Human Foreskin Fibroblast Cells Research Articles

An in vitro examination of the antioxidant, cytoprotective and anti-inflammatory properties of chrysin-loaded nanofibrous mats for potential wound healing applications

Chrysin (Chr) is a naturally occurring flavone with a wide spectrum of biological functions including anti-cancer, anti-inflammatory and anti-oxidant properties. Due to the low bioavailability and in vivo stability of Chr at therapeutic levels for wound-healing applications, Chr-loaded PCL/PEG nanofibrous mats were successfully fabricated by optimizing the electrospinning parameters and characterized using FE-SEM and FTIR. Results of MTT showed that Human foreskin fibroblast cells (HFF-1) have more than 80% viability on Chr-loaded nanofibers. The antioxidant activity of Chr-loaded PCL/PEG electrospun nanofibers was demonstrated applying an ORAC assay and by the capability of the nanofibers to maintain the viability of HFF-1 cells on the mats under an oxidative stress condition. The Chr-blended PCL/PEG nanofibrous mats also reduced overexpression of IL-6, IL-1β, TNF-α and excessive production of nitric oxide (NO) in J774A1 following stimulation by lipopolysaccharide (LPS). These results suggest that the proposed natural substance based nanofibrous mats can accelerate wound healing process with cell proliferation, antioxidative and anti-inflammatory activities.

<i>In vitro</i> and <i>in vivo</i> antiplasmodial activity of extracts from <i>Polyalthia suaveolens, Uvaria angolensis and Monodora tenuifolia</i> (Annonaceae)

The present study aimed at investigating the in vitro and in vivo susceptibility of malaria parasites to crude extracts and fractions from Polyalthia suaveolens, Uvaria angolensis, and Monodora tenuifolia. The ethanolic extracts were partitioned using water, dichloromethane, hexane, and methanol. The most promising fraction was subjected to column chromatography. The antiplasmodial effect of extracts, fractions and subfractions against P . falciparum Chloroquine resistant (PfK1) strain was determined using SYBR green florescence assay. The promising fraction was assessed for cytotoxicity against Human Foreskin Fibroblast (HFF) cells and further for safety in Swiss albino mice and suppressive effect against P. berghei. The methanol sub-fraction of P. suaveolens [PStw(Ace)] showed the highest potency with IC50 of 3.24 μg/mL. Sub-fraction PS8 from PStw(Ace) was the most active with IC50 of 4.42 μg/mL. Oral administration of PStw(Ace) at 5000 mg/kg b.w in mice showed no signs of toxicity. Also, it exerted the highest suppressive effect against P. berghei at 400 mg/kg b.w throughout the 4 days experiment. Overall, the results achieved supported the use of the three plants in the traditional treatment of malaria in Cameroon. More interestingly, the PStw(Ace) fraction might be of interest in future development of an antimalarial phytodrug. Keywords : Polyalthia suaveolens, Uvaria angolensis, Monodora tenuifolia, Plasmodium falciparum K1, Toxicity, Antimalarial.

Transcriptomic profiles of human foreskin fibroblast cells in response to orf virus.

Orf virus has been utilized as a safe and efficient viral vector against not only diverse infectious diseases, but also against tumors. However, the nature of the genes triggered by the vector in human cells is poorly characterized. Using RNA sequencing technology, we compared specific changes in the transcriptomic profiles in human foreskin fibroblast cells following infection by the orf virus. The results indicated that orf virus upregulates or downregulates expression of a variety of genes, including genes involved in antiviral immune response, apoptosis, cell cycle and a series of signaling pathways, such as the IFN and p53-signaling pathways. The orf virus stimulates or inhibits immune gene expression such as chemokines, chemokine receptors, cytokines, cytokine receptors, and molecules involved in antigen uptake and processing after infection. Expression of pro-apoptotic genes increased at 8 hours post-infection. The p53 signaling pathway was activated to induce apoptosis at the same time. However, the cell cycle program was promoted after infection, which may be due to the immunomodulatory genes of the orf virus. This presents the first description of transcription profile changes in human foreskin fibroblast cells after orf virus infection and provides an in-depth analysis of the interaction between the host and orf virus. These data offer new insights into the understanding of the mechanisms of infection by orf virus and identify potential targets for future studies.

