Cancer Cell Lines

The role of brown adipose tissue (BAT) in human metabolism and its potential as an antiobesity target organ have recently received much renewed attention. Following radiological detection of substantial amounts of BAT in adults by several independent research groups, an increasing number of studies are now dedicated to uncover BAT's genetic, developmental, and environmental determinants. In contrast to murine BAT, human BAT is not present as a single major fat depot in a well-defined location. The distribution of BAT in several areas in the body significantly limits its availability to research. A human brown adipocyte cell line is therefore critical in broadening the options available to researchers in the field. The human BAT-cell line PAZ6 was created to address such a need and has been well characterized by several research groups around the world. In the present review, we discuss their findings and propose potential applications of the PAZ6 cells in addressing the relevant questions in the BAT field, namely for future use in therapeutic applications.

The chemical composition of essential oils of six Stachys species, S. cretica L. ssp. vacillans Rech. fil., S. germanica L., S. hydrophila Boiss., S. nivea Labill., S. palustris L. and S. spinosa L., obtained by hydrodistillation, was studied by GC and GC–MS. All the oils have in common a great percentage of fatty acids and esters (24.2–58.5%) and a high

Two new compounds, a cyclic peptide desmocyclopeptide (1) and a special flavone desmorostratone were isolated from the stem bark of Desmos rostrata, along with two known compounds, desmosdumotins B (3) and C (4). Their structures were established on the basis of the spectral data, including mass spectrometry and 2D NMR. The total synthesis of desmorostratone (2) was performed in order to confirm its structure as well as that of desmosdumotin C (4), which was a tautomeric mixture in the solution. Finally, cytotoxity of these compounds were evaluated. Desmosdumotin C (4) displayed a moderate inhibition activity against KB cell line with an IC 50 of 19.2 mM, whereas the other products showed a weak inhibition against the same cell line target. Tetrahedron j o u r n a l h o m e p a g e : w w w . e l s e v i e r. c om / l oc a t e / t e t molecular formula itself and from elucidation of its NMR spectra. Analysis of the 1 H and 13 C NMR spectral data for 1, with the aid of 1 H-1 H-COSY, HSQC, HMBC and NOESY spectra, indicated the presence of an alanine, a glycine, a proline, an isoleucine and a phenylalanine ). The determination of the peptide sequence was established from the connectivity of the carbonyl of residue (i) and the NH proton of residue (iþ1) on the HMBC spectrum as follows: the correlations of the carbonyl at d C 173.5 of the Ala 1 with its Ha proton at d H 4.37, as well as to the NH proton of the Gly 2 at d H 7.78 were observed. The carbonyl group of the Gly 2 at d C 168.5 was not correlated to an amide proton, suggesting its connection with the Pro 3 . Cross-peaks of the carbonyl of the Pro 3 at d C 172.1 with its Ha proton at d H 4.21 and NH proton (d H 7.12) of the Ile 4 were presented. In addition, the carbonyl of Ile 4 at d C 172.3 was correlated with its Ha proton at d H 4.18 and NH proton of Phe 5 at d H 7.56. Finally, 2 J-HMBC correlation of the carbonyl of Phe 5 at d C 172.3 with its Ha proton (d H 4.64) and NH proton of Ala 1 (d H 8.25) were noted. Complete analysis of the HMBC spectrum determined the structure of 1 as drawn in . The NOESY spectrum of 1 depicted the strong spatial interactions of Ha of Gly 2 with Hd of Pro 3 , indicating that the amide bond between Gly 2 -Pro 3 was trans. 4 The d NN(i,iþ1) and d aN(i,iþ1) sequential connectivity from Ala 1 to Phe 5 was clearly observed in the NOESY spectrum as indicated in . The amino acids residue and the sequence of 1 were confirmed by fragmentation of [MþH] þ ion at m/z 486. A series of adjacent a n ion peaks were observed at m/z 311, 240, 183 and 86 corresponding to the successive loss of Phe, Ala, Gly and Pro. A second series of peaks at m/z 339, 268, 211 and 114 were assigned to adjacent b n ion (loss of Phe, Ala, Gly and Pro). This new pentacyclopeptide 1 is named desmocyclopeptide. The structure of 1 is very close to that of pseudostellarin A which has been previously reported from the roots of Pseudostellaria heterophylla. 5

The identification of signaling pathways that are involved in gliomagenesis is crucial for targeted therapy design. In this study we assessed the biological and therapeutic effect of ingenol-3-dodecanoate (IngC) on glioma. IngC exhibited dose-time-dependent cytotoxic effects on large panel of glioma cell lines (adult, pediatric cancer cells, and primary cultures), as well as, effectively reduced colonies formation. Nevertheless, it was not been able to attenuate cell migration, invasion, and promote apoptotic effects when administered alone. IngC exposure promoted S-phase arrest associated with p21CIP/WAF1 overexpression and regulated a broad range of signaling effectors related to survival and cell cycle regulation. Moreover, IngC led glioma cells to autophagy by LC3B-II accumulation and exhibited increased cytotoxic sensitivity when combined to a specific autophagic inhibitor, bafilomycin A1. In comparison with temozolomide, IngC showed a mean increase of 106-fold in efficacy, wit...

Skip Navigation Links Home > February 1960 - Volume 239 - Issue 2 > Cell and Tissue Culture. ... BOOK REVIEWS AND NOTICES: PDF Only. Cell and Tissue Culture. Paul, John MB, Ch.B., Ph.D., MRCP Ed. Collapse Box Abstract. An abstract is unavailable. ...

