Human Alveolar Basal Epithelial Cells Research Articles

Fluorophore Tagged Mixed Ligand Copper(II) Complexes: Synthesis, Structural Characterization, Protein Binding, DNA Cleavage and Anticancer Activity

AbstractTwo fluorophore tagged copper(II) complexes [Cu(phen)(L)(ClO4)2] (1) and [Cu(bpy)(L)(H2O)(ClO4)](ClO4) (2), (where L=2‐amino‐1H‐benzo[de]isoquinoline‐1,3‐(2H)dione (L), phen=1,10‐phenanthroline and bpy=2,2′‐bipyridine) have been synthesized and structurally characterized. Structures of the copper complexes 1 and 2 were confirmed by single‐crystal X‐ray structure determination. The coordination geometry around the copper center of complexes 1 and 2 is distorted octahedral. The plasmid DNA cleavage activity of the complexes has been investigated by agarose gel electrophoresis and the study reveals that both the complexes have high plasmid DNA photo‐cleavage activity. The binding interaction ability of the metal complexes with bovine serum albumin (BSA) was investigated using fluorescence spectroscopy. The cytotoxicity of the complexes has been evaluated by MTT (3‐[4,5‐dimethylthiazole‐2‐yl]‐2,5‐diphenyltetrazolium bromide) assay against A549 (adenocarcinoma human alveolar basal epithelial cells) and MCF‐7 (breast cancer cell line) cell lines in comparison with cis‐platin. Complexes 1 and 2 have exhibited better cytotoxic activity than cis‐platin against A549 and MCF‐7 cell lines. The cellular uptake study and localization of the complexes within the cells have been investigated by fluorescence microscopy. The cell staining and flow cytometry experiments suggest that complexes induce an apoptotic mode of cell death.

Cytotoxic activity of organotin compounds containing non-steroidal anti-inflammatory drugs

Two series of organotin (IV) complexes based on diclofenac L1Na and aspirin L2H of formulae Me3SnL1 (1); Ph3SnL1 (2); Bu2Sn(L1)2 (3); R2Sn(L1)2 (4, R = 3,5-di-tert-butyl-4-hydroxyphenyl); Me3SnL2 (5); Me3Sn(2-hydroxybenzoate) (6) and Me2Sn(L2)2 (7) were synthesized and characterized by 1H, 13C, 119Sn NMR, IR, ESI-MS and elemental analysis. It was found by X-ray diffraction analysis of compound 1 that carboxyl group of diclofenac binds two tin atoms each being in the trigonal bipyramid geometry and having coordination number 5. Compounds 6 and 7 were found to be monomers in the solid state with tetrahedron and octahedron geometry around Sn center, respectively. Cytotoxicity in vitro of compounds 1–7 and initial ligands was evaluated on human colon cancer (HCT-116), human breast cancer (MCF-7) and adenocarcinomic human alveolar basal epithelial (A-549) cells. The IC50 values varied in 0.17–200 µM range for compounds 1–4 and in 3.3–200 µM range for compounds 5–7 depending on the substituents at Sn center and ligand structure. It was found that compound 2 possesses the maximal cytotoxic activity and significantly induces apoptosis of HCT-116 cells.

A small molecule targeting CHI3L1 inhibits lung metastasis by blocking IL-13Rα2-mediated JNK-AP-1 signals.

Our previous big data analyses showed a high level of association between chitinase 3 like1 (CHI3L1) expression and lung tumor development. In the present study, we investigated whether a CHI3L1‐inhibiting chemical, 2‐({3‐[2‐(1‐cyclohexen‐1‐yl)ethyl]‐6,7‐dimethoxy‐4‐oxo‐3,4‐dihydro‐2‐quinazolinyl}sulfanyl)‐N‐(4‐ethylphenyl)butanamide (K284), could inhibit lung metastasis and studied its mechanism of action. We investigated the antitumor effect of K284 both in vitro and in vivo. K284 (0.5 mg·kg−1 body weight) significantly inhibited lung metastasis in in vivo models after injection of murine melanoma cells (B16F10) or adenocarcinomic human alveolar basal epithelial cells (A549). K284 significantly and concentration‐dependently also inhibited cancer cell proliferation and migration in the A549 and H460 lung cancer cell lines. We found that the binding of K284 to the chitin‐binding domain (CBD) of CHI3L1 prevented the binding of CHI3L1 to its receptor, interleukin‐13 receptor subunit alpha‐2 (IL‐13Rα2), thereby suppressing the CHI3L1 signal. This blocking of the CHI3L1‐IL‐13Rα2 signal caused the inhibition of c‐Jun N‐terminal kinase (JNK)‐activator protein 1 (AP‐1) signals, resulting in the prevention of lung metastasis and cancer cell growth. Our data demonstrate that K284 may serve as a potential candidate anticancer compound targeting CHI3L1.

