PA Cell Research Articles

A bacterial sense of touch: T4P retraction motor as a means of surface sensing by Pseudomonas aeruginosa PA14.

Most microbial cells found in nature exist in matrix-covered, surface-attached communities known as biofilms. This mode of growth is initiated by the ability of the microbe to sense a surface on which to grow. The opportunistic pathogen Pseudomonas aeruginosa (Pa) PA14 utilizes a single polar flagellum and type 4 pili (T4P) to sense surfaces. For Pa, T4P-dependent "twitching" motility is characterized by effectively pulling the cell across a surface through a complex process of cooperative binding, pulling, and unbinding. T4P retraction is powered by hexameric ATPases. Pa cells that have engaged a surface increase production of the second messenger cyclic AMP (cAMP) over multiple generations via the Pil-Chp system. This rise in cAMP allows cells and their progeny to become better adapted for surface attachment and activates virulence pathways through the cAMP-binding transcription factor Vfr. While many studies have focused on mechanisms of T4P twitching and regulation of T4P production and function by the Pil-Chp system, the mechanism by which Pa senses and relays a surface-engagement signal to the cell is still an open question. Here we review the current state of the surface sensing literature for Pa, with a focus on T4P, and propose an integrated model of surface sensing whereby the retraction motor PilT senses and relays the signal to the Pil-Chp system via PilJ to drive cAMP production and adaptation to a surface lifestyle.

A novel multifunctional C3-symmetric triphenylamine discotic liquid crystal: synthesis, columnar self-assembly, organogel behavior, selective detection of PA and application in Si solar cells

A novel C3-symmetric triphenylamine discotic liquid crystal with fluorescent gel properties has important applications in detection of PA and Si hybrid solar cells.

FRI126 Autoreactive T Cells To Angiotensin II Type 1 Receptor In Salt-dependent Hypertension Due To Primary Aldosteronism

Abstract Disclosure: R. Scarpa: None. M. Piazza: None. B. Caroccia: None. S. Carraro: None. F. Vianello: None. F. Cinetto: None. C. Agostini: None. T.M. Seccia: None. G. Rossi: None. Compelling evidence suggested a role of acquired immunity in hypertension and hypertension-mediated organ damage (HMOD), but the pathogenic role of this acquired immunity is still unknown. Primary Aldosteronism, the paradigm of salt-dependent hypertension, is characterised by a more prominent HMOD than primary (‘essential’) hypertension and by high titer of autoantibodies against the type 1 angiotensin receptor (AT1R). As T cells are broadly recognised as HMOD-mediators, we aim to investigate: 1) the presence of AT1R-autoreactive T cells in PA; 2) aldosterone rapid and chronic effect on AT1R-autoreactive T lymphocytes; 3) the presence and functional role of know aldosterone receptors on T lymphocytes. The presence of AT1R-autoreactive T cells in 11 PA patients was investigated by flow cytometry after stimulation with pool of peptides spanning the full length of AT1R. The expression of mineralocorticoid receptor (MR) and G Protein-Coupled Estrogen Receptor (GPER) on T cells was tested by measuring mRNA copy number by droplet digital PCR and immunoblotting from healthy donors (HD) subjects and PA patients. Peripheral blood mononuclear cells (PBMCs) from PA patients were exposed to aldosterone 10-10 M with or without MR antagonist (canrenone) and GPER agonist (G1) and antagonist (G36). We evaluated the effect of aldosterone on CD8+ T cells by measuring IFNɣ release with flow cytometry. PA patients were found to have CD8+ AT1R-autoreactive T cells after activation of PBMC with AT1R peptides. MR and GPER were detected both at mRNA and protein level in CD4+ and CD8+ T cells with the following rank of expression GR>MR>>GPER. Aldosterone significantly increased IFNɣ release in CD8+ T-cell both under chronic and acute stimulation. Chronic exposure to aldosterone increased IFN-γ production by CD8+ T cells by acting via the MR, as it was prevented (p < 0.004) by canrenone, while the rapid aldosterone action was MR and GPER mediated, as it was reduced by canrenone (p < 0.003), but also by G36 (p < 0.02), and mimicked by G1. These results showed the presence of AT1R-autoreactive T cells in patients with PA. Moreover, upon of MR and GPER receptor stimulation with aldosterone, AT1-R specific CD8+ T cells undergo profound changes that mediated a strong cytotoxic response towards AT1-R-expressing organs. Presentation: Friday, June 16, 2023

Tumor-Associated Fibroblast-Derived Exosomal circDennd1b Promotes Pituitary Adenoma Progression by Modulating the miR-145-5p/ONECUT2 Axis and Activating the MAPK Pathway.

