MRC-5 Cells Research Articles

Physicochemical characterization and cancer cell antiproliferative effect of silver-doped magnesia nanoparticles

Silver-doped magnesia nanoparticles (Ag/MgO) were synthesized using the precipitation method and characterized by various techniques such as X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), thermal gravimetric analysis (TGA), Brunner-Emmett-Teller (BET) surface area measurements, and dispersive X-ray spectroscopy (EDX). The morphology of Ag/MgO nanoparticles was determined by transmission and scanning electron microscopy, which revealed cuboidal shaped nanoparticles with sizes ranging from 31 to 68 nm and an average size of 43.5 ± 10.6 nm. The anticancer effects of Ag/MgO nanoparticles were evaluated on human colorectal (HT29) and lung adenocarcinoma (A549) cell lines, and their caspase-3, -8, and -9 activities, as well as Bcl-2, Bax, p53, cytochrome C protein expressions were estimated. Ag/MgO nanoparticles showed selective toxicity towards HT29 and A549 cells while remaining relatively innocuous towards the normal human colorectal, CCD-18Co, and lung, MRC-5 cells. The IC50 values of Ag/MgO nanoparticles on the HT29 and A549 cells were found to be 90.2 ± 2.6 and 85.0 ± 3.5 μg/mL, respectively. The Ag/MgO nanoparticles upregulated caspase-3 and -9 activities, downregulated Bcl-2, upregulated Bax and p53 protein expressions in the cancer cells. The morphology of the Ag/MgO nanoparticle treated HT29 and A549 cells was typical of apoptosis, with cell detachment, shrinkage, and membrane blebbing. The results suggest that Ag/MgO nanoparticles induce apoptosis in cancer cells and exhibit potential as a promising anticancer agent.

Temporal inhibition of the electron transport chain attenuates stress-induced cellular senescence by prolonged disturbance of proteostasis in human fibroblasts.

We previously developed a stress-induced premature senescence (SIPS) model in which normal human fibroblast MRC-5 cells were treated with either the proteasome inhibitor MG132 or the vacuolar-type ATPase inhibitor bafilomycin A1 (BAFA1). To clarify the involvement of mitochondrial function in our SIPS model, MRC-5 cells were treated with MG132 or BAFA1 along with an inhibitor targeting either the electron transport chain complex I or complex III, or with a mitochondrial uncoupler. SIPS induced by MG132 or BAFA1 was significantly attenuated by short-term co-treatment with the complex III inhibitor, antimycin A (AA), but not the complex I inhibitor, rotenone or the mitochondrial uncoupler, carbonyl cyanide 3-chlorophenylhydrazone. By co-treatment with AA, mitochondrial and intracellular reactive oxygen species levels, accumulation of protein aggregates and mitochondrial unfolded protein responses (UPRmt ) were remarkably suppressed. Furthermore, AA co-treatment suppressed the hyperpolarization of the mitochondrial membrane and the induction of mitophagy observed in MG132-treated cells and enhanced mitochondrial biogenesis. These findings provide evidence that the temporal inhibition of mitochondrial respiration exerts protective effects against the progression of premature senescence caused by impaired proteostasis.

Derrone Targeting the TGF Type 1 Receptor Kinase Improves Bleomycin-Mediated Pulmonary Fibrosis through Inhibition of Smad Signaling Pathway.

Transforming growth factor-β (TGF-β) has a strong impact on the pathogenesis of pulmonary fibrosis. Therefore, in this study, we investigated whether derrone promotes anti-fibrotic effects on TGF-β1-stimulated MRC-5 lung fibroblast cells and bleomycin-induced lung fibrosis. Long-term treatment with high concentrations of derrone increased the cytotoxicity of MRC-5 cells; however, substantial cell death was not observed at low concentrations of derrone (below 0.05 μg/mL) during a three-day treatment. In addition, derrone significantly decreased the expressions of TGF-β1, fibronectin, elastin, and collagen1α1, and these decreases were accompanied by downregulation of α-SMA expression in TGF-β1-stimulated MRC-5 cells. Severe fibrotic histopathological changes in infiltration, alveolar congestion, and alveolar wall thickness were observed in bleomycin-treated mice; however, derrone supplementation significantly reduced these histological deformations. In addition, intratracheal administration of bleomycin resulted in lung collagen accumulation and high expression of α-SMA and fibrotic genes-including TGF-β1, fibronectin, elastin, and collagen1α1-in the lungs. However, fibrotic severity in intranasal derrone-administrated mice was significantly less than that of bleomycin-administered mice. Molecular docking predicted that derrone potently fits into the ATP-binding pocket of the TGF-β receptor type 1 kinase domain with stronger binding scores than ATP. Additionally, derrone inhibited TGF-β1-induced phosphorylation and nuclear translocations of Smad2/3. Overall, derrone significantly attenuated TGF-β1-stimulated lung inflammation in vitro and bleomycin-induced lung fibrosis in a murine model, indicating that derrone may be a promising candidate for preventing pulmonary fibrosis.

