Lung Cell Lines Research Articles

Toxicity assessment of green synthesized Cu nanoparticles by cell-free extract of Pseudomonas silesiensis as antitumor cancer and antimicrobial

Spherical homogeneous 32 nm, protein coated Pseudomonas silesiensis strain A3 CuNPs was investigated for their cytotoxicity effect as well as antimicrobial and antitumor activity. CuNPs cytotoxicity was estimated using human normal lung cell lines (Wi38) against CuNPs with concentrations ranging from 25 to 150 μg/mL using neutral red uptake assay. The cytotoxicity study revealed that the bacterial CuNPs had an impact on Wi38 cell viability at concentrations of 25, 50, 100 and 150 μg/mL CuNPs were 95.8, 91.1, 89.2 and 82.3%, respectively, with a strong correlation coefficient (r = 0.94) and a CuNPs IC50 value of 1057.0 μg/mL. CuNPs exhibit a broad-spectrum antimicrobial activity against various microorganism species, including fungi and Gram positive and negative bacteria using the agar-well diffusion method. The findings revealed that the most sensitive pathogens were Staphylococcus aureus ATCC5638 and Aspergillus flavus ATCC 9643 which tended to have a high inhibition zone diameter (50 and 47 mm, respectively). The minimum inhibitory concentration (MIC) of CuNPs was 50 μg/mL. The minimum lethal concentration (MLC) values were 50 and 75 μg/mL for S. aureus ATCC 5638 and A. flavus ATCC 9643, respectively. MLC/MIC ratio was ≤2, suggesting the CuNPs had a bactericidal or fungicidal effect on both pathogenic strains. Results also indicated that bacterial CuNPs at varying concentrations of 25, 50, 100 and 150 μg/mL were such a good antitumor agent against A549 lung carcinoma cell lines with an IC50 value of 137.5 μg/mL and a cell viability of 89.3, 79.6, 64.9 and 44.1%, respectively. The results also suggested that the biosynthesized-CuNPs were an antimicrobial and anticancer agent that could be used in future in food preservation, biomedicine and pharmaceutical fields.

Design, synthesis and biological evaluation of 2,3-dihydro-[1,4]dioxino[2,3-f]quinazoline derivatives as EGFR inhibitors

Epidermal growth factor receptor (EGFR) is the most attractive target for drug research in non-small cell lung cancer (NSCLC). The first-generation EGFR tyrosine kinase inhibitors (TKIs) Gefetinib and Elotinib showed good clinical efficacy on lung adenocarcinoma tumors, but almost all patients developed resistance to these inhibitors over time. Quinazoline and quinoline derivatives are common targeted inhibitors of EGFR kinase, and their structural optimization is an important direction for the development of effective targeted anticancer drugs. Based on these facts, a series of heterocyclic 2,3-dihydro-[1,4]dioxino[2,3-f]quinazoline derivatives have been designed and synthesized and their structures were confirmed by spectral analyses. The cytotoxic activity of the newly synthesized compounds was evaluated against the human kidney epithelial T293 cell line, normal lung cell lines WI-38, non-small cell lung cancer A549 and NCI-H157 cell lines using MTT. The tested compounds showed an evident anticancer activity against the tested cell lines, especially compound 13c, which was the most potent anticancer agent with half maximal inhibitory concentrations (IC50) between 8.82 and 10.24 μM. Docking study showed that compound 13b could be nicely bound to the ATP binding pocket of EGFR. In addition, the inhibitory activity of the target compounds against epidermal growth factor receptor tyrosine kinase (EGFR-TK) was evaluated. Results indicated the ability of the target compounds to inhibit EGFR-TK with half maximal inhibitory concentrations (IC50) in the range of 10.29 nM to 652.3 nM. In view of the reported compound activity, the structure deserves further optimization as cancer treatment agents.

Determination of Genotoxicity Attributed to Diesel Exhaust Particles in Normal Human Embryonic Lung Cell (WI-38) Line.