TAF participated in the progression of various cancers, including PA via the release of soluble factors. Exosomes belonged to extracellular vesicles, which were revealed as a crucial participator in intercellular communication. However, the expression pattern and effect of TAF-derived exosomes remained largely unknown in PA. In the present study, we performed in silico analysis based on public RNA-seq datasets to generate the circRNA/miRNA regulatory network. The qRT-PCR, Western blotting, RNA pull-down, and luciferase assay were performed to investigate the effect of TAF-derived exosomes. TAF-derived exosomal circDennd1b was significantly upregulated in PA and promoted the proliferation, migration, and invasion of PA cells via sponging miR-145-5p in PA cells. In addition, miR-145-5p directly regulated One Cut homeobox 2 (ONECUT2/OC2) expression and inhibited the promoting effect of ONECUT2 on PA. We further demonstrated that ONECUT2 transcriptionally increased fibroblast growth factor receptor 3 (FGFR3) expression, which further activates the mitogen-activated protein kinases (MAPK) pathway, thus promoting PA progression. Moreover, the suppression of TAFs by ABT-263 and ONECUT2 by CSRM617 inhibited the growth of PA. In conclusion, our study illustrated that TAF-derived exosomal circDennd1b affected PA progression via regulating ONECUT2 expression, which provides a potential therapeutic strategy against aggressive PA.

LGG-03. EXPLORING THE SIGNALING NETWORK INVOLVED IN MAPK PATHWAY INHIBITION BY THE MEK INHIBITOR TRAMETINIB IN BRAF-FUSION-DRIVEN PEDIATRIC PILOCYTIC ASTROCYTOMA

Abstract INTRODUCTION Pilocytic astrocytomas (PA) are the most common pediatric brain tumors. They are characterized by MAPK pathway alterations, leading to its constitutive activation and modulating the balance between proliferation and oncogene-induced senescence (OIS) sustained by senescence-associated secretory phenotype (SASP). Little is known about the molecular implications of MAPK pathway inhibition in the proliferating and senescent tumor compartments. METHODS DKFZ-BT66 cells derived from a primary KIAA:BRAF-fusion positive PA cell line and BT40 cells derived from pleomorphic xanthoastrocytoma with a BRAFV600E mutation and CDKN2A/B deletion, were used as model systems. RNA-sequencing and phospho-/proteomic datasets were generated in both the proliferative and senescent cells, and treated with the MEKi trametinib for different time-spans. A multi-omics factor analysis tool (MEFISTO) was used to identify key OIS/proliferation effectors. RESULTS Differential gene expression analysis revealed that MEK inhibition leads to the inhibition of the OIS/SASP gene program in senescent DKFZ-BT66. In addition, the protein level of several SASP factors was decreased. This translated in reduced sensitivity towards senolytics drugs, indicating inhibition of senescence features upon MEKi. MEFISTO analysis allowed to identify key transcription factors, genes and proteins involved in MAPK-induced OIS in the senescent PA cells, that were mapped using a prior knowledge network approach. Finally, single sample geneset enrichment analysis showed that most MAPK-related signatures were downregulated upon MEKi treatment, while pathways related to upstream MAPK activators (including several RTK pathways) were predicted to be upregulated, in both proliferating and senescent cells. CONCLUSION This data suggests that MAPKi reverses OIS in senescent PA cells, while inducing the activation of MAPK upstream regulators, identifying putative co-targets for the treatment of PA. Further validation of the targetability of these pathways is pending. Furthermore, the identification of the MAPK-related OIS/SASP genes provide insight about the regulation of OIS/SASP by the MAPK pathway.