Hydroxyapatite/poly(vinyl alcohol)/chitosan coating with gentamicin for orthopedic implants

Electrophoretic deposition (EPD) was employed to fabricate composite coatings aimed for orthopedic implants. Hydroxyapatite (HAP), poly(vinyl alcohol) (PVA) and chitosan (CS) coatings, with or without antibiotic gentamicin (Gent) were deposited in a single-step process from aqueous suspension, using EPD. Polymer/mineral composite coatings were formed with the aim to mimic the natural composition of bone tissue, along with improved coatings antibacterial properties, at the same time serving as a drug carrier, allowing local administration of antibiotic. Structural and morphological characterization of HAP/PVA/CS and HAP/PVA/CS/Gent coatings was investigated by Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), thermogravimetric analysis (TG/DTG), and scanning electron microscopy (SEM) proving carbonate-substituted HAP. The presence of gentamicin contributed to the bactericidal activity of composite HAP/PVA/CS/Gent coating against Staphylococcus aureus and Escherichia coli. Cytotoxicity studies were performed towards human MRC-5 and mice L929 fibroblast cell lines, and both investigated samples (HAP/PVA/CS and HAP/PVA/CS/Gent) expressed good biocompatibility, without causing toxic effect on tested cells. Biomineralization potential was investigated by measuring the alkaline phosphatase activity, demonstrating high potential of HAP/PVA/CS/Gent to induce the growth of new HAP layer. Therefore, the present investigation proposes the production of composite coatings from non-toxic (aqueous) suspensions, consisting of biocompatible and bioactive components, along with antibiotic gentamicin, improving the antibacterial coatings' properties.

A comparative study on the metal complexes of an anticancer estradiol-hydroxamate conjugate and salicylhydroxamic acid

Hydroxamic acids bearing an (O,O) donor set are well-known metal-chelating compounds with diverse biological activities including anticancer activity. Since steroid conjugation with a pharmacophoric moiety may have the potential to improve this effect, a salicylhydroxamic acid–estradiol hybrid molecule (E2HA) was synthesized. Only minimal effect of the conjugation on the proton dissociation constants was observed in comparison to salicylhydroxamic acid (SHA).The complexation with essential metal ions (iron, copper) was characterized, since E2HA may exert its cytotoxicity through the binding of these ions in cells. UV–visible spectrophotometric and pH-potentiometric titrations revealed the formation of high-stability complexes, while the Fe(III) preference over Fe(II) was proved by cyclic voltammetry and spectroelectrochemical measurements.Complex formation with half-sandwich Rh(III)(η5-Cp*) and Ru(II)(η6-p-cymene) organometallic cations was also studied as it may improve the anticancer effect and the pharmacokinetic profile of the ligand. At equimolar concentration the speciation is complicated because of the presence of mononuclear and binuclear complexes. The complexes readily react with small molecules e.g. glutathione, 1-methylimidazole and nucleosides, having major effect on solution speciation, namely mixed-ligand complex formation and ligand displacement occur. These processes serve as models for the interactions with biomolecules in the body.E2HA exerted moderate anticancer activity (IC50 = 25–59 μM) in the tested three human cancer cell lines (Colo205, Colo320 and MCF-7), while being non-toxic on non-cancerous MRC-5 cells. Meanwhile, SHA was inactive in the same cells. Complexation with half-sandwich Rh(III) and Ru(II) cations had only a minor improvement on the cytotoxic effect of E2HA.

Mammalian Cell-Growth Monitoring Based on an Impedimetric Sensor and Image Processing within a Microfluidic Platform.

In recent years, advancements in microfluidic and sensor technologies have led to the development of new methods for monitoring cell growth both in macro- and micro-systems. In this paper, a microfluidic (MF) platform with a microbioreactor and integrated impedimetric sensor is proposed for cell growth monitoring during the cell cultivation process in a scaled-down simulator. The impedimetric sensor with an interdigitated electrode (IDE) design was realized with inkjet printing and integrated into the custom-made MF platform, i.e., the scaled-down simulator. The proposed method, which was integrated into a simple and rapid fabrication MF system, presents an excellent candidate for the scaled-down analyses of cell growths that can be of use in, e.g., optimization of the cultivated meat bioprocess. When applied to MRC-5 cells as a model of adherent mammalian cells, the proposed sensor was able to precisely detect all phases of cell growth (the lag, exponential, stationary, and dying phases) during a 96-h cultivation period with limited available nutrients. By combining the impedimetric approach with image processing, the platform enables the real-time monitoring of biomasses and advanced control of cell growth progress in microbioreactors and scaled-down simulator systems.

