Girl meets girl: sexual sitings in lesbian romantic comedies

Hollywood romantic comedies are, by and large, an ideologically conservative genre. Based around gender stereotypes and the idealised pursuit, however disguised, of heteropatriarchal monogamy, Hollywood romantic comedies offer countless variations of heteronormative ‘intimacy’. How, then, does the shift from ‘boy meets girl’ to ‘girl meets girl’ in lesbian romantic comedies—a genre that emerged in 1994 with the release of films like Bar Girls and Go Fish—effect the representation of intimacy? This chapter focuses on Better than Chocolate to investigate how lesbian intimacies, and lesbian sex in particular, occupy space. Where are lesbian intimacies sited and what, if any, negotiations of space are triggered through the embodiment of those intimacies? Ultimately, this chapter argues that through an unusually explicit emphasis on sex, Better than Chocolate draws attention to the limited public mobility of lesbian intimacies through a consistent siting of lesbian sex as a site of spatial negotiation

To lift or recut: changing trends in LASIK enhancement.

PURPOSE: To report serious complications caused by recutting laser in situ keratomileusis (LASIK) flaps for enhancement and reconsider the current preferred method of LASIK enhancement. SETTING: Multiple surgeon practices. METHODS: This retrospective noncomparative nonconsecutive case series comprised LASIK patients in the private practices of 9 experienced refractive surgeons and those reported in a survey of refractive surgeons. Case histories, refractions, corneal topographies, slitlamp photographs, and measurements of uncorrected and best corrected (BCVA) visual acuity after recutting LASIK flaps were collected. Surveys of refractive surgeons and an analysis of changing practice trends among the authors and these surgeons were assessed. RESULTS: In 12 cases, significant loss of BCVA and subjective visual difficulties resulted from recutting LASIK flaps. Most surveyed surgeons had changed their practice from recutting to lifting flaps even 9 to 10 years postoperatively with good results. CONCLUSION: Recutting flaps for enhancement should be avoided unless other alternatives are unavailable

Wnt isoform-specific interactions with coreceptor specify inhibition or potentiation of signaling by LRP6 antibodies.

β-Catenin-dependent Wnt signaling is initiated as Wnt binds to both the receptor FZD and coreceptor LRP5/6, which then assembles a multimeric complex at the cytoplasmic membrane face to recruit and inactivate the kinase GSK3. The large number and sequence diversity of Wnt isoforms suggest the possibility of domain-specific ligand-coreceptor interactions, and distinct binding sites on LRP6 for Wnt3a and Wnt9b have recently been identified in vitro. Whether mechanistically different interactions between Wnts and coreceptors might mediate signaling remains to be determined. It is also not clear whether coreceptor homodimerization induced extracellularly can activate Wnt signaling, as is the case for receptor tyrosine kinases. We generated monoclonal antibodies against LRP6 with the unexpected ability to inhibit signaling by some Wnt isoforms and potentiate signaling by other isoforms. In cell culture, two antibodies characterized further show reciprocal activities on most Wnts, with one antibody antagonizing and the other potentiating. We demonstrate that these antibodies bind to different regions of LRP6 protein, and inhibition of signaling results from blocking Wnt binding. Antibody-mediated dimerization of LRP6 can potentiate signaling only when a Wnt isoform is also able to bind the complex, presumably recruiting FZD. Endogenous autocrine Wnt signaling in different tumor cell lines can be either antagonized or enhanced by the LRP6 antibodies, indicating expression of different Wnt isoforms. As anticipated from the roles of Wnt signaling in cancer and bone development, antibody activities can also be observed in mice for inhibition of tumor growth and in organ culture for enhancement of bone mineral density. Collectively, our results indicate that separate binding sites for different subsets of Wnt isoforms determine the inhibition or potentiation of signaling conferred by LRP6 antibodies. This complexity of coreceptor-ligand interactions may allow for differential regulation of signaling by Wnt isoforms during development, and can be exploited with antibodies to differentially manipulate Wnt signaling in specific tissues or disease states

Spatiotemporally Regulated Protein Kinase A Activity Is a Critical Regulator of Growth Factor-Stimulated Extracellular Signal-Regulated Kinase Signaling in PC12 Cells▿

PC12 cells exhibit precise temporal control of growth factor signaling in which stimulation with epidermal growth factor (EGF) leads to transient extracellular signal-regulated kinase (ERK) activity and cell proliferation, whereas nerve growth factor (NGF) stimulation leads to sustained ERK activity and differentiation. While cyclic AMP (cAMP)-mediated signaling has been shown to be important in conferring the sustained ERK activity achieved by NGF, little is known about the regulation of cAMP and cAMP-dependent protein kinase (PKA) in these cells. Using fluorescence resonance energy transfer (FRET)-based biosensors localized to discrete subcellular locations, we showed that both NGF and EGF potently activate PKA at the plasma membrane, although they generate temporally distinct activity patterns. We further show that both stimuli fail to induce cytosolic PKA activity and identify phosphodiesterase 3 (PDE3) as a critical regulator in maintaining this spatial compartmentalization. Importantly, inhibition of PDE3, and thus perturbation of the spatiotemporal regulation of PKA activity, dramatically increases the duration of EGF-stimulated nuclear ERK activity in a PKA-dependent manner. Together, these findings identify EGF and NGF as potent activators of PKA activity specifically at the plasma membrane and reveal a novel regulatory mechanism contributing to the growth factor signaling specificity achieved by NGF and EGF in PC12 cells

Derivation of sarcomas from mesenchymal stem cells via inactivation of the Wnt pathway

Malignant fibrous histiocytoma (MFH), now termed high-grade undifferentiated pleomorphic sarcoma, is a commonly diagnosed mesenchymal tumor, yet both the underlying molecular mechanisms of tumorigenesis and cell of origin remain unidentified. We present evidence demonstrating that human mesenchymal stem cells (hMSCs) are the progenitors of MFH. DKK1, a Wnt inhibitor and mediator of hMSC proliferation, is overexpressed in MFH. Using recombinant proteins, antibody depletion, and siRNA knockdown strategies of specific Wnt elements, we show that DKK1 inhibits hMSC commitment to differentiation via Wnt2/β-catenin canonical signaling and that Wnt5a/JNK noncanonical signaling regulates a viability checkpoint independent of Dkk1. Finally, we illustrate that hMSCs can be transformed via inhibition of Wnt signaling to form MFH-like tumors in nude mice, and conversely, MFH cells in which Wnt signaling is appropriately reestablished can differentiate along mature connective tissue lineages. Our results provide mechanistic insights regarding the cell of origin of MFH, establish what we believe is a novel tumor suppressor role for Wnt signaling, and identify a potential therapeutic differentiation strategy for sarcomas

Deconstructing the ßcatenin destruction complex: mechanistic roles for the tumor suppressor APC in regulating Wnt signaling

APC is a key tumor suppressor and Wnt signaling regulator, but its mechanism of action remains mysterious. We combined parallel assays in Drosophila and cultured human colon cancer cell lines to test hypotheses regarding APC function and to develop novel hypotheses, using mutants altering its structure in specific ways