LGG-02. DRUG-CLASS SPECIFIC GENE-BASED MAPK SENSITIVITY SCORES (MSS) PREDICT SENSITIVITY TO MAPK INHIBITORS AND IDENTIFY IMMUNE INFILTRATION AS PUTATIVE TARGET IN PEDIATRIC LOW-GRADE GLIOMAS

Abstract INTRODUCTION Pediatric low-grade gliomas (pLGG), the most common brain tumors in children, are driven by alterations in the MAPK pathway. Several clinical trials have shown the potential for MAPK inhibitors (MAPKi) treatment in pLGG. However, the range of response is broad, even within entities sharing the same driving genetic MAPK alteration. A predictive stratification tool is needed to identify patients that will be more likely to benefit from MAPKi therapy. METHODS We generated gene-expression-based MAPKi sensitivity scores (MSS) for each MAPKi class (BRAFi, MEKi, ERKi), based on MAPK-related genes differentially regulated between MAPKi sensitive and non-sensitive cell lines from the Genomics of Drug Sensitivity in Cancer (GDSC) dataset. Single sample Gene Set Enrichment Analysis (ssGSEA) was used to measure and validate our MSSs in the GDSC dataset and an independent PDX dataset (XevaDB). The validated signatures were tested in a pLGG-specific background, using gene expression data from PA cell lines and primary pLGG samples. RESULTS Our MSS could differentiate MAPKi sensitive cells in the GDSC dataset, and significantly correlated with MAPKi response in the XevaDB PDX dataset. The MSS were able to differentiate glioma entities with differing MAPK alterations from non-MAPK altered entities, and showed the highest scores in pLGG. The MSSs were heterogeneous within pLGG entities with a common MAPK alteration, as observed in MAPKi clinical studies. Intriguingly, a strong correlation between our MSS and the predicted immune cell infiltration rate, as determined by the Estimate score, was observed and confirmed in a pLGG scRNA sequencing dataset. CONCLUSION These data demonstrate the relevance of gene-expression signatures to predict response to MAPKi treatment in pLGG, and will be further investigated in a prospective manner in upcoming clinical trials. In addition, our data could suggest a role of immune infiltration in the response to MAPKi in pLGG that warrants further validation.

Optimization of Photoacoustic Cell for Trace Acetylene Detection in Transformer Oil

This paper presents the development of a highly sensitive gas detection system based on a resonant photoacoustic cell for detecting dissolved gases in transformer oil. A simulation model of the resonant photoacoustic cell was studied and optimized the buffer chamber volume while ensuring signal enhancement. The volume of the photoacoustic cell was reduced by about 80% compared to the classical model. A resonant photoacoustic cell was then fabricated based on the optimized simulation optimization. The dual-resonance photoacoustic system was constructed by combining the resonant PA cell with a handmade cantilever fiber acoustic sensor. The system’s sensitivity was further improved by using an erbium-doped fiber amplifier, wavelength modulation, and harmonic detection technology. The experimental results showed that the system achieved a detection limit of 6 ppb and an excellent linear range under 1000 ppm for C2H2 gas. The developed gas detection system has potential applications for monitoring the condition of power transformers in power grids.

Wavelength-modulated photoacoustic spectroscopy sensor for multi-gas measurement of acetone, methane, and water vapor based on a differential acoustic resonator

Precise assessment of breath acetone and methane is significant to the medical diagnosis process. A multi-gas sensing system based on wavelength-modulated differential photoacoustic spectroscopy was developed for simultaneous measurement of acetone and methane. A distributed feedback diode laser emitting in the range from 3363 to 3371 nm was employed to scan the absorption lines of acetone and methane. The cross sensitivities in terms of spectral interference among acetone, methane, and water vapor are effectively eliminated by using a linear combination method of reference spectra for accurately determining the concentration of acetone and methane. The positive effect of water vapor on photoacoustic signal resulting from the light absorption of acetone and methane was precisely evaluated. To improve the instrument performance, a differential PA cell companying with a differential amplifier circuit is experimentally demonstrated in providing a better performance of noise suppression compared with a single acoustic resonator. With a low detection limit down to 0.43 ppm and 12 ppb (integration time of 10 s) for acetone and methane, the sensor shows a great potential for medical diagnosis in simultaneous measurement of acetone and methane.