Three canine cell lines, K1, K6 and DH82, derived from canine malignant neoplasms, were characterised. They were examined for expression of surface antigens, cytokines, neuropeptide receptors, matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs). The growth characteristics of the cell lines were established and bioassays used to detect production of TNF-a, IL-1 and IL-6. In the DH82 cell line, production of TNF-a and IL-6 was readily detected. Neither K1 or K6 cell lines produced any measurable amounts of TNF-a, IL-1 or IL-6. At a molecular level, using reverse transcription-polymerase chain reaction (RT-PCR) to detect speci®c mRNA, the DH82 cell line expressed TNF-a, IL-1 and IL-6, whereas the K1 and K6 cell lines expressed TNF-a. Canine IL-5, IL-8 and IL-10 mRNA were detected in the DH82 cell line but only IL-5 and IL-8 mRNA were detected in the K1 and K6 cell lines. Gelatin zymography was used for the detection of MMP-2 and MMP-9 and all three cell lines produced MMP-2 but only the DH82 cell line produced MMP-9. Reverse zymography was used to detect TIMP-1 and TIMP-2 and all three cell lines produced both proteins. The presence of these MMPs and TIMPs was con®rmed at a molecular level using RT-PCR. Canine MMP-14 mRNA was detected in all three cell lines. For this investigation several genes for canine in¯ammatory molecules were cloned and sequenced for molecular detection; these included IL-1, IL-6, IL-8, TNF-a, MMP-9, MMP-14, TIMP-1, TIMP-2 and b-actin. Of all the cell surface antigens tested, only CD14 was expressed on the DH82 cell line although CD5 and CD45 was partially expressed. The K1 and K6 cell lines were negative for all of the CD markers tested. K1 and K6 were negative for Neurokinin 1 receptor (NK1-R) but positive for Calcitonin gene related peptide receptor type 1 (CGRP-1R) and Calcitonin gene related peptide Veterinary Immunology and Immunopathology 75 (2000) 9±25 : S 0 1 6 5 -2 4 2 7 ( 0 0 ) 0 0 1 8 4 -7 receptor component protein (CGRP-RCP). The DH82 cell line expressed neither NK1-R or CGRP-1R; however, it did express CGRP-RCP. Generally the DH82 cell line exhibited considerable similarity to canine monocytes, but all three cell lines will be useful as standards and for the puri®cation of various immunological and in¯ammatory mediators in the dog. #

The aim of this study was to investigate the adhesive phenotype of the human intestinal isolate Bifidobacterium bifidum MIMBb75 to human colon carcinoma cell lines. We have previously shown that the adhesion of this strain to Caco-2 cells is mediated by an abundant surface lipoprotein named BopA. In this study, we found that this strain adheres to Caco-2 and HT-29 cells, and that its adhesion strongly depends on the environmental conditions, including the presence of sugars and bile salts and the pH. Considerably more adhesion to a Caco-2 monolayer occurred in the presence of fucose and mannose and less when MIMBb75 grew in Oxgall bile salts compared to standard environmental conditions. In particular, growth in Oxgall bile salts reduced the adhesion ability of MIMBb75 and modified the SDS-PAGE profile of the cell wall associated proteins of the strain. The pH markedly affected both adhesion to Caco-2 and bacterial autoaggregation. Finally, experiments with sodium metaperiodate suggested that not only proteinaceous determinants are involved in the adhesion process of B. bifidum. In conclusion, it seems that the colonization strategy of this bacterium can be influenced by factors varying along the gastrointestinal tract, such as the presence of specific sugars and bile salts and the pH, possibly limiting the adhesion of B. bifidum to only restricted distal sites of the gut.

Vero cells growth and rabies production in IPT-AF medium, a property animal-component-free medium are described in this work. Kinetics of cell growth and rabies virus (strain LP 2061) production were first conducted in spinner flasks. Over eight independent experiments, Vero cell growth in IPT-AF medium, on 2 g/l Cytodex 1 was consistent. An average Cd (cell division number) of 3.3+/-0.4 and a specific growth rate micro of 0.017+/-0.006 h(-1) were achieved. Such performances were comparable to those obtained in serum-containing medium (MEM+10% FCS). Rabies virus production on Vero cells in IPT-AF medium was also optimised in spinner flasks. The effects of multiplicity of infection (MOI), regulation of glucose level at 1 g/l and cell washing step, were investigated. The highest virus titer was achieved when the cells were infected at an MOI of 0.1; this level was equal to 10(7) FFU/ml. The step of medium exchange before cell infection can be omitted; nevertheless in this case glucose...

Adansonia digitata leaves, fruit-pulp and seeds were acquired and extracted with three different solvents, water, methanol and DMSO. The extracts were compared quantitatively for antiviral MIC 100 (minimal inhibitory concentration) values against influenza virus, herpes simplex virus and respiratory syncytial virus and for their effects on cytokine secretion (IL-6 and IL-8) in human epithelial cell cultures. The leaf extracts had the most potent antiviral properties, especially the DMSO extracts and influenza virus was the most susceptible virus. Pulp and seed extracts were less active but significant. Cytotoxic activities were only evident at much higher concentrations of extract. Several of the extracts, especially leaf extracts, were also active as cytokine modulators, some being pro-inflammatory and others being anti-inflammatory. The results overall indicated the presence of multiple bioactive compounds in different parts of the plant and these activities could explain some of the medical benefits attributed to traditional leaf and pulp preparations, in the treatment of infectious diseases and inflammatory conditions.

In the present research, synthesis and characterization of novel heterocyclic compounds substituted chitosan moieties was achieved. Bis Schiff base [1] was utilized as a commencing material for the synthesis method. Schiff base[1] was achieved from the reaction of two mole of 2-amino-5-mercapto-1,3-thiadiazole with one mole of terephthalaldehyde in ethanol absolute. Also, the reaction of compound [1] with chloroacetyl chloride, thioglycolic acid, sodium azide or various anhydrides to give azetidinone [2], thiazolidinone [3], tetrazole [4], and 1,3-oxazepine derivatives [5-7] respectively, then compounds[2-7] were reacted with Na 2 CO 3 of distilled H 2 O, then chloroacetic acid was added to yield [8-13]. Also, compounds [8-13] reacted with thionyl chloride in benzene to procure [14-19]. O-chitosan derivatives[20-25] were synthesized by reaction of chitosan with compounds [8-13] in acidic media in distilled water according to the steps of Fischer and Speier. O,N-Chitosan derivatives [26-31] had been synthesized by reaction of chitosan with compounds [14-19] in trichloromethane and pyridine. The synthesized compounds were identified via spectral analysis techniques, including FT-IR, 1 H-NMR, UV-Vis Spectroscoy and the elemental analysis (C.H.N-S). Insertion of heterocyclic units to chitosan molecule were expected to improve the selectivity in the potent biological activities, therefore the toxicity study and anticancer activity, were examined for some organic compounds and chitosan derivatives.

