Loss of CEACAM1 leads to elevated Syk activation and enhanced inflammasome activation.

<p>(<b>A</b>) Immunoblot analysis of WT and Ceacam1^−/− (KO) neutrophils with p-Syk (Y525/526), Syk and GAPDH antibodies. (<b>B</b>) Immunoblot analysis of TLR4, Syk, p-Syk and CEACAM1 in WT and Ceacam1^−/− (KO) neutrophils with or without LPS treatment (100 ng/ml) after immunoprecipitation with TLR4 antibody. (<b>C</b>) Confocal microscopy showing LPS induced lysosomal destabilization in WT and Ceacam1^−/− neutrophils (DQ-Ovalbumin, 10 mg/ml; green) with or without LPS (100 ng/ml), cell membranes were stained with fluorescent cholera toxin B-subunit (red). (<b>D</b>) Quantification of lysosomal destabilization of WT and Ceacam1^−/− neutrophils as measured by MFI of LysoSensor Green using FACS. (<b>E</b>) IL-1β production in the supernatants of LPS treated WT and Ceacam1^−/− neutrophils under different treatment conditions. (<b>F</b>) Immunoblot analysis showing caspase-1 activation of LPS treated WT and Ceacam1^−/− neutrophils under different treatment conditions. (<b>G</b>) IL-1β production in the supernatants of LPS treated WT neutrophils with or without glibenclamide treatment (250 µM). Data are representative of 3 different experiments and p values (<b>C</b>) were calculated by a 2-tailed T-test *0.01</p

LPS triggered IL-1β production in WT and Ceacam1^−/− neutrophils is partially dependent on cathepsin B.

<p>(<b>A</b>) Confocal microscopy showing cathepsin B activity in LPS treated WT and Ceacam1^−/− (KO) neutrophils. (<b>B</b>) Immunoblot analysis showing cathepsin B amounts in the supernatants of LPS treated WT and Ceacam1^−/− neutrophils. (<b>C</b>) IL-1β production in the supernatants of LPS treated WT and Ceacam1^−/− neutrophils after treatment with CA-074-Me. (<b>D</b>) Immunoblot analysis showing caspase-1 activation of LPS treated WT and Ceacam1^−/− neutrophils after CA-074-Me treatment. Data are representative of 3 different experiments (<b>A</b>, <b>B</b> and <b>D</b>) and p values (<b>C</b>) were calculated by a 2-tailed T-test ***p≤0.001.</p

Model for the inhibition of the inflammasome in neutrophils by CEACAM1.

<p>(<b>A</b>) LPS binds to TLR (the usual downstream effects such as activation of NFκB are not shown for clarity). (<b>B</b>) The complex recruits and activates Syk (pSyk) which in the absence of CEACAM1 fully activates the inflammasome that includes ROS production from the mitochondrion and cathepsin B from the lysosome. The activated inflammasome converts pro-caspase-1 to active caspase-1, which in turn converts pro-IL-1β to active IL-1β. (<b>C</b>) In the presence of CEACAM1, both Syk and CEACAM1 are phosphorylated when LPS binds to TLR4. CEACAM1 recruits SHP1 via its phosphorylated ITIM. SHP1 dephosphorylates pSyk, reducing the production of ROS and lysosome disruption, which in turn, reduces the activity of the inflammasome.</p

LPS induced neutrophil inflammasome activation depends on ROS production and lysosome destabilization.

<p>(<b>A</b>) IL-1β production in the supernatants of wild type (WT), P2X7^−/− and Cybb^−/− neutrophils under different treatment conditions: LPS (100 ng/ml), KN62 (2 µM, neutrophils were pre-treated for 30 minutes before LPS treatment), APDC (100 µM), bafilomycin A (125 nM), z-YVAD-fmk (1 mM). (<b>B</b>) Immunoblot analysis showing caspase-1 activation of WT, P2X7^−/− and Cybb^−/− neutrophils under different treatment conditions. (<b>C</b>) ROS production by LPS treated WT and CEACAM1^−/− neutrophils with or without APDC (100 µM) as measured by MFI of fluorescent probe H2DCFDA using FACS. Data are representative of 3 different experiments (<b>B</b>) and p values (<b>A</b> and <b>C</b>) were calculated by a 2-tailed T-test, **0.001</p

CEACAM1 down-regulates Syk activation through ITIM recruitment of SHP-1.