Miniaturized nonresonant photoacoustic gas analyzer for CO2 detection

AbstractIn this letter, a portable trace carbon dioxide (CO2) measurement instrument based on photoacoustic spectroscopy (PAS) was proposed and designed. The nonresonance PA cell (PAC) including a cooper tube with 251.2 μl was designed. Two condenser microphones were used to measure the PA signal simultaneously. The center wavelength of the distributed feedback laser was chosen to 2004 nm. Within 1 s integral time, the detection limit for CO2 reached 12.7 ppm. The normalized noise‐equivalent absorption coefficient was calculated to be 4.8468 × 10−9 cm−1W/Hz1/2. With Allan‐Werle deviation analysis, the measurement detection limit for CO2 improved to 4 ppm with 30 s integration time. As a result, the trace CO2 instrument based on PAS shows a good performance in long‐term detection.

MicroRNA-149 Regulates Proliferation, Migration, and Invasion of Pituitary Adenoma Cells by Targeting ADAM12 and MMP14

ObjectivePituitary adenomas (PAs) can adapt an aggressive phenotype by invading adjacent brain structures with rapid cellular proliferation. Previous studies demonstrated that excessive expression of metalloproteases ADAM12 and MMP-14 is instrumental for the active proliferation and invasiveness of PA cells in vitro and of tumors in vivo. However, the mechanisms regulating ADAM12 and MMP-14 expression in PAs remain unclear.MethodsTarget gene prediction and transcriptomic profiling of invasive vs. noninvasive human PA samples were performed to identify miRNA species potentially involved in the regulation of ADAM12 and MMP14. For cellular analyses of miRNA functions, two mouse PA cell lines (AtT20 and TtT/GF) were transfected with miR-149-3p and miR-149-5p, respectively. The effects of miR-149 (3p and 5p) on expression levels of ADAM12 and MMP14 were determined by Western blotting followed by an analysis of proliferation and colony formation assays, scratch migration assays, and invasion assays.ResultsA significant downregulation of miRNA-149 was observed in invasive vs. noninvasive PA (0.32 vs. 0.09, P<0.0001). In AtT-20 and TtT/GF mouse PAs cells, transfection of mimic miRNA-149 (3p and 5p) caused a significantly reduced cell proliferation and matrigel invasion, whilst the effect on cell migration was less pronounced. Both strands of miRNA-149 (3p and 5p) markedly reduced protein levels of ADAM12 and MMP-14 by at least 40% in both cell lines.ConclusionThis study proved that the invasiveness of PA cells is, at least partly, regulated by miRNA-149-dependent expression of ADAM12 and MMP-14.

A sensor based on high-sensitivity multi-pass resonant photoacoustic spectroscopy for detection of hydrogen sulfide

A high-sensitive tunable diode laser sensor based on photoacoustic spectroscopy and multi-pass absorption spectroscopy is developed for hydrogen sulfide (H2S) detection using a target absorption line at 6287.005 cm−1. Two spherical concave mirrors are utilized as the reflectors to make the laser beam pass through the photoacoustic cell for 24 times. The wavelength modulation spectroscopy technique with second harmonic detection (WMS-2f) method is employed to reduce external interference and increase the detection sensitivity. With the enhancement of the effective laser power in the multi-pass PA cell, the photoacoustic 2f signal of the target H2S absorption line has a ∼ 8 times improvement compared to the sensor system without multi-pass cell. Based on the results of Allan variance analysis, the minimum detection concentration of the system could reach 0.68 ppm with a 10-s average time and it can be further lowered down to ∼ 0.1 ppm when a 1-minute averaging time is used. The linear responsivity of the multi-pass PA spectroscopy sensor is estimated to be ∼ 0.00112 mV/ppm by linear fitting for a continuous flow gas measurement results. The excellent performance validates the potential of the sensor for field application.

