Human Laryngeal Epidermoid Carcinoma Research Articles

Facile synthesis of silver nanoparticles using Averrhoa bilimbi L and Plum extracts and investigation on the synergistic bioactivity using in vitro models

AbstractThe bacterial communities (Gram-negative and Gram-positive) form the biofilms which oppose the mode of action of antibiotics and affecting the immune system of the human. These chronic infections related to biofilm are always hard to be cured because of their inherent resistance to both antimicrobial agents and host defense. The present study is devoted to the synthesis of silver nanoparticles (AgNPs) using aqueous extracts ofAverrhoa bilimbileaf and Plum fruit (Prunus bokharensis) and its inhibitory effect onStaphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, andSalmonella typhi. Here, AgNPs) were successfully prepared by green synthesis method obtained by the reduction of silver nitrate into silver ions. The synthesized AgNPs were characterized by UV-Vis Spectroscopy, XRD, FTIR, FESEM-EDAX and DLS analysis. The antimicrobial effects of the two extract were evaluated by the disk diffusion method. The green synthesized AgNPs exhibited good antibacterial activity against both Gram-negative and Gram-positive. They were also analyzed for their significant antibacterial activities against all the test cultures exposed whereasEscherichia coliandSalmonella typhitopped among other test cultures. The anti-proliferative activity of phytochemical mediated synthesis of Ag NPs was investigated for their cytotoxicity in Vero and Human epidermoid larynx carcinoma cell lines (HEp-2) based on their viability using MTT assay. The present study also represents the synergistic bioactivity of silver nanoparticles using in vitro models.

Bio-fabrication of pigment-capped silver nanoparticles encountering antibiotic-resistant strains and their cytotoxic effect towards human epidermoid larynx carcinoma (HEp-2) cells.

Bacterial biomolecule-mediated nanoparticle (NP) synthesis constitutes a reliable, eco-friendly approach that ameliorates green-chemistry principles. In this study, stable silver nanoparticles were synthesized by exposing aqueous silver ions to an extracellular diffusible pigment produced by Pseudomonas aeruginosa (PA6) under optimized laboratory conditions. Spectroscopic and microscopic analyses showed the typical characteristics of silver with an average size of ∼28.30 nm and spherical shape. The particles were polydispersed and showed no definite agglomeration with a zeta potential of −32.3 mV, conferring stability. Antimicrobial studies were carried out using 5, 15, 25 and 50 μg mL−1 concentrations of pcAgNPs, which showed significant antibacterial activity toward clinically important pathogens at all concentrations compared to with the control sample. The bactericidal effect induced by pcAgNPs associated with cell damage was well demonstrated using electron microscopic studies. ROS production was measured using the DCFH-DA method and the oxidative stress was assessed by measuring the reduced glutathione (GSH) levels. Cytotoxicity studies on HEp-2 (Human Epidermoid Larynx Carcinoma) cells exposed to pcAgNPs showed dose-dependent cytotoxic effect with IC50 of 14.8 μg mL−1 compared to with IC50 of 7.38 μg mL−1 for the Vero cell control. Mechanistically, the pcAgNPs activated p53 that induced catalase, leading to apoptosis and DNA fragmentation via a p53 transcriptional pathway and electron transport arrest, which resulted in cell death. This synergistic efficacy of pigment-AgNPs demonstrated excellent antimicrobial and anti-proliferative activities, providing a potential lead for developing a broad-spectrum antibacterial agent and improving the therapeutic modalities targeting carcinoma cells at the gene level.

Mechanisms underlying the cytotoxic effect of propolis on human laryngeal epidermoid carcinoma cells

Propolis has been used as a traditional remedy for centuries because of its beneficial effects, including anticancer properties. The aim of this study was to compare the cytotoxic mechanism of Cuban red propolis (CP) and Brazilian green propolis (BP) on human laryngeal carcinoma (HEp-2) cells. Cell viability, leakage of lactate dehydrogenase, fluorescence staining, mitochondrial membrane potential (ΔΨm) and the expression of pro/anti-apoptotic genes were assessed. Cell viability and cytotoxic assays suggested a dose-dependent effect of CP and BP extracts with a possible association of intracellular reactive oxygen species production and decreased ΔΨm. Both samples induced apoptosis via activation of TP53, CASP3, BAX, P21 signalling, and downregulation of BCL2 and BCL-XL. CP exerted a higher cytotoxic effect than BP extract. Our findings suggest further investigation of the main components of each propolis sample, what may lead to the development of strategies for the treatment of laryngeal cancer.

