Becoming the Bimboy (Bimbo Transformation Story, Gay Alpha Male M/M Romance)

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Active targeting of nanoparticles to receptor-overexpressing cancer cells has great potential for enhancing the cellular uptake of nanoparticles and for reducing fast clearance of the nanoparticles from the body. Herein, we present a preparation method of a porous silicon (PSi)-based nanodelivery system for breast cancer targeting, by covalently conjugating a synthesized amide-modified hyaluronic acid (HA +) derived polymer on the surface of undecylenic acid-modified thermally hydrocarbonized PSi (UnTHCPSi) nanoparticles. The resulting UnTHCPSi–HA + nanoparticles showed relatively small size, reduced polydispersibility, high biocompatibility, improved colloidal and human plasma stability, as well as enhanced cellular interactions and internalization. Moreover, we demonstrated that the enhanced cellular association of UnTHCPSi–HA + relies on the capability of the conjugated HA + to bind and consequently target CD44 receptors expressed on the surface of breast cancer cells, thus making the HA +-functionalized UnTHCPSi nanoparticles a suitable and promising nanoplatform for the targeting of CD44-overexpressing breast tumors and for drug delivery.

As with all newbies, I started on general TikTok: dance routines, funny home videos, viral skits and pet videos.K. K. Upadhyay, A. K. Mishra, K. Chuttani, A. Kaul, C. Schatz, J.-F. Le Meins, A. Misra and S. Lecommandoux, Nanomedicine, 2012, 8, 71–80 CrossRef CAS PubMed. R. E. Serda, B. Godin, E. Blanco, C. Chiappini and M. Ferrari, Biochim. Biophys. Acta, 2011, 1810, 317–329 CrossRef CAS PubMed. N. Hasan, A. Mann, M. Ferrari and T. Tanaka, Methods Mol. Biol., 2013, 1049, 481–493 Search PubMed.

The physical properties of bare UnTHCPSi, namely the specific surface area, total pore volume and average pore diameter, were determined by N 2-sorption at −196 °C, using a TriStar 3000 equipment (Micromeritics Inc., USA). The specific surface area was calculated using the Brunauer–Emmett–Teller (BET) theory. 56 The total pore volume was considered to be the total adsorbed amount at a relative pressure p/ p 0 = 0.97. 57 The average pore diameter was calculated from the values of specific surface area and total pore volume, assuming the pores of the UnTHCPSi nanoparticles as cylindrical. S. H. Kim, H. C. Shum, J. W. Kim, J. C. Cho and D. A. Weitz, J. Am. Chem. Soc., 2011, 133, 15165–15171 CrossRef CAS PubMed. K. Hettiarachchi, S. Zhang, S. Feingold, A. P. Lee and P. A. Dayton, Biotechnol. Prog., 2009, 25, 938–945 CrossRef CAS PubMed. This is the long awaited Anthology for Animated Movies for Crossovers. Usual all smut, I take requests and write original content, and take contributions. I want these Anthologies to contain works from as many authors as possible and I will credit each one as I go. Language: English Words: 31,875 Chapters: 8/? Comments: 27 Kudos: 66 Bookmarks: 15 Hits: 27,604 Electronic supplementary information (ESI) available: ESI 1: morphological changes over time of the THCPSiMPs–lipid vesicles during release experiments. ESI 2: cell viability experiments. See DOI: 10.1039/c3lc51260fP. Kinnari, E. Mäkilä, T. Heikkilä, J. Salonen, J. Hirvonen and H. A. Santos, Int. J. Pharm., 2011, 414, 148–156 CrossRef CAS PubMed. C.-F. Wang, E. M. Mäkilä, M. H. Kaasalainen, D. Liu, M. P. Sarparanta, A. J. Airaksinen, J. J. Salonen, J. T. Hirvonen and H. A. Santos, Biomaterials, 2014, 35, 1257–1266 CrossRef CAS PubMed. Introduction Cancer is a complex disease that has been estimated to have caused more than 8.2 million deaths, counted with approximately 14.1 million new cases in 2012, and forecasted to reach over 25 million new cases in the next two decades, representing one of the leading causes of death worldwide. 1 The modern-day bimbo is a fresh approach to intersectional feminism. There is, actually, careful thought behind bimbology, and it could be a way to reach true liberation.

