Conditioned media from adipocytes promote proliferation,migration, and invasion in melanoma and colorectal cancer cells

Epidemiological evidence suggests that obesity can significantly increase the risk of various cancers, although the mechanisms underlying this link are completely unknown. Here, we analyzed the effect of adipocytes on melanoma and colon cancer cells proliferation, migration, and invasion. The potential effects of conditioned media (CM) obtained from differentiated mouse 3T3-L1 cells and human adipose tissue-derived mesenchymal stem cells (hAMSC) on the proliferation, migration, and invasion of B16BL6 melanoma and colon 26-L5 cancer cells were investigated. The 3T3-L1 and hAMSC CM increased cell proliferation, migration, and invasion in both the cell lines. In addition, adipocytes CM increased matrix metalloproteinase 9 (MMP-9) and MMP-2 activity in both B16BL6 and colon 26-L5 cells. These effects were found to be associated with an increased expression of various oncogenic proteins in B16BL6 and colon 26-L5 cells. Also, adipocyte CM induced Akt and mTOR activation in both tumor cell lines, and the pharmacological inhibition of Akt and mTOR blocked the CM induced Akt as well as mTOR activation and CM-stimulated melanoma and colon cancer cell proliferation, migration, and invasion. These data suggest that adipocyte promotes melanoma and colon cancer progression through modulating the expression of diverse proteins associated with cancer growth and metastasis as well as modulation of the Akt/mTOR signaling.     

Roche Susceptibility Test (RST) medium,a defined formulation for susceptibility testing: I. Nutrient interaction and optimization studies

Roche Susceptibility Test (RST) Medium represents the most completely optimized and convenient fully defined medium described. It requires no post-autoclaving supplementation with vitamins, supports good growth of all common aerobic and anaerobic pathogens and may be used as a broth or agar gel on which the swarming of Proteus spp. is virtually eliminated. The broth as a superior buffering capacity to most complex media and an osmolality and pH close to those of human serum. RST is highly satisfactory for the susceptibility testing of commonly used antibiotics and meets the requirements of the National Committe for Clinical Laboratory Standards of the U.S.A. in almost every respect.     

Direct measurement of water availability in gelled plant tissue culture media

Summary Water constitutes nearly 100% of the volume and 95% of the mass of gelled plant tissue culture media. Even so, plant growth and development responses observed to occur with relatively small changes in gelling agent concentration (0.1% of media total mass) have been attributed to changes in media water availability. Measurements with three alternative direct techniques, specific for measuring physiochemical water availability indicated the effects of a change of this magnitude in gelling agent concentration negligibly affected the media water potential and water conductivity. Sensitive pressure membrane measurements indicated that incremental gelling agent concentration increases of 0.1% (of media total mass) within the range normally used for plant tissue culture media, depressed water matric potential only 1–2cm H₂O (1–2×10⁻⁴ MPa (mega pascal.)); these values were confirmed with equally sensitive tensiometer measurements. Moreover, no effect of concentration on water movement could be detected with a precise constant-head permeameter over a broader range of gelling agent concentrations. These results indicate that eitherin vitro plants are extremely sensitive to subtle shifts in water status, or other physiochemical factors that also change with gelling agent concentration are contributing to the reported effects on plant growth and development.     

Genetic essentialism,mana , and the meaning of DNA

Concerns about the commercialization of genetics have spawned a debate over the symbolic logic and meaning of DNA. The assumption is that different meanings for DNA have social and ethical consequences. Genetic essentialism as an interpretive meaning for DNA is argued to encapsulate values of materialism and autonomy that make it compatible with capital accumulation. Whether or not genetic commerce actually requires genetic essentialism is an empirical question and this study proposes that it is not difficult to find non-essentialist genetics. Two paths of inquiry are adopted. First, the history and origins of the distinction between genotype and phenotype is revisited. This history of gene theory, in particular the effort to purge vitalism, is linked to DNA and the central dogma of molecular biology. Secondly, a rather specialized debate within anthropology about the meaning of mana is introduced. An analysis of definitions for genotype and phenotype reveals a structure commensurate with the metaphysics of mana. Parallels are established between how the meaning of mana has been essentialized and the current efforts to fix the symbolic logic of DNA.     