Bioactivity guided isolation and characterization of antimicrobial principle from Hydrocotyle javanica Thunb.

Hydrocotyle javanica Thunb. (Family Araliaceae) has been used in traditional herbal medicine as a substitute of Centella asiatica (L.) Urban. Although, the latter has been studied extensively and various biologically active constituents have been reported, the former has not been studied for its active constituents. The current study followed activity guided isolation and identification of the fraction with antimicrobial activity from H. javanica. Plant extracts were prepared by extracting successively with petroleum ether, ethyl acetate and ethanol. Antimicrobial activity was evaluated using gram positive and gram negative bacteria and Candida albicans. Bioactivity-guided isolation was carried out by preparative thin layer chromatography and bioautography. Cytotoxicity of the most active antimicrobial principle was evaluated against Human embryonic kidney (HEK293) and Adenocarcinomic Human Alveolar Basal Epithelial (A549) cell lines. The tentative structure of the most active antimicrobial principle was constructed from the data obtained from FTIR, MS, H-NMR and C-NMR.

Effect of cyanide ions (CN-) extracted from cassava (Manihotesculenta Crantz) on Alveolar Epithelial Cells (A549 cells)

Cassava (Manihotesculenta Crantz) is one of the most important root crops in tropical countries. It is a major source of cyanogenic glycosides viz. linamarin and lotaustralin, and these on breakdown liberate HCN and ketone. Cassava cyanide extract (CCE) from cassava leaves and tuber rinds were formulated as a biopesticide against certain borer insect pests of horticultural crops. Adenocarcinomic human alveolar basal epithelial cells (A549) were treated with three different concentrations (100, 200, 400 ppm) of CCE. The MTT and NRU assays showed dose-dependent cytotoxicity. The DCFH-DA assay does not show any free radical scavenging activity, whereas the NRR assay showed a reduction in the nitrile radicals with an increase in the concentration of the bioactive compound. A negative correlation was found between the concentration of the bioactive principles and mitochondrial and lysosomal functions. Various cellular assays demonstrated the cellular response of the CCE, and it was found that at higher concentration (400 ppm), the CCE exert a significant necrotic cell death rather than apoptosis. The results of the study indicated that the CCE have a remarkable tendency of anti-proliferative ability.

Anticancer activity of secondary metabolites from Bauhinia tomentosa Linn. leaf – An in silico approach

Introduction and Aim: Plants and plant derived products are used for human healthcare since the dawn of human civilization. About 80% of modern drugs are from natural origin. Many dreadful diseases like cancer are treated using drugs of natural origin. In the present study, a medicinal plant B. tomentosa Linn. leaves were investigated for its anticancer activity using A549 (Human Adenocarcinomic Alveolar Basal Epithelial Cells) cell lines. The active components were identified using GC-MS (Gas Chromatography- Mass Spectrophotometry) analysis. The aim of the present study was to identify potential lead compounds against various protein targets that are involved in lung carcinogenesis using molecular docking approach.
 
 Materials and Methods: 3D structures of compounds reported from GCMS analysis of B. tomentosa were built using Chemsketch software. All the compounds analyzed exhibited antioxidant, anticancer, antimicrobial, anti-inflammatory, and chemo preventive properties. Docking studies were performed using Molegro virtual docker (MVD).
 
 Results: The docking studies revealed that the ligands either activate or inhibit the selected target proteins appropriately. This shows that the phytochemicals of B. tomentosa leaf was found to have appreciable anticancer activity.
 
 Conclusion: The presence of various bioactive phytoconstituents justifies the use of the leaf for various ailments by traditional practitioners.

Adsorptive Microtiter Plates As Solid Supports in Affinity Purification Workflows.

