Effect of cell shape change on the function and differentiation of rabbit mammary cells in culture

We examined the role of cell shape, cytodifferentiation, and tissue topography on the induction and maintenance of functional differentiation in rabbit mammary cells grown as primary cultures on two-dimensional collagen surfaces or in three-dimensional collagen matrices. Mammary glands from mid-pregnant rabbits were dissociated into single cells, and epithelial cells were enriched by isopycnic centrifugation. Small spheroids of epithelial cells (approximately 50 cells) that formed on a rotary shaker were plated on or embedded in collagen gels. The cells were cultured for 1 d in serum-containing medium and then for up to 25 d in chemically defined medium. In some experiments, epithelial monolayers on gels were mechanically freed from the dishes on day 2 or 5. These gels retracted and formed floating collagen gels. On attached collagen gels, flat monolayers of a single cell type developed within a few days. The cells synthesized DNA until the achievement of confluence but did not accumulate milk proteins. No morphological changes were induced by prolactin (PRL). On floating gels, two cell types appeared in the absence of cell proliferation. The cells in direct contact with the medium became cuboidal and developed intracellular organelles typical of secretory cells. PRL-induced lipogenesis, resulting in large fat droplets filling the apical cytoplasm and accumulation of casein and α-lactalbumin in vesicles surrounding the fat droplets. We detected tranferrin in the presence or absence of PRL intracellularly in small vesicles but also in the collagen matrix in contact with the cell layer. The second cell type, rich in microfilaments and reminiscent of the myoepithelial cells, was situated between the secretory cell layer and the collagen matrix. In embedding gels, the cells formed hollow ductlike structures, which grew continuously in size. Secretory cells formed typical lumina distended by secretory products. We found few microfilament-rich cells in contact with the collagen gels. Storage and secretion of fat, caseins and alpha-lactalbumin required the presence of PRL, whereas the accumulation and vectorial discharge of transferrin was prolactin independent. There was no differentiation gradient between the tip and the cent of the outgrowth, since DNA synthesis and milk protein storage were random along the tubular structures. These results indicate that establishment of functional polarity and induction of cytodifferentiation are influenced by the nature of the interaction of the cells with the collagen structure. The morphological differentiation in turn plays an important role in the synthesis, storage, and secretion of fat and milk proteins

Francesco 4. per la grazia di Dio duca di Modena, Reggio, Mirandola ec. ec. ec. arciduca d'Austria, principe reale d'Ungheria e Boemia

1 manifesto : 60 cm Incipit del testo: Fra gli oggetti più importanti, che fissarono la nostra attenzione nel sistemare i diversi rami della pubblica amministrazione, .. Data di emanazione in calce: Modena 3 ottobre 1825 Data di stampa presunta: 1825 (emanazione) Firmatario in calce In testa stemma del ducato di Modena e Reggio xilogr Testo stampato su due colonn

Conoscere è partecipare: digital public history, wiki e citizen humanities

La citizen science (CS), “scienza dei cittadini” o “scienza partecipata”, indica la partecipazione e il coinvolgimento attivo dei cittadini in attività di ricerca scientifica. Mentre la CS include le scienze naturali, come la biologia, la chimica e la fisica e la citizen social science si occupa delle società, le citizen humanities si applicano alle discipline storiche, letterarie, linguistiche e filosofiche. La digital public history, per la sua vocazione pubblica, partecipativa e collaborativa in un contesto digitale, si presenta come uno dei campi di applicazione privilegiata dalle citizen humanities. Il valore aggiunto delle citizen humanities dipende primariamente da due condizioni alla base della digital public history: la partecipazione degli storici che non provengono dall’accademia alla creazione di progetti collaborativi e la comunicazione dei risultati della ricerca storica al pubblico della rete. L’articolo intende riflettere su come la scrittura collaborativa, resa possibile dagli strumenti wiki, possa costituire un ottimo punto di partenza per una democrazia partecipata che si situi nel solco delle citizen humanities.Citizen science (CS), "citizen science" or "participatory science", refers to the participation and active involvement of citizens in scientific research activities. While CS includes natural sciences, such as biology, chemistry and physics, and citizen social science deals with societies, citizen humanities apply to historical, literary, linguistic and philosophical disciplines. Digital public history, due to its public, participatory and collaborative vocation in a digital context, presents itself as one of the privileged fields of application of citizen humanities. The added value of citizen humanities depends primarily on two conditions underlying the digital public history: the participation of historians who do not come from the academy in the creation of collaborative projects and the communication of the results of historical research to the public on the network. The article intends to reflect on how collaborative writing, made possible by wiki tools, can be an excellent starting point for a participatory democracy that is located in the wake of citizen humanities