Several epidemiological studies concluded that inhalation of diesel exhaust particles (DEP) is associated with an increase in the relative risk of lung cancer. In vitro research evaluating the genetic damage and/or changes in gene expression have been attempted to explain the relationship between DEP exposure and carcinogenicity. However, to date, investigations have been largely confined to studies in immortalized or tumorigenic epithelial cell models. Few studies have investigated damage at the chromosomal level to DEP exposure in normal cell lines. Here, we present the genotoxic effects of DEP in normal cells (embryonic human lung fibroblasts) by conventional genotoxicity testing (micronuclei (MN) and comet assay). We show the differentially expressed genes and enriched pathways in DEP-exposed WI-38 cells using RNA sequencing data. We observed a significant increase in single-strand DNA breaks and the frequency of MN in DEP-exposed cells in a dose-dependent manner. The differentially expressed genes following DEP exposure were significantly enriched in the pathway for responding to xenobiotics and DNA damage. Taken together, these results show that DEP exposure induced DNA damage at the chromosomal level in normal human lung cells and provide information on the expression of genes associated with genotoxic stress.

Brilacidin Demonstrates Inhibition of SARS-CoV-2 in Cell Culture.

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), the newly emergent causative agent of coronavirus disease-19 (COVID-19), has resulted in more than two million deaths worldwide since it was first detected in 2019. There is a critical global need for therapeutic intervention strategies that can be deployed to safely treat COVID-19 disease and reduce associated morbidity and mortality. Increasing evidence shows that both natural and synthetic antimicrobial peptides (AMPs), also referred to as Host Defense Proteins/Peptides (HDPs), can inhibit SARS-CoV-2, paving the way for the potential clinical use of these molecules as therapeutic options. In this manuscript, we describe the potent antiviral activity exerted by brilacidin—a de novo designed synthetic small molecule that captures the biological properties of HDPs—on SARS-CoV-2 in a human lung cell line (Calu-3) and a monkey cell line (Vero). These data suggest that SARS-CoV-2 inhibition in these cell culture models is likely to be a result of the impact of brilacidin on viral entry and its disruption of viral integrity. Brilacidin demonstrated synergistic antiviral activity when combined with remdesivir. Collectively, our data demonstrate that brilacidin exerts potent inhibition of SARS-CoV-2 against different strains of the virus in cell culture.

RET inhibition in novel patient-derived models of RET-fusion positive lung adenocarcinoma reveals a role for MYC upregulation.

ABSTRACTMulti-kinase RET inhibitors, such as cabozantinib and RXDX-105, are active in lung cancer patients with RET fusions; however, the overall response rates to these two drugs are unsatisfactory compared to other targeted therapy paradigms. Moreover, these inhibitors may have different efficacies against RET rearrangements depending on the upstream fusion partner. A comprehensive preclinical analysis of the efficacy of RET inhibitors is lacking due to a paucity of disease models harboring RET rearrangements. Here, we generated two new patient-derived xenograft (PDX) models, one new patient-derived cell line, one PDX-derived cell line, and several isogenic cell lines with RET fusions. Using these models, we re-examined the efficacy and mechanism of action of cabozantinib and found that this RET inhibitor was effective at blocking growth of cell lines, activating caspase 3/7 and inhibiting activation of ERK and AKT. Cabozantinib treatment of mice bearing RET fusion-positive cell line xenografts and two PDXs significantly reduced tumor proliferation without adverse toxicity. Moreover, cabozantinib was effective at reducing growth of a lung cancer PDX that was not responsive to RXDX-105. Transcriptomic analysis of lung tumors and cell lines with RET alterations showed activation of a MYC signature and this was suppressed by treatment of cell lines with cabozantinib. MYC protein levels were rapidly depleted following cabozantinib treatment. Taken together, our results demonstrate that cabozantinib is an effective agent in preclinical models harboring RET rearrangements with three different 5′ fusion partners (CCDC6, KIF5B and TRIM33). Notably, we identify MYC as a protein that is upregulated by RET expression and downregulated by treatment with cabozantinib, opening up potentially new therapeutic avenues for the combinatorial targetin of RET fusion- driven lung cancers. The novel RET fusion-dependent preclinical models described here represent valuable tools for further refinement of current therapies and the evaluation of novel therapeutic strategies.

