LGR5+ epithelial tumor stem-like cells generate a 3D-organoid model for ameloblastoma

Ameloblastoma (AM) is a benign but locally aggressive tumor with high recurrences. Currently, underlying pathophysiology remains elusive, and radical surgery remains the most definitive treatment with severe morbidities. We have recently reported that AM harbors a subpopulation of tumor epithelial stem-like cells (AM-EpiSCs). Herein, we explored whether LGR5+ epithelial cells in AM possess stem-like cell properties and their potential contribution to pathogenesis and recurrence of AM. We found that LGR5 and stem cell-related genes were co-expressed in a subpopulation of AM epithelial cells both in vivo and in vitro, which were enriched under 3D-spheroid culture. As compared to LGR5− counterparts, LGR5+ AM epithelial cells showed increased expression of various EMT- and stemness-related genes, and functionally, exhibited increased capacity to form 3D-spheroids and generate human tumor 3D organoids, which recapitulated the histopathologic features of distinct subtypes of solid AM, thus, contributing a useful human tumor platform for targeted therapeutic screening. Treatment with a selective BRAFV600E inhibitor, vemurafenib, unexpectedly enriched the subpopulation of LGR5+ AM-EpiSCs in tumor 3D organoids, which may have explained therapeutic resistances and recurrences. These findings suggest that LGR5+ AM-EpiSCs play a pivotal role in pathogenesis and progression of AM and targeted inhibition of both BRAF and LGR5 potentially serves a novel nonsurgical adjuvant therapeutic approach for this aggressively benign jaw tumor. © 2020, The Author(s)

Roles of the DYRK Kinase Pom2 in Cytokinesis, Mitochondrial Morphology, and Sporulation in Fission Yeast

Pom2 is predicted to be a dual-specificity tyrosine-phosphorylation regulated kinase (DYRK) related to Pom1 in Schizosaccharomyces pombe. DYRKs share a kinase domain capable of catalyzing autophosphorylation on tyrosine and exogenous phosphorylation on serine/threonine residues. Here we show that Pom2 is functionally different from the well-characterized Pom1, although they share 55% identity in the kinase domain and the Pom2 kinase domain functionally complements that of Pom1. Pom2 localizes to mitochondria throughout the cell cycle and to the contractile ring during late stages of cytokinesis. Overexpression but not deletion of pom2 results in severe defects in cytokinesis, indicating that Pom2 might share an overlapping function with other proteins in regulating cytokinesis. Gain and loss of function analyses reveal that Pom2 is required for maintaining mitochondrial morphology independently of microtubules. Intriguingly, most meiotic pom2Δ cells form aberrant asci with meiotic and/or forespore membrane formation defects. Taken together, Pom2 is a novel DYRK kinase involved in regulating cytokinesis, mitochondrial morphology, meiosis, and sporulation in fission yeast

Datacenter-Network-Aware Online Load Balancing in MapReduce

MapReduce has emerged as a powerful tool for distributed and scalable processing of voluminous data. Different from earlier heuristics in the very late reduce stage or after seeing all the data, we address the skew from the beginning of data input, and make no assumption about a priori knowledge of the data distribution nor require synchronized operations. We show that the optimal strategy is a constrained version of online minimum makespan and, in the MapReduce context where pairs with identical keys must be scheduled to the same machine, we propose an online algorithm with a provable 2-competitive ratio. We further suggest a sample-based enhancement, which, probabilistically, achieves a 3/2-competitive ratio with a bounded error. Examine the project again, we found that the datacenter network could be a bottleneck in the shuffle subphase of MapReduce. This could potentially lead to a poor overall performance even with a balanced workload and thus needed to be addressed. Earlier studies either assume the network inside a datacenter is of negligible delay and infinite capacity, or use a hop count as the network cost measurement. We consider the realistic bandwidth constraints in real world datacenter networks and propose an effective solution toward near optimal datacenter-network-aware load balancing

Online Load Balancing for MapReduce with Skewed Data Input

Abstract—MapReduce has emerged as a powerful tool for distributed and scalable processing of voluminous data. In this paper, we, for the first time, examine the problem of accommodating data skew in MapReduce with online operations. Different from earlier heuristics in the very late reduce stage or after seeing all the data, we address the skew from the beginning of data input, and make no assumption about a priori knowledge of the data distribution nor require synchronized operations. We examine the input in a continuous fashion and adaptively assign tasks with a load-balanced strategy. We show that the optimal strategy is a constrained version of online minimum makespan and, in the MapReduce context where pairs with identical keys must be scheduled to the same machine, there is an online algorithm with a provable 2-competitive ratio. We further suggest a sample-based enhancement, which, probabilistically, achieves a 3/2-competitive ratio with a bounded error. I

