Normal Human Fibroblast Cells Research Articles

The Antiproliferation Activity of Ganoderma formosanum Extracts on Prostate Cancer Cells

Androgen-independent prostate cancer accounts for mortality in the world. In this study, various extracts of a medical fungus dubbed Ganoderma formosanum were screened for inhibition of DU145 cells, an androgen-independent prostate cancer cell line. Results demonstrated that both hexane (GF-EH) and butanol (GF-EB) fraction of G. formosanum ethanol extract inhibited DU145 cell viability in a dose-dependent manner. GF-EH induced cell-cycle arrest in G1 phase of DU145 cells via downregulation of cyclin E2 protein expression. In addition, GF-EB triggered extrinsic apoptosis of DU145 cells by activating caspase 3 gene expression resulting in programed cell death. Above all, both GF-EH and GF-EB show lower toxicity to normal human fibroblast cell line compared to DU145 cell, implying that they possess specific drug action on cancer cells. This study provides a molecular basis of G. formosanum extract as a potential ingredient for treatment of androgen-independent prostate cancer.

Antimicrobial Activity and DNA/BSA Binding Affinity of Polynuclear Silver(I) Complexes with 1,2-Bis(4-pyridyl)ethane/ethene as Bridging Ligands.

1,2-Bis(4-pyridyl)ethane (bpa) and 1,2-bis(4-pyridyl)ethene (bpe) were used for the synthesis of polynuclear silver(I) complexes, {[Ag(bpa)]NO3}n (1), {[Ag(bpa)2]CF3SO3.H2O}n (2) and {[Ag(bpe)]CF3SO3}n (3). In complexes 1–3, the corresponding nitrogen-containing heterocycle acts as a bridging ligand between two Ag(I) ions. In vitro antimicrobial activity of these complexes, along with the ligands used for their synthesis, was evaluated against the broad panel of Gram-positive and Gram-negative bacteria and fungi. The silver(I) complexes 1–3 showed selectivity towards Candida spp. and Gram-negative Escherichia coli in comparison to the other investigated bacterial strains, effectively inhibiting the growth of four different Candida species with minimal inhibitory concentrations (MICs) between 2.5 and 25 μg/mL and the growth of E. coli, with MIC value being 12.5 μg/mL. Importantly, complex 2 significantly reduced C. albicans filamentation, an essential process for its pathogenesis. Antiproliferative effect on the normal human lung fibroblast cell line MRC-5 was also evaluated with the aim of determining the therapeutic potential of the complexes 1–3. The interactions of these complexes with calf thymus DNA (ctDNA) and bovine serum albumin (BSA) were studied to evaluate their binding activities towards these biomolecules for possible insights on their mode of action.

Molecular mechanisms of apoptosis induction in K562 and KG1a leukemia cells by a water-soluble copper(II) thiosemicarbazone complex

Thiosemicarbazones (TSCs) and their metal complexes exhibit pronounced and selective cytotoxic potential against a broad span of cancers. Here, we assessed the anti-cancer activity of a water-soluble copper(II) complex of thiosemicarbazone (Cu-TSC) against two cancer cell lines of human leukemia. Our analysis revealed that Cu-TSC treatment results in a time and dose-dependent growth inhibition in K562 and KG1a cells while sparing normal human fibroblast (HFF2) cells. The IC50 values for the Cu-TSC treatment were measured to be 21.7 ± 1.5µM and 50.25 ± 2.5µM for K562 and KG1a cells, respectively. Cell cycle analysis indicated that Cu-TSC induces the accumulation of cells in the sub-G1 fraction as well as the reversible arrest in G0/G1 and G2/M phases in K562 and KG1a cells, respectively. Furthermore, the occurrence of apoptosis as the prime mode of cell death was verified through apoptotic body formation, phosphatidylserine externalization, and caspase-3 activation. Additionally, the real-time quantitative PCR analysis revealed that Cu-TSC triggers apoptosis in both cell lines via the upregulation of caspases-8, -9, and the changing of Bax/Bcl2 ratio. Finally, flow cytometric analysis confirmed that Cu-TSC treatment causes the enhancement of reactive oxygen species formation in both K562 and KG1a cells. Altogether, these findings suggest that Cu-TSC is a promising inducer of apoptosis in leukemia cells and carries potential as an anti-cancer compound.