L.-S. Wang, L.-C. Wu, S.-Y. Lu, L.-L. Chang, I.-T. Teng, C.-M. Yang and J.-A. A. Ho, ACS Nano, 2010, 4, 4371–4379 CrossRef CAS PubMed. The TEM images elucidated that the association and internalization of the bare UnTHCPSi nanoparticles with both cell lines was negligible, which can be possibly explained by the negative charge, as well as by the deficient colloidal stability of these nanoparticles. In contrast, when functionalized with HA +, and despite the less pronounced negative ζ-potential, the nanoparticles were massively associated with the cell membrane and further internalized by both MDA-MB-231 and MCF-7 cells, after which they seemed to be mainly enclosed in the endosomes of the cells. Although the aforementioned improved colloidal stability of UnTHCPSi–HA + nanoparticles favors their internalization by the cancer cells, the HA +-mediated targeting of CD44 receptor might be a driving force for the enhanced interaction and internalization of the nanoparticles by the breast cancer cells. These results represent clear evidence of the enhanced cellular association of UnTHCPSi–HA + when compared with bare UnTHCPSi nanoparticles. Flow cytometric analysis of CD44 expression and cellular association The expression of CD44 receptor in both MDA-MB-231 and MCF-7 breast cancer cells was evaluated by flow cytometry, after staining the cells with PE-CF594-labeled anti-human CD44 antibody (ESI, Fig. S2 †). When compared with the respective negative controls, both the cell lines studied were shown to express the CD44 receptor, as elucidated by the increase in the mean fluorescence intensity of the stained cells. However, the incubation of MCF-7 cancer cells with up to 4-fold increased anti-human CD44 antibody concentration was not reflected by the intensification of the mean fluorescence intensity, which experienced an approximately 9-fold increase in comparison with unstained cells regardless of the antibody concentration, thus indicating the saturation of the receptors promptly available for interacting with the complementary antibody. In contrast, in the case of MDA-MB-231 cancer cells, the successive increase of the anti-human CD44 antibody concentration resulted in the consecutive intensification of the fluorescence signal up to approximately 670-fold compared to the controls, suggesting significantly higher levels of CD44 expression in MDA-MB-231 than in MCF-7 breast cancer cells. 50 Justin Cord, The Unexpected Evolution of Language: Discover the Surprising Etymology of Everyday Words Hayes Adams Media, Sep 18, 2012 A. Abbaspourrad, N. J. Carroll, S. H. Kim and D. A. Weitz, J. Am. Chem. Soc., 2013, 135, 7744–7750 CrossRef CAS PubMed. The size and morphology of the UnTHCPSi and UnTHCPSi–HA + nanoparticles were analyzed by transmission electron microscopy (TEM). The TEM pictures were captured using a Jeol JEM-1400 microscope (Jeol Ltd., Tokyo, Japan) at an 80 kV voltage. The nanoparticle suspensions were centrifuged, re-dispersed in ethanol at a concentration of 10 μg mL −1, and dropped on a carbon-coated copper TEM grid, followed by drying for 48 h at room temperature. Stability studies in human plasma For performing these experiments, 300 μg of UnTHCPSi and UnTHCPSi–HA + were dispersed in 200 μL of 1× PBS (pH 7.4), exposed to 1500 μL of human plasma, and left under stirring at 800 rpm and 37 °C for 2 h. Sample aliquots of 200 μL were withdrawn at pre-determined time points (1, 5, 10, 15, 30, 60, 90, and 120 min), and the corresponding size, ζ-potential, and PdI were subsequently determined by DLS and ELS. The results are shown as the average of at least three independent measurements. Anonymous human plasma donors were obtained from the Finnish Red Cross Blood Service, with the permission from the respective institutional ethical committee. Cell lines and culturing conditions For the in vitro studies, MCF-7 and MDA-MB-231 breast cancer cells were cultured according to the protocols described in detail in the ESI. † In vitro cytotoxicity studies The in vitro cytotoxicity of both UnTHCPSi and UnTHCPSi–HA + was evaluated by a CellTiter-Glo ® Luminescent Cell Viability assay, as previously described elsewhere. 26,29 MCF-7 and MDA-MB-231 breast cancer cells were suspended in the corresponding cell culture media at a concentration of 2 × 10 5 cells per mL, and approximately 2 × 10 4 cells per well were seeded in 96-well plates (Corning Inc. Life Sciences, USA). The cells were allowed to attach overnight at 37 °C, after which the cell culture medium was removed and replaced with 100 μL of UnTHCPSi and UnTHCPSi–HA + nanoparticle suspensions at concentrations of 25, 50, and 100 μg mL −1, with 1× HBSS (pH 7.4) and 1% Triton X-100 as positive and negative controls, respectively. After incubating for 6 and 24 h at 37 °C, 100 μL of the assay reagent was added to each well and the number of viable cells was determined by measuring the luminescence from the living cells using a Varioskan Flash fluorometer (Thermo Fisher Scientific Inc., USA). The results presented correspond to the average of at least three independent measurements. Cellular internalization The cellular uptake and intracellular localization of the UnTHCPSi and UnTHCPSi–HA + nanoparticles was assessed by TEM. Approximately 10 5 cells per well of MCF-7 and MDA-MB-231 breast cancer cells were seeded in 24-well plates (Corning Inc. Life Sciences, USA) with each well containing a 13 mm round shaped coverslip and allowed to attach overnight at 37 °C. After removing the cell culture media, 500 μL per well of the nanoparticle suspensions at a concentration of 50 μg mL −1 were added to the wells. After an incubation period of 6 h at 37 °C, the nanoparticle suspensions were carefully removed and the samples were rinsed twice with HBSS–HEPES (pH 7.4). Then, the cells were fixed with 2.5% glutaraldehyde in 0.1 M PBS buffer (pH 7.4) for 1 h at room temperature, and subsequently washed twice with HBSS–HEPES (pH 7.4) and sodium cacodylate buffer (NaCac) for 3 min, prior to post-fixation with 1% osmium tetroxide in 0.1 M NaCac buffer (pH 7.4). Thereafter, the cells were dehydrated with 30–100% ethanol for 10 min and embedded in epoxy resin. Ultrathin sections with approximately 60 nm were sliced parallel to the coverslips, post-stained with uranyl acetate and lead citrate, and finally analyzed by TEM as described above. Flow cytometric analysis of CD44 expression and cellular association The expression of CD44 receptor in MCF-7 and MDA-MB-231 breast cancer cell lines was evaluated by flow cytometry according to a protocol adapted from the manufacturer's instructions, as described in the ESI. †