RETRACTED ARTICLE: Empirical Modeling of Physiochemical Immune Response of Multilayer Zinc Oxide Nanomaterials under UV Exposure to Melanoma and Foreskin Fibroblasts

Carcinogenesis is a complex molecular process starting with genetic and epigenetic alterations, mutation stimulation, and DNA modification, which leads to proteomic adaptation ending with an uncontrolled proliferation mechanism. The current research focused on the empirical modelling of the physiological response of human melanoma cells (FM55P) and human foreskin fibroblasts cells (AG01518) to the multilayer zinc oxide (ZnO) nanomaterials under UV-A exposure. To validate this experimental scheme, multilayer ZnO nanomaterials were grown on a femtotip silver capillary and conjugated with protoporphyrin IX (PpIX). Furthermore, PpIX-conjugated ZnO nanomaterials grown on the probe were inserted into human melanoma (FM55P) and foreskin fibroblasts cells (AG01518) under UV-A light exposure. Interestingly, significant cell necrosis was observed because of a loss in mitochondrial membrane potential just after insertion of the femtotip tool. Intense reactive oxygen species (ROS) fluorescence was observed after exposure to the ZnO NWs conjugated with PpIX femtotip model under UV exposure. Results were verified by applying several experimental techniques, e.g., ROS detection, MTT assay, and fluorescence spectroscopy. The present work reports experimental modelling of cell necrosis in normal human skin as well as a cancerous tissue. These obtained results pave the way for a more rational strategy for biomedical and clinical applications.

Time-resolved Global and Chromatin Proteomics during Herpes Simplex Virus Type 1 (HSV-1) Infection

Herpes simplex virus (HSV-1) lytic infection results in global changes to the host cell proteome and the proteins associated with host chromatin. We present a system level characterization of proteome dynamics during infection by performing a multi-dimensional analysis during HSV-1 lytic infection of human foreskin fibroblast (HFF) cells. Our study includes identification and quantification of the host and viral proteomes, phosphoproteomes, chromatin bound proteomes and post-translational modifications (PTMs) on cellular histones during infection. We analyzed proteomes across six time points of virus infection (0, 3, 6, 9, 12 and 15 h post-infection) and clustered trends in abundance using fuzzy c-means. Globally, we accurately quantified more than 4000 proteins, 200 differently modified histone peptides and 9000 phosphorylation sites on cellular proteins. In addition, we identified 67 viral proteins and quantified 571 phosphorylation events (465 with high confidence site localization) on viral proteins, which is currently the most comprehensive map of HSV-1 phosphoproteome. We investigated chromatin bound proteins by proteomic analysis of the high-salt chromatin fraction and identified 510 proteins that were significantly different in abundance during infection. We found 53 histone marks significantly regulated during virus infection, including a steady increase of histone H3 acetylation (H3K9ac and H3K14ac). Our data provide a resource of unprecedented depth for human and viral proteome dynamics during infection. Collectively, our results indicate that the proteome composition of the chromatin of HFF cells is highly affected during HSV-1 infection, and that phosphorylation events are abundant on viral proteins. We propose that our epi-proteomics approach will prove to be important in the characterization of other model infectious systems that involve changes to chromatin composition.

Determination of the Cell Permissiveness Spectrum, Mode of RNA Replication, and RNA-Protein Interaction of Zika Virus

BackgroundTwo lineages of Zika virus (ZIKV) have been classified according to the phylogenetic analysis: African and Asian lineages. It is unclear whether differences exist between the two strains in host cell permissiveness, this information is important for understanding viral pathogenesis and designing anti-viral strategies.MethodsIn the present study, we comparatively studied the permissive spectrum of human cells for both the African (MR766) and Asian strains (PRVABC59) using an RNA in situ hybridization (RISH) to visualize RNA replication, an immunofluorescence technology, and a western blot assay to determine viral protein production, and a real-time RT-PCR to examine viral RNA multiplication level. The experiments were undertaken in the condition of cell culture.ResultsWe identified several human cell lines, including fibroblast, epithelial cells, brain cells, stem cells, and blood cells that are susceptible for the infection of both Asian and African strains. We did not find any differences between the MR766 and the PRVABC59 in the permissiveness, infection rate, and replication modes. Inconsistent to a previous report (Hamel et al. JVI 89:8880–8896, 2015), using RISH or real-time RT-PCR, we found that human foreskin fibroblast cells were not permissive for ZIKV infection. Instead, human lung fibroblast cells (MRC-5) were fully permissive for ZIKV infection. Surprisingly, a direct interaction of ZIKV RNA with envelop (E) protein (a structure protein) was demonstrated by an RNA chromatin immunoprecipitation (ChIP) assay. Three binding sites were identified in the ZIKV RNA genome for the interaction with the E protein.ConclusionOur results imply that the E protein may be important for viral RNA replication, and provide not only the information of ZIKV permissiveness that guides the usage of human cells for the ZIKV studies, but also the insight into the viral RNA-E protein interaction that may be targeted for intervention by designing small molecule drugs.