Mechanisms involved in the cytotoxic action of Brazilian propolis and caffeic acid against HEp-2 cells and modulation of P-glycoprotein activity

The effects of propolis and phenolic compounds (caffeic acid - Caf; dihydrocinnamic acid - Cin; p-coumaric acid - Cou) in the same quantity found in our propolis sample were investigated on human laryngeal epidermoid carcinoma (HEp-2) cells. Cell viability, apoptosis/necrosis and cell cycle arrest, P53 and CASPASE-3 gene expression, generation of reactive oxygen species (ROS) and the ability of propolis to induce doxorubicin (DOX) efflux using a P-glycoprotein (P-gp) inhibitor (verapamil) were assayed. Propolis exerted a cytotoxic effect on HEp-2 cells, whereas isolated compounds had no effect on cell viability. Higher concentrations were tested and Caf induced late apoptosis or necrosis in HEp-2 cells, while propolis induced apoptosis, both probably due to ROS generation. P53 expression was downregulated by propolis but not by Caf. CASPASE-3 expression was correlated with induction of both early and late apoptosis, with both propolis and Caf alone upregulating its expression. Propolis induced cell cycle arrest at G2/M phase and Caf at S phase. Propolis but not Caf may act as a P-gp inhibitor by modulating P-gp activity and inhibiting DOX efflux. Propolis exerted cytotoxic effects on HEp-2 cells, and the mechanisms are discussed, showing its potential as an antitumour drug.

ANTIMICROBIAL AND ANTICANCER ACTIVITY OF SILVER NANOPARTICLES FROM EDIBLE MUSHROOM: A REVIEW

ABSTRACTBiologically inspired nanoparticle synthesis is currently a rapid expanding area of research in nanotechnology. Nanoparticle synthesis utilizing thebioresources such as plants and microbes appears to be a viable, low-cost, and eco-friendly approach. Especially mushrooms can be used for largescalesynthesis of silver nanoparticles as mushroom produces many proteins that reduce the silver nitrate during the biosynthesis. Silver nanoparticlescan be characterized using ultraviolet-visible (UV-VIS) spectroscopy, fourier transform infrared spectroscopy, X-ray diffraction, scanning electronmicroscopy, energy dispersive X-ray, and transmission electron microscope. Silver nanoparticles possess high antibacterial activity since silver indifferent forms has been extensively used as a medicine for curing diseases and promote wound healing. Silver nanoparticles have high surfacespecific area, which will lead to excellent antimicrobial activity as compared with bulk metallic silver. Further, the silver nanoparticles show anticanceractivity against various cell lines such as human epidermoid larynx carcinoma (HEP-2), colon adenocarcinoma (HCT-116), breast adenocarcinoma(MCF-7), liver carcinoma (Hep-G2), and intestinal adenocarcinoma (Caco2) were well documented. This review intends to present green synthesis ofsilver nanoparticles and their application as antimicrobial and anticancer agents.Keywords: Silver nanoparticles, Bioresources, Mushroom, Antimicrobial activity, Anticancer property.

Cytotoxicity comparison on different parts of three Digitalis species and isolation of potential bioactive compounds