Overexpression of the Rybp Gene Inhibits Differentiation of Bovine Myoblasts into Myotubes

RING1 and YY1 binding protein (Rybp) genes inhibit myogenesis in mice, but there are no reports on the effects of these genes in cattle. The aim of this study is to investigate the roles of the Rybp gene on bovine skeletal muscle development and myoblast differentiation. In the present study, the Rybp gene was overexpressed in bovine myoblasts via adenovirus. RNA-seq was performed to screen differentially expressed genes (DEGs). The results showed that overexpressing the Rybp gene inhibits the formation of myotubes. The morphological differences in myoblasts began on the second day and were very significant 6 days after adenovirus induction. A total of 1311 (707 upregulated and 604 downregulated) DEGs were screened using RNA-seq between myoblasts with added negative control adenoviruses (AD-NC) and Rybp adenoviruses (AD-Rybp) after 6 days of induction. Gene ontology (GO) and KEGG analysis revealed that the downregulated DEGs were mainly involved in biological functions related to muscle, and, of the 32 pathways, those associated with muscle development were significantly enriched for the identified DEGs. This study can not only provide a theoretical basis for the regulation of skeletal muscle development in cattle by exploring the roles of the Rybp gene in myoblast differentiation, but it can also lay a theoretical foundation for molecular breeding of beef cattle

Ultrasensitive Resonance Scattering (RS) Spectral Detection for Trace Tetracycline in Milk Using Aptamer-Coated Nanogold (ACNG) as a Catalyst

This paper reports an ultrasensitive resonance scattering (RS) method to detect tetracycline (TET) in milk based on the competition of aptamers between nanogold and TET, aggregation of naked nanogold, nanocatalytic Fehling reaction, and RS signals of catalytic product Cu₂O cubic. The detection principle was confirmed by the nanoparticle size analyzer (NANOS), scanning electron microscopy (SEM), and X-ray diffraction (XRD). The variations of RS intensity had good linear correlation with TET concentrations, and the limit of detection was calculated as 11.6 nM. The proposed method was successfully applied for analysis of TET in milk, with total recoveries ranging from 105 to 109%

Redistribution of the tight junction protein ZO-1 during physiological shedding of mouse intestinal epithelial cells

We questioned how tight junctions contribute to intestinal barrier function during the cell shedding that is part of physiological cell renewal. Intravital confocal microscopy studied the jejunal villus epithelium of mice expressing a fluorescent zonula occludens 1 (ZO-1) fusion protein. Vital staining also visualized the cell nucleus (Hoechst staining) or local permeability to luminal constituents (Lucifer Yellow; LY). In a cell fated to be shed, ZO-1 redistributes from the tight junction toward the apical and then basolateral cell region. ZO-1 rearrangement occurs 15 ± 6 min (n = 28) before movement of the cell nucleus from the epithelial layer. During cell extrusion, permeation of luminal LY extends along the lateral intercellular spaces of the shedding cell only as far as the location of ZO-1. Within 3 min after detachment from the epithelial layer, nuclear chromatin condenses. After cell loss, a residual patch of ZO-1 remains in the space previously occupied by the departed cell, and the size of the patch shrinks to 14 ± 2% (n = 15) of the original cell space over 20 min. The duration of cell shedding measured by nucleus movement (14 ± 1 min) is much less than the total duration of ZO-1 redistribution at the same sites (45 ± 2 min). In about 15% of cell shedding cases, neighboring epithelial cells also undergo extrusion with a delay of 5–10 min. With the use of normal mice, ZO-1 immunofluorescent staining of fixed tissue confirmed ZO-1 redistribution and the presence of ZO-1 patches beneath shedding cells. Immunostaining also showed that redistribution of ZO-1 occurred without corresponding mixing of apical and basolateral membrane domains as marked by ezrin or E-cadherin. ZO-1 redistribution is the earliest cellular event yet identified as a herald of physiological cell shedding, and redistribution of tight junction function along the lateral plasma membrane sustains epithelial barrier during cell shedding