Structural Mimicry in Class A G Protein-coupled Receptor Rotamer Toggle Switches

In this study, we tested the hypothesis that a CB1 TMH3-4-5-6 aromatic microdomain, which includes F3.25(190), F3.36(201), W5.43(280), and W6.48(357), is centrally involved in CB1 receptor activation, with the F3.36(201)/W6.48(357) interaction key to the maintenance of the CB1-inactive state. We have shown previously that when F3.36(201), W5.43(280), and W6.48(357) are individually mutated to alanine, a significant reduction in ligand binding affinity is observed in the presence of WIN 55,212-2 and SR141716A but not CP55,940 and anandamide. In the work presented here, we report a detailed functional analysis of the F3.36(201)A, F3.25(190)A, W5.43(280)A, and W6.48(357)A mutant receptors in stable cell lines created in HEK cells for agonist-stimulated guanosine 5′-3-O-(thio)triphosphate (GTPγS) binding and GIRK1/4 channel current effects in Xenopus oocytes where the mutant proteins were expressed transiently. The F3.36(201)A mutation showed statistically significant increases in ligand-independent stimulation of GTPγS binding versus wild type CB1, although basal levels for the W6.48(357)A mutant were not statistically different from wild type CB1. F3.36(201)A demonstrated a limited activation profile in the presence of multiple agonists. In contrast, enhanced agonist activation was produced by W6.48(357)A. These results suggest that a F3.36(201)/W6.48(357)-specific contact is an important constraint for the CB1-inactive state that may need to break during activation. Modeling studies suggest that the F3.36(201)/W6.48(357) contact can exist in the inactive state of CB1 and be broken in the activated state via a χ1 rotamer switch (F3.36(201) trans, W6.48(357) g+) → (F3.36(201) g+, W6.48(357) trans). The F3.36(201)/W6.48(357) interaction therefore may represent a “toggle switch” for activation of CB1

[Carta al rey Felipe IV [Manuscrito] : relatándole los atropellos y abusos cometidos en el Reino y ciudad de Valencia por el Capitán General D. Luigi Reggio y Branciforte, Principe de Campofiorito]

Alcance y contenido: Copia ms. de la carta enviada por D. Luis Cortés y Cantos, a Felipe IV, narrándole los desmanes y delitos llevados a cabo por el entonces Cap. General del Reino de Valencia y sus ministros y asesores, tanto a la Hacienda Real como a los habitantes del reino.Nombre del Capitan General del Reino de Valencia, deducido de las fechas que aparecen en la carta.La carta original se dato en: "Villareal y julio 20 de 1734"Documento guillotinado en el corte superior aunque sin afectar al text

Cancer cells adapt FAM134B/BiP mediated ER-phagy to survive hypoxic stress

In the tumor microenvironment, cancer cells experience hypoxia resulting in the accumulation of misfolded/unfolded proteins largely in the endoplasmic reticulum (ER). Consequently, ER proteotoxicity elicits unfolded protein response (UPR) as an adaptive mechanism to resolve ER stress. In addition to canonical UPR, proteotoxicity also stimulates the selective, autophagy-dependent, removal of discrete ER domains loaded with misfolded proteins to further alleviate ER stress. These mechanisms can favor cancer cell growth, metastasis, and long-term survival. Our investigations reveal that during hypoxia-induced ER stress, the ER-phagy receptor FAM134B targets damaged portions of ER into autophagosomes to restore ER homeostasis in cancer cells. Loss of FAM134B in breast cancer cells results in increased ER stress and reduced cell proliferation. Mechanistically, upon sensing hypoxia-induced proteotoxic stress, the ER chaperone BiP forms a complex with FAM134B and promotes ER-phagy. To prove the translational implication of our mechanistic findings, we identified vitexin as a pharmacological agent that disrupts FAM134B-BiP complex, inhibits ER-phagy, and potently suppresses breast cancer progression in vivo

Crucial Positively Charged Residues for Ligand Activation of the GPR35 Receptor

GPR35 is a G protein-coupled receptor expressed in the immune, gastrointestinal, and nervous systems in gastric carcinomas and is implicated in heart failure and pain perception. We investigated residues in GPR35 responsible for ligand activation and the receptor structure in the active state. GPR35 contains numerous positively charged amino acids that face into the binding pocket that cluster in two distinct receptor regions, TMH3-4-5-6 and TMH1-2-7. Computer modeling implicated TMH3-4-5-6 for activation by the GPR35 agonists zaprinast and pamoic acid. Mutation results for the TMH1-2-7 region of GPR35 showed no change in ligand efficacies at the K1.32A, R2.65A, R7.33A, and K7.40A mutants. However, mutation of arginine residues in the TMH3-4-5-6 region (R4.60, R6.58, R3.36, R(164), and R(167) in the EC2 loop) had effects on signaling for one or both agonists tested. R4.60A resulted in a total ablation of agonist-induced activation in both the β-arrestin trafficking and ERK1/2 activation assays. R6.58A increased the potency of zaprinast 30-fold in the pERK assay. The R(167)A mutant decreased the potency of pamoic acid in the β-arrestin trafficking assay. The R(164)A and R(164)L mutants decreased potencies of both agonists. Similar trends for R6.58A and R(167)A were observed in calcium responses. Computer modeling showed that the R6.58A mutant has additional interactions with zaprinast. R3.36A did not express on the cell surface but was trapped in the cytoplasm. The lack of surface expression of R3.36A was rescued by a GPR35 antagonist, CID2745687. These results clearly show that R4.60, R(164), R(167), and R6.58 play crucial roles in the agonist initiated activation of GPR35

Folla real que en celebridad de los años de la magestad de la reina Isabela ... mando executar el ... principe de Campoflorido ... Capitan General del reino de Valencia, &c. en ... su palacio el dia 25 de octubre de este año 1728 : [sainetes]

Texto en español e italianoSubtít. y menciónes de resp. tomados de p. [3] y 1Fecha de 1728 tomada del títSign.: []2, [A]-G4Port. con orla ti