In the Flora of Turkey, the genus Digitalis is represented by nine species [1]. Digitalis species contain biologically active compounds such as cardenolides, phenylethanoid glycosides, flavonoids, and anthraquinones. In this research three Digitalis species [Plantaginaceae]; D. davisiana Heywood, D. grandiflora Miller and D. viridiflora Lindley were studied in the point of cytotoxicity. To determine the bioactive part of the plant, root and aerial parts were extracted with methanol separately. Cytotoxic activity was performed by MTT method through 2 cancer cell lines HEp-2 (Human larynx epidermoid carcinoma), HepG2 (Human hepatocelular carcinoma) and a non-cancerous cell line 3Y1 (rat fibroblast cell) [2]. Methanol extracts of the aerial parts and the roots of three plants were used for cytotoxic activity tests. It was found that these three plants have different potentials for cytotoxicity. Aerial parts were found to be more active than the roots on tested cancer cell lines. In addition to this, D. davisiana aerial parts extract (DD-APE) was found to be the most active among all. IC50 values of DD-APE were 50.8, 211.4 and 94.7 µg/mL respectively for HEp-2, HepG2 and 3Y1 cell lines. The tested extracts are more cytotoxic on HEp-2 cell line than HepG2 and 3Y1 cell lines. This selectivity is important for the discovery of new anticancer agents. After evaluating the bioactivity test results, isolation studies started from active, phenylethanoid glycosides rich fractions. Repeated column chromatography studies resulted in the isolation of five phenylethanoid glycosides. Deacyllugrandoside (1), lugrandoside (2), isolugrandoside (3), maxoside (4) and 3,4-dihydroxy-β-phenylethoxy-O-[β-glucopyranosyl-(1→3)]-O-[α-rhamnopyranosyl-(1→6)]-4-O-(E)-feruloyl-β-glucopyranoside (5) were identified by extensive NMR techniques. Isolation and cytotoxic activity studies on isolated compounds are still in progress.

Combinatorial effects of geopropolis produced by Melipona fasciculata Smith with anticancer drugs against human laryngeal epidermoid carcinoma (HEp-2) cells.

The identification of natural products exerting a combined effect with therapeutic agents could be an alternative for cancer treatment, reducing the concentration of the drugs and side effects. Geopropolis (Geo) is produced by some stingless bees from a mixture of vegetable resins, gland secretions of the bees and soil. It has been used popularly as an antiseptic agent and to treat respiratory diseases and dermatosis. To determine whether Geo enhances the anticancer effect of carboplatin, methotrexate and doxorubicin (DOX), human laryngeal epidermoid carcinoma (HEp-2) cells were treated with Geo alone or in combination with each drug. Cell growth, cytotoxicity and apoptosis were evaluated using 3-(4,5-dimethyl thiazol-2yl)-2,5-diphenyltetrazolium bromide (MTT) assay, lactate dehydrogenase (LDH) release, and flow cytometry. Scratch assay was used to analyze cell migration and transmission electron microscopy to observe morphologic alterations. The influence of Geo on drug resistance was also investigated assessing P-glycoprotein (P-gp) action. Geo inhibited cell proliferation and migration. The combination Geo+DOX led to the highest cytotoxic activity and induced apoptosis, leading to loss of membrane integrity. Geo had no effect on P-gp-mediated efflux of DOX. Data indicate that Geo combined with DOX could be a potential clinical chemotherapeutic approach for laryngeal cancer treatment.

Synthesis and antitumor activity of some new pyrazolo[3,4-d]pyrimidine and pyrazolo[3,4-b]pyridine derivatives

A new series of pyrazolo[3,4-d]pyrimidine-3-carbonitrile and pyrazolo[3,4-b]pyridine-3-carbonitrile derivatives was synthesized by the reaction of 5-amino-1-tosyl-1H-pyrazole-3,4-dicarbonitrile as a key starting material with various electrophilic and nucleophilic reagents. All the newly synthesized compounds were structurally confirmed by various modern analytical methods (IR, 1H NMR and MS). All the title compounds have been evaluated for their potential cytotoxicity against human laryngeal epidermoid carcinoma cells (Hep2). Among the screened compounds, 3 and 4 exhibited the highest significant effect.

Characterization and cytotoxicity studies of thiol-modified polystyrene microbeads doped with [{Mo6X8}(NO3)6]2-(X = Cl, Br, I)

Halide octahedral molybdenum clusters [(Mo6X8)L6]n- possess luminescence properties that are highly promising for biological applications. These properties are rather dependent on the nature of both the inner ligands X (i.e. Cl, Br, or I) and the apical organic or inorganic ligands L. Herein, the luminescence properties and the toxicity of thiol-modified polystyrene microbeads (PS-SH) doped with [(Mo6X8)(NO3)6]2- (X=Cl, Br, I) were studied and evaluated using human epidermoid larynx carcinoma (Hep2) cell cultures. According to our data, the photoluminescence quantum yield of (Mo6I8)@PS-SH is significantly higher (0.04) than that of (Mo6Cl8)@PS-SH (6Br8)@PS-SH (6X8)@PS-SH showed that all three types of doped microbeads had no significant effect on the viability and proliferation of the cells.