<p>(<b>A</b>) Immunoblot analysis of p-Tyr, Syk, SHP-1 and CEACAM1 in WT and Ceacam1^−/− (KO) neutrophils with or without LPS treatment (100 ng/ml) after IP with anti-CEACAM1 antibody. (<b>B</b>) Immunoblot analysis of Syk, SHP-1 in WT and Ceacam1^−/− neutrophils with or without LPS after IP with anti-SHP-1 antibody. (<b>C</b>) Immunoblot analysis of p-Syk, Syk and GAPDH in neutrophils from Ceacam1^−/− mice reintroduced with empty vector (KO/(emp)), CEACAM1-2L (KO/(CC1-2L)), CEACAM1-4L (KO/(CC1-4L)), CEACAM1-2S (KO/(CC1-2S)), CEACAM1-4S (KO/(CC1-4S)), ITIMs mutated CEACAM1-2L (KO/(CC1-2 Lm)) and ITIMs mutated CEACAM1-4L (KO/(CC1-4 Lm)) with or without LPS treatment. (<b>D</b>) Immunoblot analysis of the p-Tyr, Syk, SHP-1 and CEACAM1 in neutrophils from KO/(emp), KO/(CC1-2L), KO/(CC1-4L), KO/(CC1-2S), KO/(CC1-4S), KO/(CC1-2 Lm) and KO/(CC1-4 Lm) chimeras after IP with CEACAM1 antibody. (<b>E</b>) IL-1β production in the supernatants of LPS treated neutrophils from KO/(emp), KO/(CC1-2L), KO/(CC1-4L), KO/(CC1-2S), KO/(CC1-4S), KO/(CC1-2 Lm) and KO/(CC1-4 Lm) chimeras. (<b>F</b>) Immunoblot analysis showing caspase-1 activation of LPS treated neutrophils KO/(emp), KO/(CC1-2L), KO/(CC1-4L), KO/(CC1-2S), KO/(CC1-4S), KO/(CC1-2 Lm) and KO/(CC1-4 Lm) chimeras. Data are representative of 3 different experiments and p values (<b>E</b>) were calculated by a 2-tailed T-test ***p≤0.001.</p

RNAi reduction of SHP-1 leads to elevated Syk activation and augmented inflammasome activation.

<p>(<b>A</b>) Immunoblot analysis of p-Syk, Syk, SHP-1 and GAPDH of LPS treated neutrophils from WT mice reconstituted with shRNA control retroviral vector transduced WT BM cells (WT/(shcontrol)), WT mice reconstituted with shSHP-1 retroviral vector transduced WT BM cells (WT/(shSHP-1)) compared with WT mice. (<b>B</b>) ROS production by LPS treated neutrophils from WT mice, WT/(shcontrol) and WT/(shSHP-1) chimera measured by MFI of fluorescent probe H2DCFDA using FACS. (<b>C</b>) Quantification of lysosome destabilization of LPS treated neutrophils from WT mice, WT/(shcontrol) and WT/(shSHP-1) chimera measured by MFI of LysoSensor Green using FACS. (<b>D</b>) Immunoblot analysis showing capase-1 activation of LPS treated neutrophils from WT mice, WT/(shcontrol) and WT/(shSHP-1). (<b>E</b>) IL-1β production of LPS treated neutrophils from WT mice, WT/(shcontrol) and WT/(shSHP-1) chimera. Data are representative of 3 different experiments (<b>A</b> and <b>D</b>) and p values (<b>B, C</b> and <b>E</b>) were calculated by a 2-tailed T-test *0.01</p

