Lysine demethylase 1A exacerbates LPS-induced inflammation of vascular smooth muscle cells through modulation of NF-κB activation

Purpose: To study the effect of lysine demethylase 1A (LSD1) on inflammatory responses of vascular smooth muscle cells (VSMCs), and investigate the mechanism. Methods: VSMCs were treated with lipopolysaccharide (LPS). Overexpression and knockdown of LSD1 in VSMCs were performed by transfecting with LSD1 overexpression plasmid and small interfering RNAs (siRNAs), respectively. Western blot and quantitative real-time polymerase chain reaction (qRTPCR) were used to measure protein and mRNA levels. Enzyme-linked immunosorbent (ELISA) assay was used to determine the levels of inflammatory cytokines. Results: Phosphorylation of LSD1 (p-LSD1) was significantly increased in LPS-induced VSMCs. Monocyte chemoattractant protein-1 and IL-6 levels were also increased by LPS, but attenuated by LSD1 knockdown in VSMCs. Activation of NF-κB was increased by LPS, but was also decreased by LSD1 knockdown. Level of methylated p65 (p65-me) in VSMCs was increased by treatment with SET7/9 (p65 methyltransferase), but this effect was attenuated by overexpression of LSD1. Besides, the increased levels of MCP-1 and IL-6 induced by overexpression of LSD1 were reversed by NF-κB signaling inhibitor, PDTC. Conclusion: LSD1 exacerbates LPS-induced inflammation of VSMCs through NF-κB activation via p65 demethylation, which indicates that LSD1 might be a potential target for the treatment of cardiovascular diseases. Keywords: Vascular smooth muscle cells, Lysine demethylase 1A, Phosphorylation, NF-κB, p65, Demethylatio

Rapid FRD determination for multiplexed fibre systems -- I. The quasi-near field model and its uncertainties

Focal Ratio Degradation (FRD) in fibres is a crucial factor to control in astronomical instruments in order to minimize light loss. As astronomical instrumentation has advanced, the integration of large populations of fibres has become common. However, determining FRD in multiplexed fibre systems has become a challenging and time-consuming task. The Integral Field Unit for the Fiber Arrayed Solar Optical Telescope (FASOT-IFU) represents the most densely arranged fibre-based IFU in a single unit. Due to the close packing of fibres in the V-groove of the slit end, measuring FRD is particularly challenging as the output spots are prone to overlapping with adjacent fibres. In this paper, a novel method based on the quasi-near field model is proposed to enable rapid FRD measurement in highly multiplexed fibre systems like IFUs and multi-object observation systems. The principle and uncertainties associated with the method are investigated. The method's validity is demonstrated by applying it to determine the FRD in FASOT-IFU, with the achieved FRD performance meeting the acceptable requirements of FASOT-IFU, where the output focal ratio primarily falls within the range of 5.0-7.0. The results indicate that the proposed method offers several advantages, including the simultaneous and rapid measurement of FRD in multiple fibres with high accuracy (error smaller than 0.35 in F-ratio). Furthermore, besides FRD, the method exhibits potential for extensive measurements of throughput, scrambling, and spectral analysis.Comment: 10 pages, 12 figures, submitted to MNRA

The small subunit of DNA polymerase D (DP1) associates with GINS-GAN complex of the thermophilic archaea in Thermococcus sp. 4557.

The eukaryotic GINS, Cdc45, and minichromosome maintenance proteins form an essential complex that moves with the DNA replication fork. The GINS protein complex has also been reported to associate with DNA polymerase. In archaea, the third domain of life, DNA polymerase D (PolD) is essential for DNA replication, and the genes encoding PolDs exist only in the genomes of archaea. The archaeal GAN (GINS-associated nuclease) is believed to be a homolog of the eukaryotic Cdc45. In this study, we found that the Thermococcus sp. 4557 DP1 (small subunit of PolD) interacted with GINS15 in vitro, and the 3’-5’ exonuclease activity of DP1 was inhibited by GINS15. We also demonstrated that the GAN, GINS15, and DP1 proteins interact to form a complex adapting a GAN-GINS15-DP1 order. The results of this study imply that the complex constitutes a core of the DNA replisome in archaea

Two-Dimensional Boronate Ester Covalent Organic Framework Thin Films with Large Single Crystalline Domains for a Neuromorphic Memory Device