Characterizing the Role of the Na+‐H+ Exchanger Isoform 1 (NHE1) in Pulmonary Fibrosis

Pulmonary fibrosis is a chronic, progressive lung disease characterized by stiffening and thickening of the lung tissue leading to reduced respiratory function. When lung tissue becomes damaged, the response is to increase recruitment and proliferation of fibroblasts and increase deposition of extracellular matrix (ECM) proteins such as collagen. This initiates the release of transforming growth factor beta 1 (TGF‐β1), a cytokine known to enhance fibrosis. TGF‐β1 induces myofibroblast differentiation and further stiffening of the ECM by stimulating formation of cytoplasmic microfilament bundles and increasing expression of alpha‐smooth muscle actin (α‐SMA). Signal molecules such as serotonin (5‐HT), and lysophosphatidic acid (LPA) contribute to this by binding to G‐protein coupled receptors that support increased release of TGF‐β1. Fibrosis ultimately results in a state of low blood flow and hypoxia in the lung tissue, a microenvironment associated with upregulation of the sodium‐hydrogen exchanger isoform 1 (NHE1). NHE1 is a ubiquitously expressed protein that regulates intracellular pH. Upregulation of NHE1 supports increased cell proliferation and migration, indicating that it may play a role in the behavior of fibroblasts in pulmonary fibrosis. The proliferation, collagen deposition, and myofibroblast differentiation of two fibroblast cell lines will be characterized. CCL39, Chinese hamster lung fibroblasts, and WI‐38, normal human fibroblast cells, will both be studied. CCL39 cells treated with EIPA, a chemical inhibitor of NHE1, show reduced proliferative rates. This suggests that proliferation in these cells is NHE1‐dependent. In addition to proliferation, it is hypothesized that the collagen deposition and myofibroblast differentiation of this cell line will be NHE1‐dependent. Furthermore, it is hypothesized that WI‐38 cells will display similar NHE1‐dependent proliferation, collagen deposition and myofibroblast differentiation as the CCL39 cells. This suggests that CCL39 cells can serve as a predictor of human fibroblast cell behaviors.

Low-level laser irradiation potentiates anticancer activity of p-coumaric acid against human malignant melanoma cells.

p-Coumaric acid (PCA) is a kind of phenolic compound, and as one of the cinnamic acid derivatives, it has many biological functions such as antioxidants, anti-inflammatory, antiplatelet, and anticancer activity. Low-level laser irradiation has received increasing interest in the fields of tissue regeneration and wound healing. In this study, the effect of low-level laser irradiation on human fibroblast cells (human dermal fibroblast) and human melanoma cancer cells (A375 and SK-MEL-37) treated with PCA was investigated. The human dermal fibroblast, A375, and SK-MEL-37 cells were exposed to low-level laser at 660-nm wavelength with 3 J/cm for 90 s, and then the cells were treated with different concentrations of PCA (0-1000 μg/ml for 24 h), separately. In another experiment, first the cells were treated by PCA and then irradiated with low-level laser as described before. The effect of various irradiation energy (1-6 J/cm) on the melanoma cells, which were then treated by PCA, was studied. The cell viability using MTT assay and lactate dehydrogenase assay was determined. Morphological changes owing to apoptosis induction by irradiation and PCA were detected by fluorescence microscopy using acridine orange/ethidium bromide double staining. The results showed that pretreatment with low-level laser irradiation and then PCA reduced the survival and growth of melanoma cells more than the early treatment with PCA and then low-level laser irradiation. Lactate dehydrogenase activity was reduced significantly by preirradiation and then PCA treatment in comparison with the dark group in melanoma cells. The cell cytotoxicity at different irradiation energy and then IC50 concentration of PCA was increased up to 3 J/cm and then decreased following increasing irradiation energy. The morphology study with light microscopy and apoptotic assay using acridine orange/ethidium bromide dual staining confirmed the MTT results. This study showed that low-level laser irradiation alone is not able to kill human normal fibroblast and human melanoma cancer cells. Preirradiation followed by treatment with PCA did not change the cell viability in human fibroblast significantly but reduced the cell viability in melanoma cells presumably through the apoptosis pathway.