Identification of a Lung Cancer Biomarker Using a Cancer Cell Line and Screening of Olfactory Receptors for Biomarker Detection

Several studies investigated the development of a non-invasive and early diagnostic method for cancer. Recently, the volatile organic compounds (VOCs) exhaled from cancer patients have been shown to act as a potential biomarker for cancer. However, the gas profile of breath is highly variable and not conducive to identification of biomarkers accurately. Accordingly, we identified a cancer-specific VOC as a biomarker using an in vitro cancer system in order to exclude the variables from patient condition. VOCs were collected from the headspace gas of lung cancer cell line (SK-MES) and normal lung cell line (MRC-5), followed by analysis using solid-phase microextraction (SPME)-GC/MS. As a result, 2-ethyl-1-hexanol was detected from headspace gas of SK-MES and decided as a potential biomarker for lung cancer. By screening human olfactory receptor libraries to detect a lung cancer biomarker, human olfactory receptor 4D11P (hOR4D11P) was identified as a receptor specific for 2-ethyl-1-hexanol and expressed heterologously in HEK293 cells. Additionally, we generated nanovesicles containing hOR4D11P from HEK293 cells and confirmed the expression and functionality of hOR4D11P in nanovesicles.

Long noncoding RNA LINC00520 accelerates lung adenocarcinoma progression via miR-1252-5p/FOXR2 pathway.

It has been corroborated that long noncoding RNA (lncRNA) played fundamental function in various human malignancies development including lung adenocarcinoma (lung ADC). In our study, LINC00520 roles in lung ADC tumorigenesis were explored. We found that LINC00520 level was elevated in lung ADC tissues and cell lines. Besides, the LINC00520 expression had a negative connection with miR-1252-5p level in lung ADC tissues. Additionally, our results demonstrated the reciprocal repression influence between LINC00520 and miR-1252-5p. Moreover, luciferase reporter assays, RIP (RNA-binding protein immunoprecipitation) and pull down assays revealed that miR-1252-5p regulated LINC00520 in RISC-dependent. Furthermore, knockdown of LINC00520 inhibited lung ADC cells proliferation, migration and invasion, while co-transfection with a miR-1252-5p inhibitor inverted these influences. Additionally, the findings also demonstrated that FOXR2 was a target of miR-1252-5p; thus, LINC00520 could regulate FOXR2 level. Moreover, LINC00520 silencing suppressed the tumor growth of lung ADC in vivo. In summary, our data indicated that LINC00520 may act as a ceRNA to modulated FOXR2 level by sponging miR-1252-5p, which might bring a potential and effective biomarker to lung ADC treatment.

Genotoxic stress-activated DNA-PK-p53 cascade and autophagy cooperatively induce ciliogenesis to maintain the DNA damage response

The DNA-PK maintains cell survival when DNA damage occurs. In addition, aberrant activation of the DNA-PK induces centrosome amplification, suggesting additional roles for this kinase. Here, we showed that the DNA-PK-p53 cascade induced primary cilia formation (ciliogenesis), thus maintaining the DNA damage response under genotoxic stress. Treatment with genotoxic drugs (etoposide, neocarzinostatin, hydroxyurea, or cisplatin) led to ciliogenesis in human retina (RPE1), trophoblast (HTR8), lung (A459), and mouse Leydig progenitor (TM3) cell lines. Upon genotoxic stress, several DNA damage signaling were activated, but only the DNA-PK-p53 cascade contributed to ciliogenesis, as pharmacological inhibition or genetic depletion of this pathway decreased genotoxic stress-induced ciliogenesis. Interestingly, in addition to localizing to the nucleus, activated DNA-PK localized to the base of the primary cilium (mother centriole) and daughter centriole. Genotoxic stress also induced autophagy. Inhibition of autophagy initiation or lysosomal degradation or depletion of ATG7 decreased genotoxic stress-induced ciliogenesis. Besides, inhibition of ciliogenesis by depletion of IFT88 or CEP164 attenuated the genotoxic stress-induced DNA damage response. Thus, our study uncovered the interplay among genotoxic stress, the primary cilium, and the DNA damage response.

