Cytotoxic potential of novel N-formyl pyrazolines derived from vanillin

Deficiency in apoptosis is one of the key factors that plays a pivotal role in cancer cell growth and proliferation. A procedure used in the treatment of cancer is the triggering of apoptosis in cancer cells. The current study aims to investigate the anticancer property of ethanolic extract of Amomum subulatum Roxb. against HeLa cell line. The MTT 3-(4, 5-di-methylthiazol-2-yl)-2,5-diphenyl-2H tetrazolium bromide assay is the quantitative cytotoxicity assay used, maximal inhibitory concentration (IC50) value was selected as the cytotoxicity parameter. The IC50 value of A. subulatum Roxb. extract is 62.5 μg/ mL and for 5-fluorouracil it is 7.8 μg/mL which indicates anticarcinogenic properties against HeLa cells. The apoptotic morphological changes in HeLa cells were observed using an inverted microscope and changes in HeLa cells were noticed after treatment with 62.5 mg/mL of ethanolic extracts, followed by acridine orange and ethidium bromide staining. The induction of apoptosis by extract of A. subulatum Roxb. was determined using the DNA fragmentation study. The results of the DNA fragmentation study, which exhibits prototypical intrinsic apoptotic characterization, also included fragmentation of nuclear DNA. We also found that the expression of Bcl2 and p53 mRNA were measured using RT-PCR. Overall, the current study results suggest that the

Antimicrobial, anticoagulant, and cytotoxic evaluation of multidrug resistance of new 1,4-dihydropyridine derivatives.

A new series of pyrano[2,3-d:6,5-d']dipyrimidine derivatives were synthesized and evaluated for their in vitro anticancer activity. The structures of all the synthesized compounds were confirmed by 1H NMR, 13C NMR, 15N NMR, HR-MS and FT-IR spectral analyses. The cytotoxic activities of these compounds against four human cancer (HeLa, SKBR-3, HepG2, and Caco-2) cell lines were determined. The synthesized compounds showed high selectivity, and four compounds (5e, 5f, 5g and 5i) showed excellent potent cytotoxicity against HeLa, SKBR-3, and HepG2 cancer cell lines. Furthermore, four other compounds (5a, 5c, 5b and 5d) have exhibited significant cytotoxicity activity in the SKBR-3 and HepG2 cell lines respectively, with moderate cytotoxicity seen in the HeLa cell line. Additionally, a molecular docking study was conducted to predict the anti-cancer behavior of the synthesized compounds via inhibition of the allosteric site of Human Kinesin Eg5.

A series of novel N-(5-benzyl-4-(tert-butyl) thiazol-2-yl)-2-(piperazin-1-yl)acetamides were designed, synthesized and evaluated for their antitumor activities in vitro. The structures of the synthesized compounds were characterized by 1H NMR, 13C NMR and elemental analysis. In general, compounds 5a, 5c and 6a showed potent antiproliferative activity against HeLa (human cervical cancer) and A549 (human lung cancer) cell lines. Compound 6a, with the best inhibitory activity against HeLa cells (IC50 = 1.6 ± 0.8 μM), was selected to investigate the induced changes of cell morphology in the HeLa cell line by means of acridine orange (AO)/ethidium bromide (EB) double staining and cell cycle analysis using flow cytometry. The results indicated that compound 6a could induce cell apoptosis and cause G1-phase arrest in the cell division cycle.