Study on Bioactivity of Opuntia Ficus-Indica Fruit Extracts fermented Low Temperature by Anti pollutants

Air pollutants can trigger various disorders in humans and induce skin aging effects, related cell death, and inflammation. This study demonstrated the effects of low temperature-fermented Opuntia Ficus-Indica Fruit extracts (LTOE) treated total suspended particles (TSP). LTOE induced the expression of pro-inflammatory cytokines such as tumor necrosis factor (TNF)-α, interleukin (IL)-8 without cytotoxicity. To investigate the anti-aging effect, LTOE inhibited the production of matrix metalloproteinase (MMP)-1 (collagenase) more than Opuntia Ficus-Indica Fruit extracts (OE) in TSP-treated Normal human fibroblast cells. This study reports the significant improvement of anti-inflammation and decrease of MMP-1 related to the LTOE, which will be helpful in processing cosmetic ingredients.

Design and chemical behavior of novel pyrimidine derivatives and their evaluation of cytotoxicity

Diphenyl-4-thioxo-1,4-dihydropyrimidin-5-yl)ethan-1-one 4 was obtained upon the reaction of (phenylamino)but-2-enethioyl)benzamide 3 with sodium hydroxide. The reactions of 4 with several active species such as benzaldehyde, urea, malononitrile, ethyl cyanoacetate, hydrazine, and carbon disulfide were studied. The antitumor activities of some selective compounds were examined against two cell lines: Mammalian cell lines: A-549 cells (human lung cancer cell line) and MRC-5 cells (normal human lung fibroblast cell line); some of the compounds were highly efficient; compound 13 has the most effective anticancer activity.

Silver(I) metallodrugs of thiosemicarbazones and naproxen: biocompatibility, in vitro anti-proliferative activity and in silico interaction studies with EGFR, VEGFR2 and LOX receptors.

Four new heteroleptic silver(I) complexes with the general formula [Ag(L1-4)(nap)] (1-4), where L1-4= 2-(1-(4-substitutedphenyl)ethylidene)hydrazinecarbothioamide and nap=naproxen, have been synthesized and characterized. The geometric parameters determined from density functional theory and UV-Vis studies indicate distorted tetrahedral geometry around silver(I) ion. Fourier transform infrared (FT IR) spectra evidenced asymmetric bidentate coordination mode of carboxyl oxygen atoms of naproxen with silver(I) ion. The complexes are stable for 72h and biocompatibility was analysed towards normal human dermal fibroblast cells, which showed non-toxic nature up to 100ng/ml. In vitro anti-proliferative activity of the complexes by MTT assay was tested against three human cancerous cell lines and one non-tumorigenic human breast epithelial cell line (MCF-10a) in which the complex 4 exhibited enhanced activity. The morphological changes observed by acridine orange/ethidium bromide and Hoechst 33258 staining method reveal apoptosis-inducing ability of the complexes. The molecular docking studies suggest hydrogen bonding, hydrophobic and π-pair interactions with the active site of epidermal growth factor receptor, vascular endothelial growth factor receptor 2 and lipoxygenase receptors.