Cadmium stress studies: Media development, enrichment, consortia analysis, and environmental relevance

The effects of Cadmium (Cd) toxicity on bacterial consortia originating from an-aerobic sewage sludge and cultivated under differing enrichment conditions were studied. Cultures were enriched in minimal media developed specifically for Cd stress studies. At inoculation all Cd was soluble in free ion or chelated form. Electron donors and acceptors were varied to obtain each physiological enrichment type. Adaptation leading to higher levels of Cd resistance of the consortia over time was observed under all physiological conditions. Initial and increased Cd tolerances were consistently greatest in multiphysiological enrichments (MPH). Sulfate reducing (SRB), methanogenic (MET), and fermentative (FRM) enrichments had less tolerance however, the level of tolerance to the Cd varied from one inoculation to the next. The Cd remained soluble as free Cd in MPH and FRM conditions and was precipitated significantly in SRB and moderately in MET conditions. Denaturing gradient gel electrophoresis (DGGE) of polymerase chain reaction (PCR) amplified 16S rRNA of the SRB, MPH, and FRM enrichments were followed over time. The consortia underwent succession under all physiological conditions when compared with the profile of the inoculum. Microbial population diversity decreased as the consortia were subcultured. The effects of chelators in the MPH medium were also evaluated. The addition of chelators transiently decreased toxicity. Effects of MPH medium on the Cd sorption capacity of soil were evaluated. Microbial growth decreased the amount of Cd left in solution.     

Ferguson and the death of Michael Brown on Twitter: #BlackLivesMatter, #TCOT,and the evolution of collective identities

Recent events related to police brutality and the evolution of #BlackLivesMatter provides an empirical case to explore the vitality of social media data for social movements and the evolution of collective identities. Social media data provide a portal into how organizing and communicating generate narratives that survive over time. We analyse 31.65 million tweets about Ferguson across four meaningful time periods: the death of Michael Brown, the non-indictment of police officer Darren Wilson, the Department of Justice report on Ferguson, and the one year aftermath of Brown’s death. Our analysis shows that #BlackLivesMatter evolved in concert with protests opposing police brutality occurring on the ground. We also show how #TCOT (Top Conservatives on Twitter) has operated as the primary counter narrative to #BlackLivesMatter. We conclude by discussing the implications our research has for the #BlackLivesMatter movement and increased political polarization following the election of Donald Trump.     

Effect of media composition,including gelling agents,on isolation of previously uncultured rumen bacteria

The aim of this study was to develop novel anaerobic media using gellan gum for the isolation of previously uncultured rumen bacteria. Four anaerobic media, a basal liquid medium (BM) with agar (A‐BM), a modified BM (MBM) with agar (A‐MBM), an MBM with phytagel (P‐MBM) and an MBM with gelrite (G‐MBM) were used for the isolation of rumen bacteria and evaluated for the growth of previously uncultured rumen bacteria. Of the 214 isolates composed of 144 OTUs, 103 isolates (83 OTUs) were previously uncultured rumen bacteria. Most of the previously uncultured strains were obtained from A‐MBM, G‐MBM and P‐MBM, but the predominant cultural members, isolated from each medium, differed. A‐MBM and G‐MBM showed significantly higher numbers of different OTUs derived from isolates than A‐BM (P < 0·05). The Shannon index indicated that the isolates of A‐MBM showed the highest diversity (H′ = 3·89) compared with those of G‐MBM, P‐MBM and A‐BM (H′ = 3·59, 3·23 and 3·39, respectively). Although previously uncultured rumen bacteria were isolated from all media used, the ratio of previously uncultured bacteria to total isolates was increased in A‐MBM, P‐MBM and G‐MBM.     