Waterpipe smoke condensate influences epithelial to mesenchymal transition and interferes with the cytotoxic immune response in non-small cell lung cancer cell lines.

Waterpipe tobacco smoking (WPS) continues to spread globally and presents serious health hazards. The aim of the present study was to investigate the effects of treatment with WPS condensate (WPSC) on lung cell proliferation and plasticity as well as tumor cell recognition and killing by natural killer (NK) cells using cytotoxicity assays. The results indicated that exposure of normal and cancer lung cell lines to WPSC resulted in a decrease in their in vitro growth in a dose-dependent manner and it induced tumor senescence. In addition, WPSC selectively caused DNA damage as revealed by an increase in γH2AX and 53BP1 in tumor lung cells. To gain further insight into the molecular mechanisms altered by WPSC, we conducted a global comprehensive transcriptome analysis of WPSC-treated tumor cells. Data analysis identified an expression profile of genes that best distinguished treated and non-treated cells involving several pathways. Of these pathways, we focused on those involved in epithelial to mesenchymal transition (EMT) and stemness. Results showed that WPSC induced an increase in SNAI2 expression associated with EMT, ACTA2 and SERPINE2 were involved in invasion and CD44 was associated with stemness. Furthermore, WPSC exposure increased the expression of inflammatory response genes including CASP1, IL1B, IL6 and CCL2. While immune synapse formation between NK and WPSC-treated lung cancer target cells was not affected, the capacity of NK cells to kill these target cells was reduced. The data reported in the present study are, to the best of our knowledge, the first in vitro demonstration of WPSC effects on lung cellular parameters providing evidence of its potential involvement in tumor physiology and development.

Design, synthesis and biological evaluation of water soluble and non-aggregated silicon phthalocyanines, naphthalocyanines against A549, SNU-398, SK-MEL128, DU-145, BT-20 and HFC cell lines as potential anticancer agents

Cancer has become an important public problem in worldwide since cancer incidence and mortality are growing rapidly. In this study, water soluble and non-aggregated silicon (IV) phthalocyanines and naphthalocyanines containing (3,5-bis{3-[3-(diethylamino)phenoxy]propoxy}phenyl)methoxy groups have been synthesized and characterized to investigate their anticancer potential. Their DNA binding/nuclease, topoisomerases inhibition were investigated using UV–Vis absorption, thermal denaturation and agarose gel electrophoresis. The in vitro cytotoxic properties of the compounds on human lung (A549), breast (BT-20), liver (SNU-398), prostate (DU-145), melanoma (SK-Mel 128) carcinoma and human fibroblast (HFC) normal cell lines were evaluated by using MTT assay. In order to determine the mechanism of cancer cell growth suppression, cell cycle analysis was carried out using flow cytometer on A549 cell line. The Kb values of SiPc1a and SiNc2a were 6.85 ± (0.35) × 106 and 1.72 ± (0.16) × 104 M−1 and Tm values of ct-DNA were calculated as 82.02 °C and 78.07 °C, respectively in the presence of both compounds. The ΔTm values of SiPc1a and SiNc2a were calculated as 6.45 and 2.50 °C, respectively. The nuclease effects of SiPc1a and SiNc2a with supercoiled plasmid pBR322 DNA demonstrated that both compounds did not trigger any DNA nuclease effects at the lowest concentrations without irradiation whereas both compounds in the presence of activating agent (H2O2) showed significant plasmid DNA nuclease actions under irradiation (22.5 J/cm2). SiPc1a and SiNc2a inhibited to topoisomerase I on increasing concentrations whilst they had lower inhibition action toward topoisomerase II that of topoisomerase I. The in vitro cytotoxicity studies displayed that SiPc1a had the highest cytotoxic effects among the tested compounds against A549, SNU-398, SK-MEL128, DU-145, BT-20 and HFC cell lines with CC50 values ranged from 0.49 to 2.99 µM. Furthermore, SiPc1a inhibited cell proliferation by cell cycle arrest in G0/G1 phase. All of these results suggested that SiPc1a is a promising candidate as an anticancer agent.