Affinity ligands such as antibodies are widely used in (bio)medical research for purifying proteins from complex biological samples. These ligands are generally immobilized onto solid supports which facilitate the separation of a captured protein from the sample matrix. Adsorptive microtiter plates are commonly used as solid supports prior to immunochemical detection (e.g., immunoassays) but hardly ever prior to liquid chromatography–mass spectrometry (LC-MS-)-based detection. Here, we describe the use of adsorptive microtiter plates for protein enrichment prior to LC-MS detection, and we discuss opportunities and challenges of corresponding workflows, based on examples of targeted (i.e., soluble receptor for advanced glycation end-products (sRAGE) in human serum) and discovery-based workflows (i.e., transcription factor p65 (NF-κB) in lysed murine RAW 264.7 macrophages and peptidyl-prolyl cis–trans isomerase FKBP5 (FKBP5) in lysed human A549 alveolar basal epithelial cells). Thereby, we aim to highlight the potential usefulness of adsorptive microtiter plates in affinity purification workflows prior to LC-MS detection, which could increase their usage in mass spectrometry-based protein research.

Synthesis, Bioactivity, Pharmacokinetic and Biomimetic Properties of Multi-Substituted Coumarin Derivatives.

A series of novel multi-substituted coumarin derivatives were synthesized, spectroscopically characterized, and evaluated for their antioxidant activity, soybean lipoxygenase (LOX) inhibitory ability, their influence on cell viability in immortalized human keratinocytes (HaCaT), and cytotoxicity in adenocarcinomic human alveolar basal epithelial cells (A549) and human melanoma (A375) cells, in vitro. Coumarin analogues 4a–4f, bearing a hydroxyl group at position 5 of the coumarin scaffold and halogen substituents at the 3-phenyl ring, were the most promising ABTS•+ scavengers. 6,8-Dibromo-3-(4-hydroxyphenyl)-4-methyl-chromen-2-one (4k) and 6-bromo-3-(4,5-diacetyloxyphenyl)-4-methyl-chromen-2-one (3m) exhibited significant lipid peroxidation inhibitory activity (IC50 36.9 and 37.1 μM). In the DCF-DA assay, the 4′-fluoro-substituted compound 3f (100%), and the 6-bromo substituted compounds 3i (80.9%) and 4i (100%) presented the highest activity. The 3′-fluoro-substituted coumarins 3e and 4e, along with 3-(4-acetyloxyphenyl)-6,8-dibromo-4-methyl-chromen-2-one (3k), were the most potent lipoxygenase (LOX) inhibitors (IC50 11.4, 4.1, and 8.7 μM, respectively) while displaying remarkable hydroxyl radical scavenging ability, 85.2%, 100%, and 92.9%, respectively. In silico docking studies of compounds 4e and 3k, revealed that they present allosteric interactions with the enzyme. The majority of the analogues (100 μΜ) did not affect the cell viability of HaCaT cells, though several compounds presented over 60% cytotoxicity in A549 or A375 cells. Finally, the human oral absorption (%HOA) and plasma protein binding (%PPB) properties of the synthesized coumarins were also estimated using biomimetic chromatography, and all compounds presented high %HOA (>99%) and %PPB (60–97%) values.

Nuclear-localized human respiratory syncytial virus NS1 protein modulates host gene transcription.

SUMMARYHuman respiratory syncytial virus (RSV) is a common cause of lower respiratory tract infections in the pediatric, elderly, and immunocompromised individuals. RSV non-structural protein NS1 is a known cytosolic immune antagonist, but how NS1 modulates host responses remains poorly defined. Here, we observe NS1 partitioning into the nucleus of RSV-infected cells, including the human airway epithelium. Nuclear NS1 coimmunoprecipitates with Mediator complex and is chromatin associated. Chromatin-immunoprecipitation demonstrates enrichment of NS1 that overlaps Mediator and transcription factor binding within the promoters and enhancers of differentially expressed genes during RSV infection. Mutation of the NS1 C-terminal helix reduces NS1 impact on host gene expression. These data suggest that nuclear NS1 alters host responses to RSV infection by binding at regulatory elements of immune response genes and modulating host gene transcription. Our study identifies another layer of regulation by virally encoded proteins that shapes host response and impacts immunity to RSV.