The evaluation of tumorigenicity and characterization of colonies in a soft agar colony formation assay using polymerase chain reaction

In regenerative medicine, the tumorigenic potency of cells in cellular therapy products (CTPs) is a major concern for their application to patients. This study presents a method—the soft agar colony formation assay using polymerase chain reaction (PCR)—to evaluate tumorigenicity. MRC-5 cells, contaminated with HeLa cells, were cultured for up to 4 weeks in soft agar medium. Cell-proliferation-related mRNAs, Ki-67 and cyclin B, could be detected in 0.01% of HeLa cells after 5 days of culture, whereas cyclin-dependent kinase 1 (CDK1) could be detected after 2 weeks. On the other hand, CDK2, proliferating cell nuclear antigen (PCNA), and minichromosome maintenance protein 7 (MCM7) were not useful to detect HeLa cells even after 4 weeks of culture. The cancer stem cell (CSC) markers, aldehyde dehydrogenase 1 (ALDH1) and CD133 in 0.01% of HeLa cells, could be detected 2 and 4 weeks after culture, respectively. However, another CSC marker CD44 was not useful because its expression was also detected in MRC-5 cells alone. This study suggests that the application of the PCR method to the soft agar colony formation assay could evaluate not only the tumorigenic potency in the short-term but also characterize the colonies, eventually improving the safety of CTPs.

MP14-10 ENHANCED ANTITUMOR EFFECT OF THE COMBINATION OF BACILLE CALMETTE-GUÉRIN AND AN IMMUNE CHECKPOINT INHIBITOR IN BLADDER CANCER-ON-A-CHIP

You have accessJournal of UrologyCME1 Apr 2023MP14-10 ENHANCED ANTITUMOR EFFECT OF THE COMBINATION OF BACILLE CALMETTE-GUÉRIN AND AN IMMUNE CHECKPOINT INHIBITOR IN BLADDER CANCER-ON-A-CHIP Se Young Choi, Jong Hyun Tae, Byung Hoon Chi, In Ho Chang, Tae-Hyoung Kim, Soon Chul Myung, Jung Hoon Kim, Jin Wook Kim, and Yong Seong Lee Se Young ChoiSe Young Choi More articles by this author , Jong Hyun TaeJong Hyun Tae More articles by this author , Byung Hoon ChiByung Hoon Chi More articles by this author , In Ho ChangIn Ho Chang More articles by this author , Tae-Hyoung KimTae-Hyoung Kim More articles by this author , Soon Chul MyungSoon Chul Myung More articles by this author , Jung Hoon KimJung Hoon Kim More articles by this author , Jin Wook KimJin Wook Kim More articles by this author , and Yong Seong LeeYong Seong Lee More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003234.10AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: We upgraded preexisting bladder cancer-on-a-chip (BCOC) by adding T cells and evaluated the antitumor effect of a combination of intravesical Bacille Calmette-Guérin (BCG) and pembrolizumab. METHODS: We fabricated bioprinted BCOC with microfluids, incorporating HT1376, MRC-5, HUVEC, THP-1 and Jurkat cells. We evaluated the effector-to-target cytotoxicity, cytokine, and cell viability in 2D culture, live/dead assay, migration assay, and cytokine assay in BCOC. Additionally, we evaluated the antitumor efficacy of the combination of BCG and pembrolizumab in an orthotopic mouse model. RESULTS: The combination group showed the most effective reduction compared to the control in 2D culture (100.0±0.8% vs. 36.4±0.8%, p<0.001). In BCOC, cancer cell viabilities were decreased at 3 days in the BCG group (70.1±9.8%, p=0.013) and combination group (49.3±8.1%, p<0.001). The combination group showed the highest immune reaction in the cytokine assay (interferon-γ, p=0.045; interleukin-6, p=0.037) and migration assay (fold change 1.3±0.1, p<0.001), whereas in the in vivo model, it showed lower signal intensities from days 10 to 14 compared to that in the control group (p=0.031 and p=0.014, respectively). No significant weight changes were observed among the groups. CONCLUSIONS: We developed a 3D bioprinted BCOC via use of the monocytic THP-1 cells and Jurkat T cells to assess the efficacy of immunotherapy. The combination of BCG and pembrolizumab showed the best antitumor efficacy in BCOC and animal models. Source of Funding: This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT & Future Planning (NRF-2019R1C1C1005170 & NRF-2022R1A2C2008207) © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e186 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Se Young Choi More articles by this author Jong Hyun Tae More articles by this author Byung Hoon Chi More articles by this author In Ho Chang More articles by this author Tae-Hyoung Kim More articles by this author Soon Chul Myung More articles by this author Jung Hoon Kim More articles by this author Jin Wook Kim More articles by this author Yong Seong Lee More articles by this author Expand All Advertisement PDF downloadLoading ...

