No description available.

No description available.

Cell contractility is mainly imagined as a force dipole-like interaction based on actin stress fibers that pull on cellular adhesion sites. Here, we present a different type of contractility based on isotropic contractions within the actomyosin cortex. Measuring mechanosensitive cortical contractility of suspended cells among various cell lines allowed us to exclude effects caused by stress fibers. We found that epithelial cells display a higher cortical tension than mesenchymal cells, directly contrasting to stress fiber-mediated contractility. These two types of contractility can even be used to distinguish epithelial from mesenchymal cells. These findings from a single cell level correlate to the rearrangement effects of actomyosin cortices within cells assembled in multicellular aggregates. Epithelial cells form a collective contractile actin cortex surrounding multicellular aggregates and further generate a high surface tension reminiscent of tissue boundaries. Hence, we suggest this intercellular structure as to be crucial for epithelial tissue integrity. In contrast, mesenchymal cells do not form collective actomyosin cortices reducing multicellular cohesion and enabling cell escape from the aggregates.

Baggalútar are well-rounded spheroids that typically measures 16-18 mm in size and have a brownreddish appearance. They can be found in the Hvalfjörður bay in SW Iceland. There are manydescriptions in literature and on the internet that Baggalútar are volcanic spherulites formed by quartzor cristobalite spheres growing out of a common centre and there is also a broadly accepted consensusof this theory. However, despite this consensus that Baggalútar are volcanic spherulites there exist nodetailed investigation of their origin. The aim of the thesis is to investigate what Baggalútar is exactlyand how they form. This is achieved by using a combination of petrographic observations with apolarization microscope, mineral chemistry from electron microprobe analysis and measurements oftheir magnetic properties. The results are compared with different geological and anthropological spheroids, spherulites,nodules and concretions. The petrographic observations show that they predominantly consist of finegrained basaltic tephra (groundmass) together with zeolites infilling voids. Analyses of mineralchemistry indicate that the groundmass consists of augitic pyroxene, plagioclase, and two differentoxides where one classifies as titanomagnetite. The magnetic measurements support this by showing aCurie temperature at approximately 460-470 °C which is likely to be titano-magnetite. Although thedifferent geological and anthropological processes that typically results spheroidal shapes have somesimilarities that could explain the formation of baggalútar, most of these can be excluded for differentreasons. The internal textures of baggalútar strongly indicate that the shape is controlled by externalfactors, like weathering or erosion from beach outcrops. This could explain the spherical shape of asingle baggalút, but it fails to explain the spheroidal shapes of individual baggalútar joined together inclusters. / Baggalútar är väl rundade sfäroider som vanligtvis mäter 16–18 mm i storlek och har ett brunt rödaktigtutseende. De hittas i Hvalfjörðurbukten i SW Island. Det finns många beskrivningar i litteraturen ochpå internet som säger att Baggalútar är vulkaniska sfäruliter som bildas av kvarts- eller kristobalitsfärersom växer fram ur ett gemensamt centrum och det finns också en allmänt accepterad konsensus omdenna teori. Men trots denna konsensus om att Baggalútar är vulkaniska sfäruliter finns det ingendetaljerad undersökning av deras ursprung. Syftet med avhandlingen är att undersöka exakt vadBaggalútar är och hur de bildas. Detta uppnås genom att använda en kombination av petrografiskaobservationer med ett polarisationsmikroskop, mineralkemi från elektronmikrosondanalys ochmätningar av deras magnetiska egenskaper. Resultaten jämförs med olika geologiska och antropologiska sfäroider, sfäruliter, noduler ochkonkretioner. De petrografiska observationerna visar att de till övervägande del består av finkornigbasaltisk tefra (grundmassa) tillsammans med zeoliter som fyller ut tomrum. Analyser av mineralkemivisar att grundmassan består av augitisk pyroxen, plagioklas och två olika oxider där den enaklassificeras som titanomagnetit. De magnetiska mätningarna stödjer detta genom att visa en Curietemperatur på cirka 460–470 °C som sannolikt är titanomagnetit. Även om de olika geologiska ochantropologiska processerna som vanligtvis resulterar i sfäroida former har vissa likheter som kanförklara bildandet av baggalútar, kan de flesta av dessa uteslutas av olika anledningar. Baggalútars inretexturer indikerar starkt att formen styrs av yttre faktorer, som väderpåverkan eller erosion frånstrandhällar. Detta kan förklara den sfäriska formen av en enda baggalút, men det misslyckas med attförklara de sfäriska formerna av individuella baggalútar i sammanfogade kluster.