In recent years, nanoparticles have been increasingly used in several industrial, consumer and medical applications because of their unique physico-chemical properties. However, in vitro and in vivo studies have demonstrated that these properties are also closely associated with detrimental health effects. There is a serious lack of information on the potential nanoparticle hazard to human health, particularly on their possible toxic effects on the endocrine system. This topic is of primary importance since the disruption of endocrine functions is associated with severe adverse effects on human health. Consequently, in order to gather information on the hazardous effects of nanoparticles on endocrine organs, we reviewed the data available in the literature regarding the endocrine effects of in vitro and in vivo exposure to different types of nanoparticles. Our aim was to understand the potential endocrine disrupting risks posed by nanoparticles, to assess their underlying mechanisms of action and identify areas in which further investigation is needed in order to obtain a deeper understanding of the role of nanoparticles as endocrine disruptors. Current data support the notion that different types of nanoparticles are capable of altering the normal and physiological activity of the endocrine system. However, a critical evaluation of these findings suggests the need to interpret these results with caution since information on potential endocrine interactions and the toxicity of nanoparticles is quite limited.

The pituitary gland is an important component of the endocrine system, and together with the hypothalamus, exerts considerable influence over the functions of other endocrine glands. The hypothalamus either positively or negatively regulates hormonal productions in the pituitary through its release of various trophic hormones which act on specific cell types in the pituitary to secrete a variety of pituitary hormones that are important for growth and development, metabolism, reproductive and nervous system functions. The pituitary is divided into three sections-the anterior lobe which constitute the majority of the pituitary mass and is composed primarily of five hormone-producing cell types (thyrotropes, lactotropes, corticotropes, somatotropes and gonadotropes) each secreting thyrotropin, prolactin, ACTH, growth hormone and gonadotropins (FSH and LH) respectively. There is also a sixth cell type in the anterior lobe-the non-endocrine, agranular, folliculostellate cells. The intermediate lobe produces melanocyte-stimulating hormone and endorphins, whereas the posterior lobe secretes anti-diuretic hormone (vasopressin) and oxytocin. Representative cell lines of all the six cell types of the anterior pituitary have been established and have provided valuable information on genealogy of the various cell lineages, endocrine feedback control of hormone synthesis and secretions, intrapituitary interactions between the various cell types, as well as the role of specific transcription factors that determine each differentiated cell phenotype. In this review, we will discuss the morphology and function of the cell types that make up the anterior pituitary, and the characteristics of the various functional anterior pituitary cell systems that have been established to be representative of each anterior pituitary cell lineage. Crown

This paper details the derivation of a list of 60 reference chemicals for the development of alternatives to animal testing in ecotoxicology with a particular focus on fish. The chemicals were selected as a prerequisite to gather mechanistic information on the performance of alternative testing systems, namely vertebrate cell lines and fish embryos, in comparison to the fish acute lethality test. To avoid the need for additional experiments with fish, the U.S. EPA fathead minnow database was consulted as reference for whole organism responses. This database was compared to the Halle Registry of Cytotoxicity and a collation of data by the German EPA (UBA) on acute toxicity data derived from zebrafish embryos. Chemicals that were present in the fathead minnow database and in at least one of the other two databases were subject to selection. Criteria included the coverage of a wide range of toxicity and physico-chemical parameters as well as the determination of outliers of the in vivo/in vitro correlations. While the reference list of chemicals now guides our research for improving cell line and fish embryo assays to make them widely applicable, the list could be of benefit to search for alternatives in ecotoxicology in general. One example would be the use of this list to validate structure-activity prediction models, which in turn would benefit from a continuous extension of this list with regard to physico-chemical and toxicological data.

The use of animal cell cultures as tools for studying the microsporidia of insects and mammals is briefly reviewed, along with an in depth review of the literature on using fish cell cultures to study the microsporidia of fish. Fish cell cultures have been used less often but have had some success. Very short-term primary cultures have been used to show how microsporidia spores can modulate the activities of phagocytes. The most successful microsporidia/fish cell culture system has been relatively long-term primary cultures of salmonid leukocytes for culturing Nucleospora salmonis. Surprisingly, this system can also support the development of Enterocytozoon bienusi, which is of mammalian origin. Some modest success has been achieved in growing Pseudoloma neurophilia on several different fish cell lines. The eel cell line, EP-1, appears to be the only published example of any fish cell line being permanently infected with microsporidia, in this case Heterosporis anguillarum. These ce...

The development of immortalized cell lines has been a significant benefit to the study of human disease, due to limitations in using primary cells and the availability of tissue. The immortalization of cells from cystic fibrosis (CF) patients as well as cells from non-CF individuals from tissues relevant to CF has been critical to enhancing our understanding of the physiological, biochemical and genetic mechanisms underlying CF and for the development of therapeutic strategies designed to manage CF pathology. A comprehensive list of immortalized cells from various tissue and species, with an emphasis on epithelial cells, is presented and discussed here.