Despite the recent progress in the synthesis of crystalline boronate ester covalent organic frameworks (BECOFs) in powder and thin-film through solvothermal method and on-solid-surface synthesis, respectively, their applications in electronics, remain less explored due to the challenges in thin-film processability and device integration associated with the control of film thickness, layer orientation, stability and crystallinity. Moreover, although the crystalline domain sizes of the powder samples can reach micrometer scale (up to ≈1.5 μm), the reported thin-film samples have so far rather small crystalline domains up to 100 nm. Here we demonstrate a general and efficient synthesis of crystalline two-dimensional (2D) BECOF films composed of porphyrin macrocycles and phenyl or naphthyl linkers (named as 2D BECOF-PP or 2D BECOF-PN) by employing a surfactant-monolayer-assisted interfacial synthesis (SMAIS) on the water surface. The achieved 2D BECOF-PP is featured as free-standing thin film with large single-crystalline domains up to ≈60 μm2 and tunable thickness from 6 to 16 nm. A hybrid memory device composed of 2D BECOF-PP film on silicon nanowire-based field-effect transistor is demonstrated as a bio-inspired system to mimic neuronal synapses, displaying a learning–erasing–forgetting memory process. © 2020 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA

Protection effect of gut microbiota composition and acetate absorption against hypertension-induced damages on the longevity population in Guangxi, China

IntroductionRecent evidence supports a role for the gut microbe-metabolites in longevity. However, the phenomenon of hypertension is more common in the longevity area and whether hypertension is associated with longevity remains unclear. Here, we hypothesize that the levels of gut microbiota, SCFAs, and urine metabolites were different between hypertension elderly and hypertension longevity.MethodsWe recruited 46 elderly volunteers from Donglan County, Guangxi, and 32 were selected and included in the experiment. The subjects with hypertension were divided into two groups according to age, Hypertension Elderly (HTE, aged 70.5 ± 8.59, n = 19) and Hypertension Longevity (HTL, aged 100 ± 5.72, n = 13). The gut microbiota, SCFAs, and urine metabolites were determined by three-generation 16S rRNA full-length sequencing, GC-MS, and 1H-NMR, respectively.ResultsCompared with the HTL group, the HTE group had higher levels of hypertension-related genera Klebsiella and Streptococcus, while having lower levels of the SCFA-producing genera Bacteroides, Faecalibacterium, and Alistipes. Based on LEFse analysis, Klebsiella pneumoniae, Lactobacillus gasseri, Streptococcus salivarius, Ruminococcus, Actinomyces, Rikenellaceae, f_Saccharimonadaceae, Clostridium perfringens, and Bacteroids, Faecalibacterium prausnitzii, Parabacteroides, Alistipes were biomarkers that showed significant differences between the groups. In addition, the microbial pathways associated with K. pneumoniae and E. coli may promote hypertension, while A. muciniphila may play a role in reversing the development of hypertension in long-lived elderly. Metabolomics revealed that HTL contained a lower concentration of fecal acetate and propionate than HTE, while it contained a higher concentration of serum acetate and urine acetate. Furthermore, their immune cells exhibited no significant changes in SCFAs receptors.ConclusionAlthough long-lived elderly have extremely high systolic blood pressure, their unique gut microbiota composition and efficient acetate absorption in the colon may offset the damages caused by hypertension and maintain healthy homeostasis

Imaging-guided synergistic targeting-promoted photo-chemotherapy against cancers by methotrexate-conjugated hyaluronic acid nanoparticles

Abstract(#br)A combination of chemotherapy and photothermal therapy (PTT) against cancer, overcoming the intrinsic limitations of single-modal chemotherapy or PTT, has emerged as a promising strategy to achieve synergistic therapeutic effect. However, the lack of precise drug delivery and intelligent drug release based on photo-chemotherapy at specific tumor sites remained a challenge. Hence, the both tumor-specific targeting molecule (methotrexate) and ligand (hyaluronic acid)-introduced, glutathione-responsive amphiphiles (deoxycholic acid-hyaluronic acid-methotrexate, DA-SS-HA-MTX) were developed for synchronous delivery of indocyanine green (ICG) and doxorubicin (DOX). The as-synthesized DOX/ICG@DSHM remarkably improved the intracellular drug uptake and accumulation owing to both the CD44/folate receptors-mediated synergistic targeting and the glutathione-triggered rapid drug release. Moreover, DOX/ICG@DSHM efficiently accumulated at the tumor sites, realizing the notable tumor ablation under the guidance of dual-modal optical imaging. Taken together, this study provided a promising nanotheranostic agent for imaging-guided chemo-photothermal combination therapy

Inhibition of acute allogaft rejection - experimental transplant studies with biological and chemical immunosuppressants