Characterization and Anticancer Activity of Biosynthesized Au/Cellulose Nanocomposite from Chlorella vulgaris.

Therapeutic selectivity is a critical issue in cancer therapy. As a result of its adjustable physicochemical characteristics, the Au/cellulose nanocomposite currently holds a lot of potential for solving this challenge. This work was designed to prepare a Au/cellulose nanocomposite with enhanced anticancer activity through the regulation of the mitogen-activated protein kinases (MAPK) signaling pathway. Nanocellulose, nanogold (AuNPs), and a Au/cellulose nanocomposite were biosynthesized from microgreen alga Chlorella vulgaris. Using UV-Vis absorption spectroscopy, transmission electron microscope (TEM), zeta potential analyzer, and Fourier transform infrared spectroscopy (FTIR), the synthesized nanoparticles were confirmed and characterized. In human alveolar basal epithelial cells (A549 cells), the selectivity and anticancer activity of the produced nanoparticles were evaluated. The cytotoxicity results revealed that the inhibitory concentration (IC50) of the Au/cellulose nanocomposite against A549 cancer lung cells was 4.67 ± 0.17 µg/µL compared to 182.75 ± 6.45 µg/µL in the case of HEL299 normal lung fibroblasts. It was found that treatment with nanocellulose and the Au/cellulose nanocomposite significantly increased (p < 0.05) the relative expression of tumor suppressor 53 (p53) in comparison to control cells. They also significantly (p < 0.05) decreased the relative expression of the Raf-1 gene. These findings indicate that nanocellulose and the Au/cellulose nanocomposite regulate cell cycles mostly via the motivation of p53 gene expression and reduction of Raf-1 gene expression.

IL-17 stimulates neutrophils to release S100A8/A9 to promote lung epithelial cell apoptosis in Mycoplasma pneumoniae–induced pneumonia in children

Mycoplasma pneumoniae-induced pneumonia (MPP) is a common cause of community-acquired respiratory tract infections, increasing risk of morbidity and mortality, in children. However, diagnosing early-stage MPP is difficult owing to the lack of good diagnostic methods. Here, we examined the protein profile of bronchoalveolar lavage fluid (BALF) and found that S100A8/A9 was highly expressed. Enzyme-linked immunosorbent assays used to assess protein levels in serum samples indicated that S100A8/A9 concentrations were also increased in serum obtained from children with MPP, with no change in S100A8/A9 levels in children with viral or bacterial pneumonia. In vitro, S100A8/A9 treatment significantly increased apoptosis in a human alveolar basal epithelial cell line (A549 cells). Bioinformatics analyses indicated that up-regulated S100A8/A9 proteins participated in the interleukin (IL)−17 signaling pathway. The origin of the increased S100A8/A9 was investigated in A549 cells and in neutrophils obtained from children with MPP. Treatment of neutrophils, but not of A549 cells, with IL-17A released S100A8/A9 into the culture medium. In summary, we demonstrated that S100A8/A9, possibly released from neutrophils, is a new potential biomarker for the clinical diagnosis of children MPP and involved in the development of this disease through enhancing apoptosis of alveolar basal epithelial cells.

Ugonin J Acts as a SARS-CoV-2 3C-like Protease Inhibitor and Exhibits Anti-inflammatory Properties.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection causes severe “flu-like” symptoms that can progress to acute respiratory distress syndrome (ARDS), pneumonia, renal failure, and death. From the therapeutic perspective, 3-chymotrypsin-like protein (3CLpro) is a plausible target for direct-acting antiviral agents because of its indispensable role in viral replication. The flavonoid ugonin J (UJ) has been reported to have antioxidative and anti-inflammatory activities. However, the potential of UJ as an antiviral agent remains unexplored. In this study, we investigated the therapeutic activity of UJ against SARS-CoV-2 infection. Importantly, UJ has a distinct inhibitory activity against SARS-CoV-2 3CLpro, compared to luteolin, kaempferol, and isokaempferide. Specifically, UJ blocks the active site of SARS-CoV-2 3CLpro by forming hydrogen bonding and van der Waals interactions with H163, M165 and E166, G143 and C145, Q189, and P168 in subsites S1, S1′, S2, and S4, respectively. In addition, UJ forms strong, stable interactions with core pharmacophore anchors of SARS-CoV-2 3CLpro in a computational model. UJ shows consistent anti-inflammatory activity in inflamed human alveolar basal epithelial A549 cells. Furthermore, UJ has a 50% cytotoxic concentration (CC50) and a 50% effective concentration (EC50) values of about 783 and 2.38 µM, respectively, with a selectivity index (SI) value of 329, in SARS-CoV-2-infected Vero E6 cells. Taken together, UJ is a direct-acting antiviral that obstructs the activity of a fundamental protease of SARS-CoV-2, offering the therapeutic potential for SARS-CoV-2 infection.