HPLC Profile, Anticancer, Anti-inflammatory and Antioxidant Activities of Euphorbia inarticulata Ethanolic Extract

Antioxidants, in addition to protection against oxidative stress, also possess anti-inflammatory and anticancer potentials. In this work, the ethanolic extract (EE) of Euphorbia inarticulata has been evaluated for antioxidant, anti-inflammatory and anticancer activities. The antioxidant activity showed that EE significantly scavenges DPPH (IC₅₀ = 54.56 ± 2.38 µg/ml) and H₂O₂(77.44 ± 2.76 µg/ml), whereas ferric reducing antioxidant power (FRAP) and total antioxidant capacity (TAC) were found to be 100.93 ± 4.15 µg/ml and 46.72 ± 3.86 mg gallic acid equi/gm, respectively. The EE exhibited significant inhibition on COX-1 (IC₅₀ 79.84µg/ml) and protein denaturation (IC₅₀57.6 µg/ml), compared to indomethacin and diclofenac, respectively. Furthermore, EE was found to be effective in killing Huh-7 and HeLa cancerous cells with IC₅₀ 104.62 (SI 2.36) and 145.11 µg/ml (SI 1.7), respectively, while doxorubicin displayed IC₅₀ 0.71 (Huh-7) and 2.63 µg/ml (HeLa), whereas it was safe on normal MRC-5 cells. The HPLC analysis showed the presence of ellagic acid, caffeic acid, gallic acid, syringic acid, cinnamic acid and catechol which may be responsible for the promising antioxidant, anti-inflammatory and anticancer potentials exerted by the ethanolic extract of E. inarticulata. These results conclude that Euphorbia inarticulata ethanolic extract has anti-inflammatory and anticancer effects by triggering oxidative stress.

Cytotoxic activity of Ru(II)/DPEPhos/N,S-mercapto complexes (DPEPhos = bis-[(2-diphenylphosphino)phenyl]ether)

We report here on three new ruthenium(II) complexes, [Ru(DPEPhos)(mtz)(bipy)]PF6 (Ru1), [Ru(DPEPhos)(mmi)(bipy)]PF6 (Ru2) and [Ru(DPEPhos)(dmp)(bipy)]PF6 (Ru3). DPEPhos = bis-[(2-diphenylphosphino)phenyl]ether, mtz = 2-mercapto-2-thiazoline, mmi = 2-mercapto-1-methylimidazole, dmp = 4,6-diamino-2-mercaptopyrimidine and bipy = 2,2′-bipyridine. The compounds were characterized by several spectroscopic techniques, and the molecular structure of Ru1 complex was determined by single-crystal X-ray diffraction. The cytotoxicity of Ru1 – Ru3 complexes were tested against the A549 (human lung) and the MDA-MB-231 (human breast) cancer cell lines and against MRC-5 (non-tumor lung) and MCF-10A (non-tumor breast) cell lines through the MTT assay. All three complexes are cytotoxic against the cell lines studied, with IC50 values lower than those found for the cisplatin. Among them, the Ru2 complex has shown the best selectivity against MDA-MB-231 cancer cell lines, with an IC50 value 12 times lower than that on MCF-10A. The complex Ru2 was capable to induce changes in MDA-MB-231 cells morphology, with loss of cellular adhesion, inhibited colony formation and induce an accumulation of cells at the sub-G1 phase, with an increase in S-phase and decrease of cells at G2 phase. Viscosity, electrochemical and Hoechst 33258 displacement experiments for Ru1 – Ru3 complexes with calf thymus DNA (CT-DNA) showed an electrostatic and groove binding mode of interaction. Additionally, the complexes interact with the protein Human Serum Albumin (HSA) by static mechanism. The negative values for ΔH and ΔS indicate that van der Waals forces and hydrogen bonding may occurs between the complexes and HSA. Therefore, this class of complexes are promising anticancer candidates and may be selected to further detailed studies.