In clinical transplantation immunosuppressive agents are used in combination with the objective to achieve additive effects and less toxicity. A large number of novel drugs are candidates for clinical introduction at this time and experiments in animal models have an important role to play. The aim of this study was in brief to evaluate the capacity to inhibit the acute rejection process with biological and chemical immunosuppressants including anti-CD4 monoclonal antibody (mAb), and leflunomide and its analogues malononitrilamides (MNA) 279 and 715, given alone or in combination with cyclosporine or tacrolimus. A rat cardiac transplant model was used and immunosuppressive drugs given as 10 days of induction. Graft survival was the most important variable. Flow cytometry was used for analyses of donor-specific antibody production and histopathology for evaluation of acute rejection changes at various time points posttransplant. Unresponsiveness was tested with retransplantation and with harvesting of peritoneal excudate cells (PEC) following intraperitoneal challenge with donor-specific or third party cells. Various methods for analysis of combined drug effects were used including enhancement of rejection with an immunomodulator, Linomide, and with presensitisation, and iso-effect curves of drugs given alone or in combination. Major findings of this study was as follows. A single administration of anti-CD4 mAb induced unresponsiveness in a low-responder rat strain combination, whereas the effect was minimal in high-responders. Level of response was demonstrated to depend on selection of recipient as well as donor. Unresponsiveness induced by a low dose of CsA induction was donor-specific and PEC revealed a corresponding pattern of cytotoxicity. Exogenous administration of interleukin-2 could restore cytotoxicity against donor target cells suggesting a state of anergy. Leflunomide was shown to prolong graft survival and in combination with CsA to overcome the challenge of Linomide. The MNA 279 and 715 were successfully used to reverse on-going acute rejection and inhibit allospecific antibody production. Combined therapy with CsA and MNA, given on the day of retransplantation in presensitised rats, prevented accelerated acute rejection. Using iso-effect curves for tacrolimus and MNA, alone or in combination, additive effects of the two drugs were demonstrated

Implantation with SHED sheet induced with homogenate protein of spinal cord promotes functional recovery from spinal cord injury in rats

Introduction: After spinal cord injury (SCI) occurs, the lesion is in a growth inhibitory microenvironment that severely hinders neural regeneration. In this microenvironment, inhibitory factors are predominant and factors that promote nerve regeneration are few. Improving neurotrophic factors in the microenvironment is the key to treating SCI.Methods: Based on cell sheet technology, we designed a bioactive material with a spinal cord‐like structure –SHED sheet induced with homogenate protein of spinal cord (hp–SHED sheet). Hp–SHED sheet was implanted into the spinal cord lesion for treating SCI rats with SHED suspensions as a control to investigate the effects on nerve regeneration.Results: Hp–SHED sheet revealed a highly porous three–dimensional inner structure, which facilitates nerve cell attachment and migration. Hp-SHED sheet in vivo restored sensory and motor functions in SCI rats by promoting nerve regeneration, axonal remyelination, and inhibiting glial scarring.Discussion: Hp–SHED sheet maximally mimics the microenvironment of the natural spinal cord and facilitate cell survival and differentiation. Hp–SHED sheet could release more neurotrophins and the sustained action of neurotrophins improves the pathological microenvironment, which effectively promotes nerve regeneration, axonal extension, and inhibits glial scarring, thereby promoting the in situ centralis neuroplasticity. Hp–SHED sheet therapy is a promising strategy for effective treatment of SCI based on neurotrophins delivery

Additive effects of leflunomide and tacrolimus in prevention of islet xenograft rejection

Leflunomide is a low molecular weight immunosuppressive drug which inhibits the enzymes dehydroorotate dehydrogenase and protein tyrosine kinase, both of which are important components in the immune response. As the mechanisms of action of leflunomide and tacrolimus are different, we postulated an additive or synergistic effect of the two drugs and investigated the effects of leflunomide alone, or in combination with a suboptimal dose of tacrolimus, on xenogeneic islet transplantation in a rat-to-mouse model. A total of 1200-1500 rat islets were transplanted under the left kidney capsule of streptozotocin-induced diabetic BALB/c mice. The median survival time (MST) of the untreated group was 6 days. Leflunomide at 5, 10 and 20 mg/kg/d administrated for 10 days significantly prolonged MST to 10, 16 and 20 days. A dose of tacrolimus (2 mg/kg/d) was associated with a graft survival of 9 (range 6-12) days; most grafts rejected during ongoing therapy. When tacrolimus (2 mg/kg/d) was combined with leflunomide (10 mg/kg/d), the survival time of the islet xenografts was increased further to 22 days, significantly longer than with leflunomide or tacrolimus alone. In summary, our findings demonstrate that leflunomide prolonged xenogeneic islet graft survival, and that its immunosuppressive effect was improved when combined with tacrolimus