Bio-fabrication of multifunctional quasi-spherical green α-Fe2O3 nanostructures for paracetamol sensing and biomedical applications

Green metal oxide nanostructures with various diverse properties are synthesized to find solutions to various problems arising in the field of medicine and technology. The present study includes cost-effective, eco-friendly synthesis of α-Fe2O3 nanoparticles (ISD NPs), using varied volumes of extract of Scoparia dulsis as a reducing agent via Solution combustion synthesis. The extract influences the size, shape of quasi-spherical, bandgap positions, surface area to volume ratio of the nanoparticles prepared. The structural purity is ascertained via p-XRD studies, with nano regime size in the range of 27–37 nm. The optical energy gap (1.90–2.23 eV) and the mesoporous nature of the biogenic NPs have attributed by UV-DRS and surface area studies. The characterization findings reveal the “symbiotic association” of bio reducing agents and metal to influence the pronounced activity in biomedical and electrochemical studies. The controlled size and morphology give explicit results in in-vitro assays performed. The free radical scavenging hunt was maximum with DPPH inhibition of 71.88% and also exhibited depreciated cell viability of adenocarcinoma human alveolar basal epithelial cells to 39.88% with a lower IC50 value. The inherent stabilization behavior of ISD NPs towards human RBC's is appreciated (71.43%) owing to its bio-compatibility nature. The antidiabetic property via pancreatic α-amylase inhibition was in the range of 60–82%. The distinct electrochemical activities of the ISD NPs showed a higher proton diffusion coefficient (D) with 8.21 × 10−6 cm2s−1, low charge resistance (85.27 Ω) and increased capacitance (5.3 × 10−4 F). The electrochemical studies indicate that the ISD NPs as bio electrodes are a promising sensor for paracetamol drug detection.

Stereo‐ and Regioselective [3+3] Annulation Reaction Catalyzed by Ytterbium: Synthesis of Bicyclic 1,4‐Dihydropyridines

AbstractAn ytterbium catalyzed formal [3+3] cycloaddition of cyclic enamines and α,β‐unsaturated ketones catalyzed is reported. The reaction proceeds with a ‘head to tail’ regioselectivity through a conjugate addition of the enamine moiety followed by an amine‐carbonyl condensation. In addition the use of chiral enamines provided a high degree of stereoselectivity, driven by a possible balance between steric and π‐stacking effects. The resulting bicyclic 1,4‐dihydropyridines were evaluated as antiproliferative agents against A549 (carcinomic human alveolar basal epithelial cell) and SKOV3 (human ovarian carcinoma) human tumor cell lines. Good toxicities were found for some of the compounds against A549 and SKOV3 cell lines, with best IC50 values of 0.89 μM for A549 and 6.69 μM for SKOV3, and a very good selectivity was observed towards MRC5 (non‐malignant) cell lines.magnified image

The Chemokine CCL5 Inhibits the Replication of Influenza A Virus Through SAMHD1 Modulation.