Phytochemical Screening and Cytotoxic Properties of Ethanolic Extract of Young and Mature Khat Leaves

The khat plant has been culturally used in many parts of Africa and the Arabian Peninsula for many years to induce psycho-stimulating effect. Because of the global wide-spreading nature, khat chewing is being considered as a universally growing problem. Catha abbottii, Catha edulis, and Catha transvaalensis are the three species of khat commonly chewed in Saudi Arabia and nearby regions. Khat users usually prefer to chew young leaves over mature ones due to the diverse effects produced by both. Though many of the constituents of khat leaves have been identified, the complete phytochemical profile of young and mature leaves was not performed or compared; also, no evidence is available to affirm the cytotoxicity of young or mature leaves. Therefore, this study aimed to investigate the phytochemical basis of the differential response of the young and mature leaves and to assess the cytotoxicity of young and mature khat leaves. Ethanolic extracts of young and mature leaves of three khat cultivars were subjected to GC-MS. Hierarchical cluster analysis revealed the existence of two major clusters. The extracts of young leaves were found to contain the maximum content of cathinone; however, methoxyamphetamine was found in only one extract of young leaves. Cytotoxicity investigations were also conducted on both types of leaves using three cancer cell lines, human breast adenocarcinoma, human ovary adenocarcinoma, and human colon adenocarcinoma and also normal human fetal lung fibroblast cell line was used. All extracts showed comparable cytotoxicity, IC50 ranging from 22–59 μg/mL on the cancer cells; however, we observed more cytotoxicity against normal cells (IC50: 6–41 μg/mL). The predominant cytotoxicity on normal cells may pose many health hazards to khat consumers.

Zinc L-Carnosine Protects CCD-18co Cells from L-Buthionine Sulfoximine-Induced Oxidative Stress via the Induction of Metallothionein and Superoxide Dismutase 1 Expression.

Zinc L-carnosine (ZnC) is the chelate form of zinc and L-carnosine and is one of the zinc supplements available in the market. This study aims to determine the protective effects of ZnC against L-buthionine sulfoximine (BSO)-induced oxidative stress in CCD-18co human normal colon fibroblast cell line. CCD-18co cells were pretreated with ZnC (0-100μM) for 24h before the induction of oxidative stress by BSO (1mM) for another 24h. Results from this present study demonstrated that ZnC up to the concentration of 100μM was not cytotoxic to CCD-18co cells. Induction with BSO significantly increased the intracellular reactive oxygen species (ROS) levels and reduced the intracellular glutathione (GSH) levels in CCD-18co cells. Pretreatment with ZnC was able to attenuate the increment in intracellular ROS level in CCD-18co cells significantly in a concentration-dependent manner. However, ZnC did not have any effects on intracellular GSH levels and Nrf2 activation. Mechanistically, pretreatment with ZnC was able to upregulate the expression of metallothionein (MT) and superoxide dismutase 1 (SOD1) in CCD-18co cells. Results from dual-luciferase reporter gene assay reported that ZnC was able to increase the MRE-mediated relative luciferase activities in a concentration-dependent manner, suggesting that the induction of MT expression by ZnC was due to the activation of MTF-1 signaling pathway. Taken together, our current findings suggest that ZnC can protect CCD-18co cells from BSO-induced oxidative stress via the induction of MT and SOD1 expression.

Synergistic antibacterial activity of alpha mangostin and resveratrol loaded polymer-based films against bacteria infected wound

In general, a wound infection is caused by Staphylococcus aureus (S. aureus) which can be treated by antibacterial agents. Alpha mangostin (M) and resveratrol (R) have been reported to possess antimicrobial properties. However, the M and R combination has not been studied. Thus, this study aimed to develop polymer-based films for the co-delivery of M and R for the treatment of S. aureus infection. M and R were studied for their synergistic antibacterial activity against S. aureus. Various single (ethyl cellulose (EC) and Eudragit® L 100 (L)) and combined polymers (polyvinyl alcohol and chitosan (PVA:CS)) were used to prepare the films. The M and R loaded films were characterized for their physical, chemical and mechanical properties. The M and R release was investigated using Franz diffusion cell. The antibacterial efficacy of the films was assessed against S. aureus. The in vitro cytotoxicity in normal human fibroblast (NHF) was evaluated as well. The results showed that the M and R combination provided synergistic antibacterial activity. The combined PVA:CS polymer showed good mechanical properties and thus was selected to load M and R. The M and R loaded PVA:CS (at a weight ratio of 1:2)-based films provided satisfactory physical, chemical and mechanical properties. Films wetted, swelled and then M and R released at the physiological pH of skin. The rapid bactericidal effect of the developed films was even faster than that of a commercial dressing (Bactigras®). In addition, the films provided low cytotoxicity on NHF cells. Thus, the developed films may serve as a promising drug delivery carrier for the treatment of bacteria infected wound.