&lt;i&gt;TLR9&lt;/i&gt; gene expression in immune cells of patients with psoriasis

Background. Psoriasis (skin lesions and psoriatic arthritis PA) is a chronic inflammatory autoimmune disease that can be triggered by excessive activation of endosomal toll-like receptors (TLRs), mainly TLR9. Elevated TLR9 levels are observed in both PA and psoriasis. Therefore, studying the expression pattern of the TLR9 gene may help select therapies for patients with PA and psoriasis.&#x0D; Aim. To study the expression pattern of the TLR9 gene in blood mononuclear cells of PA and psoriasis patients without joint involvement for possible use in the transition to targeted therapy.&#x0D; Materials and methods. Mononuclear cells were isolated from the peripheral blood of 31 patients with psoriasis vulgaris without joint involvement, 45 patients with PA, and 20 healthy volunteers. TLR9 gene expression was analyzed by real-time polymerase chain reaction.&#x0D; Results. A comparison of expression levels in PA patients and healthy volunteers showed that the expression level of TLR9 in PA patients was 591 times higher than that in healthy volunteers. The expression level of TLR9 in psoriasis patients without joint involvement was also significantly (423-fold) higher than that in healthy subjects.&#x0D; Conclusion. TLR9 gene expression in mononuclear cells of psoriasis patients with severe skin lesions is similar to that in PA patients. High levels of TLR9 gene expression may be a marker of possible joint involvement in patients with psoriasis and indicate the need to reconsider the therapeutic approach to a particular patient.

LBODP025 A Novel Drug Discovery For Primary Aldosteronism Targeting Aldosterone Synthase (cyp11b2) And Kcnj5 Mutant

Abstract Primary aldosteronism (PA) is a common cause of secondary hypertension. PA induces damage to cardiovascular and renal blood vessels. Therefore, PA is regarded as a problem because it increases the risk of stroke, cardiomyopathy, kidney disease, etc. Aldosterone-producing adenomas (APA) are one of the major causes of PA, and gene mutations are often identified in tumor cells. KCNJ5 mutation is well known as a gene mutation that causes the loss of ion selectivity in APA. KCNJ5 is an inwardly rectifying potassium channel. KCNJ5 induces sodium ion influx due to mutations in APA. The sodium ion influx induces depolarization, and which overexpresses CYP11B2. CYP11B2 is the rate-determining enzyme for aldosterone synthesis, overexpression of CYP11B2 induces aldosterone overproduction. Examples of therapeutic drugs for PA and APA include calcium channel blockers, aldosterone antagonist and β-blockers. However, therapeutic drugs for PA and APA targeting cell membrane depolarization have not been developed yet. We aim to find compounds that suppress CYP11B2 expression by membrane depolarization to develop novel therapeutic drugs for PA and APA. By developing a therapeutic drug that targets depolarization, we hope to establish an effective therapeutic method for APA caused by gene mutations such as KCNJ5 mutation. We performed high-throughput screening (HTS) using Tohoku University Compound Library (6080 compounds). First, we generated H295R-CYP11B2 N-Luc cells in which the luciferase gene is inserted downstream of the CYP11B2 locus. The cells H295R-CYP11B2 N-Luc were stimulated with KCl to cause depolarization, after which candidate compounds were added and HTS was performed by Luc assay. As a result, 101 compounds were selected. Furthermore, 25 of 101 compounds were selected by examining the cytotoxicity and dose response. Also, 11 of the 25 compounds suppressed CYP11B2 mRNA expression in KCl stimulated H295R cells. Secondary, we generated KCNJ5 mutant cells that expressed the KCNJ5 mutant (L168R) in a Doxycycline-inducible manner. 3 of 11 compounds that suppressed the expression of CYP11B2 mRNA were selected by examination using KCNJ5 mutant cells. After that, 2 of 3 compounds decreased the aldosterone production. These compounds have a different structure from existing therapeutic drugs such as calcium channel blockers. Since these can be novel therapeutic drugs for PA and APA caused by depolarization such as KCNJ5, we are examining the detailed mechanism of action of the compounds. Presentation: No date and time listed

Photoacoustic Detection of Pollutants Emitted by Transportation System for Use in Automotive Industry