Identifying connexin expression and determining gap junction intercellular communication in rainbow trout cells.

Gap junctions are groups of membrane-bound channels that allow the passage of small molecules and ions between cells, permitting cell-cell communication. Because of their importance in cell homeostasis, gap junction presence and function were characterized in three commonly studied rainbow trout cell lines, namely RTgill-W1, RTgutGC, and RTG-2. Firstly, gap junction presence was determined by screening for gap junction protein alpha 7 and alpha 1 (GJA7 and GJA1) presence at the transcript level and GJA7 at the protein level. GJA7 was successfully identified at both the transcript and protein levels, and GJA1 was detected at the transcript level in all three cell lines. This is the first report of a GJA7 full-length transcript sequence in rainbow trout cells. Gap junction function, as determined by gap junction intercellular communication (GJIC), was examined using Lucifer yellow dye migration with the scrape and load technique; visualized by fluorescence microscopy. Phorbol 12-myristate 13-acetate (PMA), a gap junction inhibitor, was used to confirm the presence of functional gap junctions. Effects of serum deprivation on GJIC were also monitored; 24-h serum deprivation resulted in greater dye migration compared with 30-min serum deprivation. Both RTG-2 and RTgill-W1 showed significant dye migration that was inhibited by PMA while RTgutGC did not. Human foreskin fibroblast (HFF-1) cells were used as a positive control for gap junction presence and function. Taken together, our study shows that rainbow trout cells express connexin transcripts and proteins, and RTG-2 and, to a lesser extent, RTgill-W1 cells are able to perform GJIC.

Diastereoselective Flexible Synthesis of Carbocyclic C-Nucleosides.

Carbocyclic C-nucleosides are quite rare. Our route enables flexible preparation of three classes of these nucleoside analogs from common precursors-properly substituted cyclopentanones, which can be prepared racemic (in six steps) or optically pure (in ten steps) from inexpensive norbornadiene. The methodology allows flexible manipulation of individual positions around the cyclopentane ring, namely highly diastereoselective installation of carbo- and heterocyclic substituents at position 1', orthogonal functionalization of position 5', and efficient inversion of stereochemistry at position 2'. Newly prepared carbocyclic C-analog of tubercidine, profiled in MCF7 (breast cancer) and HFF1 (human foreskin fibroblasts) cell cultures, is less potent than tubercidine itself, but more selectively toxic toward the tumorigenic cells.

Protective effects of tetrahydrocurcumin (THC) on fibroblast and melanoma cell lines in vitro: it's implication for wound healing.

The aim of this study was to investigate the role of tetrahydrocurcumin (THC) against various skin health parameters using in vitro human foreskin fibroblast and melanoma cell lines (i.e. HFF-1 and B16-F10). The study was assessed using cell viability by MTT assay, identification of extracellular matrix component in HFF-1 cell line (i.e. collagen, elastin and hyaluronic acid), melanin synthesis in B16-F10 cells, cell viability against UVB-induced stress in HFF-1 cells, and in vitro wound healing by the scratch assay. THC was found to be safe and nontoxic up to the concentration of 10µg/mL with improved level of collagen (37.90%), elastin (90.1%), and hyaluronic acid (74.19%) at 1µg/mL. Besides, melanin was significantly inhibition by 78.5% at the lowest THC concentration of 0.1µg/mL. UVB-protection rate was significantly improved by 61.2% and improved cell viability by THC in HFF-1 cells, which indicated protection from photoaging. In addition, THC showed significant wound healing activity (78.51%) and greater migration of fibroblast in HFF-1 cells at different time period. It can be concluded from the study that THC can protect the skin matrix with improved extracellular component synthesis and would healing via collagen synthesis in the skin, which improved the skin elasticity and tightness. Overall, it might be suggested that THC can be used as a safe skin whitening agent, wounds management, cosmetic applications, and treating various skin-related disorders.