Hope and Sorrow: Uncivil Religion,Tibetan Music Videos,and YouTube

Tibetan activists and their supporters are interpreting the lyrical and visual symbolism of contemporary Tibetan music videos from China as a call for Tibetans to return to a shared Tibetan identity, centered around religious piety and implied civil disobedience, in order to counter fears of cultural assimilation. As the popularity of some videos on social-networking sites dovetailed with the 2008 protests in Tibet, viewers employed a progressive hermeneutical strategy which demanded a sectarian political interpretation of the lyrics and imagery of the most popular videos out of Tibet. Within China, Tibetans have begun to add these videos to the growing canon of an emerging uncivil religion, which emphasizes Tibetan cultural, linguistic, and religious autonomy within China. Through comparing online and offline ethnography, this article explores the relationship between offline and online worlds and the connections between Tibetans in China and their supporters.     

Prevalence of hicAB, lav, traA, and hifBC among Haemophilus influenzae middle ear and throat strains

Nontypeable Haemophilus influenzae (NTHi) is an important cause of illness among children. To further understand the role of laterally transferred genes in NTHi colonization and otitis media, the prevalence of hicAB, lav, tnaA, and hifBC was determined among 44 middle ear and 35 throat NTHi isolates by dot-blot hybridization.     

Laminin provides a better substrate than fibronectin for attachment,growth, and differentiation of 1003 embryonal carcinoma cells

Summary Culture of cells in hormonally defined media has allowed (a) the demonstration of physiological responses from cells usually unable to express them in vitro and (b) the study of the effects on growth and differentiation of diffusible factors and attachment factors. The embryonal carcinoma line 1003 forms multidifferentiated tumors in vivo but is unable to differentiate in vitro when grown in serum-containing medium. In a defined medium containing insulin, transferrin, selenium, and fibronectin as attachment factors, 1003 cells grow for several generations and differentiate into neurons and embryonic mesenchyme (Darmon et al., 1981, Dev. Biol. 85: 463–473). In the present work the effects of fibronectin and laminin were compared. In the presence of laminin the cells attached and spread better, grew faster, and could be plated at lower densities. Neurite extension was also better under these conditions and most importantly, it was found that laminin induced an important formation of muscular tissue when the cells had been seeded at low densities. Multinucleated myotubes could be stained with antibodies directed against embryonic muscular myosin. Coating the dishes with polylysine or adding FGF or serum-spreading factor to the medium allowed growth of low-density cultures with fibronectin instead of laminin but muscular differentiation was not detected under these conditions. Addition of fibronectin to laminin-containing medium did not inhibit muscular differentiation. Presented in the symposium on Plant and Animal Physiology in Vitro at the 33rd Annual Meeting of the Tissue Culture Association, San Diego, California, June 6–10, 1982. This research was supported in part by grants from the “Centre National de la Recherche Scientifique” (LA 269), the “Délégation Générale à la Recherche Scientifique et Technique,” the Fondation pour la Recherche Médicale Fran?aise,” the “Institut National de la Santé et de la Recherche Medicale,” the “Ligue Nationale Fran?aise centre le Cancer,” and the “Fondation André Meyer.” This symposium was supported in part by the following organizations: Bellco Glass, Inc., California Branch of the Tissue Culture Association, Collaborative Research, Hana Media, Hybridtech, K C Biological, Inc., and Millipore Corporation.     