In the present work, a concise library of 1,3,5-triaryl-2-pyrazolines (2a–2q) was designed and synthesized by employing a multistep strategy, and the newly synthesized compounds were screened for their urease and α-glucosidase inhibitory activities. The compounds (2a–2q) were characterized using a combination of several spectroscopic techniques including FT-IR, 1H NMR, 13C NMR, and EI-MS. All the synthesized compounds, except compound 2i, were potent against urease as compared to the standard inhibitor thiourea (IC50 = 21.37 ± 0.26 μM). These analogs disclosed varying degrees of urease inhibitory activities ranging from 9.13 ± 0.25 to 18.42 ± 0.42 μM. Compounds 2b, 2g, 2m, and 2q having IC50 values of 9.36 ± 0.27, 9.13 ± 0.25, 9.18 ± 0.35, and 9.35 ± 0.35 μM, respectively, showed excellent inhibitory activity as compared to standard thiourea (IC50 = 21.37 ± 0.26 μM). A kinetic study of compound 2g revealed that compound 2g inhibited urease in a competitive mode. Among the synthesized pyrazolines, the compounds 2c, 2k, 2m, and 2o exhibited excellent α-glucosidase inhibitory activity with the lowest IC50 values of 212.52 ± 1.31, 237.26 ± 1.28, 138.35 ± 1.32, and 114.57 ± 1.35 μM, respectively, as compared to the standard acarbose (IC50 = 375.82 ± 1.76 μM). The compounds (2a–2q) showed α-glucosidase IC50 values in the range of 114.57 ± 1.35 to 462.94 ± 1.23 μM. Structure–activity relationship revealed that the size and electron-donating or -withdrawing effects of substituents influenced the activities, which led to the urease and α-glucosidase inhibiting properties. Compound 2m was a dual potent inhibitor against urease and α-glucosidase due to the presence of 2-CF3 electron-withdrawing functionality on the phenyl ring. To the best of our knowledge, these synthetic compounds were found to be the most potent dual inhibitors of urease and α-glucosidase with minimum IC50 values. The cytotoxicity of the compounds (2a–2q) was also investigated against human cell lines MCF-7 and HeLa. Compound 2l showed moderate cytotoxic activity against MCF-7 and HeLa cell lines. Moreover, in silico studies on most active compounds were also performed to understand the binding interaction of most active compounds with active sites of urease and α-glucosidase enzymes. Some compounds exhibited drug-like characteristics due to their lower cytotoxic and good ADME profiles.

Electrically regulated morphological and cytoskeletal changes of HeLa cells were studied on an optically transparent electrode (OTE), on which potential-applied surface cellular behavior and morphogenesis were easily observed. Upon application of a potential, HeLa cells in an OTE exhibited remarkable morphological changes above +0.5 V (vs. Ag/AgCl) and below 0 V. At each potential in this potential range, change in F-actin distribution was observed using a fluorescent probe (rhodamine phalloidin). These results suggest that an electrical field induces a subcellular cytoskeletal change. Electrostimulation of cells with OTE can be a valuable strategy for the manipulation of cultured human cells.

Investigation of potential antibiofilm properties of Antimicrobial Peptide (AMP) from Linckia laevigata against Candida albicans: An in vitro and in vivo study

Adenovirus was first recovered from tissue cultures of healthy human adenoidal tissue in 1953. The etiological roles of Adenovirus have been elucidated in many human diseases thereafter. On the way of these studies, characteristical features of this virus had attracted a great deal of attention of many investigators. Although many data on these characteristics have been accumulated, many problems still remain to be solved in the future.This paper reports the results of our experiment which tried to detect the localization of growth site of Adenovirus in HeLa cells by fractionation of the cells into nucleic and cytoplasmic ones. Three fractions obtained, nucleic, cytoplasmic and culture fluid, were titrated their infectivity and complement fixing-antigen successively in time. At the sane time, morphological changes of the infected cells were observed.Almost the same experiment was independently reported by Ginsberg, but he did not show growth curve of three cellular fractions timmsequentially.Experimental materials and method: Adenovirus types 1 (Ad, 71) and 3 (G, B) and HeLa cells were used as experimental materials. HeLa cells infected with Adenovirus type 1 or 3 were sequentially fractionated into nucleic and cytoplasmic fractions with treatment of 2.5% citric acid and centrifugation; of these fractions the infectivity and complement-fixing antigenicity were titrated. Paralleled to these procedures, infected HeLa cells, cultured on cover slips, were stained with hematoxylin and eosin after fixation with 95% methanol and observed the morphological changes under a light microscope.Summary:1) At the first time, influences of fractionating procedures upon infectivity, complement-fixing antigenicity of Adenovirus and upon HeLa cells themselves were tested. It was revealed that there was no influence of the procedures upon infectlvlty and complement-fixing antigenicity and that loss of number of HeLa cells by such procedures were under 10% of the starting number.2) Infectivity and comlement-fixing antigenicity in HeLa cells infected with Adenovirus type, 1 or 3 were always higher in cytoplasmic than in nucleic fractions. In all cases, infectivity appeared earlier than complement-fixing antigenicity in both cytoplasmic and nucleic fractions. In fluid phase, infectivity and complement-fixing antigenicity were detected later than intracellular ones, and increased after the time when intracellular antigenicities began to decrease.3) Morphological changes of HeLa cells infected with Adenovirus type 1 were mostly predominent in the nucleus. According to the sequential features of nucleus, they were classified into first, middle and late stages.