A chitin-synthesizing cockroach cell line (UMBGE-4) previously shown to secrete ecdysteroids was analyzed for its ability to metabolize potential precursors of ecdysone (e.g., 2-deoxyecdysonet 2,22-dideoxyecdysone, 2,22,25trideoxyecdysone, and cholesterol). All, except cholesterol, were actively metabolized by UMBGE-4 cells. However, all but 2-deoxyecdysone were converted to polar and hydrolyzable metabolites, and not to ecdysone. Labeling with cholesterol was unsuccessful. Labeling experiments with molting hormones, i.e., ecdysone and 20-hydroxyecdysone, confirmed that this cell line can metabolize ecdysteroids and allowed identification of some of the products. Molting hormones were converted into acetate conjugates and polar conjugates which were often double-conjugates, i.e., polar conjugates of acetate conjugates. Labeling experiments with ecdysone demonstrated that this cell line possesses a low ecdysone 20-hydroxylase activity. The capacity of UMBGE-2 cells, which do not synthesize chitin or ecdysteroids, was also examined. Neither ecdysone nor 20-hydroxyecdysone was significantly metabolized by UMBCE-2 cells. 2-Deoxyecdysone and 2,22-dideoxyecdysone were very slowly metabolized respectively to more polar compounds.

Diabetes mellitus is a metabolic disorder and the management is an important criterion for pharmacotherapy. The medicinal plants play very important role in preventing the progress of the disease. Present study deals with screening of polyherbal extracts using in-vitro techniques for its antidiabetic activity. The plants used in the study are Eugenia jambolana and Cinnamomum zeylenicum. The polyherbal extracts were subjected to in-vitro evaluation using cell culture assays, α-glucosidase inhibition and antioxidant property. The cell lines 3T3L1 C2C12 and HepG2 were used to measure the uptake of glucose. The results showed that the glucose uptake was more in APKJ-004 polyherbal extract when compared with other extracts. α-glucosidase inhibitory activity was found in all the extracts and they were comparable to the standard acarbose and the highest inhibitory activity was found in the polyherbal extract APKJ004 (IC 50 147.2µg/ml). In-vitro antioxidant potential revealed that the extract APKJ004 is more potent and is comparable to the standard. Based on in-vitro evaluations, unique combination of glucose uptake activity and α glucosidase inhibition potential of extract APKJ-004 with prominent hypoglycemic activity will aid in the treatment of diabetes mellitus.

Labeling nonquiescent cells with carboxyfluorescein succinimidyl ester (CFSE)-like dyes gives rise to a population width exceeding the threshold for resolving division peaks by flow cytometry. Width is a function of biological heterogeneity plus extrinsic and intrinsic error sources associated with the measurement process. Optimal cytometer performance minimizes extrinsic error, but reducing intrinsic error to the point of facilitating peak resolution requires careful fluorochrome selection and fluorescent cell sorting. In this study, we labeled the Jurkat and A549 cell lines with CFSE, CellTrace-Violet (CTV), and eFluor 670 proliferation dye (EPD) to test if we could resolve division peaks in culture after reducing the labeled input widths by cell sorting. Reanalysis of the sorted populations to ascertain the level of reduction achieved always led to widths exceeding the gated limits due to the contribution of errors. Measuring detector-specific extrinsic error by sorting uniform fluorescent particles with similar spectral properties to the tracking dyes allowed us to determine the intrinsic error for each dye and cell type using a simple mathematical approach. We found that cell intrinsic error ultimately dictated whether we could resolve division peaks, and that as this increased, the required sort gate width to resolve any division peaks decreased to the point whereby issues with yield made A549 unsuitable for this approach. Finally, attempts to improve yields by setting two concurrent sort gates on the fluorescence distribution enriched for cells in different stages of the cell cycle that had nonequivalent proliferative properties in culture and thus should be practiced with caution. V C 2013 International Society for Advancement of Cytometry

The synthesis of the adamantane phenylalkylamines 2a−d, 3a−c, and 4a−e is described. These compounds exhibited significant antiproliferative activity, in vitro, against eight cancer cell lines tested. The σ 1 , σ 2 , and sodium channel binding affinities of compounds 2a, 3a, 4a, and 4c−e were investigated. The most interesting analogue, 4a, exhibited significant in vivo anticancer profile on pancreas, prostate, leukemia, and ovarian cancer cell line xenografts together with apoptosis and caspase-3 activation. Inhibition of the cancer cells cycle at the sub-G1 level was also obtained with 4a. Finally, encouraging results were observed with 4a in vivo on mice, suggesting putative antimetastatic and analgesic activities of this compound.

Jatropha curcas L., a multipurpose shrub has acquired significant economic importance for its seed oil which can be converted to biodiesel, is emerging as an alternative to petro-diesel. The deoiled seed cake remains after oil extraction is toxic and cannot be used as a feed despite having best nutritional contents. No quantitative and qualitative differences were observed between toxic and non-toxic varieties of J. curcas except for phorbol esters content. Development of molecular marker will enable to differentiate non-toxic from toxic variety in a mixed population and also help in improvement of the species through marker assisted breeding programs. The present investigation was undertaken to characterize the toxic and non-toxic varieties at molecular level and to develop PCR based molecular markers for distinguishing non-toxic from toxic or vice versa. The polymorphic markers were successfully identified specific to non-toxic and toxic variety using RAPD and AFLP techniques. Totally 371 RAPD, 1,442 AFLP markers were analyzed and 56 (15.09%) RAPD, 238 (16.49%) AFLP markers were found specific to either of the varieties. Genetic similarity between non-toxic and toxic verity was found to be 0.92 by RAPD and 0.90 by AFLP fingerprinting. In the present study out of 12 microsatellite markers analyzed, seven markers were found polymorphic. Among these seven, jcms21 showed homozygous allele in the toxic variety. The study demonstrated that both RAPD and AFLP techniques were equally competitive in identifying polymorphic markers and differentiating both the varieties of J. curcas. Polymorphism of SSR markers prevailed between the varieties of J. curcas. These RAPD and AFLP identified markers will help in selective cultivation of specific variety and along with SSRs these markers can be exploited for further improvement of the species through breeding and Marker Assisted Selection (MAS).