Cellular internalisation, bioimaging and dark and photodynamic cytotoxicity of silica nanoparticles doped by {Mo6I8}4+ metal clusters.

Silica nanoparticles (SNPs) doped by hexanuclear molybdenum cluster complexes [{Mo6X8}L6]n (X = Cl, Br, or I; L = various inorganic or organic ligands) have been recently suggested as materials with high potential for biomedical applications due to both their outstanding photoluminescence properties and their ability to efficiently generate singlet oxygen upon photoirradiation. However, no studies were undertaken so far to prove this concept. Therefore, here we examined the potential of photoluminescent SNPs doped by {Mo6I8}4+ for applications such as bioimaging and photodynamic therapy using the human epidermoid larynx carcinoma (Hep-2) cell line as a model. Our results demonstrated both: (i) significant luminescence from cells with internalised molybdenum cluster-doped SNPs combined with the low cytotoxicity of particles in the darkness and (ii) significant cytotoxicity of the particles upon photoirradiation. Thus, this research provides strong experimental evidence for high potential of molybdenum-cluster-doped materials in biomedical applications such as optical bioimaging, biolabeling and photodynamic therapy.

Antitumor Activity of Extracts from Medicinal Basidiomycetes Mushrooms.

Aqueous extracts from the vegetative submerged mycelia of cultivated Basidiomycetes Ganoderma lucidum, Lentinus edodes, and Grifola frondosa, as well as from the fruiting bodies of G. lucidum, were found to have antitumor activity. The antitumor effect of the mycelial extracts from all 3 fungal species was ascertained in vivo in rats with implanted kidney cancer. Dystrophic changes in tumor cells and tumor necrosis (up to 90%) were noted. In vitro cytotoxicity studies of the A549 human lung adenocarcinoma cell line and HEp-2 human laryngeal epidermoid carcinoma cells showed that the extracts from the G. lucidum fruiting bodies and from the L. edodes vegetative mycelium were the most effective. The animals' immune systems were activated, and the fungal extracts displayed no toxicity when administered orally.

Biogenic synthesis, characterization of antibacterial silver nanoparticles and its cell cytotoxicity

The advanced research and development of silver nanoparticles (AgNPs) is vast due to their incredible applications today. In this work, AgNPs were synthesized using soil derived Pseudomonas putida MVP2. The AgNPs formation on the P. putida cell membrane and its cell free supernatant was studied. The synthesized AgNPs were characterized by UV–visible spectroscopy, scanning transmission electron microscopy (STEM), X-ray diffraction (XRD), energy dispersive X-ray (EDAX) and Fourier transform infrared (FTIR) spectrum analysis. The mode of action of AgNPs on the bacteria was studied against clinically isolated bacterial pathogens, Staphylococcus aureus, Escherichia coli, Bacillus cereus, Pseudomonas aeruginosa and Helicobacter pylori by membrane integrity, and protein leakage using confocal and electron microscopy. Interestingly, AgNPs had no cytotoxicity under 25μg/mL and it was toxic at above 50μg/mL on human epidermoid larynx carcinoma (HEp-2) cells. This study evidenced that biogenic nanoparticles could affect the bacterial replication, protein leakage and eventually cell death. This might be used for active antimicrobial agents for the chronic infections.

Ocular biocompatibility of polyquaternium 10 gel: functional and morphological results

This paper deals with the characterization study of topical and intraocular biocompatibility and toxicity of cationic hydroxyethylcellulose Polyquaternium 10 (PQ10). It also evaluates the rheological properties of gels. The cytotoxicity assays were done in two cell lines: HEp-2 and VERO (human larynx epidermoid carcinoma cell and African green monkey kidney cells respectively). For the in vivo study, New Zealand albino rabbits were used. The in vitro cytotoxic activity of PQ10 shows no statistically significant differences in relation to the control of hydroxypropylmethylcellulose (HPMC) in any of the cell lines used in this study. Similarly, the signs of inflammation observed after treatment showed no significant difference between the groups of animals treated with the polymer compared to the control group. Normal histological characteristics were seen in both groups with no histological inflammatory reaction. After 1 month of the intracameral application of 2% PQ10 (treatment group) or 0.3% HPMC (control group), electroretinograms showed similar levels of a- and b-waves latencies and amplitude. In summary, PQ10 gel was well tolerated in these experiments, with proper monitoring, it could stand as a new alternative in the development of ophthalmic viscosurgical devices.