Self‐Healing, High Adherent, and Antioxidative LbL Multilayered Film for Enhanced Cell Adhesion

AbstractCell adhesion acts as a complex process that is hardly promoted through surfaces with a single performance. Controlling the cell's adhesion on the interfaces between the synthetic materials and the biological environments remains a real challenge in the biomedical fields. Herein, based on RGD peptide modified carboxymethyl chitosan (CCS‐R) and dopamine modified oxidized alginate (OALG‐D), a layer‐by‐layer (LbL) self‐assemble multilayered (CCS‐R/OALG‐D)15 film is developed. The multifunctional (CCS‐R/OALG‐D)15 film with a self‐healing, highly adherent, biocompatible, and antioxidative biomatrix property for the cell adhesion is fabricated with the Schiff‐base bond, electrovalent bond, and hydrogen bond. The LbL multilayered film exhibits the significant cell adhesion ability for the normal human dermal fibroblast cells and the coral Acropopa formosa cells. Furthermore, it holds high self‐healing efficiency (88.8%), excellent adhesion strength (adhesive stress: 61.1 MPa), and outstanding antioxidation (free radical scavenging rate: 86.8%). Furthermore, it is demonstrated that the LbL multifunctional film, as a culture matrix, has prolonged the survival of the coral Acropopa formosa. Therefore, the incorporation of a self‐healing, high adhesion, superior cell adhesion, and antioxidation property into a single LbL self‐assembly film provides a promising platform for regenerative medicine.

Cytotoxic and Apoptotic Induction Potential of Extracts from Fermented Citrullus vulgaris Thunb. Seeds on Cervical and Liver Cancer Cells

The anti-cancer activities of many fermented foods and beverages are now scientifically established. Ogiri-egusi is a condiment prepared from fermentation of Citrullus vulgaris (melon) seeds and consumed in many countries of West Africa. Its anti-oxidative and anti-hyperlipidaemic properties have been reported. This study investigated the anti-cancer activities of the aqueous and methanolic extracts from ogiri-egusi. Cytotoxicity was investigated using the MTT and colony-formation inhibition assays while flow-cytometer based Apopercentage assay was used to quantify apoptosis in extracts-treated cervical and liver cancer and normal human fibroblast cells. The inhibitory concentration responsible for killing 50% of cells after 24 h by the aqueous extract in KMST-6, HeLa, and Hep-G2 cells were estimated at 1.610, 1.020, and 1.507 mg/mL respectively. While these values reduced with increasing incubation time in cancer cells it increased in the non-cancer cell. Furthermore, the extract significantly induced apoptosis in HeLa (97 ± 0.18%) and Hep-G2 (73 ± 6.73%) cells. These findings were corroborated by cells morphologic presentations and inhibition of colony formation assay. These findings suggest that the aqueous extract from fermented Citrullus vulgaris seeds might be a nutraceutical with potential anti-cancer properties.

Pristimerin inhibits the proliferation of HT1080 fibrosarcoma cells by inducing apoptosis.