Recombinant Expression of Cec-B Peptide in Escherichia coli with a Significant Anticancer Effect on Hepatocellular Carcinoma.

Cecropin-B (Cec-B) is an Antimicrobial Peptide (AMP) found in insects. Recombinant production of Cec-B peptide in Escherichia coli (Rosetta™ DE3), and studying its anticancer effect on Hepatocellular Carcinoma Cell line (HCC). The Cec-B gene of Drosophila melanogaster was synthesized by PCR assembly using the Simplified Gene Synthesis (SGS) method. To express the recombinant peptide in E. coli (Rosetta™ DE3); the synthesized gene was cloned into pET-15b expression vector. The recombinant peptide was expressed as insoluble aggregates called Inclusion Bodies (IBs) using 2mM lactose inducer. IBs were solubilized in a denatured form using 8 M urea followed by in-vitro protein refolding using rapid dilution method. The refolded Cec-B was purified using cation-exchange SP-FF column. Cytotoxicity of recombinant Cec-B (rCec-B) was reported on normal human lung cell line (WI-38), and Hepatocellular carcinoma cell line (HepG2). The Cec-B gene was expressed and purified at concentration 1.212±0.1 mg/ml which represents 48.49±4% of the total proteins injected to the column (2.5±0.2 mg/ml). The safe dose of purified rCec-B on normal WI-38 cells was calculated to be 1.57 mg/ml. The half-maximal Inhibitory Concentration (IC50) of rCec-B on HepG2 cell line was calculated to be 25 μg/ml. Scanning Electron Microscope (SEM) showed that untreated and treated HepG2 cells had cell diameters from 11-12.92 μm and 14.18-21.58 μm, respectively. The results of this study revealed a successful expression of the rCec-B peptide using a pET-based expression system with a simple purification step. The purified peptide could be considered as a hopeful anticancer drug against HCC.

Anticancer, Enhanced Antibacterial, and Free Radical Scavenging Potential of Fucoidan- (Fucus vesiculosus Source) Mediated Silver Nanoparticles.

The present research displays the green synthesis of stable silver nanoparticles (AgNPs). The aqueous solution of Fucoidan from Fucus vesiculosus source (brown marine algae) is used as a reducing and capping agent. UV-Vis spectroscopy, XRD, FT-IR, SEM, EDX, and TEM with selected area electron diffraction are used to characterize the synthesized silver nanoparticles (AgNPs). The synthesized AgNPs exhibit a surface plasmon resonance at 430 nm after 24 h. The characterization results showed that AgNPs are crystalline in nature and exhibit mostly spherical shapes with an average diameter of 4-45 nm. Silver nanoparticles showed effective antibacterial activity against representative pathogens of bacteria. The activities of commercial antibiotics were enhanced by impregnation with the synthesized AgNPs. It also shows good fungicidal and anticancer activity against liver and lung cell lines and shows significant antioxidant efficacy (84%) at 10 μg/ml AgNP concentration against DPPH. The utilization of environmentally synthesized AgNPs offers numerous benefits of ecofriendliness and compatibility for biomedical applications.

What’s Testosterone Got to Do with It? A Critical Assessment of the Contribution of Testosterone to Gender Disparities in COVID-19 Infections and Deaths