Migration,clogging, and carbon source release of nano emulsified vegetable oil in porous media,evaluated by column experiments

To effectively solve the problem of aquifer clogging in the process of in situ bioremediation of groundwater pollution by micron emulsified vegetable oil (Micron EVO), Nano emulsified vegetable oil (Nano EVO) was prepared to replace conventional micron EVO, and three one-dimensional laboratory columns packed with medium or fine sands were conducted to simulate migration, clogging, and carbon source release of EVO in porous media. Column experiment results show that micron and nano EVO resulted in a 20.40% and 3.20% reduction in permeability of medium sand, respectively. Correspondingly, the interception of micron and nano EVO in medium sand were 28.51% and 20.15%, respectively. Obviously, EVO interception is an important reason for permeability loss, and reducing EVO droplet size can effectively alleviate permeability loss in porous media. The COD ratios (dissolved COD/total COD) of micron and nano EVO in medium sand were 87.61% and 61.95%, respectively. The release effect of nano EVO was better than that of micron EVO. Effective longevity of micron and nano EVO were 243.17 d and 98.80 d, respectively. The effect of fine sand media on EVO indicated nano EVO can be used in a finer granular media, and its longevity can also be extended in this media.     

Growth and Protein Content in Colletotrichum circinans, Fusarium solani and Rhizoctonia solani in Liquid Culture

The phytopathogenic fungi Colletotrichum circinans, Fusarium solani, and Rhizoctonia solani were incubated in aerated (0, 0.5, 1 dm³ min^–1) potato dextrose broth (PDB) or Czapek-Dox broth (CDB), under 0-, 12- or 24-h photoperiods. Greater dry mass was produced in PDB. Higher air flows improved dry mass of F. solani and R. solani. The 24-h photoperiod improved F. solani dry mass. Except for F. solani, which was not affected, incubation in PDB increased protein content. The no air treatment increased protein content in F. solani, 0.5 dm³ min^–1 produced the highest protein content in R. solani, but air flow-rate did not affect C. circinans. Incubation in the dark produced the lowest protein content in C. circinans, the highest under the 24-h photoperiod for R. solani, and photoperiod did not affect protein content in F. solani. Protein content in R. solani, incubated in CDB, was lowest at the 0 dm³ min^–1 air flow at all photoperiods. Molecular masses of most proteins were under 30 kDa, and numbers of bands in SDS-PAGE gels varied with air flow. In CDB, especially under 12- or 24-h photoperiods, proteins in F. solani were between 1.6 and 310 kDa. For R. solani in PDB, at 0.5 dm³ min^–1 air flow and 12-h light, proteins were between 6 and 81 kDa.     

Colonization and turnover of Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis in otitis-prone children

Recurrent otitis media are frequently intractable during childhood. It is unclear whether recurrent otitis media is caused by etiological bacteria colonization or by new infections. Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis were isolated from the nasopharynx of 7 otitisprone and 2 non-prone children with recurrent otitis media. Plural bacterial species and strains were found in all children while affected by otitis media. The same strain was repeatedly isolated from all otitisprone children even after administration of antibiotics but was not from the non-prone children. Antibiotic susceptibility did not differ significantly among the same repeatedly isolated strains. This pilot study suggests that the etiological bacteria tend to colonize and is hard to eliminate in otitis-prone children.     

Advanced Granulation Technology (AGTTM). An alternate format for serum-free, chemically-defined and protein-free cell culture media

To overcome limitations of conventional milling technology, we investigated the application of fluid bed granulation for the production of dry-form nutrient media. Serum-free, protein-free and chemically-defined specialty media were produced in granulated format and compared with identical formulations manufactured by conventional methods. HPLC analysis of multiple lots of granulated materials demonstrated that biochemical constituents were precisely and homogeneously distributed throughout the granules and that nutrient levels were comparable to conventional formats. Comparison of medium performance in cell proliferation and biological production assays demonstrated equivalence with reference media. The fluid bed granulation process meets pharmaceutical quality requirements and may be applied to a broad range of nutrient formulations required for bioproduction applications. This revised version was published online in July 2006 with corrections to the Cover Date.     