Fibrosarcoma is a soft tissue sarcoma that is classified as a rare cancer. Therefore, no standard anti-tumor drug therapy has been established for fibrosarcoma. Although pristimerin (PM) has been reported to exert an anti-tumor effect on various types of cancer, no studies have examined the therapeutic effect of PM on soft tissue sarcoma. The purpose of the current study was to investigate the anti-tumor effect of PM on human fibrosarcoma cells (HT1080). The present study examined the cell viability, IC50 values and ability to induce apoptosis of PM in HT1080 and normal human dermal fibroblast (aHDF) cells. The effect of PM on the following signaling pathways associated with cell proliferation was also evaluated: AKT and mitogen-activated protein kinase (MAPK). Using mice subcutaneously transplanted with fibrosarcoma cells, the effect of PM treatment was investigated on tumor growth inhibition, body weight and liver and renal function. The results revealed that PM administration reduced cell viability and induced apoptosis in a dose-dependent matter. In HT1080 cells, the IC50 value of PM was 0.16 µM at 24 h and 0.13 µM at 48 h. PM treatment also decreased the levels of phosphorylated AKT, mTOR, NF-κB and phosphorylated ERK in a dose-dependent manner. In the PM injection group, the increase in tumor volume was significantly reduced and the effect on weight loss and liver and renal function were revealed to be insignificant. PM exerted little effect on normal human dermal fibroblasts and was highly effective against human fibrosarcoma cells. The results indicated that PM may be used as a potential therapeutic agent against fibrosarcoma.

Abstract P3-10-21: Preclinical evaluation of a novel phospholipid drug conjugate with a combretastatin A-4 analogue for improved breast cancer therapy

Abstract Purpose: Breast cancer (BrCa) is the most common invasive cancer in women globally and the second most common cause of death from cancer in women in the US. Combretastatin A-4 (CA4) and associated analogues have been shown to be potent cytotoxins and inhibitors of tumor blood flow. However, CA4 and its analogues have not progressed rapidly through clinical trials. It has been shown that with some of the CA4 analogues, incomplete disruption of the vasculature and/or proangiogenic factors around the tumor result in survival of the cap portion of the tumor and future progression. Additionally, the over expression of MDR-1 gene which encodes for P-glycoprotein (Pgp) efflux pumps which is overexpressed in many cancer cells has been shown to limit the activity of these molecules. Phospholipid-drug conjugates (PDCs) are a modality designed to selectively deliver molecules to the tumor or tumor microenvironment, significantly increasing intracellular or local concentrations. In the current study, we established a series of PDCs using a single CA4 analogue (CA4A) conjugated to our phospholipid ether targeting platform (PDC-CA4A) utilizing different peptide linkers that allow for facile targeting and release of the CA4A payload. In this study, we evaluated the in vitro and in vivo therapeutic potential of different PDC-CA4A molecules. Methods: In vitro assessment of cellular internalization, accumulation and release of the payload was evaluated via mass spectrometry in A549 and normal human dermal fibroblast (NHDFs) cells. Additionally, in vitro assessment of cytotoxicity was performed in A549, MCF7, HCT116 and NHDFs with fifty percent inhibitory concentrations (IC50s) being determined for each. Subcutaneous HCT116 and MCF7 xenograft tumor models were used to evaluate the antitumor efficacy of several PDC-CA4A molecules compared with the free CA4A molecule. Results: All PDC-CA4A molecules showed excellent uptake, accumulation and release of the CA4A payload in the A549 cell line. Uptake was detected at the first time point tested (30 minutes) after exposure and continued through 24 hours. A steady state of release of the payload and new PDC-CA4A molecules entering the cell was reached 3 hours post exposure. Cytotoxicity tests demonstrated IC50s ranging from nanomolar to micromolar concentration depending upon the linker being utilized. In vivo, PDC-CA4A-109 (specific molecule) demonstrated statistically significant benefit over both free CA4A and the positive control group. Conclusion: Our results suggest that PDC-CA4A-109 is a promising antitumor agent to treat both breast cancer and colorectal cancers. Further clinical study is warranted to evaluate the safety and feasibility of PDC-CA4A-109 as a novel targeted therapy. Citation Format: Jarrod Longcor, Randall Hoover, Mark Wolf. Preclinical evaluation of a novel phospholipid drug conjugate with a combretastatin A-4 analogue for improved breast cancer therapy [abstract]. In: Proceedings of the 2019 San Antonio Breast Cancer Symposium; 2019 Dec 10-14; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2020;80(4 Suppl):Abstract nr P3-10-21.