Automated and manual solid phase peptide synthesis techniques were combined with chemical ligation to produce a 37-residue peptide toxin derivative of iberiotoxin which contained: (i) substitution of Val16 to Ala, to facilitate kinetic feasibility of native chemical ligation, and; (ii) substitution of Asp19 to orthogonally protected Cys-4-MeOBzl for chemical conjugate derivatization following peptide folding and oxidation. This peptide ligation approach

Zerumbone (ZER), a potential anticancer compound, isolated from the fresh rhizomes of Zingiber zerumbet. In this investigation, the cytotoxic properties of ZER were evaluated, on cancer cells of human cervix (HeLa), breast and ovary, and normal cells of Chinese Hamster ovary, using MTT assay. Apoptogenic effects of ZER on HeLa were studied using fluorescence microscopy (AO/PI double staining), scanning and transmission electron microscopy (SEM and TEM), and colorimetric assay of the apoptosis promoter enzyme, caspase-3. The results of MTT assay showed that ZER has less effect on normal cells compared to cancer cells. The lowest IC 50 of ZER was observed on HeLa cells. Cytological observations showed nuclear and chromatin condensation, cell shrinkage, multinucleation, abnormalities of mitochondrial cristae, membrane blebbing, holes, cytoplasmic extrusions and formation of apoptotic bodies as confirmed collectively by double staining of AO/PI, SEM and TEM. Statistical analysis (twotailed t-test) of differential counting of 200 cells under fluorescence microscope revealed significant difference in apoptotic cells populations between treated and untreated HeLa cells. In addition, ZER has increased the cellular level of caspase-3 on the treated HeLa cells. It could be concluded that ZER was able to produce distinctive morphological features of cell death that corresponds to apoptosis.

Renal cell carcinoma (RCC) is the most common kidney cancer in adults. Although several genes have been found to be involved in carcinogenesis of RCC, more great efforts are needed to identify new genes which are responsible for the process. Clear cell RCC, originated from proximal tubule cells, is the most common pathological type of RCC. Alkaline phosphatase (ALP) is a marker enzyme of brush border membrane of proximal tubular cells. Our previous studies showed a significant decreased activity of Liver/Bone/Kidney (L/B/K) alkaline phosphatase in RCC. In the present study, we explored the molecular basis of the decreased activity of ALP in RCC. Immunohistochemistry, immunofluorescence and flow cytometry analysis showed decreased ALP protein in RCC. Additionally, real time PCR documented significantly reduced ALP gene expression (P ¼ 0.009). Moreover, RCC cell lines (ACHN and A498) transfected with full length L/B/K cDNA showed decreased migratory property as well as viability of these cells as compared with controls (P ¼ 0.000). Further, L/B/K ALP cDNA transfected cells (ACHN and A498) showed significant increased apoptosis as compared to control (P ¼ 0.000).

Vero cells are commonly used for the growth of avian metapneumovirus subtype C (aMPV-C). This study was conducted to evaluate 17 different cell types for the growth of a Minnesota strain of aMPV-C. The virus was inoculated into these cell types and virus growth was monitored by the development of cytopathic effects (cpe) and immunofluorescence. Virus growth was obtained in 6 of 17 cell types tested with the highest virus titers observed in BGM and DF-1 cells. The flow cytometric analysis of cells at 72 h post inoculation found the highest number of infected cells in BGM cells followed by QT-35 cells. At 48 h post inoculation, DF-1 and BGM cells showed the highest number of infected cells. These results suggest that BGM, QT-35, and DF-1 cells can be used for high titer propagation of aMPV-C.

In the yeast two-hybrid library screening, the heart-specific FHL2 protein was found to interact with hCDC47. In vitro interaction study between FHL2 protein and hCDC47 was demonstrated. From the results of domain studies by the yeast two-hybrid assay, the second and third LIM domains in conjunction with the first half LIM domain of FHL2 were identified to be important in binding with hCDC47. Besides, in Northern blot hybridization of human cancer cell lines, the highest FHL2 mRNA expression was detected in colorectal adenocarcinoma SW480 and HeLa cell S3. Our results imply that FHL2 protein may associate with cancer development and may act as a molecular adapter to form a multicomplex with hCDC47 in the nucleus, thus it plays an important role in the specification or maintenance of the terminal differentiated phenotype of heart muscle cells.

Dear Sir/Madam

We are currently hosting the Oncologist & Diagnostics Conference  which is going to be organized during August 28-30, 2017 Brussels, Belgium. With great pride and honour, we would like to invite you to participate in Oncologist & Diagnostics Conference in Brussels, Belgium

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ConferenceSeries Ltd invites all the participants across the globe to attend the Oncologist & Diagnostics Conference slated on August 28-30, 2017 at Brussels, Belgium.

Cancer Diagnostics 2017 directs towards addressing main issues as well as future strategies of Cancer. This is going to be the largest and most promising international conference where the program includes Clinical Oncologists, Registered Nurses, Nurse practitioners and the entire medical team involved in patient care, researchers, professional, early career individuals and patient advocates who wish to learn principles of tumor immunology and immunotherapy well as decision makers will come to discuss and debate on various aspects of the challenges, risks and investment opportunities throughout the complete information of Cancers. Cancer Diagnostics 2017 will also provide the three days of discussions on methods and strategies related to management and quality improvement of Diagnostics device developing and Cancer Research and as well as to explore the new ideas and concepts on global scale and the topics include lung cancer, breast cancer, bone cancer, leukemia, blood cancer, cervical cancer, colon-rectum cancer, pediatric cancer, surgical cancer, Prostate cancer, thyroid cancer.