An investigation of electrospun Henna leaves extract-loaded chitosan based nanofibrous mats for skin tissue engineering

Wound healing characteristics of some plant extracts have been well known for many years, and they have been utilized for such applications in traditional way. Recently electrospun nanofibrous mats showed promising properties for tissue engineering and especially for skin repair. It is expected that incorporation of plant extracts into such structures could provide higher performance and synergistic effect for biomedical and wound healing applications. The final purpose of this study is to fabricate chitosan based nanofiber mats loaded with a traditional plant extract of Lawsonia inermis (Henna) leaves to enhance the antibacterial efficacy and wound healing of the precursor nanofibers. The morphology, structure, mechanical properties and swelling and weight loss degree of the electrospun nanofibers have been investigated in this study. Antibacterial activity, cell biocompatibility evaluations and in vivo wound healing activity of the abovementioned mats were also studied. The FESEM images of Henna leaves extract-loaded nanofibers proved that homogeneous, smooth and defect free nanofibers of 64–87nm in diameter have been prepared. Presence of Henna extract in the electrospun fibers was approved by Fourier Transform Infrared spectroscopy. Incorporation of Henna extract into the nanofiber mats exhibited significant synergistic antibacterial activity against bacterial cells. It was well supported by the results of cell viability and proliferation of human foreskin fibroblast cells on the prepared scaffolds. Therefore, the results of this work showed that Henna leaves extract incorporated chitosan nonwoven mats have a great potential to be used as the biodegradable, biobased and antibacterial wound healing dressings.

Antioxidant, cytotoxic and DNA protective properties of Achillea eriophora DC. and Achillea biebersteinii Afan. extracts: A comparative study.

Achillea is a traditional medicinal herb which contains different phenol and flavonoid compounds that are responsible for Achillea pharmacological effects. We aimed to determine phenol and flavonoid contents, besides antioxidant activities of different extracts from Achillea eriophoraa (A. eriophora) DC. and Achillea biebersteinii (A. biebersteinii) Afan. (endemic species in Iran) and to investigate their effects on human cells. Achillea extracts, were prepared by maceration and shaking methods, from different parts (aerial parts, stem, leaves and inflorescence) of two species using methanol and ethanol as solvents. Total phenol and flavonoid contents were measured by spectrophotometry, and antioxidant activities of the extracts were determined by DPPH radical scavenging, BCB and TBARS assays. Cytotoxicity and antioxidant activities of the extracts were investigated in Human Foreskin Fibroblast (HFF3) cells using MTT, comet and H2O2 assays. Methanol extracts of A. biebersteinii prepared from leaves and inflorescence by maceration method exhibited maximum phenol (1657.58 ± 36.45 mg GAE/100 g DW) and flavonoid (264.00 ± 62.16 mg QUE/100 g DW) contents. Leaf methanol extract showed significantly higher antioxidant activity (0.0276 ± 0.003, 0.16 ± 0.016 and 13.96 ± 0.26 mg/ml for DPPH, BCB and TBARS IC50s, respectively) than those of the other extracts. Leaf extract of A. biebersteinii was not cytotoxic even at the highest examined dose (512 µg/ml) and inhibited cell toxicity induced by H2O2 (98% viability for the cells pretreated with plant extract in the presence of H2O2). Comet assay also confirmed high DNA protective activity of leaf extracts. Achillea extracts possess remarkable antioxidant activity, and could be good natural alternatives to synthetic antioxidants in pharmaceutical and food industries.

Effects of Thai piperaceae plant extracts on Neospora caninum infection

Neosporosis has a worldwide distribution and causes economic losses in farming, particularly by increasing the risk of abortion in cattle. This study investigated the effects of Thai piperaceae (Piper betle, P. nigrum, and P. sarmentosum) extracts on Neospora caninum infections in vitro and in vivo. In an in vitro parasite growth assay based on the green fluorescent protein (GFP) signal, P. betle was the most effective extract at inhibiting parasite growth in human foreskin fibroblast cells (IC50 of GFP-expressing N. caninum parasites, 22.1μg/ml). The P. betle extract, at 25μg per ml, inhibited parasite invasion into host cells. Furthermore, in two independent experiments, treating N. caninum-infected mice with the P. betle extract for 7days post-infection increased their survival. In trial one, the anti-N. caninum effects of the P. betle extract reduced the mouse clinical scores for 30days post-infection (dpi). The survival rate of the mice treated with 400mg/kg was 100% compared with 66.6% for those treated with 100mg/kg and the non-treated controls. In trial two, treating the infected mice with the P. betle extract increased their survival at 50dpi. All mice in the non-treatment group died; however, the survival rates of the 400mg/kg-treated and 100mg/kg-treated mice were 83.3% and 33.3%, respectively. Also, a trend towards a reduced parasite burden was noted in the brains of the P. betle extract-treated mice, compared with the control mice. Therefore P. betle extract has potential as a medicinal plant for treating neosporosis.