Discovery of Novel Dimeric Pyridinium Bromide Analogues Inhibits Cancer Cell Growth by Activating Caspases and Downregulating Bcl-2 Protein.

Flexible dimeric substituted pyridinium bromides with primary and tertiary amines are prepared by conventional and solvent-free methods. The formation of compounds 2 and 4 is much easier than that of compounds 1 and 3 because of the benzyl carbon which is more electropositive than the primary alkyl carbon. The newly synthesized dimeric pyridinium compounds are optimized using DFT and B3LYP 6-31 g(d,p). The in vitro antiproliferative activity is studied in lung (A549) and breast cancer cell lines (MDA-MB 231). Among the four compounds, 1,1'-(1,3-phenylene bis(methylene)bis 2-aminopyridinium bromide 4 showed potent anticancer activity when compared to the standard drug 5-fluorouracil. 1,1'-(1,3-Phenylene bis(methylene)bis 2-aminopyridinium bromide 4 is not toxic to normal cell lines 3T3-L1 and MRC-5 cell lines. Also, 1,1'-(1,3-phenylene bis(methylene)bis 2-aminopyridinium bromide 4-induced apoptosis in cancer cell lines is examined using AO/EB and Hoechst staining, which is further supported by cell cycle analysis. Western blot analysis showed that 1,1'-(1,3-phenylene bis(methylene)bis 2-aminopyridinium bromide 4 induces apoptosis through the extrinsic apoptotic pathway by upregulating caspase 3 and caspase 9. This compound also downregulates intrinsic apoptotic proteins, including Bcl-2, Bcl-x, and Bad. From the present study results, it is confirmed that 1,1'-(1,3-phenylene bis(methylene)bis 2-aminopyridinium bromide 4 has potent anticancer activity when compared to other compounds.

Flavonol-Glycoside and Rare Triterpenoid Derivatives Isolated from Leaves of Combretum glutinosum Perr. Ex Dc. with In Vitro Cytotoxic Activity

Combretaceae plants are used traditionally by many cultures, especially in Sudanese patients for the treatment of diverse ailments such as anti-inflammatory, antimicrobial, antitumor, and antioxidant disorders. Of these plants, the genus Combretum are traditional medicinal plants. Thus, they are formed from the non-polar or polar extracts of many isolated phytochemicals. Of these necessities, the use of Combretum extracts for their medicinal properties can be found in the earliest of myths and traditions used to document the plants’ ability to treat diseases. Combretum glutinosum Perr. Ex Dc. is a common shrub native to the African continent, especially Sudan. Currently, there are no published data regarding its cytotoxic activity. Additionally, there are few chemical and biological reports of C. glutinosum. Therefore, the current study aimed to isolate the chemical bioactive compounds (1–6) from the ethyl acetate (EtOAc) extract of C. glutinosum. A new flavonoid compound, namely, glutosinumoside (4), was afforded, and five known compounds were obtained: three oleanane-glycosides (1–3) and two phenolic acids (5,6). The structures of the six compounds were determined by spectroscopic analysis, including one- and two-dimensional (1D and 2D) NMR, mass spectrometry, and chromatographic analysis. Moreover, an in vitro cytotoxic evaluation of the successive extracts and the bioactive EtOAc fractions of C. glutinosum against MCF7 (breast), HT29 (colon), HepG2 (liver), and MRC5 (normal lung) cell lines was performed. The isolated compounds showed comparable cytotoxic activities with the crude EtOH extract and doxorubicin against the tested cell lines. Compounds (1) and (6) exhibited the highest cytotoxicity against MCF7 (1.37 ± 0.21 and 1.48 ± 0.34 µg/mL, respectively) and HepG2 (3.30 ± 0.02 and 2.10 ± 0.22 µg/mL, respectively) in the MTT assay. In addition, compounds (1) and (3) demonstrated a significant upregulation of cancer’s two important hallmarks (caspase 3 and bax genes) by inducing apoptosis and perturbing the MCF7 cell cycle.