A new pimarane-type diterpene obtained by biotransformation inhibits human HCT-116 colorectal carcinoma through inhibition of LTA4H activity

Chronic inflammation mediated by several markers promotes cancer progression. Leukotriene A4 hydrolase (LTA4H), an inflammatory marker, is highly expressed in colorectal cancer. Therefore, inhibition of LTA4H could reduce the incidence and progression of this cancer type. Several studies supported the valuable effects of naturally occurring diterpenes against several diseases. In this study, a new pimarane-type diterpene, namely, 8,15R-epoxy,16-hydroxy-pimaran-19-oic acid 1 was obtained from the biotransformation of 8β-hydroxypimar-15-en-19-oic acid 2 using a filamentous fungus, Cordyceps sinensis. Both compounds were in vitro tested for their cytotoxic effects against human colorectal carcinoma (HCT-116) cells. In addition, their selectivity was examined using the normal human fibroblast cells, HF-19 and TIG-1. The new metabolite 1 exhibited potent cytotoxic activity against HCT-116 (IC50 7.53 μM) with no cytotoxicity against TIG-1 and HF-19. Thus, LTA4H inhibitory activity including both aminopeptidase (AP) and epoxide hydrolase (EH) functionalities were assessed in comparison with bestatin and 4-(4-Benzylphenyl)-thiazol-2-amine (4BSA, ARM1), respectively, as positive controls. Interstingly, the new metabolite 1 showed higher AP, and EH inhibitory activities (IC50 11.21 and 6.64 μM, respectively), confirming the impact of the achieved structure modification on the related anticancer activity. To gain further understanding of the mode of binding of the new metabolite 1 within the active binding site of LTA4H, docking experiments were conducted. The results indicated the significance of pimarane-type diterpenes as a new candidate for the design of promising LTA4H inhibitors.

Synthesis, Structural, and Cytotoxic Properties of New Water-Soluble Copper(II) Complexes based on 2,9-Dimethyl-1,10-Phenanthroline and Their One Derivative Containing 1,3,5-Triaza-7-Phosphaadamantane-7-Oxide.

A series of water-soluble copper(II) complexes based on 2,9-dimethyl-1,10-phenanthroline (dmphen) and mixed-ligands, containing PTA=O (1,3,5-triaza-7-phosphaadamantane-7-oxide) have been synthesized and fully characterized. Two types of complexes have been obtained, monocationic [Cu(NO3)(O-PTA=O)(dmphen)][PF6] (1), [Cu(Cl)(dmphen)2][PF6] (2), and neutral [Cu(NO3)2(dmphen)] (3). The solid-state structures of all complexes have been determined by single-crystal X-ray diffraction. Magnetic studies for the complex 1–3 indicated a very weak antiferromagnetic interaction between copper(II) ions in crystal lattice. Complexes were successfully evaluated for their cytotoxic activities on the normal human dermal fibroblast (NHDF) cell line and the antitumor activity using the human lung carcinoma (A549), epithelioid cervix carcinoma (HeLa), colon (LoVo), and breast adenocarcinoma (MCF-7) cell lines. Complexes 1 and 3 revealed lower toxicity to NHDF than A549 and HeLa cells, meanwhile compound 2 appeared to be more toxic to NHDF cell line in comparison to all cancer lines. Additionally, interactions between the complexes and human apo-transferrin (apo-Tf) using fluorescence and circular dichroism (CD) spectroscopy were also investigated. All compounds interacted with apo-transferrin, causing same changes of the protein conformation. Electrostatic interactions dominate in the 1/2 – apo- Tf systems and hydrophobic and ionic interactions in the case of 3.