In the short time since severe acute respiratory syndrome coronavirus 2 (henceforth referred to as coronavirus disease 2019 [COVID-19]) appeared, numerous articles have suggested that testosterone (T) may be a major contributor to infection and death since more men than women die, and two proteins involved in viral host entry are thought to be upregulated by androgens. We investigated whether the available data supported this supposition. A MEDLINE search was performed with keywords of COVID-19 variations and androgens or T. Data regarding COVID-19 infections and deaths were obtained from the literature, the World Health Organization, and the U.S. Center for Disease Control, GLOBAL HEALTH5050 and the Harvard School of Public Health Gender Science Laboratory. Studies with T measurements in COVID-19 patients were reviewed. Studies investigating the relationship between T and angiotensin converting enzyme 2 (ACE2) and the transmembrane protease serine 2 (TMPRSS2) expression were reviewed. Global and U.S. data reveal that infection rates in men and women are similar. Men accounted for 58% and women 42% of global deaths. U.S. data revealed a ratio of 54% male deaths to 46% female deaths. However, this finding was inconsistent, as several countries reported greater numbers of female deaths, for example, Canada, Portugal, Finland, and Vietnam. In the United States, 23.5% of states and territories reported more deaths among females. Highest death rates for men and women occurred among the elderly, when serum T is at its lifetime nadir, and low death rates were observed in young adults when serum T is at its peak. All four studies reporting T measurements in COVID-19 patients indicated that low T levels were associated with adverse outcomes, that is, transfer to intensive care unit or death. Although several studies did show androgenic upregulation of TMPRSS2 and ACE2 in prostate, and cancer cell lines of prostate and lung, human and murine lung tissue fails to show a difference in expression between males and females. Observed data fail to support the popular notion that androgens contribute meaningfully to COVID-19 infection and severity of illness. On the contrary, these data raise the possibility that low T may be responsible for disease severity. There is no evidence that androgens upregulate key proteins involved with COVID-19 infection in lung. The possibility that T therapy may aid management of hospitalized COVID-19 patients merits investigation.

Polyphenol rich horseradish root extracts and juice: In vitro antitumor activity and mechanism of action

Background/Aim. Plant polyphenols are well known to show antimutagenic, anticarcinogenic, antiviral and antioxidative activity. The aim of this study was to investigate bio-active potential of Armoracia rusticana root juice and extracts: their polyphenol content, as well as in vitro antitumor activity and cell-death mechanism. Methods. Liquid-liquid extraction of polar and non-polar compounds was used and poly-phenolic compounds were identified and quantified by high performance liquid chomatography (HPLC) analysis. Anti-proliferative activity was examined in vitro on human cervix carcinoma (HeLa), breast adenocarcinoma (MCF7, MDA-MB-231), colon adenocarcinoma (HT-29), lung adenocarcinoma (A549), prostate adenocarcinoma (PC-3), melanocyte carcinoma (Hs 294T), hepatocyte carcinoma (Hep G2), as well as rat hepatocyte carcinoma (H-4-II-E), and normal human fetal lung (MRC-5) cell line using sulforhodamine B assay. The mechanism of cell-death in cell line was determined using Cell Death Detection ELISAPLUS kit. Results. Dichloromethane extracts had the highest content of cate-chin, p-hydroxybenzoic, syringic and gallic acid (pulp, E1), and epicatechin (juice, E3). The results showed strong and non-selective antiproliferative activity of chloroform and di-chloromethane extracts and root juice ? highest being to-wards liver, breast and lung tissue cells. IC50 values of ex-tracts and juice had low range of concentrations (IC50 = 3.49?26.5 ?g/mL) and high range of dilutions (IC50 = 418?1,590). High and unfavorable potential of horseradish juice and chloroform juice extract (E4) to induce necrotic cell death was detected. Conclusion. Strong and non-selective in vitro antiproliferative activity of chloroform and dichloromethane extracts and root juice of horseradish was detected, with necrosis as a main mechanism of induced cell death. In order to utilize horseradish root bioactive potential further investigations that will pinpoint active components with more favourable apoptosis/necrosis inducing properties are needed.

Implementation of the microdosimetric kinetic model using analytical microdosimetry in a treatment planning system for proton therapy.

