted via progesterone-induced and GR-mediated MKP-1 activation to repress IL-1β-driven COX-2 and that although the interaction between p65 and PR-B might be involved in the repression of progesterone-driven gene expression, it did not seem to be responsible for progesterone repression of IL-1β-induced COX-2 expression. Finally, the cDNA microarray analysis showed that the PR and GR regulated distinct gene networks and cellular functions in the absence or presence of ligand. However, the ability of progesterone to modulate gene expression can be mediated via both PR and GR. These data broaden our view of progesterone action and suggest alternative roles for PR and GR in human parturition.
With the rapid development of engineering component with integration,high-speed and multi-parameter,traditional techniques haven＇t met practical needs in extreme service environment.Laser welding,a ...new welding technology,has been widely used.However,it would generate the drop of mechanical properties for laser welded joint due to its thermal effect.Laser shock processing（LSP） is one of the most effective methods to improve the mechanical properties of laser welded ANSI 304 stainless steel joint.In this paper,the effects of LSP on the mechanical properties of laser welded ANSI 304 stainless steel joint have been investigated.The welded joint on the front of the tensile samples is treated by LSP impacts,and the overlapping rate of the laser spot is 50%.The tensile test of the laser welded joint with and without LSP impacts is carried out,and the fracture morphology of the tensile samples is analyzed by scanning electron microscope（SEM）.Compared with the yield strength of 11.70 kN,the tensile strength of 37.66 kN,the yield-to-tensile strength ratio of 0.310 7,the elongation of 25.20%,the area reduction of 32.68% and the elastic modulus of 13 063.876 MPa,the corresponding values after LSP impacts are 14.25 kN,38.74 kN,0.367 8,26.58%,42.29% and 14 754.394 MPa,respectively.Through LSP impacts,the increasing ratio of the yield strength and tensile strength are 121.79% and 102.87%,respectively;the elongation and area reduction are improved by 5.48% and 29.38%,respectively.By comparing with coarse fracture surface of the welded joint,the delamination splitting with some cracks in the sharp corner of the welded joint and asymmetric dimples,LSP can cause brighter fracture surface,and finer and more uniform dimples.Finally,the schematic illustration of dimple formation with LSP is clearly described.The proposed research ensures that the LSP technology can clearly improve the yield strength,tensile strength,yield-to-tensile strength ratio,elongation,area reduction and elastic modulus of the welded joint.The enhancement mechanism of LSP on laser welded ANSI 304 stainless steel joint is mainly due to the fact that the refined and uniform dimples effectively delay the fracture of laser welded joints.
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