Nonreciprocal Pancharatnam-Berry Metasurface for Unidirectional Wavefront Manipulation

Optical metasurfaces have been widely regarded as a new generation of building blocks for designing multifunctional devices to manipulate electromagnetic (EM) waves due to their low intrinsic loss and easy fabrication. The Pancharatnam-Berry (PB) geometric phase has been extensively exploited in metasurfaces to realize multiple spin-dependent functionalities, such as the circularly polarized (CP) beam steering, focusing, holography, etc. However, the ever-increasing demand for PB metasurfaces in complex environments has simultaneously brought about one challenging problem, i.e., the interference of multiple wave channels, which limits the performance of various PB metadevices. One possible solution is developing nonreciprocal PB metasurfaces with the optical isolation property. Here, we present a magnetic-field-biased nonreciprocal PB metasurface composed of elliptical dielectric cylinders and a thin YIG layer, which can achieve nearly 92% and 81% isolation rates of CP lights at 5.5 GHz and 6.5 GHz, respectively. The high isolation of this metasurface is attributed to the enhancement of the Faraday-magnetic-optical (FMO) effect by the resonant coupling between the dielectric Mie modes and the Fabry-P\'erot (FP) cavity mode. We apply the PB phase and nonreciprocity of the metasurface to achieve unidirectional manipulation of the wavefront of CP light beams. This gives rise to two types of metadevices: the metadeflector with nonreciprocal beam steering, and the metalens with nonreciprocal beam focusing. The proposed PB metasurface can enable efficient wavefront control while isolating the undesired wave channels, thus promising highly useful applications in optical communications, optical sensing, and quantum information processing

Protective Effect of Metalloporphyrins against Cisplatin-Induced Kidney Injury in Mice

<div><p>Oxidative and nitrative stress is a well-known phenomenon in cisplatin-induced nephrotoxicity. The purpose of this work is to study the role of two metalloporphyrins (FeTMPyP and MnTBAP), water soluble complexes, in cisplatin-induced renal damage and their ability to scavenge peroxynitrite. In cisplatin-induced nephropathy study in mice, renal nitrative stress was evident by the increase in protein nitration. Cisplatin-induced nephrotoxicity was also evident by the histological damage from the loss of the proximal tubular brush border, blebbing of apical membranes, tubular epithelial cell detachment from the basement membrane, or intra-luminal aggregation of cells and proteins and by the increase in blood urea nitrogen and serum creatinine. Cisplatin-induced apoptosis and cell death as shown by Caspase 3 assessments, TUNEL staining and DNA fragmentation Cisplatin-induced nitrative stress, apoptosis and nephrotoxicity were attenuated by both metalloporphyrins. Heme oxygenase (HO-1) also plays a critical role in metalloporphyrin-mediated protection of cisplatin-induced nephrotoxicity. It is evident that nitrative stress plays a critical role in cisplatin-induced nephrotoxicity in mice. Our data suggest that peroxynitrite is involved, at least in part, in cisplatin-induced nephrotoxicity and protein nitration and cisplatin-induced nephrotoxicity can be prevented with the use of metalloporphyrins.</p></div

Supplemental Material - Ropivacaine represses the ovarian cancer cell stemness and facilitates cell ferroptosis through inactivating the PI3K/AKT signaling pathway

Supplemental Material for Ropivacaine represses the ovarian cancer cell stemness and facilitates cell ferroptosis through inactivating the PI3K/AKT signaling pathway by Yi Lu, Jinbao Mao, Yanbing Xu, Hao Pan, Yu Wang and Wei Li in Human & Experimental Toxicology</p

Effects of metalloporphyrins on Cisplatin induced tubular apoptosis.

<p>TUNEL-positive apoptotic cell numbers were also increased in cisplatin-treated mice and attenuated by FeTMPyP or MnTBAP treatments.</p