Germline mutations in the tumor suppressor gene PTEN cause PTEN Hamartoma Tumor Syndrome (PHTS). Pediatric patients with PHTS frequently develop lipomas. Treatment attempts with the mTORC1 inhibitor rapamycin were unable to reverse lipoma growth. Recently, lipomas associated with PIK3CA-related overgrowth syndrome were successfully treated with the novel PI3K inhibitor alpelisib. Here, we tested whether alpelisib has growth-restrictive effects and induces cell death in lipoma cells. We used PTEN-haploinsufficient lipoma cells from three patients and treated them with alpelisib alone or in combination with rapamycin. We tested the effect of alpelisib on viability, proliferation, cell death, induction of senescence, adipocyte differentiation, and signaling at 1–100 M alpelisib. Alpelisib alone or in combination with rapamycin reduced proliferation in a concentrationand time-dependent manner. No cell death but an induction of senescence was detected after alpelisib incubation for 72 h. Alpelisib treatment led to a reduced phosphorylation of AKT, mTOR, and ribosomal protein S6. Rapamycin treatment alone led to increased AKT phosphorylation. This effect could be reversed by combining rapamycin with alpelisib. Alpelisib reduced the size of lipoma spheroids by attenuating adipocyte differentiation. Since alpelisib was well tolerated in first clinical trials, this drug alone or in combination with rapamycin is a potential new treatment option for PHTS-related adipose tissue overgrowth.

No description available.

No description available.

It was postulated that 3D cell culture models more accurately reflect the complex tissue physiology and morphology in comparison to 2D cell monolayers. Currently, there is a shortage of well-characterized and easily maintainable high-throughput experimental models of the human placenta. Here, we characterized three different 3D cultures (e.g., spheroids) derived from trophoblast cell lines and studied their functionality in comparison to primary fetal trophoblasts and placental tissue. The spheroid growth rates of JEG3, BeWo and HTR8/SVneo cell lines were similar among each other and were significantly larger in comparison to primary trophoblast spheroids. All spheroids exhibited migratory properties and shortest distances were registered for JEG3 spheroids. Even though all spheroids displayed invasive capabilities, only the invasive features of HTR8/SVneo spheroids resulted in specific branching. This was in agreement with the invasive properties of the spheroids obtained from primary trophoblasts. Human chorionic gonadotropin production was highest in JEG3 spheroids and only increased when stimulated with cAMP and forskolin in BeWo, but not HTR8/SVneo spheroids. The gene expression analysis confirmed that 3D trophoblast cell cultures and especially HTR8/SVneo spheroids showed considerable similarities with the gene expression profile of primary placental tissue. This study offers a broad characterization of 3D trophoblast spheroids that, in turn, can help in selecting the best model depending on the scientific question that needs to be answered.