According to 2012 statistics approximately 14.1 million cases are reported and 8.2 million deaths are reported. Lung, Liver, Stomach and bowel cancers cause the most common deaths among the worldwide that accounts nearly half of all the cancer deaths. About 70% increase in the cases are to be expected over the next 2 decades. In males 5 most common sites of cancer are diagnosed in 2012 were lung, prostate, colorectal and liver cancers. In females 5 most common sites diagnosed were breast, colorectal, cervix, lung and stomach cancer. Nearly 60% of the world’s total new cases occur in the Africa, Asia and Central and South America regions. All over the world Denmark ranks 1st in the cancer deaths USA is accompanied next to it.

ConferenceSeries Ltd  which is comprised of 3000+ Global Events with over 600+ Conferences, 1200+ Symposiums and 1200+Workshops on diverse Medical, Pharmaceutical, Clinical, Engineering, Science, Technology, Business and Management field is organizing conferences all over the globe. It hosts numerous Cancer Conferences, Cancer meeting, global cancer conference, European Oncology Conferences, Surgical oncology like skin cancer conference, breast cancer conference, liver cancer conference, lung cancer meetings, and Oncology Conferences where knowledge transfer takes place through debates, round table discussions, poster presentations, workshops, symposia and exhibitions and which helps get possibilities for treating the diseases with new era, technology and research.



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Brussels, a region of Belgium comprising 19 municipalities, is a city of contrasts with enormous diversity, experiencing cool climates all throughout the year. Brussels is also becoming known as a mecca of style, art, and design having bilingualism of Modern French and Old Dutch.

Down syndrome (DS) in humans, or trisomy of autosome 21, represents the hyperdiploidy that most frequently survives gestation, reaching an incidence of 1 in 700 live births. The condition is associated with multisystemic anomalies, including those affecting the central nervous system (CNS), determining a characteristic mental retardation. At a neuronal level, our group and others have shown that the condition determines marked alterations of action potential and ionic current kinetics, which may underlie abnormal processing of information by the CNS.

The present study focuses on evaluating the potential of flattened AgClBr fiber optic evanescent wave spectroscopy (FTIR-FEWS) technique for detection and identification of cancer cells in vitro using cell culture as a model system. The FTIR-FEWS results are compared to those from FTIR-Microspectroscopy (FTIR-MSP) method extensively used to identify spectral properties of intact cells. Ten different samples of control and malignant cells were measured in parallel by the above two methods.

The lack of cell lines from mononuclear phagocytes of salmonid ®sh has impeded the study of immune function at a cellular level in these economically and ecologically important animals. Here, we report on the further characterization of RTS11, a previously described macrophage-like cell line from the rainbow trout spleen, with regard to its expression of a number of immunologically relevant genes. Analysis of gene expression by reverse transcription-polymerase chain reaction, using rainbow-trout-speci®c primers demonstrated that RTS11 cells express the b chain of the class II major histocompatibility complex, the cytokines transforming growth factor-b (TGF-b) and interleukin-1b (IL-1b), and cyclo-oxygenase-2 (COX-2). Inducing the cells with lipopolysaccharide led to increased expression of IL-1b and COX-2, as determined by Northern blotting. These results together suggest that RTS11 retains many of the characteristics expected of mature macrophages, and should be a valuable tool for further study of the expression and function of these immunomodulatory proteins in ®sh. 7

p53 has a determinant role in cancer prevention and is among the most studied proteins in the world. The majority of studies devoted to this protein are carried out in cell lines because they are easy to use and have naturally emerged as the main research tool in laboratories. However, the p53 pathway is commonly deregulated in cancer cells, from which the experimental cell lines are generally derived. The fact that the pathway is deregulated challenges the relevance of using cancer-derived cell lines to study wild-type p53 activities, or, in a broader sense, to study any normal cellular process. In the present article, we identify and discuss a number of limitations of cell lines using examples related to p53. Finally, we point out the general limitations of cell lines and propose solutions as alternatives to these cells.

<p><b>A:</b> Effect of M<sub>4</sub>N on the contents of glutathione-related metabolites. This data is a selection from <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0148685#pone.0148685.s003" target="_blank">S1 Table</a>. LNCaP, AsPC-1, and L428 cells were treated with M<sub>4</sub>N for 8 h and metabolite contents of the samples were measured by LC/GC mass spectroscopy by Metabolon. 'M/C' indicates the ratio of metabolite contents for the samples treated with M<sub>4</sub>N vs. the control. For paired comparisons between the control and M<sub>4</sub>N-treated samples, strongly shaded cells indicate statistically significant differences at p≤0.05 while lightly shaded cells indicate them at 0.05B: Effect of M<sub>4</sub>N on the expression of CHAC1. The LNCaP cells were treated with M<sub>4</sub>N (80 μM) for either 5 or 18 h and the expression of the protein was measured by the western blotting. β-Actin was used as the control. <b>C:</b> A schema for M<sub>4</sub>N-regulation of glutathione metabolism. The upward pointing arrows indicate that the contents of the metabolites associated with these arrows were significantly (p≤0.05) induced by M<sub>4</sub>N in at least two out of three cell lines (LNCaP, AsPC-1, and L428 cells) under the additional condition that the effect of M<sub>4</sub>N on the contents of these metabolites in the third cell line was an induction without statistical significant difference, no significant change, or not examined. Meanwhile the downward pointing arrows indicate that the contents of the metabolites associated with these arrows were significantly (p≤0.05) suppressed by M<sub>4</sub>N in at least two out of three cell lines (LNCaP, AsPC-1, and L428 cells) under the additional condition that the effect of M<sub>4</sub>N on the contents of these metabolites in the third cell line was a suppression without statistical significant difference, no significant change, or not examined. The exception to these rules was cystine (whose content was statistically significantly induced by M<sub>4</sub>N only in LNCaP cells. In other two cell lines the data are not available). The effect of M<sub>4</sub>N on ATP content in the whole cells is shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0148685#pone.0148685.g006" target="_blank">Fig 6A</a>. 'N.A.' indicates that the data on these particular metabolites are not available. The blue arrow indicates that CHAC1 expression was enhanced by M<sub>4</sub>N treatment (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0148685#pone.0148685.g006" target="_blank">Fig 6B</a>). The effect of M<sub>4</sub>N on ATP content in the whole cells is shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0148685#pone.0148685.g006" target="_blank">Fig 6A</a>. <b>D:</b> Effect of combination treatments on ROS production in LNCaP cancer cells. The cells were treated with M<sub>4</sub>N (80 μM), etoposide (30 μM), rapamycin (30 μM), or UCN-01 (2 μM). ROS content was measured at 5 h after treatment. C: control, E: etoposide, R: rapamycin, U: UCN-01, M: M<sub>4</sub>N, ME: M<sub>4</sub>N+etoposide, MR: M<sub>4</sub>N+rapamycin, MU: M<sub>4</sub>N+UCN-01.</p