Chemical Characterization, Antioxidant Activity, and Cytotoxity of Wild-Growing and In Vitro Cultivated Rindera umbellata (Waldst. and Kit.) Bunge

The aim of this study was to comparatively analyze chemical composition and biological activity of wild- and in vitro grown Rindera umbellata. Explants were cultivated on 0.003–0.3 M sucrose, fructose, or glucose. HPLC-DAD for quantifying rosmarinic (RA) and lithospermic B (LAB) acids and GC-MS/FID for qualitative pyrrolizidine alkaloids (PAs) detection were used. Antioxidant activity (DPPH and ABTS assays) and cytotoxicity (MTT test) were monitored. Identified PAs were 7-angeloyl heliotridane, lindelofine, 7-angeloyl heliotridine, 7-angeloyl-9-(+)-trachelanthylheliotridine, punctanecine, and heliosupine, with higher variability reported in wild-growing samples. Total phenolic contents (TPCs) were comparable in wild-growing and in vitro samples, but total flavonoid (TFC) and RA levels were multifold higher in in vitro samples. Notably, high concentration of LAB was detected in wild-growing roots. Amounts of 0.3 M and 0.1 M of sucrose were optimal for TFC and RA production, while maximal antioxidant activity was monitored in plants grown on 0.3 M sucrose. The MTT test indicated colorectal HT-29 as more sensitive than A549 lung adenocarcinoma and normal MRC-5 cells, showing selective sensitivity to wild-growing and 0.3 M sucrose samples. In conclusion, PAs in vitro, as well as TPC, TFC, RA, and LAB in both growing conditions were detected for the first time in R. umbellata.

Physico-Chemical Study of Mn(II), Co(II), Cu(II), Cr(III), and Pd(II) Complexes with Schiff-Base and Aminopyrimidyl Derivatives and Anti-Cancer, Antioxidant, Antimicrobial Applications.

A new class of biologically active mineral complexes was synthesized by reacting the following metal salts: MnCl2·4H2O, CoCl2·6H2O, CuCl2·2H2O, CrCl3·6H2O, and PdCl2 respectively with 2-amino-4,6-dimethyl pyrimidine (ADMPY) and Schiff's base resulting from the condensation reaction between benzaldehyde with p-phenylenediamine and 2-hydroxy-1-naphthaldehyde as ligands have been synthesized and characterized on the basis of their CHN, thermal analysis, XRD, SEM and magnetic measurements along with their FT-IR and UV-vis spectra. The scanning electron microscope SEM measurements and the calculations on the powder XRD data indicate the nano-sized nature of the prepared complexes (average size 32-88 nm). The spectral data confirmed the coordinated ligand (HL) via a nitrogen atom of an azomethine group (-C=N-) and phenolic -OH group and NH2-ADMPY ligand with the metal ions. An octahedral geometry for all complexes has been proposed based on magnetic and electronic spectral data except Pd(II) complex, which has a tetrahedral geometry. Molecular modeling was performed for Cu(II) complex using the density functional method DFT/B3LYP to study the structures and the frontier molecular orbitals (HOMO and LUMO). The antioxidant of the complexes was studied using the 2,2-diphenyl-1-picrylhydrazyl (DPPH)-free radical-scavenging assays. The metal complexes were tested in vitro for anticancer activities against two cancer lines A-549 and MRC-5 cells. Cu(II) and Pd(II) complexes showed the highest cytotoxicity effect, comparable to that of other cis-platinum-based drugs. The complexes showed significant activity against fungi and bacteria.

DAB2 promotes pulmonary fibrosis and may act as an intermediate between IGF‑1R and PI3K/AKT signaling pathways.

Idiopathic pulmonary fibrosis (IPF) is a heterogeneous lung disease associated with high mortality. Disabled-2 (DAB2), an adapter protein, regulates cell-fibrinogen adhesion and fibrinogen uptake. DAB2 is differentially expressed in mouse fibrotic lungs induced by bleomycin according to a genome microarray analysis based on Gene Expression Omnibus database. However, the role of DAB2 in IPF has not been revealed. A bleomycin-induced mouse model of pulmonary fibrosis was constructed in the present study. It found that the expression of DAB2 was upregulated in bleomycin-induced fibrotic lung tissue with collagen fiber deposition and pulmonary interstitium thickening. Colocalization of DAB2 with α-smooth muscle actin (SMA) was observed in lung tissue sections. In vitro, human lung fibroblast MRC-5 cells were treated with TGF-β1 and the expression of DAB2 was increased. Knockdown of DAB2 suppressed cell proliferation and the expression of α-SMA, collagen I, collagen IV and fibronectin in TGF-β1-treated MRC-5 cells. The phosphorylation levels of PI3K and AKT were suppressed in DAB2-knockdown cells. IGF-1/IGF-1R has been reported to promote pulmonary fibrosis and activate the PI3K/Akt signaling. In the present study, the activation of IGF-1/IGF-1R signaling pathways in bleomycin-induced fibrotic lung tissues were positively associated with DAB2 expression. The phosphorylation level of IGF-1R was increased in MRC-5 cells with TGF-β1 treatment, and DAB2 expression was decreased by silencing of IGF-1R. This suggested that DAB2 might be a downstream target of the IGF-1R pathway and thus induced PI3K/AKT signaling activation and fibrogenesis. The current study demonstrated the importance of DAB2 in pulmonary fibrosis and suggested the potential of IGF-1R/DAB2/PI3K in the pathogenesis of IPF.