In photoacoustic spectroscopy, the signal is inversely proportional to the resonant cell volume. Photoacoustic spectroscopy (PAS) is an absorption spectroscopy technique that is suitable for detecting gases at low concentrations. This desirable feature has created a growing interest in miniaturizing PA cells in recent years. In this paper, a simulation of a miniaturized H-type photoacoustic cell consisting of two buffer holes and a resonator was performed in order to detect CO, NH3, NO, and CH4 pollutants. These gases are the main components of the air pollutants that are produced by the automotive industry. The linear forms of the continuity, Navier–Stokes equations, and the energy equation were solved using the finite element method in a gaseous medium. The generated pressure could be measured by a MEMS sensor. Photoacoustic spectroscopy has proven to be a sensitive method for detecting pollutant gases. The objectives of the measurements were: determining the proper position of the pressure gauge sensor; measuring the frequency response; measuring the frequency response changes at different temperatures; studying the local velocity at the resonant frequency; and calculating the quality factor. The acoustic quality coefficient, acoustic response (pressure), local velocity, frequency response, and the effect of different temperatures on the frequency response were investigated. A frequency response measurement represents the fact that different gases have different resonance frequencies, for which CO and NO gases had values of 23.131 kHz and 23.329 kHz, respectively. The difference between these gases was 200 Hz. NH3 and CH4 gases with values of 21.206 kHz and 21.106 kHz were separable with a difference of 100 Hz. In addition, CO and NO gases had a difference of 2000 Hz compared to NH3 and CH4, which indicates the characteristic fingerprint of the designed cell in the detection of different gases. Better access to high-frequency acoustic signals was the goal of the presented model in this paper.

EC-QCL based photoacoustic spectroscopy for detection of SF6 decomposition components

The detection of multiple SF6 decomposition gases based on photoacoustic spectroscopy (PAS) has been a research hotspot in recent years. A highly sensitive sensor for the detection of sulfuryl fluoride (SO2F2), carbon tetrafluoride (CF4), and sulfur dioxide (SO2) in sulfur hexafluoride (SF6) was demonstrated by using PAS. The PA sensor is composed of an external cavity quantum cascade laser (EC-QCL) emitting between 1278 and 1360 cm−1, a Helmholtz differential resonant PA cell isolating the target gas from the surrounding environment, and an acousto-optic modulator (AOM) modulating the laser intensity. When the noise level was evaluated based on the PA signal noise of pure SF6 gas, minimum detection limits of 510 ppb, 9 ppb, and 69 ppb with 1 s integration time for SO2F2, CF4, and SO2 in SF6 were achieved at the characteristic absorption wavenumbers of 1282.8 cm−1, 1283.0 cm−1, and 1348.4 cm−1, respectively. Further, if the integration time was increased to the optimal integration time determined via Allan-Werle deviation analysis, the minimum detection limits were further improved to 70 ppb, 1.5 ppb, and 7 ppb for SO2F2, CF4, and SO2, respectively. The multilinear regression (MLR) statistical algorithm based on the least square method was utilized for discrimination of three-gas spectra and retrieving of three-gas concentrations from a three-gas mixture in SF6 despite the significant overlap between their absorption features.

LGG-27. Molecular implications of mitogen-activated protein kinase pathway inhibition by the MEK inhibitor trametinib in BRAF-fusion-driven pediatric pilocytic astrocytoma

Pilocytic astrocytomas (PA) are the most common pediatric brain tumors. They are characterized by driving alterations in the mitogen-activated protein kinase (MAPK) pathway, leading to its constitutive activation and modulating the balance between cell proliferation and oncogene-induced senescence (OIS) sustained by senescence-associated secretory phenotype (SASP) factors. This makes PA susceptible to MAPK inhibitor (MAPKi) therapies, which show encouraging results in phase 1/2 clinical trials. However, little is known about the molecular implications of MAPK inhibition in PA. The DKFZ-BT66 cell line, derived from a primary KIAA:BRAF-fusion positive PA, was used as a model system. DKFZ-BT66 were treated with the MEKi trametinib for different durations in both proliferative and senescent states. Gene expression was analyzed by gene expression profiling and protein expression/phospho-regulation by data-dependent mass spectrometry followed by label-free quantitative analysis. A time course analysis based on differentially expressed genes and phosphorylated proteins was performed, followed by a single-sample gene set enrichment analysis (ssGSEA) and kinase substrate enrichment analysis, respectively. Differential gene expression analysis revealed that MEK inhibition led to the inhibition of the OIS/SASP gene programs in senescent DKFZ-BT66, with downregulation of key OIS/SASP partners such as IL1B on the protein level. This functionally translated into a de-sensitization of these cells towards the senolytic agent navitoclax. ssGSEA showed that most MAPK-related signatures were downregulated upon MEKi treatment, while pathways related to upstream MAPK activators (including FGFR, NTRK and TGFB pathways) were upregulated, in both proliferating and senescent DKFZ-BT66. This data indicates that MAPKi reverses OIS in senescent PA cells, while inducing the activation of MAPK upstream regulators in proliferating and senescent PA cells, identifying putative co-targets that could help increase treatment’s efficacy. Validation of these targets by post-translational modification enrichment analysis of the phospho-proteomics dataset is ongoing.