The chemical composition and pharmacological activities of geopropolis produced by Melipona fasciculata Smith in northeast Brazil

Aim: Geopropolis is produced by stingless bees from resinous materials of plants, adding salivary secretions, wax, soil, and has been used in folk medicine for the treatment of respiratory diseases and dermatoses. Therefore, it is important to study its antibacterial, antitumor and immunomodulatory properties to confirm the ethnopharmacological applications. This work aimed to evaluate its chemical composition, and its antimicrobial, anti-tumoral and immunomodulatory activities. Methods: Geopropolis composition was investigated using GC-MS analysis. Antibacterial tests were performed to determine the minimum inhibitory concentration. HEp-2 cells viability was determined by the reduction of MTT, and cytokine production by human monocytes was determined by ELISA. Results: The major constituents of geopropolis compounds were carbohydrates and their derivatives, triterpenes, anacardic acid, alkylresorcinols, and sugar alcohols. Geopropolis alone showed no antibacterial activity against Staphylococcus aureus and Escherichia coli, but its combination with chloramphenicol exerted a greater action against S. aureus than chloramphenicol alone. Geopropolis exhibited a cytostatic action toward human laryngeal epidermoid carcinoma cells and stimulated tumor necrosis factor alpha and interleukin-10 production by human monocytes, showing an activator profile for human monocytes. Conclusion: The synergistic effect of geopropolis and chloramphenicol deserves further investigation due to bacterial resistance to antibiotics. Geopropolis also displayed antitumoral and immunomodulatory activity, and its biological properties may be due to triterpenes - one of its major chemical constituents.

Antiproliferative efficacy of Tabernaemontana divaricata against HEP2 cell line and Vero cell line.

Background:Laryngeal cancer may also be called cancer of the larynx or laryngeal carcinoma. Conventional plants are a precious source of novel anticancer agents and are still in performance better role in health concern. The study was intended to estimation of the anticancer activity of the chloroformic extract of Tabernaemontana divaricata on the human epidermoid larynx carcinoma cell line (Hep 2).Materials and Method:The aerial parts (leaves, stem, and flowers) of T. divaricata were tested for its inhibitory effect in 96 microplate formats against Hep 2 cell line. The anticancer activity of samples on Hep 2 and Vero was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and various enzymatic parameters like catalase, reduced glutathione (GSH), GSH peroxidase, and superoxide anion scavenging activity. Viable cells were determined by the absorbance at 540 nm. Measurements were performed, and the concentration required for a 50% inhibition of viability (IC50) was determined graphically. The effect of the samples on the proliferation of Hep 2 and Vero cells was expressed as the % cell viability.Results:The extract on Hep 2 cell line up to 7.8 μg/ml and that IC50 value on Hep 2 cell line was 112 μg whereas 94 μg for Vero cell line. Hence, T. divaricata has lesser significant action on Vero cell line.Conclusion:Medicinal plant drug discovery continues to provide new and important leads against various pharmacological targets including cancer. Our results clearly indicate the anticancer property of the medicinal plant T. divaricata against the human laryngeal carcinoma cell lines (Hep 2 cell line).