Anti-Zika Virus Activity and Isolation of Flavonoids from Ethanol Extracts of Curatella americana L. Leaves.

The ethnomedicinal plant Curatella americana L. (Dilleniaceae) is a common shrub in the Brazilian Cerrado, whose ethanolic extract showed significant in vitro anti-Zika virus activity by the MTT colorimetric method. Currently, there is no drug in clinical use specifically for the treatment of this virus; therefore, in this work, the antiviral and cytotoxic properties of the ethanolic extract, fractions, and compounds were evaluated. The ethanolic extract of the leaves showed no cytotoxicity for the human MRC-5 cell and was moderately cytotoxic for the Vero cell (CC50 161.5 ± 2.01 µg/mL). This extract inhibited the Zika virus multiplication cycle with an EC50 of 85.2 ± 1.65 µg/mL. This extract was fractionated using the liquid-liquid partition technique, and the ethyl acetate fraction showed significant activity against the Zika virus with an EC50 of 40.7 ± 2.33 µg/mL. From the ethyl acetate fraction, the flavonoids quercetin-3-O-hexosylgallate (1), quercetin-3-O-glucoside (2), and quercetin (5) were isolated, and in addition to these compounds, a mixture of quercetin-3-O-rhamnoside (3) and quercetin-3-O-arabinoside (4) was also obtained. The isolated compounds quercetin and quercetin-3-O-hexosylgallate inhibited the viral cytopathic effect at an EC50 of 18.6 ± 2.8 and 152.8 ± 2.0, respectively. Additionally, analyses by liquid chromatography coupled to a mass spectrometer allowed the identification of another 24 minor phenolic constituents present in the ethanolic extract and in the ethyl acetate fraction of this species.

Cardamonin as a p38 MAPK Signaling Pathway Activator Inhibits Human Coronavirus OC43 Infection in Human Lung Cells.

A natural chalcone, cardamonin (2',4'-dihydroxy-6'-methoxychalcone; CDN) was isolated from the seeds of Alpinia katsumadai Hayata, which has been traditionally used to treat stomach aches. CDN has been reported to possess various pharmacological properties, including anticancer and anti-inflammatory effects. This study evaluated the antiviral activity of CDN against human coronavirus HCoV-OC43 and determined the mode of action in HCoV-OC43-infected human lung cell lines (MRC-5 and A549 cells). CDN significantly inhibited HCoV-OC43-induced cytopathic effects with an IC50 of 3.62 μM and a CC50 of >50 μM, resulting in a selectivity index of >13.81. CDN treatment reduced the level of viral RNA and the expression of spike and nucleocapsid proteins in HCoV-OC43-infected cells as determine through qRT-PCR and Western blot analysis. Additionally, the activation of p38 mitogen-activated protein kinase (MAPK) by anisomycin decreased viral protein expression, whereas an inhibitor of p38 MAPK signaling, SB202190, increased viral protein expression. CDN also amplified and extended the p38 MAPK signaling pathway in HCoV-OC43-infected cells. In conclusion, CDN inhibited HCoV-OC43 infection by activating the p38 MAPK signaling pathway and has potential as a therapeutic agent against human coronavirus.

Structural and Biological Properties of Heteroligand Copper Complexes with Diethylnicotinamide and Various Fenamates: Preparation, Structure, Spectral Properties and Hirshfeld Surface Analysis