Functional Characterization of Rhoptry Kinome in the Virulent Toxoplasma gondii RH Strain.

Toxoplasma gondii is an obligatory intracellular apicomplexan protozoan which can infect any warm-blooded animal and causes severe diseases in immunocompromised individuals or infants infected in utero. The survival and success of this parasite require that it colonizes the host cell, avoids host immune defenses, replicates within an appropriate niche, and exits the infected host cell to spread to neighboring non-infected cells. All of these processes depend on the parasite ability to synthesis and export secreted proteins. Amongst the secreted proteins, rhoptry organelle proteins (ROPs) are essential for the parasite invasion and host cell manipulation. Even though the functions of most ROPs have been elucidated in the less virulent T. gondii (type II), the roles of ROPs in the highly virulent type I strain remain largely un-characterized. Herein, we investigated the contributions of 15 ROPs (ROP10, ROP11, ROP15, ROP20, ROP23, ROP31, ROP32, ROP33, ROP34, ROP35, ROP36, ROP40, ROP41, ROP46, and ROP47) to the infectivity of the high virulent type I T. gondii (RH strain). Using CRISPR-Cas9, these 15 ROPs genes were successfully disrupted and the effects of gene knockout on the parasite’s ability to infect cells in vitro and BALB/c mice in vivo were investigated. These results showed that deletions of these ROPs did not interfere with the parasite ability to grow in cultured human foreskin fibroblast cells and did not significantly alter parasite pathogenicity for BALB/c mice. Although these ROPs did not seem to be essential for the acute infectious stage of type I T. gondii in the mouse model, they might have different functions in other intermediate hosts or play different roles in other life cycle forms of this parasite due to the different expression patterns; this warrants further investigations.

Enhancement of magnetofection efficiency using chitosan coated superparamagnetic iron oxide nanoparticles and calf thymus DNA

Superparamagnetic iron oxide nanoparticles (MNPs) were prepared and coated with chitosan (CS). The chitosan-magnetic iron oxide nanoparticles (CS-MNPs) were characterized using Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), vibrating sample magnetometry (VSM), and the morphology of the particles was studied by transmission electron microscopy (TEM). Our findings show that the magnetic particles were monodisperse (10nm mean diameter) and exhibited superparamagnetic behavior. The interaction between the particles and calf-thymus DNA (DNA) in physiological buffer was studied with UV–vis, fluorescence and circular dichroism spectroscopy and zeta potential. Spectroscopic studies were indicated DNA conformational changes in the presence of CS-MNPs. Binding and thermodynamic parameters at different temperatures were calculated using the Stern–Volmer, Hill, Scatchard and Van’t Hoff equations. The binding process was spontaneous and interactions were electrostatic with the appropriate binding constant (Kb=4.52×103M−1, 3.69×103M−1 and 3.02×103M−1 at 300K, 310K and 320K, respectively). Zeta potential measurements of DNA continually increased with the addition of CS-MNPs, supporting our thermodynamic findings. Moreover, CS-MNPs were able to quench the fluorescence of DNA-intercalated ethidium bromide (DNA-EB) by a static quenching mechanism. Cytotoxicity studies show that the DNA-CS-MNP system is biocompatible with a human foreskin fibroblast cell line, HFFF2. Collectively, these results suggest that surface cationic magnetic chitosan-iron oxide nanoparticles can potentially enhance magnetofection efficiency.