Green synthesis of silver nanoparticles using the mushroom fungus Schizophyllum commune and its biomedical applications

Silver nanoparticles, which are small metallic colloidal particles, ranging 1 ∼ 100 nm in size, have several nano biotechnological applications in medicine, manufacturing and engineering industries. Fungus-mediated synthesis of silver nanoparticles is an ecofriendly, green process. Further, extracellular enzymes and proteins elaborated by fungi are involved in the synthesis of the silver nanoparticle, which makes the downstream processing relatively simpler. In the present investigation, Schizophyllum commune, a mushroom fungus, was tested for its ability to synthesize extracellular as well as intracellular silver nanoparticles. When the fungus was challenged with 1 mM silver nitrate, a change in colour of the broth and the mycelium was observed, indicative of extracellular and intracellular synthesis of silver nanoparticles. The presence of silver nanoparticles was confirmed by studying its Surface Plasmon Resonance absorption band in the visible wavelength. FTIR spectrum analysis of the silver nanoparticles indicated the presence of biomolecules in association with the reduction of silver ions. Scanning Electron Microscopic analysis of the silver nanoparticles revealed the nanorange dimensions of both the extracellular and the intracellular silver nanoparticles. Analysis of biological activities of the silver nanoparticles disclosed their significant antibacterial activity against Escherichia coli, Bacillus subtilis, Klebsiella pneumoniae and Pseudomonas fluorescens. Additionally, the silver nanoparticles inhibited the growth of the dermatophytic fungal pathogens viz. Trichophyton simii, Trichophyton mentagrophytes and Trichophyton rubrum significantly. Anticancer activity of silver nanoparticles, assayed through MTT cytotoxicity assay, uncovered that 27.2 and 64% mortality could be obtained in Human Epidermoid Larynx Carcinoma (HEP -2) cell lines at concentrations between 10 and 100 µg/mL, respectively. The results obtained indicate that Schizophyllum commune is capable of synthesizing silver nanoparticles in shaken broth cultures (120 rpm) at 25 ± 2℃ and pH 7.

Low concentrations of trichosanthin induce apoptosis and cell cycle arrest via c-Jun N-terminal protein kinase/mitogen-activated protein kinase activation.

Trichosanthin (TCS) is a type I ribosome--inactivating protein, which inhibits cell viability in human epithelial type 2 (HEp-2) and AMC-HN-8 human laryngeal epidermoid carcinoma cells. Although TCS is a potential chemotherapeutic agent, its mechanism of action remains to be elucidated. In the present study, HEp-2 and AMC-HN-8 cells were treated with different concentrations of TCS combined with or without cisplatin. After 5 days of successive treatment, different experimental groups were detected using a cell counting kit-8 and the collected supernatants were analyzed using a lactate dehydrogenase kit. Flow cytometric assays were performed to detect apoptosis and cell cycle arrest in the HEp-2 and AMC-HN-8 cells, reverse transcription quantitative polymerase chain reaction was performed to detect the levels of p27, p21WAF and western blot analysis was performed to detect changes in c-Jun N-terminal protein kinase (JNK)/phosphorylated (phospho)-JNK, p38/phospho-p38, extracellular signal-regulated kinase (ERK)/phospho-ERK, caspase-3 and caspase-9 in the HEp-2 and AMC-HN-8 cancer cells. TCS significantly inhibited the cell viability of the HEp-2 and AMC-HN-8 cells, independently of necrosis. TCS induced apoptosis and increased the percentage of HEp-2 and AMC-HN-8 cells in the S-phase of the cell cycle. In addition, the JNK/mitogen-activated protein kinase (MAPK) pathway was activated by TCS in the HEp-2 and AMC-HN-8 cells. Low concentrations of TCS also induced apoptosis and S-phase cell cycle arrest in the HEp-2 and AMC-HN-8 cells. The antitumor effects of TCS may be associated with JNK/MAPK activation.

Phytochemical study, cytotoxic, analgesic, antipyretic and anti-inflammatory activities of Strychnos nux-vomica

The strychnine tree (Strychnos nux-vomica L.) (S. nux-vomica) belonging to family Loganiaceae has been a very promising medication for certain disorders. Different chromatographic methods were used to isolate the phenolic compounds from the aqueous methanolic extract of the S. nux-vomica leaves. Their identification was achieved through spectroscopic techniques. Cytotoxicity, analgesic, antipyretic and anti-inflammatory activities of S. nux-vomica leaves extract were evaluated. Five phenolic compounds were isolated and identified; Kaempferol-7 glucoside 1, 7-Hydroxy coumarin 2, Quercetin-3-rhamnoside 3, Kaempferol 3-rutinoside 4, and Rutin 5. Furthermore, the cytotoxic activity of the extract was evaluated against different cancer cell lines. The extract showed potential cytotoxic activity against human epidermoid larynx carcinoma cells (Hep-2) and against breast carcinoma cell line (MCF-7). Colon carcinoma cells (HCT) were the least one affected by the extract. In addition, the extract exhibited promising analgesic, antipyretic as well as anti-inflammatory activities. It is concluded that, leaves extract of S. nux vomica possess potent cytotoxic, analgesic, antipyretic and anti-inflammatory activities. These activities could be due to the presence of phenolic compounds revealed by our phytochemical investigations.