Herein, we discuss the synthesis, structural and spectroscopic characterization, and biological activity of five heteroligand copper(II) complexes with diethylnicotinamide and various fenamates, as follows: flufenamate (fluf), niflumate (nifl), tolfenamate (tolf), clonixinate (clon), mefenamate (mef) and N, N-diethylnicotinamide (dena). The complexes of composition: [Cu(fluf)2(dena)2(H2O)2] (1), [Cu(nifl)2(dena)2] (2), [Cu(tolf)2(dena)2(H2O)2] (3), [Cu(clon)2(dena)2] (4) and [Cu(mef)2(dena)2(H2O)2] (5), were synthesized, structurally (single-crystal X-ray diffraction) and spectroscopically characterized (IR, EA, UV-Vis and EPR). The studied complexes are monomeric, forming a distorted tetragonal bipyramidal stereochemistry around the central copper ion. The crystal structures of all five complexes were determined and refined with an aspheric model using the Hirshfeld atom refinement method. Hirshfeld surface analysis and fingerprint plots were used to investigate the intermolecular interactions in the crystalline state. The redox properties of the complexes were studied and evaluated via cyclic voltammetry. The complexes exhibited good superoxide scavenging activity as determined by an NBT assay along with a copper-based redox-cycling mechanism, resulting in the formation of ROS, which, in turn, predisposed the studied complexes for their anticancer activity. The ability of complexes 1–4 to interact with calf thymus DNA was investigated using absorption titrations, viscosity measurements and an ethidium-bromide-displacement-fluorescence-based method, suggesting mainly the intercalative binding of the complexes to DNA. The affinity of complexes 1–4 for bovine serum albumin was determined via fluorescence emission spectroscopy and was quantitatively characterized with the corresponding binding constants. The cytotoxic properties of complexes 1–4 were studied using the cancer cell lines A549, MCF-7 and U-118MG, as well as healthy MRC-5 cells. Complex 4 exhibited moderate anticancer activity on the MCF-7 cancer cells with IC50 = 57 μM.

IL-27 induces autophagy through regulation of the DNMT1/lncRNA MEG3/ERK/p38 axis to reduce pulmonary fibrosis

PurposePrevious studies have shown that interleukin-27 (IL-27) can reduce bleomycin (BLM)-induced pulmonary fibrosis (PF). However, the underlying mechanism by which IL-27 attenuates PF is not fully clear.MethodsIn this research, we used BLM to construct a PF mouse model, and MRC-5 cells stimulated by transforming growth factor-β1 (TGF-β1) were used to construct a PF model in vitro. The lung tissue status was observed by Masson and hematoxylin and eosin (HE) staining. To detect gene expression, RT‒qPCR was used. The protein levels were detected by western blotting and immunofluorescence staining. EdU and ELISA were used to detect cell proliferation viability and hydroxyproline (HYP) content, respectively.ResultsAberrant IL-27 expression was observed in BLM-induced mouse lung tissues, and the use of IL-27 attenuated mouse lung tissue fibrosis. TGF-β1 induced autophagy inhibition in MRC-5 cells, and IL-27 alleviated MRC-5 cell fibrosis by activating autophagy. The mechanism is inhibition of DNA methyltransferase 1 (DNMT1)-mediated lncRNA MEG3 methylation and ERK/p38 signaling pathway activation. Overexpression of DNMT1, knockdown of lncRNA MEG3, autophagy inhibitor or ERK/p38 signaling pathway inhibitors reversed the positive effect of IL-27 in a lung fibrosis model in vitro.ConclusionIn conclusion, our study shows that IL-27 upregulates MEG3 expression through inhibition of DNMT1-mediated lncRNA MEG3 promoter methylation, which in turn inhibits ERK/p38 signaling pathway-induced autophagy and attenuates BLM-induced PF, providing a contribution to the elucidation of the potential mechanisms by which IL-27 attenuates PF.

Biological Activity of Cupressus sempervirens Essential Oil.

The aim of this study was to evaluate the antioxidant, antibiofilm, antimicrobial (in situ and in vitro), insecticidal, and antiproliferative activity of Cupressus sempervirens essential oil (CSEO) obtained from the plant leaf. The identification of the constituents contained in CSEO was also intended by using GC and GC/MS analysis. The chemical composition revealed that this sample was dominated by monoterpene hydrocarbons α-pinene, and δ-3-carene. Free radical scavenging ability, performed by using DPPH and ABTS assays, was evaluated as strong. Higher antibacterial efficacy was demonstrated for the agar diffusion method compared to the disk diffusion method. The antifungal activity of CSEO was moderate. When the minimum inhibitory concentrations of filamentous microscopic fungi were determined, we observed the efficacy depending on the concentration used, except for B. cinerea where the efficacy of lower concentration was more pronounced. The vapor phase effect was more pronounced at lower concentrations in most cases. Antibiofilm effect against Salmonella enterica was demonstrated. The relatively strong insecticidal activity was demonstrated with an LC50 value of 21.07% and an LC90 value of 78.21%, making CSEO potentially adequate in the control of agricultural insect pests. Results of cell viability testing showed no effects on the normal MRC-5 cell line, and antiproliferative effects towards MDA-MB-231, HCT-116, JEG-3, and K562 cells, whereas K562 cells were the most sensitive. Based on our results, CSEO could be a suitable alternative against different types of microorganisms as well as suitable for the control of biofilms. Due to its insecticidal properties, it could be used in the control of agricultural insect pests.