Exogenous tumor necrosis factor-alpha could induce egress ofToxoplasma gondiifrom human foreskin fibroblast cells

Toxoplasma gondii is an intra-cellular protozoan parasite that can infect almost all nucleated cells, eliciting host immune responses against infection. Host tissue damage is mainly caused by cellular lysis when T. gondii egresses from infected cells. However, the effects of cytokines released by host immune cells on egression of T. gondii remain elusive. This study aimed to investigate the role of tumor necrosis factor-alpha (TNF-α) on the egress of T. gondii from infected human foreskin fibroblast (HFF) cells and to elucidate the underlying mechanisms that regulate TNF-α-induced egress. Using flow cytometry to count tachyzoites of T. gondii released into cell culture medium, we found that egress of T. gondii from infected HFF cells could be induced by 10 ng/mL TNF-α in a time-dependent manner. Pre-treatment of infected HFF cells with BAPTA-AM to chelate intra-parasitic calcium could greatly inhibit TNF-α-induced egress. Similar results were obtained when using cytochalasin D to block parasite motility before the TNF-α-induced egress assay. In addition, blocking host apoptosis by Z-VAD-FMK could decrease TNF-α induced egress, while blocking necroptosis by necrostatin-1 has little impact on TNF-α-induced egress. The egressed tachyzoites displayed a normal growth rate and lost no virulence. Our results suggest that host cytokines could influence the cellular lytic processes of T. gondii, providing new insights into the relationship between host TNF-α and T. gondii pathogenesis.

Flos Farfarae Inhibits Enterovirus 71-Induced Cell Injury by Preventing Viral Replication and Structural Protein Expression.

Enterovirus 71 (EV71) infection can cause airway symptoms, brainstem encephalitis, neurogenic shock, and neurogenic pulmonary edema with high morbidity and mortality. There is no proven therapeutic modality. Flos Farfarae is the dried flower bud of Tussilago farfara L. that has been used to manage airway illnesses for thousands of years. It has neuro-protective activity and has been used to manage neuro-inflammatory diseases. However, it is unknown whether Flos Farfarae has activity against EV71-induced neuropathy. The current study used both human foreskin fibroblast (CCFS-1/KMC) and human rhabdomyosarcoma (RD) cell lines to test the hypothesis that a hot water extract of Flos Farfarae could effectively inhibit EV71 infection. The authenticity of Flos Farfarae was confirmed by HPLC-UV fingerprint. Through plaque reduction assays and flow cytometry, Flos Farfarae was found to inhibit EV71 infection ([Formula: see text]). Inhibition of viral replication and protein expression were further confirmed by reverse transcription polymerase chain reaction (RT-PCR) and quantitative RT-PCR (qRT-PCR), and western blot, respectively. The estimated IC[Formula: see text]s were 106.3[Formula: see text][Formula: see text]g/mL in CCFS-1/KMC, and 15.0[Formula: see text][Formula: see text]g/mL in RD cells. Therefore, Flos Farfarae could be beneficial to inhibit EV71 infection by preventing viral replication and structural protein expression.

In vitro Cytotoxicity Comparison of MTA Fillapex, AH-26 and Apatite Root Canal Sealer at Different Setting Times.

Introduction:This study aimed to compare the cytotoxicity of MTA Fillapex, AH-26 and Apatite root canal sealers at different times after mixing. Methods and Materials:In this in vitro study, MTA Fillapex, AH-26 and Apatite root canal sealer were spilled uniformly by 40 µm mesh in a 96-well plate. Then, human fetal foreskin fibroblast cell line (HFFF2) were added to each sealer cell culture medium. Cytotoxicity was measured using MTT assay after 24, 48 and 72 h and seven days. Multiple comparisons were done using analysis of variances (ANOVA) and Scheffe’s post hoc test. Results:All studied sealers exhibited severe cytotoxicity (more than 70%) except for Apatite sealer (95%) at 48 h after mixing. Cytotoxicity of MTA Fillapex and AH-26 were similar (P>0.05) at 24, 48 and 72 h and 7 days after mixing of sealers. Cytotoxicity of MTA Fillapex and Apatite root canal sealer, at 24 and 48 h, were significantly different (P=0.003 and P=0.000, respectively); MTA Fillapex was more cytotoxic. However in 72 h and 7 days after mixing, the difference was not significant (P>0.05). At 24 and 48 h after mixing, AH-26 was more cytotoxic (P=0.002 and P=0.000, respectively). Same as above at 72 h and 7 days after mixing, their cytotoxicity were similar (P>0.05). Conclusion:Overall cytotoxicity of all studied materials were severe. However, it was observed that the cytotoxicity of MTA Fillapex, AH-26 and Apatite root canal sealer decreased over time. Apatite root canal sealer exhibited the least cytotoxicity. Cytotoxicity of MTA Fillapex and AH-26 were similar at different time intervals.