O diferencial da cultura de células em 3-dimensões é permitir que as células explorem as 3-dimensões do espaço, aumentando assim as interações com o ambiente e entre as células. Em estudos relacionados à biologia do câncer de mama, vem ganhando espaço a utilização de esferóides para estudos que visam à compreensão da morfogênese do espaço luminal. Neste trabalho foi mostrado que as células MCF-7 reorganizam-se em estruturas tubulares e acinares. Em ambas as situações, a formação do lúmen veio acompanhada pelo estabelecimento de uma camada de células polarizadas, arranjo este muito semelhante ao encontrado em glândulas mamárias. Os resultados apresentados apontam para a existência de uma população de células na linhagem MCF-7 que não estão totalmente comprometidas ao fenótipo tumoral. Mantidos diferenciados, os esferóides de células MCF-7 apontam como um novo modelo para estudos relacionados à formação do lúmen, permitindo assim explorar o papel de diferentes vias como as relacionadas a apoptose, autofagia, diferenciação e sobrevivência celular. / As a particularity, a 3D cell culture permits cells to explore the three dimensions of the space thereby increasing cell-cell interactions, as well as interaction with the environment. In studies related to breast cancer biology, spheroids are becoming widely used in the aim to comprehend luminal space morphogenesis. We showed that MCF-7 cells reorganize themselves in tubular and acinar structures. In both situations, lumen formation was accompanied by the establishment of a layer of polarized cells, an arrangement that is very similar to that of breast glands. The presented results suggest the existence of an MCF cell line population not completely committed to the tumor phenotype. When maintained as differentiated, MCF-7 cell spheroids can be a new model for studies regarding lumen formation, thereby exploring the role of diiferent pathways, such as those related to cell apoptosis, autophagy, differentiation and survival.

Intracellular redox potential (IRP) is a measure of how oxidising or reducing the environment is within a cell. It is a function of numerous factors including redox couples, antioxidant enzymes and reactive oxygen species. Disruption of the tightly regulated redox status has been linked to the initiation and progression of cancer. However, there is very limited knowledge about the quantitative nature of the redox potential and pH gradients that exist in cancer tumour models. Multicellular tumour spheroids (MTS) are three-dimensional cell cultures that possess their own microenvironments, similar to those found in tumours. From the necrotic core to the outer proliferating layer there exist gradients of oxygen, lactate, pH and drug penetration. Tumours also have inadequate vasculature resulting in a state of hypoxia. Hypoxia is a key player in metabolic dysregulation but can also provide cells with resistance against cancer treatments, particularly chemotherapy and radiation therapy. The primary hypoxia regulators are HIFs (Hypoxia Inducible Factors) which under low O2 conditions bind a hypoxia response element, inhibiting oxidative phosphorylation and upregulating glycolysis which has two significant implications: the first is an increase in levels of NADPH/NADH, the main electron donors found in cells which impacts the redox state, whilst the second is a decrease in intracellular pH (pHi) because of increased lactate production. Thus, redox state and intracellular pHi can be used as indicators of metabolic changes within 3D cultures and provide insight into cellular response to therapy. Surface-Enhanced Raman Spectroscopy (SERS) provides a real-time, high resolution method of measuring pHi and IRP in cell culture. It allows for quick and potentially portable analysis of MTS, providing a new platform for monitoring response to drugs and therapy in an unobtrusive manner. Redox and pH-active probes functionalised to Au nanoshells were readily taken up by prostate cancer cell lines and predominantly found to localise in the cytosol. These probes were characterised by density functional theory and spectroelectrochemistry, and their in vitro behaviour modelled by the chemical induction of oxidative and reductive stress. Next, targeting nanosensors to different zones of the MTS allowed for spatial quantification of redox state and pHi throughout the structure and the ability to map the effects of drug treatments on MTS redox biology. The magnitude of the potential gradient can be quantified as free energy (ΔG) and used as a measurement of MTS viability. Treatment of PC3 MTS with staurosporine, an apoptosis inducer, was accompanied by a decrease in free energy gradients over time, whereas treatment of MTS with cisplatin, a drug to which they are resistant, showed an increase in viability indicating a compensatory mechanism and hence resistance. Finally, using this technique the effects of ionising radiation on IRP and pHi in the tumour model was explored. Following exposure to a range of doses of x-ray radiation, as well as single and multi-fractionated regimes, IRP and pHi were measured and MTS viability assessed. Increased radiation dosage diminished the potential gradient across the MTS and decreased viability. Similarly, fractionation of a single large dose was found to enhance MTS death. This novel SERS approach therefore has the potential to not only be used as a mode of drug screening and tool for drug development, but also for pre-clinical characterisation of tumours enabling clinicians to optimise radiation regimes in a patient-specific manner.