Evaluation of antimicrobial and anticancer activity of Rosemary Rosmarinus oficinalis leaves extracts on pathogenic bacteria and cancer cell lines

The present study was carried out to detect the active constitutions extracted from Rosmarinus oficinalis leaves by 70% methanol, the dried weight of aqueous extract (F1) were 12%(w:w) contain polyphenols represented by phenols, flavonoids and coumarins as a major compounds , while the dried weight of organic extract, [chloroform(F2)] was 2.8%(w:w) represented as terpenoids, tannins and coumarins detected by qualitative chemical analysis. Antimicrobial activity of Polyphenols and terpenoids were tested against four pathogenic bacteria isolates including: Escherichia coli; Staphylococcus aureus, Pseudomonas aeruginosa and Bacillus cereus ,which grown on Mueller Hinton agar (MHA) plates and then treated with different concentrations of rosemary extract (F1) and (F2) ranged from 0.15-1.2mg/ml, the results revealed that the higher inhibition zone obtained by F1 were 35mm in S. aureus, 24mm in B. cereus and lowest inhibition were 16mm in E. coli and P. aeruginosa at concentration 1.2mg/ml, and the inhibition were depends on extract concentrations. The MIC value of F1 was 600 µg/ml against S. aureus, 1200 µg/ml against B. cereus and 2400 µg/ml against E. coli and P. aeruginosa, while the MICS values of F2 were 2400 µg/ml against S. aureus and P. aeruginosa, and 4800 µg/ml against E. coli and B. cereus. Furthermore calculating index fractional inhibitory concentration (∑FIC) of rosemary extracts (F1 or/and F2), the ∑FICA ranged from synergistic to additional on E. coli and P. aeruginosa respectively, antagonistic on S. aueras and B. cereus, while the ∑FICB index showed synergistic effects on all tested organisms except the additional effect on P. aeurginosa on the basis of resultant zone of inhibition. The anticancer effects of (F1) were used on three cell lines, human larynx epidermoid carcinoma (Hep-2), mammary adenocarcinoma (AMN-3), rhabdomyosarcoma (RD) and one normal cell line, Rat embryo fibroblast (REF) at different concentrations ranged from 62.5-1000 µg/ml. The highest inhibition rate (IR) was 82.41%, 85% on RD, 52.62%, 75.48% on Hep2, 39.14% and 49.75% on AMN-3 at concentrations 500 and 1000µg/ml after 72h exposure time respectively, while IR of Ref cell line exhibit low inhibition rate ranging from 5.8 to 15.7% after 72h in the same conditions.

In Vitro Effects of Ivermectin and Sulphadiazine on Toxoplasma gondii.

Ivermectin and sulphadiazine were tested individually to determine their in vitro effects on Toxoplasma gondii grown in human epidermoid larynx carcinoma (Hep-2) cell culture. In-vitro study. Toxoplasma growth was quantities by an enzyme immunoassay performed directly on the fixed cultures, using a rabbit anti-T. gondii immunoglobulin G as the first antibody and a phosphatase-labeled anti-rabbit immunoglobulin G as the second antibody. For each drug, regression models were used to quantify the relationship between optical density values and antimicrobial agent concentrations in the cultures. The 50% inhibitory concentrations (IC50) of ivermectin and sulphadiazine were found to be 0.2 μg/mL and 7.3 μg/mL after 48 h of exposure, respectively. None of the concentrations tested for each drugs demonstrated toxicity to Hep-2 cells after 72 h of incubation. These results indicate that ivermectin significantly inhibited replication of the tachyzoites of T. gondii RH strain.