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Medulloblastoma (MB) is a heterogeneous disease in which neoplastic cells and associated immune cells contribute to disease progression. We aimed to determine the influence of neoplastic and immune cell diversity on MB biology in patient samples and animal models. To better characterize cellular heterogeneity in MB we used single-cell RNA sequencing, immunohistochemistry and deconvolution of transcriptomic data to profile neoplastic and immune populations in patient samples and animal models across childhood MB subgroups. Neoplastic cells cluster primarily according to individual sample of origin which is influenced by chromosomal copy number variance. Harmony alignment reveals novel MB subgroup/subtype-associated subpopulations that recapitulate neurodevelopmental processes, including photoreceptor and glutamatergic neuron-like cells in molecular subgroups GP3 and GP4, and a specific nodule-associated neuronally-differentiated subpopulation in subgroup molecular SHH. We definitively chart the spectrum of MB immune cell infiltrates, which include subpopulations that recapitulate developmentally-related neuron-pruning and antigen presenting myeloid cells. MB cellular diversity matching human samples is mirrored in subgroup-specific mouse models of MB. These findings provide a clearer understanding of the diverse neoplastic and immune cell subpopulations that constitute the MB microenvironment. These findings provide a clearer understanding of the diverse neoplastic and immune cell subpopulations that constitute the MB microenvironment. Waterpipe tobacco (WT) smoking by young adults remains high and misperceptions are common. Product warnings can increase knowledge of harms and reduce use. The goal of this study was to test warning statements, including the FDA-required nicotine warning (prior to implementation), on young adults' thinking about harms of and discouragement from WT smoking. We conducted a between-subjects experiment in a nationally representative telephone survey of 1,152 young adults ages 18-29. https://www.selleckchem.com/products/anidulafungin-ly303366.html Participants were randomly assigned to hear one of five warning statements and reported how much, on a 4-point scale, the warning made them think about the harms and discouraged them from WT smoking. The sample was 36.8% female, 57.8% white, 20.2% Black, 24.1% Hispanic, with a mean age of 23.2 (SE=0.25). Under half (43.5%) had ever smoked WT. There were significant differences among the statements on both thinking about harms (p<.0001) and discouragement (p<.0001). The FDA-required 'nicotine' warning led to the lowest thinwarning is the least effective at making young adults think about the harms of and discouraging waterpipe tobacco smoking. The FDA and other countries should consider requiring warnings to cover a broader range of health harms, misperceptions, and possibly comparisons to cigarettes.Physiologically based pharmacokinetic (PBPK) models are commonly used in risk assessments to perform inter- and intraspecies extrapolations as well as to extrapolate between different dosing scenarios; however, they must first undergo quality assurance review, which can be a time-consuming process, especially when model code is not readily available. We developed and implemented (using R and MCSim) a PBPK model template capable of replicating published model results for several chemical-specific PBPK models. This model template allows for faster quality assurance review because the general model equations only need to be reviewed once, and application to a specific chemical then only requires reviewing input parameters. The model template can implement PBPK models with oral and intravenous exposure routes, varying numbers of tissue compartments, renal reabsorption, and multiple elimination pathways, including fecal, urinary, and biliary. Using the model template, we reproduced published model simulation results for perfluorohexanesulfonic acid, perfluorononanoic acid, perfluorodecanoic acid, perfluorooctanoate, and perflouorooctane sulfonate. We also show that the template can be a useful tool for identifying potential model errors. Thus, the model template allows for faster evaluation and review of published PBPK models and provides a proof of concept for using this approach with broader classes of chemical-specific PBPK models.In most flowering plants, asymmetric cell division of zygotes is the initial step to establish the apical-basal axis. In the Arabidopsis zygote, vacuolar accumulation at the basal cell end is crucial to ensure zygotic division asymmetry. Despite the importance, it was unclear whether this polar vacuolar distribution is achieved by predominant biogenesis at the basal region or by directional movement after biogenesis. Here, we found that apical and basal vacuolar contents are dynamically exchanged via tubular vacuolar network and the vacuoles gradually migrate towards the basal end. The mutant of a vacuolar membrane protein, SHOOT GRAVITROPISM2 (SGR2), failed to form tubular vacuoles, and the mutant of a putative vacuolar fusion factor, VESICLE TRANSPORT THROUGH INTERACTION WITH T-SNARES11 (VTI11), could not flexibly rearrange the vacuolar network. Both mutants failed to exchange the apical and basal vacuolar contents and to polarly migrate the vacuoles, resulting in more symmetric division of zygotes. Additionally, we observed that in contrast to sgr2, the zygotic defects of vti11 were rescued by the pharmacological depletion of phosphatidylinositol 3-phosphate (PI3P), a distinct phospholipid in the vacuolar membrane. Thus, SGR2 and VTI11 have individual sites of action in the zygotic vacuolar membrane processes. Further, a mutant of YODA (YDA) MAPKK kinase, a core component of the embryonic axis formation pathway, generated proper vacuolar network; however, it failed to migrate the vacuoles towards the basal region, which suggests impaired directional cues. Overall, we conclude that SGR2- and VTI11-dependent vacuolar exchange and YDA-mediated directional migration are necessary to achieve polar vacuolar distribution in the zygote.Calcium (Ca2+)-dependent signalling plays a well-characterized role in the response to different environmental stimuli, in both plant and animal cells. In the model organism for green algae, Chlamydomonas reinhardtii, Ca2+ signals were reported to have a crucial role in different physiological processes, such as stress responses, photosynthesis, and flagella functions. Recent reports identified the underlying components of the Ca2+ signalling machinery at the level of specific subcellular compartments and reported in vivo imaging of cytosolic Ca2+ concentration in response to environmental stimuli. The characterization of these Ca2+-related mechanisms and proteins in C. reinhardtii is providing knowledge on how microalgae can perceive and respond to environmental stimuli, but also on how this Ca2+ signalling machinery has evolved. Here, we review current knowledge on the cellular mechanisms underlying the generation, shaping, and decoding of Ca2+ signals in C. reinhardtii, providing an overview of the known and possible molecular players involved in the Ca2+ signalling of its different subcellular compartments.
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