Welcome to the Humanist republic of Bimbolands! | Bimbolands Forum". bimbo.land . Retrieved 2021-03-25. Presumed portrait of Rosalie Duthé (1748–1830), called "the first officially recorded dumb blonde." J. Lesley, R. Hyman, N. English, J. B. Catterall and G. A. Turner, Glycoconjugate J., 1997, 14, 611–622 CrossRef CAS.

As the inner water phase, we used an aqueous suspension of THCPSiMPs with typical particle sizes of ca. 15 μm. To facilitate the dispersion of the THCPSiMPs, they were first wetted with ethanol and later suspended in an aqueous solution that contained 8 wt-% polyethylene glycol (PEG, 6 kDa) and 2 wt-% polyvinyl alcohol (PVA, 13–23 kDa) at a final concentration of 3 mg mL −1. The suspension was sonicated and poured into a syringe together with a magnetic stirrer allowing vigorous shaking of the suspension, thereby avoiding microparticle aggregation and sedimentation during the injection of the suspension within the microfluidic chip. Importantly, our microfluidic approach provides a hundred percent encapsulation efficiency as all the particles in the inner water phase ended-up in the cores of the double emulsions as shown in Fig. 1b. The middle oil phase contained 4.6 mg mL −1 of 1,2-dioleyl-sn-glycero-3-phosphocholine (DOPC, Avanti) dissolved in a mixture of chloroform and hexane at a volume ratio of 1 : 1.8, eventually containing 0.25 mole-% of 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine- N-(lissamine rhodamine B sulfonyl) (DHPE-Rh, Molecular Probes) to fluorescently label the lipid bilayer. The outer aqueous phase consisted of a 10 wt-% PVA (13–23 kDa) aqueous solution. M. Sarparanta, L. M. Bimbo, J. Rytkonen, E. Mäkilä, T. J. Laaksonen, P. Laaksonen, M. Nyman, J. Salonen, M. B. Linder, J. Hirvonen, H. A. Santos and A. J. Airaksinen, Mol. Pharmaceutics, 2012, 9, 654–663 CrossRef CAS PubMed. Kogal or, more correctly, kogyaru and ganguro carry similar connotations as a Japanese version of a "valley girl" or bimbo. Q. Xu, M. Hashimoto, T. T. Dang, T. Hoare, D. S. Kohane, G. M. Whitesides, R. Langer and D. G. Anderson, Small, 2009, 5, 1575–1581 CrossRef CAS PubMed. Around the same time, another drama erupted, though less intense: the travel agency drama. Although this travel agency has been teased since 2008, mentions of it were systematically banned as well as the members bringing it up. One player famously got banned for saying that the travel agency won't open til 2010... Turns out this player was right since the travel agency finally opened in late 2010.E. C. Wu, J. S. Andrew, L. Cheng, W. R. Freeman, L. Pearson and M. J. Sailor, Biomaterials, 2011, 32, 1957–1966 CrossRef CAS PubMed. D. Liu, H. Zhang, B. Herranz-Blanco, E. Mäkilä, V. Lehto, J. Salonen, J. Hirvonen and H. A. Santos, Small, 2014, 10, 2029–2038 CrossRef CAS PubMed. This form of leveraged buyout (LBO) is used to streamline the transition from one owner to the next with little interruption in business operations. Because of this U-turn, older fans felt rejected and cringed at the staff's attempt at communicating with young players as they exited the community. Bunch of Danganronpa kink requests from my tumblr. Language: English Words: 159,088 Chapters: 80/? Comments: 118 Kudos: 1,119 Bookmarks: 54 Hits: 120,620