Four continuous cell lines were established from the embryos of the glassy-winged sharpshooter, Homalodisca coagulata (Say), an economically important insect vector of bacterial pathogens of grape, almond, citrus, oleander, and other agricultural and ornamental plantings. The cell lines were designated GWSS-Z10, GWSS-Z15, GWSS-G3, and GWSS-LH. The GWSS-Z10, GWSS-Z15, and GWSS-G3 lines were cultured in Ex-Cell 401 medium supplemented with 10% fetal bovine serum (FBS), whereas the GWSS-LH line was cultured in LH medium supplemented with 20% FBS. The cell lines were characterized in terms of their morphology, growth, protein composition, and polymerase chain reactionamplification patterns of their chromosomal deoxyribonucleic acid. The population doubling times of GWSS-Z10, GWSS-Z15, GWSS-G3, and GWSS-LH were 46.2, 90.9, 100.3, and 60.2 h, respectively. These lines should be useful for the study of insect-pathogenic viruses of leafhoppers, aphids, treehoppers, and other related insects as well as plant-pathogenic viruses that are transmitted by these insects.

Primary isolation of avian metapneumovirus (aMPV) is carried out using tracheal organ culture (TOC) or chicken embryonated eggs with subsequent adaptation in chicken embryo fibroblasts (CEF) or Vero cultures. This study was conducted to evaluate six different cell lines and two avian culture systems for the propagation of aMPV subtypes A and B. The chicken embryo related (CER) cells were used successfully for primary isolation. In addition to Vero and baby hamster kidney (BHK-21) cells, CER cells were also shown to be the most appropriate for propagation of aMPV considering high titres. Propagation of A and B subtypes in CEF and TOC remained efficient after the primary isolation and several passages of viruses in the CER cell line. The growth curves were created using CER, Vero and BHK-21 cell lines. Compared with growth, both yielded higher titres in CER cells during the first 30 h after infection, but no significant difference was observed in the results obtained from CER and Vero cells. This data show that CER cells are adequate for aMPV subtypes A and B propagation, giving similar results to Vero cells.

Background
Cyanuric acid and its derivatives belong to the group of s-triazines. They have wide industrial application, generally in the production
of pesticides, bleaching agents and disinfectants. Recent reports showed significant negative effect of cyanuric acid in combination with
melamine but low general cytotoxicity of cyanuric acid alone. However, evaluations of cyanuric acid toxicity against different human cell
types using a panel of in vitro assays have not been performed. In addition, little is known about the cytotoxicity and potential antitumor
effects of certain cyanuric acid derivatives, for example trichloroisocyanuric acid and 1,3,5-tris(2-hydroxyethyl) isocyanurate.
Aim
To investigate the toxicity of cyanuric acid, trichloroisocyanuric acid and 1,3,5-tris(2-hydroxyethyl) isocyanurate using different cancer
cell lines and normal fibroblasts.
Materials and Methods
MTT, Neutral red and clonogenic assays were used to characterize the in vitro cytotoxicity of cyanuric acid, trichloroisocyanuric acid and
1,3,5-tris(2-hydroxyethyl) isocyanurate. Different concentrations of the compounds were tested on three cancer cell lines (HeLa, A549,
CaOV) and normal human fibroblasts. IC50 values were calculated and compared. Intracellular ATP measurements and microscopic
evaluations of cellular morphology after exposure to cyanuric acid, trichloroisocyanuric acid and 1,3,5-tris(2-hydroxyethyl) isocyanurate
were performed.
Results
Among the three test-agents trichloroisocyanuric acid (TCIC) demonstrated the highest cytotoxicity in vitro. Interestingly, MTT assays
indicated that the toxic effects of TCIC were mainly directed towards cancer cells affecting mitochondrial functionality. Clonogenicity
of cancer cell lines was completely inhibited in the presence of low concentrations of the triazine. Exposition to TCIC influenced the
morphology and intracellular ATP concentration in both cancer cells and normal fibroblasts but the observed effects were stronger in
cancer cells.
Conclusion
The order of cytotoxicity of the tested compounds is as follows: trichloroisocyanuric acid>cyanuric acid>1,3,5-tris(2-hydroxyethyl)
isocyanurate. Trichloroisocyanuric acid demonstrates selective antitumor effect.

Chemotherapy uses drugs to kill cancer cells, either by preventing them from multiplying or by causing the cells to self-destruct. However, in most cases, high systemic toxicity and drug resistance limit the successful outcomes of treatment regimes. Prevention of cancer through dietary intervention recently has received an increasing interest and dietary polyphenols have become not only important potential chemopreventive, but also therapeutic, natural agents. Combination of effective chemopreventive agent (such as ginger) with chemotherapeutic agents may enhance efficacy while reducing toxicity to normal tissues, resulting in better survival. In this study, we observed that treatment of human cervical carcinoma cell line, HeLa with ethanolic ginger extract in combination with gemcitabine resulted in significant dose-dependent decrease in cell viability. It is noteworthy that use of ginger extract increased the efficacy of gemcitabine and importantly, it was found to be minimally toxic to normal cells. Together, these results suggest a novel mechanism may be involved in the synergistic effect of this combination. Thus, this combination may be an effective modality management in the treatment of cervical cancer.