Hyperosmotic pressure on HEK 293 cells during the growth phase, but not the production phase, improves adenovirus production

Hyperosmotic stress has been widely explored as a means of improving specific antibody productivity in mammalian cell cultures. In contrast, a decrease in cell-specific productivity of adenovirus production has been reported in several studies in which virus production in HEK 293 cell cultures was conducted under hyperosmotic conditions. However, production of viral vectors and, in particular, adenoviral vectors is the result of two consecutive phases: the growth phase and the virus production phase. In this study, the singular and combined effects of osmolality on the phases of cell growth and virus production were evaluated in culture media with osmolalities ranging from 250 to 410 mOsm. A two-factor, five-level full factorial design was used to investigate the effect of osmotic stress on cell physiology, as determined through the characterization of cell growth, cell metabolism, cell viability, cell cycle, cell RNA and total protein content, and total virus yield/cell-specific virus productivity. Overall, the results show that the growth of cells under hyperosmotic conditions induced favorable physiological states for viral production, and the specific virus productivity was improved by more than 11-fold when the medium's osmolality was increased from 250 to 410 mOsm during the cell growth phase. Both hypo- and hyperosmotic stresses in the virus production phase reduced virus productivity by as much as a factor of six. Optimal virus productivity was achieved by growing cells in media with an osmolality of 370 mOsm or greater, followed by a virus production phase at an osmolality of 290 mOsm. Compared to standard culture and production conditions in isotonic media, the shift from high to low osmolality between the two phases resulted in a two- to three-fold increase in virus yields. This hyperosmotic pressure effect on virus productivity was reproduced in five different commercial serum-free media.     

Empathy,social media,and directed altruistic living organ donation

In this article we explore some of the ethical dimensions of using social media to increase the number of living kidney donors. Social media provides a platform for changing non‐identifiable ‘statistical victims’ into ‘real people’ with whom we can identify and feel empathy: the so‐called ‘identifiable victim effect’, which prompts charitable action. We examine three approaches to promoting kidney donation using social media which could take advantages of the identifiable victim effect: (a) institutionally organized campaigns based on historical cases aimed at promoting non‐directed altruistic donation; (b) personal case‐based campaigns organized by individuals aimed at promoting themselves/or someone with whom they are in a relationship as a recipient of directed donation; (c) institutionally organized personal case‐based campaigns aimed at promoting specific recipients for directed donation. We will highlight the key ethical issues raised by these approaches, and will argue that the third option, despite raising ethical concerns, is preferable to the other two.     

An Interview with Juan Jose García, President of Ojo de Agua Comunicación

This is an interview with Zapotec video maker Juan Jose Garcia, president of the award-winning media organization Ojo de Agua Comunicacion, in Oaxaca, Mexico. He presents a brief history of native video in Mexico and discusses issues such as indigenous video production and circulation, thematic content, communal practices, relation to indigenous communities, funding, and current projects of his group.     

Culture of endocrine pancreatic cells in protein-free,chemically defined media

Summary Cell suspensions prepared by collagenase digestion of pancreata obtained from 21.5-d-old rat fetuses were preincubated in RPMI medium containing 10% fetal bovine serum (FBS), to ensure cell adhesion. Twenty hours later, this medium was replaced by a chemically defined medium. Dulbecco's modified Eagle's (DME)-F12 was used alone or supplemented with various combinations of transferrin, sodium selenite, or Ultroser G. The evolution of the culture and the islet ultrastructure were similar in defined and serum-containing media. However, in the defined medium, the neoformed islets seemed less numerous, and the fibroblast layer less dense, when compared to the RPMI+10% FBS control medium. At Day 7, in defined media, the total insulin content per dish was half that of control cultures. None of the tested additives improved the yield of the cultures. The fractional insulin release per day was elevated in defined media. In subsequent incubations, glucose and leucine stimulated insulin release in a way characteristic of these cells of fetal origin. The labeling index of islet cells cultured in DME-F12 reached 10.7%, which is not far from that observed in RPMI+10% FBS. Such a defined medium is useful to study B cell physiology, avoiding the possible interaction of serum components with substances to be tested. The support of the Fonds National de la Recherche Scientifique and the ‘Loterie Nationale’ of Belgium is also acknowledged.