Evaluation of Cytotoxic and Genotoxic Effects of Euphorbia Triaculeata Forssk. Extract

Objective:To evaluate the cytotoxic and genotoxic activity of Euphorbia triaculeata Forssk. plant extract from Jazan region, Saudi Arabia, in an in vitro cancer model, which could be beneficial in anticancer therapy against human breast cancer cell line (MCF-7), prostate cell line (PC-3), human hepatocellular carcinoma cell line (HEPG2) and normal breast epithelial cell line (MCF-10A). The human foreskin fibroblast cell line, (Hs68), was also included in the cell panel. Doxorubicin and 5-Flurouracil, broad-spectrum anticancer drugs, were used as the positive control.Methods:Cytotoxicity of Euphorbia triaculeata plant extract was investigated by employing MTT assay and the genotoxicity was assessed by using comet assay.Results:Both toxicity tests exhibited significant toxicity results. In the comet assay, the Euphorbia triaculeata extract exhibited genotoxic effects against MCF-7 DNA and PC 3 but not on HEPG2 cell lines in a time-dependent manner by increasing the mean percentage of DNA damage. Euphorbia triaculeata extract showed significant toxicity against cancer cells. Comparison with positive control signifies that cytotoxicity exhibited by methanol extract might have moderate activity.Conclusion:The present work confirmed the cytotoxicity and genotoxicity of Euphorbia triaculeata plant. However, the observed toxicity of this plant extract needs to be confirmed by additional studies. Based on our results, further examination of the potential anticancer properties of Euphorbia triaculeata plant species and the identification of the active ingredients of these extracts is warranted.

Effect of clinically approved HDAC inhibitors on Plasmodium, Leishmania and Schistosoma parasite growth

Malaria, schistosomiasis and leishmaniases are among the most prevalent tropical parasitic diseases and each requires new innovative treatments. Targeting essential parasite pathways, such as those that regulate gene expression and cell cycle progression, is a key strategy for discovering new drug leads. In this study, four clinically approved anti-cancer drugs (Vorinostat, Belinostat, Panobinostat and Romidepsin) that target histone/lysine deacetylase enzymes were examined for in vitro activity against Plasmodium knowlesi, Schistosoma mansoni, Leishmania amazonensis and L. donovani parasites and two for in vivo activity in a mouse malaria model. All four compounds were potent inhibitors of P. knowlesi malaria parasites (IC50 9–370 nM), with belinostat, panobinostat and vorinostat having 8–45 fold selectivity for the parasite over human neonatal foreskin fibroblast (NFF) or human embryonic kidney (HEK 293) cells, while romidepsin was not selective. Each of the HDAC inhibitor drugs caused hyperacetylation of P. knowlesi histone H4. None of the drugs was active against Leishmania amastigote or promastigote parasites (IC50 > 20 μM) or S. mansoni schistosomula (IC50 > 10 μM), however romidepsin inhibited S. mansoni adult worm parings and egg production (IC50 ∼10 μM). Modest in vivo activity was observed in P. berghei infected mice dosed orally with vorinostat or panobinostat (25 mg/kg twice daily for four days), with a significant reduction in parasitemia observed on days 4–7 and 4–10 after infection (P < 0.05), respectively.

Synthesis and Antiviral Evaluation of Octadecyloxyethyl Benzyl 9-[(2-Phosphonomethoxy)ethyl]guanine (ODE-Bn-PMEG), a Potent Inhibitor of Transient HPV DNA Amplification.

Human papillomavirus (HPV) high-risk genotypes such as HPV-16 and HPV-18 cause the majority of anogenital tract carcinomas, including cervical cancer, the second most common malignancy in women worldwide. Currently there are no approved antiviral agents that reduce or eliminate HPV and reverse virus-associated pathology. We synthesized and evaluated several alkoxyalkyl acyclic nucleoside phosphonate diesters and identified octadecyloxyethyl benzyl 9-[(2-phosphonomethoxy)ethyl]guanine (ODE-Bn-PMEG) as an active compound which strongly inhibited transient amplification of HPV-11, -16, and -18 origin-containing plasmid DNA in transfected cells at concentrations well below its cytotoxic concentrations. ODE-Bn-PMEG demonstrated increased uptake in human foreskin fibroblast cells and was readily converted in vitro to the active antiviral metabolite, PMEG diphosphate. The P-chiral enantiomers of ODE-Bn-PMEG were obtained and appeared to have equivalent antiviral activities against HPV. ODE-Bn-PMEG is a promising candidate for the local treatment of HPV-16 and HPV-18 and other high-risk types, an important unmet medical need.