Many reports have been published about the biogenesis of silver nanoparticles using several plant extracts such as Pelargonium graveolens (P.graveolens- geranium) and Azadirachta indica (neem) but the capacity of their natural reducing constituents to form silver nanoparticles has not yet been studied. In this research the synthesis of silver nanoparticles using geraniol has been investigated. We successfully synthesized uniformly dispersed silver nanoparticles with a uniform size and shape in the range of 1 to 10 nm with an average size of 6 nm. Also the cytotoxicity of the prepared silver nanoparticles was investigated using a cancer cell line (Fibrosarcoma-Wehi 164). The cytotoxicity analysis of the sample shows a direct dose-response relationship; cytotoxicity increased at higher concentrations. At concentration of 1 µg/ml, silver nanoparticles was able to inhibit the cell line's growth by less than 30%. Conversly, the presence of 5 µg/ml of silver nanoparticlse significantl...

Dihydroactinidiolide (1) and a mixture of sterols [campesterol (2), stigmasterol (3) and β-sitosterol (4)], together with the previously isolated individual compounds βsitosterol (4), 2,4-di-tert-butylphenol (5), α-tocopherol (6), phytol (7) were isolated from the active ethyl acetate fraction of Pereskia bleo (Kunth) DC. (Cactaceae) leaves. Cytotoxic activities of the above mentioned compounds against five human carcinoma cell lines, namely the human nasopharyngeal epidermoid carcinoma cell line (KB), human cervical carcinoma cell line (CasKi), human colon carcinoma cell line (HCT 116), human hormone-dependent breast carcinoma cell line (MCF7) and human lung carcinoma cell line (A549); and non-cancer human fibroblast cell line (MRC-5) were investigated. Compound 5 possessed very remarkable cytotoxic activity against KB cells, with an IC 50 value of 0.81µg/mL. This is the first report on the cytotoxic activities of the compounds isolated from Pereskia bleo.

While polyamines (PAs) have been suggested to protect cells against Reactive Oxygen Species (ROS), their catabolism is known to generate ROS. We compared the activities of several enzymes and cellular metabolites involved in the ROS scavenging pathways in two isogenic cell lines of poplar (Populus nigra  maximowiczii) differing in their PA contents. Whereas the control cell line was transformed with b-glucuronidase (GUS), the other, called HP (High Putrescine), was transformed with a mouse ornithine decarboxylase (mODC) gene. The expression of mODC resulted in several-fold increased production of putrescine as well its enhanced catabolism. The two cell lines followed a similar trend of growth over the seven-day culture cycle, but the HP cells had elevated levels of soluble proteins. Accumulation of H 2 O 2 was higher in the HP cells than the control cells, and so were the activities of glutathione reductase and monodehydroascorbate reductase; the activity of ascorbate peroxidase was lower in the former. The contents of reduced glutathione and glutamate were significantly lower in the HP cells but proline was higher on some days of analysis. There was a small difference in mitochondrial activity between the two cell lines, and the HP cells showed increased membrane damage. In the HP cells, increased accumulation of Ca was concomitant with lower accumulation of K. We conclude that, while increased putrescine accumulation may have a protective role against ROS in plants, enhanced turnover of putrescine actually can make them vulnerable to increased oxidative damage.

We determined previously that lipoproteins of Borrelia burgdorferi stimulate inflammatory and anti-inflammatory cytokines (interleukin-10 [IL-10]) in monocytes. IL-10 could have an effect on innate and acquired immune responses to B. burgdorferi and influence the magnitude of the infectious inoculum and disease outcome. To understand the mechanism(s) of IL-10 action during early infection, when innate immunity expressed chiefly by skin macrophages is key, we investigated the effect of exogenous and endogenous IL-10 on the production of the macrophage-derived cytokines IL-6, IL-1␤, IL-12, and tumor necrosis factor alpha (TNF-␣). We used the THP-1 human monocytic cell line and recombinant lipidated OspA (L-OspA) as the model target cell and stimulant, respectively. To determine the kinetics of cytokine production by THP-1 cells, we stimulated them with L-OspA and also with heat-killed B. burgdorferi cells (HBb) and lipopolysaccharide (LPS). Exogenous IL-10 dampened production of inflammatory cytokines, as elicited by lipoproteins. The inhibition of endogenous IL-10 function by anti-IL-10 antibody reduced the production of IL-12 and IL-6 but not that of IL-1␤ and TNF-␣. An inspection of the kinetics of cytokine production clarified this finding. TNF-␣ was produced prior to, and IL-␤ was produced at the same time as, IL-10, whereas IL-6 and IL-12 were produced later. HBb, LPS, and L-OspA yielded similar kinetics of cytokine production. This result reinforces the notion that lipoproteins are the functional molecules in HBb and perhaps in vivo. It indicates also that signaling pathways utilized by LPS and lipoproteins may be extensively shared. The results are consistent with a major role played by IL-10 in controlling the initial phase of infection with this spirochete.

The aims of this work were 1) to determine the bioactivities resulting from the hydrolysis of soy (Glycine max) proteins isolate (SPI) by a double enzymatic treatment of pepsin and pancreatin and 2) to investigate whether fractionation of the protein hydrolysates by two ultrafiltration techniques using either hollow fiber (HF) or spiral-wounded (SW) membrane (10 kDa cut-off), increases biological activity of the recovered peptides. The focus was on the screening of bioactivity of the peptide fractions using multiple in vitro bioassays such as the proliferation of cancer cells, destruction of murine norovirus-1 (MNV-1), anti-oxidative and immunomodulating properties, and finally glucose metabolism in a muscle cell line. Significant biological effects of the peptides for both antioxidant capacity (ORAC assay) and metabolism (muscle glucose uptake assay) were found. These results show that ultrafiltration of SPI hydrolysates is a convenient process for the recovery of large amount of bioactive peptides.