Stimulation of cytoprotective autophagy and components of mitochondrial biogenesis / proteostasis in response to ionizing radiation as a credible pro-survival strategy

The present study illustrates mitochondria-mediated impact of ionizing radiation which is paralleled by activation of several pro-adaptive responses in normal human dermal fibroblast cells. Irradiation of cells with X-rays (5 Gy) led to an increase in fragmentation and mitochondrial mass. Distinct temporal changes in cytosolic and mitochondrial reactive oxygen species (ROS) were noted in response to radiation, which was associated with depletion in mitochondrial membrane potential followed by decrease in ATP levels. Long Amplicon-Polymerase Chain Reaction (LA-PCR) analysis showed time-dependent increase in mitochondrial DNA damage that preceded mitochondrial ROS generation. Irradiation of cells led to an initial G2/M arrest at 8 h that persisted till 16 h, with subsequent block at G0/G1 measured at 48 and 72 h time points. Interestingly, cells activated autophagy as a countermeasure against radiation-mediated cellular insults and aided in removal of damaged mitochondria. Blocking autophagy using 3-methyladenine led to cell death via activation of enhanced ROS, PARP-1 and caspase 3 cleavage. Upregulation of mitochondrial biogenesis factors Nrf1/PGC-1α, following irradiation was observed. Irradiated cells exhibited an increase in the phosphorylation of GCN2, PERK and eIF2α that might be responsible for the up-regulation of ATF4 and CHOP thereby regulating autophagy and components of integrated stress response. Apart from this, we measured accumulation of mitochondrial chaperones (HSP60/HSP10) and ATF5 which is a major molecule involved in mitochondrial stress. Taken together, mitochondria are one of the major cytoplasmic targets for ionizing radiation and possibly act as an early indicator of cellular insult. The findings also show that stressed mitochondria might influence endoplasmic reticulum (ER)-related signalling leading to the activation of adaptive mechanisms like cytoprotective autophagy, and molecules responsible for mitochondrial biogenesis and protein quality control in order to replenish mitochondrial pool and maintain cellular homeostasis.

Uses of ethyl benzoylacetate for the synthesis of thiophene, thiazole, pyridine, and pyran derivatives with antitumor activities

AbstractThe reaction of ethyl benzoylacetate with malononitrile in an oil bath at 120°C gave the condensation product 3. The latter compound underwent a series of heterocyclization to give thiophene, thiazole, pyridine, and pyran derivatives. The structures of the synthesized products were established on the basis of analytical and spectral data. The antitumor evaluation of the newly synthesized products against the six cancer cell lines namely human gastric cancer (NUGC and HR), human colon cancer (DLD1), human liver cancer (HA22T and HEPG2), human breast cancer (MCF), nasopharyngeal carcinoma (HONE1), and normal fibroblast cells (WI38) indicated that many compounds expressed high inhibition against the six cancer cell lines. Compounds 3, 8a, 8c, 14b, 16b, 16c, 16d, 19a, 19b, 20a, 22a, 27b, and 28a were the most cytotoxic compounds among the tested compounds.

Detection of hydrogen sulfide using BODIPY based colorimetric and fluorescent on-off chemosensor

A colorimetric and fluorescence turn-off probe 10-(4-azido phenyl)-5,5-difluoro-5h-dipyrrolo[1,2-c:1’,2’-f][1,3,2] diazaborinin-4-ium-5-uide, 1, for selective detection of H2S is reported. The probe displayed a robust decrease in fluorescence intensity with high sensitivity, specificity and least toxicity to detect exogenous H2S and also in live normal human oral fibroblast cells loaded with probe 1. The detection limit of probe 1 being 0.17 µm for H2S. A colorimetric and fluorescence turn-off BODIPY probe 1, for selective detection of exogenous H2S in live normal human oral fibroblast cells loaded with probe 1 and poisonous H2S gas formed under biodegradation in sewage waters and water tanks and corroded pipe lines. The detection limit of probe 1 being 0.17 μM