In this work, three N'-arylidene-5-phenyl-1H-pyrazole-3-carbohydrazide (3a-3c), derivatives have been synthesized and fully characterized by IR, 1H & 13C NMR and ESI-MS using experimental and theoretical methods. Theoretical calculations were carried out by DFT/B3LYP method using 6-311++G(d,p) basis set. For theoretical IR, 1H & 13C NMR (with GIAO method), MEP, HOMO-LUMO energies analyses of compounds 3a-3c were performed over the optimized structures. All newly synthesized compounds were screened for their cytotoxicity, proylenpeptidase and α-glucosidase inhibitory activities. Compounds 3c showed significant anticancer activity against lung cancer cell line (H460) with the IC50 value 0.15 ± 0.01 µM. The other compounds exhibited prolyl endopeptidase (PEP) activity. Compounds 3a and 3c have demonstrated a potent α-glucosidase inhibitory activity with the IC50 value 360.4 ± 0.7 µM and 370.3 ± 1.17 µM, respectively. Furthermore, in-silico based molecular docking study was performed between the title ligands and lung cancer cell line-H460 protein: 1M17. The best affinity bond was observed between compound 3c and the protein 1M17 with an energy of −8.6 (kcal/mol), an inhibition constant of 0.496769 μM and an active hydrogen bond to three.

Acylation/deacylation reactions represent a basic requirement of triglyceride as well as phospholipid metabolism, and maintenance of membrane lipid composition. In order to examine enzymes participating in these pathways, we synthesized 18-(4'-azido-2'-hydroxybenzoylamino)-oleic acid, an iodinable photoaffinity analogue of oleic acid as a new tool for analyzing enzymes, especially those binding unsaturated fatty acids or acyl-CoAs. For the synthesis of omega-amino-oleic acid, coupling two bifunctional Cg-components was used. The described synthesis scheme is also suited for the specific generation of other fatty acid analogues with distinct positions of the double bond. The functionality of 18-(4'-azido-2'-hydroxybenzoylamino)-oleic acid was investigated with the enzyme lysophosphatidylcholine:acyl-CoA-O-acyltransferase (LAT) [EC 2.3.1.23], an enzyme that shows high specificity towards (poly)unsaturated fatty acyl-CoAs. It could be shown that the photolabel, esterified with coenzyme A, acts in the dark as a reversible inhibitor of the enzyme activity, but photolysis of the label results in irreversible inactivation of LAT. This inactivation could be prevented by addition of the native substrate arachidonyl-CoA during photolysis. Several proteins could be specifically visualized using the iodinated analogue. The data indicate that this new photoaffinity label may have application to identify and characterize lipid biosynthetic enzymes using unsaturated fatty acids as well as acyl-CoA binding proteins and the active site of these proteins.

Effects of Res on proliferation and apoptosis of human cervical carcinoma cell lines C33A, SiHa and HeLa

Ginger root is one of the most widely used natural product and is excellent source for many kinds of bioactive compounds. One of them is dehydrozingerone, also very attractive biological active compound. This compound served as a starting material in the synthesis of new derivatives. The synthesized products were used in reaction with formic acid and hydrazine hydrate in order to obtain N-formyl derivative of pyrazoline. All new compounds were identified and well characterized by IR, 1H and 13C NMR spectroscopy and physical data. In vitro cytotoxic activity of pyrazolines against human cervical cancer (HeLa) was determined using MTT. Tested concentrations of substances were 100, 150 and 200 µM. Morphology changes of treated cells were visualized and compared to untreated cells using microscopy.