Influenza A virus (IAV) is the main etiological agent of acute respiratory tract infections. During IAV infection, interferon triggers the overexpression of restriction factors (RFs), the intracellular antiviral branch of the innate immune system. Conversely, severe influenza is associated with an unbalanced pro-inflammatory cytokine release. It is unclear whether other cytokines and chemokines released during IAV infection modulate RFs to control virus replication. Among the molecules enhanced in the infected respiratory tract, ligands of the CCR5 receptor play a key role, as they stimulate the migration of inflammatory cells to the alveoli. We investigated here whether ligands of the CCR5 receptor could enhance RFs to levels able to inhibit IAV replication. For this purpose, the human alveolar basal epithelial cell line (A549) was treated with endogenous (CCL3, CCL4 and CCL5) or exogenous (HIV-1 gp120) ligands prior to IAV infection. The three CC-chemokines tested reduced infectious titers between 30% to 45% upon 24 hours of infection. Eploying RT-PCR, a panel of RF mRNA levels from cells treated with CCR5 agonists was evaluated, which showed that the SAMHD1 expression was up-regulated four times over control upon exposure to CCL3, CCL4 and CCL5. We also found that IAV inhibition by CCL5 was dependent on PKC and that SAMHD1 protein levels were also increased after treatment with CCL5. In functional assays, we observed that the knockdown of SAMHD1 resulted in enhanced IAV replication in A549 cells and abolished both CCL5-mediated inhibition of IAV replication and CCL5-mediated cell death inhibition. Our data show that stimuli unrelated to interferon may trigger the upregulation of SAMHD1 and that this RF may directly interfere with IAV replication in alveolar epithelial cells.

A Multicomponent Protocol for the Synthesis of Highly Functionalized γ-Lactam Derivatives and Their Applications as Antiproliferative Agents.

An efficient synthetic methodology for the preparation of 3-amino 1,5-dihydro-2H-pyrrol-2-ones through a multicomponent reaction of amines, aldehydes, and pyruvate derivatives is reported. In addition, the densely substituted lactam substrates show in vitro cytotoxicity, inhibiting the growth of carcinoma human tumor cell lines HEK293 (human embryonic kidney), MCF7 (human breast adenocarcinoma), HTB81 (human prostate carcinoma), HeLa (human epithelioid cervix carcinoma), RKO (human colon epithelial carcinoma), SKOV3 (human ovarian carcinoma), and A549 (carcinomic human alveolar basal epithelial cell). Given the possibilities in the diversity of the substituents that offer the multicomponent synthetic methodology, an extensive structure-activity profile is presented. In addition, both enantiomers of phosphonate-derived γ-lactam have been synthesized and isolated and a study of the cytotoxic activity of the racemic substrate vs. its two enantiomers is also presented. Cell morphology analysis and flow cytometry assays indicate that the main pathway by which our compounds induce cytotoxicity is based on the activation of the intracellular apoptotic mechanism.

Development of dual crosslinked mumio-based hydrogel dressing for wound healing application: Physico-chemistry and antimicrobial activity

In this study, an antimicrobial mumio-based hydrogel dressing was developed for wound healing application. The mechanism of gel formation was achieved via a double crosslink network formation between gelatin (GT) and polyvinyl alcohol (PVA) using polyethylene glycol diglycidyl ether (PEGDE) and borax as crosslinking agents. To enhance the mechanical integrity of the hydrogel matrix, bacterial cellulose (BC) was integrated into the GT-PVA hydrogel to produce a composite gel dressing. The obtained hydrogel was characterized by FTIR, SEM, TGA, and XRD. Gel fraction, in vitro swelling and degradation as well as compressive modulus properties of the gel dressing were investigated as a function of change in PVA and BC ratios. By increasing the ratios of PVA and BC, the composite dressing showed lower swelling but higher mechanical strength. Comparing to other formulations, the gel with 4 %w/v PVA and 1 %w/v BC demonstrated to be most suitable in terms of stability and mechanical properties. In vitro cell cytotoxicity by MTT assay on human alveolar basal epithelial (A549) cell lines validated the gels as non-toxic. In addition, the mumio-based gel was compared to other formulations containing different bioactive agents of beeswax and cinnamon oil, which were tested for microbial growth inhibition effects against different bacteria (S. aureus and K. pneumoniae) and fungi (C. albicans and A. niger) strains. Results suggested that the gel dressing containing combinations of mumio, beeswax and cinnamon oil possess promising future in the inhibition of microbial infection supporting its application as a suitable dressing for wound healing.

Biological In Vitro Evaluation of PIL Graft Conjugates: Cytotoxicity Characteristics.