A Compact SiGe Stacked Common-Base Dual-Band PA With 20/18.8 dBm P sat at 36/64 GHz Supporting Concurrent Modulation

This work presents a compact, concurrent dual-band (36/64 GHz) linear power amplifier in 90 nm SiGe technology. To simultaneously overcome the gain and power limitation of conventional common-emitter (CE) silicon device at mm-wave, the stacked common-base (SCB) topology in SiGe is studied and utilized as PA cell. A highly efficient dual-band output network enables the optimum load-line matching for two widely separated frequencies. The proposed PA achieves 20/18.8 dBm <inline-formula xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink"> <tex-math notation="LaTeX">$P_{\mathrm {sat}}$ </tex-math></inline-formula> , 25/22.5% PAE at 36/64 GHz supporting two concurrent 32 QAM signals (total data rate of 9 Gb/s) with 9.5/7.8 dBm linear average power respectively, and concurrent PAE of 8.9%. To the best of the authors’ knowledge, this prototype achieves highest dual frequency output power, PAE, and one of the first concurrent dual-band modulation among silicon-based mm-wave PAs, while still maintaining a highly compact footprint.

Oxymatrine inhibits the pyroptosis in rat insulinoma cells by affecting nuclear factor kappa B and nuclear factor (erythroid-derived 2)-like 2 protein/heme oxygenase-1 pathways.

As the mechanism underlying glucose metabolism regulation by oxymatrine is unclear, this study investigated the effects of oxymatrine on pyroptosis in INS-1 cells. Flow cytometry was employed to examine cell pyroptosis and reactive oxygen species (ROS) production. Cell pyroptosis was also investigated via transmission electron microscopy and lactate dehydrogenase (LDH) release. Protein levels were detected using western blotting and interleukin (IL)-1β and IL-18 secretion by enzyme-linked immunosorbent assay. The caspase-1 activity and DNA-binding activity of nuclear factor kappa B (NF-κB) and nuclear factor (erythroid-derived 2)-like 2 protein (Nrf2) were also assessed. In the high glucose and high fat-treated INS-1 cells (HG + PA), the caspase-1 activity and LDH content, as well as Nod-like receptor family pyrin domain containing 3, Gsdmd-N, caspase-1, apoptosis-associated speck-like protein containing a CARD, IL-1β, and IL-18 levels were increased. Moreover, P65 protein levels increased in the nucleus but decreased in the cytoplasm. Oxymatrine attenuated these effects and suppressed high glucose and high fat-induced ROS production. The increased levels of nuclear Nrf2 and heme oxygenase-1 (HO-1) in the HG + PA cells were further elevated after oxymatrine treatment, whereas cytoplasmic Nrf2 and Keleh-like ECH-associated protein levels decreased. Additionally, the elevated transcriptional activity of p65 in HG + PA cells was reduced by oxymatrine, whereas that of Nrf2 increased. The results indicate that the inhibition of pyroptosis in INS-1 cells by oxymatrine, a key factor in its glucose metabolism regulation, involves the suppression of the NF-κB pathway and activation of the Nrf2/HO-1 pathway.