Three new acenaphthene derivatives cis-3-(4'-methoxyphenyl)-acenaphthene-1, 2-diol (1), trans-(1S, 2S)-3-phenyl-acenaphthene-1, 2-diol (2) and 8-(4-hydroxyphenyl)-2H-acenaphthylen-1-one (3) in company with six known compounds were isolated from the 70% ethanol extract of the rhizomes of Musa basjoo. Those chemical constituents were separated and purified by macroreticular resin, silica gel, Toyopearl HW-40F, SephadexLH-20 and other chromatographic methods, respectively. The chemical structures of new compounds were elucidated by HR-ESI-MS, 1H NMR, 13C NMR, HMQC, HMBC spectrum and specific optical rotations. Compound 4 was isolated for the first time from the genus Musa and compound 7 was firstly assigned the carbon spectrum. Furthermore, the cytotoxic activity of compounds 1–9 against WM9, MDA-MB231, HeLa, K562, DU145 and PC3 was screened with cisplatin as a positive control. Compound 9 showed promising cytotoxic activities with IC50 values of 2.65 ± 0.38 µM against the HeLa cell lines, while compound 8 possessed significant cytotoxicity with IC50 values of 6.51 ± 0.44, 18.54 ± 0.68 and 7.98 ± 1.44 µM against the HeLa, MDA-MB231 and WM9 cell lines, respectively.

Here we describe the function of a previously uncharacterized protein, named family with sequence similarity 60 member A (FAM60A) that maps to chromosome 12p11 in humans. We use quantitative proteomics to determine that the main biochemical partners of FAM60A are subunits of the Sin3 deacetylase complex and show that FAM60A resides in active HDAC complexes. In addition, we conduct gene expression pathway analysis and find that FAM60A regulates expression of genes that encode components of the TGF-beta signaling pathway. Moreover, our studies reveal that loss of FAM60A or another component of the Sin3 complex, SDS3, leads to a change in cell morphology and an increase in cell migration. These studies reveal the function of a previously uncharacterized protein and implicate the Sin3 complex in suppressing cell migration.

background: The antiproliferative studies of Titanium(II) complexes of composition TiCl2 (L)2 [L = 2,2'-bipyridine (bipy), 4,4'-dimethyl-2,2'-bipyridine (bpMe), 4,4'-dimethoxy-2,2'-bipyridine (bpoMe), 6,6'-dimethyl-2,2'-bipyridine (dpMe), adamantylamine (ada)] were done against three different HeLa (cervical), C6 (glioma) and CHO (Chinese hamster ovarian) cell lines. It has been observed that the complex with 6,6'-dimethyl-2,2'-bipyridine (dpme) ligand was more potent against HeLa cell lines with an IC50 value of 9.10 μM but less effective than the known anticancer drug Camptothecin (IC50= 6.2 μM). Furthermore, cell cycle analysis was accomplished on CHO cell lines to discover the cell death mechanism. It has been observed that all complexes induce cell death through an increase in hypo-diploid cells (Sub-G1 population), which indicates apoptosis in a dose-dependent manner. objective: Cytotoxic studies of synthesized titanium complexes on three different cell lines; MTT assay; cell cycle analysis. method: MTT assay: The cytotoxic activity of the synthesized complexes against the HeLa, C6, and CHO cell lines was determined using the colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. This assay was carried out in triplicate on 96 plates, with a well-defined procedure and minor modifications. result: Cell cycle analysis: Cells were plated at 1×106 cells/dish in 24-well plates and then grown either in the presence or absence of the complex at three different concentrations,i.e.below and above IC50 and at IC50 values. After 24 h of treatment, the cells were harvested from the dishes by collecting trypsinized cells together with floating cells in the medium. conclusion: In the present paper, we have reported the cytotoxic studies of previously reported titanium complexes against HeLa, C6, and CHO cell lines. The results have shown that titanium complex (D1) having 6,6'-dimethyl-2,2'-bipyridine ligand was more effective against the HeLa cell line. The morphological changes in CHO cells indicate characteristic features similar to apoptosis, and cell cycle analysis indicates an increase in hypo-diploid cells.