In vitro cytotoxicity of polymer-carriers, which in the side chains contain the cholinum ionic liquid units with chloride (Cl) or pharmaceutical anions dedicated for antituberculosis therapy, i.e., p-aminosalicylate (PAS) and clavulanate (CLV), was investigated. The carriers and drug conjugates were examined, in the concentration range of 3.125–100 μg/mL, against human bronchial epithelial cells (BEAS-2B) and adenocarcinomic human alveolar basal epithelial cells (A549) as an experimental model cancer cell line possibly coexisting in tuberculosis. The cytotoxicity was evaluated by MTT test and confluency index, as well as by the cytometric analyses, including Annexin-V FITC apoptosis assay. The polymer systems showed supporting activity towards the normal cells and no tumor progress, especially at the highest concentration (100 μg/mL). The analysis of cell death did not show meaningful changes in the case of the BEAS-2B, whereas in the A549 cell line, the cytostatic activity was observed, especially for the drug-free carriers, causing death in up to 80% of cells. This can be regulated by the polymer structure, including the content of cationic units, side-chain length and density, as well as the type and content of pharmaceutical anions. The results of MTT tests, confluency, as well as cytometric analyses, distinguished the polymer systems with Cl/PAS/CLV containing 26% of grafting degree and 43% of ionic units or 46% of grafting degree and 18% of ionic units as the optimal systems.

Anethum Graveolens Essential Oil Nanoemulsions (AGEO-NE) as an Exclusive Apoptotic Inducer in Human Lung Adenocarcinoma (A549) Cells

Anethum graveolens essential oil (AGEO) is used as an anticancer compound that can be made more efficacious by improving its biosolubility and biocompatibility. In the current study, we aimed to improve AGEO bioactivity using the nanoemulsion-based encapsulation technique. To formulate stable AGEO-NE, an ultrasonication method was utilized. The size, stability, and morphology of the AGEO-NE was measured, and then its cytotoxic impacts were evaluated on adenocarcinomic human alveolar basal epithelial cells (A549). The ferric reducing antioxidant power (FRAP) test was done to measure the antioxidant activity of AGEO-NE. Its cytotoxic property was analyzed by measuring the viability percentage of cancer (A549) and normal human foreskin fibroblast (HFF) cell lines after increasing AGEO-NE treatment doses. AGEO-NE apoptotic activity was evaluated by studying the flow cytometry of the cells and measuring the apoptotic gene expression profile (Cas-3 and Cas-8) in the A549 cell line. The results showed a significant correlation between the increase of AGEO-NE concentrations and decrease of cancer (A549) cell viability (p < 0.05) when AGEO-NE was compared with normal HFF cells. Thus, AGEO-NE can be an efficient novel apoptosis and antioxidant inducer for human lung cancer cells without any undesirable side effects. However, further In Vitro and In Vivo studies are needed to confirm the results.

Simulated Microgravity Effects on Human Adenocarcinoma Alveolar Epithelial Cells: Characterization of Morphological, Functional, and Epigenetic Parameters.

Background: In space, the reduction or loss of the gravity vector greatly affects the interaction between cells. Since the beginning of the space age, microgravity has been identified as an informative tool in biomedicine, including cancer research. The A549 cell line is a hypotriploid human alveolar basal epithelial cell line widely used as a model for lung adenocarcinoma. Microgravity has been reported to interfere with mitochondrial activity, energy metabolism, cell vitality and proliferation, chemosensitivity, invasion and morphology of cells and organelles in various biological systems. Concerning lung cancer, several studies have reported the ability of microgravity to modulate the carcinogenic and metastatic process. To investigate these processes, A549 cells were exposed to simulated microgravity (µG) for different time points. Methods: We performed cell cycle and proliferation assays, ultrastructural analysis of mitochondria architecture, as well as a global analysis of miRNA modulated under µG conditions. Results: The exposure of A549 cells to microgravity is accompanied by the generation of polynucleated cells, cell cycle imbalance, growth inhibition, and gross morphological abnormalities, the most evident are highly damaged mitochondria. Global miRNA analysis defined a pool of miRNAs associated with µG solicitation mainly involved in cell cycle regulation, apoptosis, and stress response. To our knowledge, this is the first global miRNA analysis of A549 exposed to microgravity reported. Despite these results, it is not possible to draw any conclusion concerning the ability of µG to interfere with the cancerogenic or the metastatic processes in A549 cells. Conclusions: Our results provide evidence that mitochondria are strongly sensitive to µG. We suggest that mitochondria damage might in turn trigger miRNA modulation related to cell cycle imbalance.