To implement RBE calculations in treatment planning systems based on the Microdosimetric Kinetic Model (MKM) upon analytical calculations of dose-mean lineal energy (yD). MKM relies on the patterns of energy deposition in sub-nuclear structures called domains, whose radii are cell-specific and need to be determined. The radius of a domain (rd) can be determined from the linear-quadratic (LQ) curves from clonogenic experiments for different cell lines exposed to X-ray and proton beams with known yD. In this work, LQ parameters for two different human lung cell lines (H1299 and H460) are used, and yD among cells is calculated through an analytical algorithm. Once rd is determined, MKM-based calculations of RBE are implemented in a treatment planning system (TPS). Results are compared to those produced by phenomenological models of RBE, such as Carabe and McNamara. Differences between model-based predictions and experimentally determined RBE are analyzed for yD=5keV/μm. For the H1299 line, mean differences in RBE are 0.13, -0.29 and -0.27 for our MKM-based calculation, Carabe and McNamara models, respectively. For the H460 line, differences become -0.044, -0.091 and -0.048, respectively. RBE is computed for these models in a simple plan, showing MKM the best agreement with the experimentally obtained RBE, keeping deviations below 0.08. Microdosimetry calculations at the TPS-level provide tools to improve predictions of RBE using the MKM with actual values of yD instead of LET. The radius of the characteristic domain needs to be determined to tailor the RBE prediction for each cell or tissue.

Phytochemical Biological and Cytotoxic Activities of Calothamnus quadrifidus Leaves Cultivated in Egypt

The hydro distilled essential oil obtained from Calothamnus quadrifidus leaves was analyzed by gas liquid chromatography-mass spectroscopy. Quantification of the oil constituents was carried out using the gas liquid chromatography-flame ionization detection device where 1,8-cineole was quantified as 525.62 μg/ml. The oil showed potential antioxidant activity when tested by 1,1-diphenyl-2-picrylhydrazine radical scavenging assay where the concentration required to scavenge 1,1-diphenyl-2-picrylhydrazine by 50 %=37.3 μg. The cytotoxic activity of the essential oil was evaluated using 3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyltetrazolium bromide assay compared with 1,8-cineole and doxorubicin. Calothamnus quadrifidus leaves essential oil cytotoxic activity was significant against human lung cell line A-549 (2.37 μg), human colon cell line HCT-116 (2.39 μg), human hepato-cellular cell line HepG-2 (2.55 μg) and intestinal epithelium carcinoma Caco-2 (5.80 μg) cell lines, respectively. The oil cytotoxicity exceeded 1,8-cineole except for intestinal epithelium carcinoma Caco-2. Additionally, the antimicrobial activity was evaluated using agar disc diffusion and minimum inhibitory concentration where the oil exhibited the best activity against Penicillium italicum and Geotrichum candidum with minimum inhibitory concentration=0.49 and 0.98 μg, respectively. Structure elucidation of the pure compounds isolated from Calothamnus quadrifidus leaves was done according to their chromatographic behavior, chemical and spectroscopic data. These compounds were isolated for the first time from Calothamnus quadrifidus leaves and identified as gallic acid, gallic acid methyl ester, 7,4`-dimethoxy-apigenin, 6,7-dimethoxy-apigenin, (Bis[3(2-methyl propan-1- one)-2, 6 dihydroxy-4- methoxy phenyl] methane, quercetin 3-O-beta-D-arbinopyranoside and quercetin 3-O-beta-D-4C1-xylopyranoside. In addition to the molecular docking of novel compound 3 was done in peroxiredoxins and the binding mode was found. Conformation 2 was found to be the most stable and may be responsible for the antioxidant effect as it showed the best free binding energy and affinity when compared to conformations.

A Method for Liposome Co-encapsulation of Phenethyl Isothiocyanate and Cisplatin and Determining Its Toxicity Against Lung and Lung Cancer Cells.

Non-small cell lung cancer (NSCLC) is one of the leading causes of cancer-related deaths in the United States. It is extremely difficult to treat, and its survival rate is low. Today, the most effective treatments are still those that implement the platinum anticancer drug cisplatin (CDDP) in combination with other drugs. We previously demonstrated that the naturally occurring compound phenethyl isothiocyanate (PEITC) could be used to sensitize NSCLC cells to CDDP. Furthermore, co-encapsulation of PEITC and CDDP in liposomes enhances their toxicity toward NSCLC cells. We have optimized a liposomal-PEITC-CDDP formulation and investigated its cytotoxicity. We determined that liposomal-PEITC-CDDP is much more toxic toward human NSCLC cell lines than it is toward human normal lung cell lines. In this chapter, we describe detailed methods for preparing liposomal-PEITC-CDDP and determining its cytotoxicity.