Liraglutide inhibits receptor for advanced glycation end products (RAGE)/reduced form of nicotinamide-adenine dinucleotide phosphate (NAPDH) signaling to ameliorate non-alcoholic fatty liver disease (NAFLD) in vivo and vitro

ABSTRACT The study was designed to investigate the effects of liraglutide and reveal its action mechanism associated with RAGE/NAPDH in NAFLD. The liver tissue was collected for HE, Masson, and ROS staining. Apoptosis levels were detected through TUNEL staining and ROS levels were evaluated through ROS staining. The expression levels of c-Jun N-terminal kinase (JNK) and transforming growth factor-β (TGF-β) were detected through Western blot. JNK and the expression of Collagenα1, Collagenα2 and connective tissue growth factor (CTGF) were detected through RT-qPCR and Western blot and the expression in mouse liver stellate cells (JS-1) cells were evaluated through immunofluorescence staining. We detected the effects of liraglutide on NAFLD in high-fat diet (HFD)-fed mice. Liraglutide treatment improved bridging fibrosis and liver function, as well as lessening ROS levels and the protein levels of RAGE, NOX1, NOX2 and NOX4. In PA and H2O2-induced AML12 cells, liraglutide treatment was able to decrease cell apoptosis, ROS levels and the levels of inflammatory factors including tumor necrosis factor (TNF)-α, interleukin (IL)-1β and IL-6, while it effects were reversed by the induction of RAGE overexpression or NOX2 overexpression. In JS-1 cells treated with medium culturing AML12 cells, liraglutide markedly suppressed cell proliferation and activation, while RAGE overexpression or NOX2 overexpression blunted these effects of liraglutide. Taken together, liraglutide exerts a protective role in improving liver injury caused by HFD, which could be related to decreased apoptosis and oxidative stress of liver cells, as well as decreased proliferation and activation of hepatic stellate cells through RAGE/NOX2.

MicroRNA-146b-5p/EPHA7 axis regulates cell invasion, metastasis, proliferation, and temozolomide-induced chemoresistance via regulation of IRAK4/TRAF6/NF-κB signaling pathway in aggressive pituitary adenoma.

Aggressive pituitary adenoma (APA) is a huge challenge for neurosurgeons. Temozolomide (TMZ) is conventionally used in chemotherapy against APA, but acquired resistance developed during long-term therapy limits its benefits. MiRNA-146b-5p has been confirmed to inhibit tumor metastasis. This study aimed to explore the underlying biological functions of miRNA-146b-5p in APA. Sixty confirmed APA tissues and corresponding adjacent normal tissues were collected. We established a TMZ-resistant cell line (GH3/TMZ) by exposing GH3 cells to gradually increasing doses of TMZ for 5 months. Cell Counting Kit-8 assay, flow cytometric analysis, RNA pull-down assay, 5-ethynyl-20-deoxyuridine assay, dual-luciferase reporter gene assay, wound healing assay, and invasion assay were used to explore the malignant biological characteristics of cells. Immunohistochemistry (IHC), western blotting analysis, and real-time quantitative PCR (qRT-PCR) were used to analyze the expression level of related proteins and nucleic acids. The expression of miRNA-146b-5p was down-regulated not only in APA tissues but also in PA cell lines compared with the matched adjacent non-tumor tissues or normal human astrocyte (NHA) cells. Low expression of miRNA-146b-5p was notably associated with poorer disease-free survival rate (P=0.032), overall survival rate (P=0.039), larger tumor size (P=0.028), poorer Knosp grade (P=0.020), and poorer Hardy grade (P=0.006) in APA patients. MiRNA-146b-5p negatively regulated cell proliferation, invasion, migration, and induced apoptosis in GH3 cells. Overexpression of miRNA-146b-5p suppressed IRAK4 and TRAF6 protein expression and negatively regulated NF-κB phosphorylation. The restoration of EPHA7 expression in GH3 cells notably reversed the inhibitory effects of miRNA-146b-5p. MiRNA-146b-5p expression was significantly down-regulated and EPHA7 gene expression was significantly up-regulated in GH3/TMZ cells, compared to the parental cell line. Similarly, EPHA7 was up-regulated, while the miRNA-146b-5p level was down-regulated in chemoresistance tissues more than in chemosensitive tissues. The autophagic activity was decreased markedly with increasing miRNA-146b-5p expression, while it was enhanced after Lv-EPHA7 treatment in GH3/TMZ cells. MiRNA-146b-5p can inhibit EPHA7 expression, suppress the IRAK4/TRAF6/NF-κB signaling pathway, and weaken PA cell invasion, metastasis, proliferation, and TMZ-induced chemoresistance in vitro.