Synergistic inhibition of SARS-CoV-2 cell entry by otamixaban and covalent protease inhibitors: pre-clinical assessment of pharmacological and molecular properties.

SARS-CoV-2, the cause of the COVID-19 pandemic, exploits host cell proteins for viral entry into human lung cells. One of them, the protease TMPRSS2, is required to activate the viral spike protein (S). Even though two inhibitors, camostat and nafamostat, are known to inhibit TMPRSS2 and block cell entry of SARS-CoV-2, finding further potent therapeutic options is still an important task. In this study, we report that a late-stage drug candidate, otamixaban, inhibits SARS-CoV-2 cell entry. We show that otamixaban suppresses TMPRSS2 activity and SARS-CoV-2 infection of a human lung cell line, although with lower potency than camostat or nafamostat. In contrast, otamixaban inhibits SARS-CoV-2 infection of precision cut lung slices with the same potency as camostat. Furthermore, we report that otamixaban's potency can be significantly enhanced by (sub-) nanomolar nafamostat or camostat supplementation. Dominant molecular TMPRSS2-otamixaban interactions are assessed by extensive 109 μs of atomistic molecular dynamics simulations. Our findings suggest that combinations of otamixaban with supplemental camostat or nafamostat are a promising option for the treatment of COVID-19.

Schiff base ligand L synthesis and its evaluation as anticancer and antidepressant agent

ObjectivesThe current study focuses on the synthesis of Schiff base ligand L [1(pyridine-2-ylimino) methyl) naphthalene-2-ol], and its applications as anticancer agent against human lung (H-460) and breast (MCF-7) cell lines, and as antidepressant agent. MethodsThe Schiff base ligand L was synthesized by aldol condensation reaction by reacting commercially available 2-hydroxy-1-napthaldehyde with 2-amino-pyridine. The synthesized ligand was characterized by different spectroscopic techniques and evaluated for its anticancer potential against H-460 and MCF-7 cell lines. The antidepressant activity was evaluated by means of elevated plus maze model using Diazepam as reference drug. ResultsA dose dependent inhibitory response was recorded against both the cell line used. For highest tested dose (500 µg/ml), 89.20% of anticancer activity with IC50 = 23.99 µg/ml against H-460 was observed while an IC50 of 31.68 µg/ml against MCF-7 indicated the potential of this compound to be used as anticancer agent. A dose dependent response was also recorded for the synthesized ligand when tested as antidepressant in elevated plus maze model. ConclusionThe synthesized Schiff base ligand L exhibited good anticancer and antidepressant activities that could be used as alternative drug to treat the related health complications. However, further toxicological and clinical studies are needed to confirm the safe use of this compound.

ATM inhibition enhances Auranofin-induced oxidative stress and cell death in lung cell lines.

Ataxia-Telangiectasia (A-T), a pleiotropic chromosomal breakage syndrome, is caused by the loss of the kinase Ataxia-telangiectasia mutated (ATM). ATM is not only involved in the response to DNA damage, but also in sensing and counteracting oxidative stress. Since a disturbed redox balance has been implicated in the pathophysiology of A-T lung disease, we aimed to further explore the interplay between ATM and oxidative stress in lung cells. Using a kinetic trapping approach, we could demonstrate an interaction between the trapping mutant TRX1-CS and ATM upon oxidative stress. We could further show that combined inhibition of thioredoxin reductase (TrxR) and ATM kinase activity, using Auranofin and KU55933 respectively, induced an increase in cellular reactive oxygen species (ROS) levels and protein oxidation in lung cells. Furthermore, ATM inhibition sensitized lung cells to Auranofin-induced cell death that could be rescued by ROS scavengers. As a consequence, targeted reduction of ATM by TRX1 could serve as a regulator of oxidative ATM activation and contribute to the maintenance of the cellular redox homeostasis. These results highlight the importance of the redox-active function of ATM in preventing ROS accumulation and cell death in lung cells.