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Dysregulation of Notch signaling in cardiac mesenchymal cells of patients with tetralogy of Fallot.

Tetralogy of Fallot (TF) is a severe congenital defect of heart development. Fine-tuned sequential activation of Notch signaling genes is responsible for proper heart chamber development. Mutations in Notch genes have been associated with TF. The aim of this study was to analyze the activity of the Notch pathway in cardiac mesenchymal cells derived from ventricular tissue of TF patients.

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Downregulated AKT-mTOR signaling pathway proteins in dorsolateral prefrontal cortex in Schizophrenia.

Abnormal neurotransmission is central to schizophrenia (SZ). Alterations across multiple neurotransmitter systems in SZ suggest that this illness may be associated with dysregulation of core intracellular processes such as signaling pathways that underlie the regulation and integration of these systems. The AKT-mTOR signaling cascade has been implicated in SZ by gene association, postmortem brain and animal studies. AKT and mTOR are serine/threonine kinases which play important roles in cell growth, proliferation, survival, and differentiation. Both AKT and mTOR require phosphorylation at specific sites for their complete activation. mTOR forms two functionally distinct multiprotein complexes, mTOR Complex 1 (mTORC1) and Complex 2 (mTORC2). mTORC1 mediates ribosome biogenesis, protein translation, and autophagy, whereas mTORC2 contributes to actin dynamics. Altered protein synthesis and actin dynamics can lead to an abnormal neuronal morphology resulting in deficits in learning and memory. Currently, there is a lack of direct evidence to support the hypothesis of disrupted mTOR signaling in SZ, and we have addressed this by characterizing this signaling pathway in SZ brain. We found a reduction in AKT and mTOR protein expression and/or phosphorylation state in dorsolateral prefrontal cortex (DLPFC) from 22 pairs of SZ and matched comparison subjects. We also found reduced protein expression of GβL, a subunit protein common to both mTOR complexes. We further investigated mTOR complex-specific subunit composition and phosphorylation state, and found abnormal mTOR expression in both complexes in SZ DLPFC. These findings provide evidence that proteins associated with the AKT-mTOR signaling cascade are downregulated in SZ DLPFC.

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Periodontal ligament cells regulate osteogenesis via miR-299-5p in mesenchymal stem cells.

The periodontal ligament contains periodontal ligament cells, which is a heterogeneous cell population, and includes progenitor cells that can differentiate into osteoblasts/cementoblasts. Mesenchymal stem cells (MSCs) can differentiate into various cells and can be used for periodontal regenerative therapy. Therefore, transplanted MSCs can be affected by humoral factors from periodontal ligament cells via the transcription factors or microRNAs (miRNAs) of MSCs. In addition, periostin (POSTN) is secreted from HPL cells and can regulate periodontal regeneration and homeostasis. To clarify the regulatory mechanism of humoral factors from periodontal ligament cells, we attempted to identify key genes, specifically microRNAs, involved in this process.

1538 related Products with: Periodontal ligament cells regulate osteogenesis via miR-299-5p in mesenchymal stem cells.

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Enhancing Hematopoiesis from Murine Embryonic Stem Cells through MLL1-Induced Activation of a Rac/Rho/Integrin Signaling Axis.

The Mixed Lineage Leukemia (MLL1, KMT2A) gene is critical for development and maintenance of hematopoietic stem cells (HSCs), however, whether this protein is limiting for HSC development is unknown due to lack of physiologic model systems. Here, we develop an MLL1-inducible embryonic stem cell (ESC) system and show that induction of wild-type MLL1 during ESC differentiation selectively increases hematopoietic potential from a transitional c-Kit/Cd41 population in the embryoid body and also at sites of hematopoiesis in embryos. Single-cell sequencing analysis illustrates inherent heterogeneity of the c-Kit/Cd41 population and demonstrates that MLL1 induction shifts its composition toward multilineage hematopoietic identities. Surprisingly, this does not occur through increasing Hox or other canonical MLL1 targets but through an enhanced Rac/Rho/integrin signaling state, which increases responsiveness to Vla4 ligands and enhances hematopoietic commitment. Together, our data implicate a Rac/Rho/integrin signaling axis in the endothelial to hematopoietic transition and demonstrate that MLL1 actives this axis.

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Anti-AICDA(Activation-ind 129 Mouse Embryonic Stem Stem Cell TF Activation P Anti AICDA(Activation ind Anti bodywall muscle cell Rabbit Anti-Integrin alph TGF beta induced factor 2 Goat Anti-Human, Mouse In gAcrp30 Adiponectin, muri ERK Signaling Phospho-Spe Rabbit Anti-Integrin alph Mouse anti human Integrin

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Evaluation of bone marrow preparations and sections of teeth prepared with modified Bouin's solution.

The quality of bone marrow preparation and sections of teeth depends on the preparation method. We investigated the posterior mandibles of male rats that were processed using a modified Bouin's solution (MBS) and stained with Harris' hematoxylin and eosin, and the results were compared to a routine decalcification process using 5% formal nitric acid. We found that MBS was applicable to both soft and hard components in bone tissues and sections of teeth. MBS provided good decalcification, which facilitated sectioning. The bone marrow specimens treated with MBS exhibited clearly distinguishable hematopoietic cells, clear tissue integrity and good cell preservation. Treatment with 5% formal nitric acid for 5 days caused degeneration, poor differentiation and poorer staining of hematopoietic components, and deteriorated soft and solid tissues in bone and sections of teeth compared to MBS treatment. MBS preserved cell and tissue integrity and good staining quality. MBS can be used for rapid preparation and diagnosis for pathology and toxicity studies.

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Comparing negative emotion differentiation in young and older individuals: A picture-based study.

Life span theories suggest that emotional experiences become more complex (i.e., nuanced and differentiated) with age. Theoretically, the cause of this increased complexity has been proposed to be age-related changes in life contexts such as goals and daily stressors. Consequently, age may not affect emotional complexity in settings where the influence of age-specific life contexts is reduced. However, this hypothesis has yet to be explored. In the present study, we investigated one aspect of emotional complexity, namely emotion differentiation. Extending previous research, we assessed age-group differences in negative emotion differentiation between young and older adults in a controlled experimental setting. A sample of 114 young and 132 older adults rated their emotional response to 34 negative pictures according to intensity of four negative emotions. Based on these ratings, two indicators of emotion differentiation were calculated. The results revealed no significant age-group differences in negative emotion differentiation. The findings indicate stability in negative emotion differentiation with increasing age when the influence of life context is reduced. The findings are consistent with life span theories suggesting that developmental changes in emotional complexity occur largely as a result of age-related changes in life contexts rather than more stable age-related changes in individual characteristics.

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Comparison of the myometrial transcriptome from singleton and twin pregnancies by RNA-Seq.

Preterm birth is recognized as the primary cause of infant mortality worldwide. Twin pregnancies are significantly more at risk of preterm birth than singleton pregnancies. A greater understanding of why this is and better modes of treatment and prevention are needed. Key to this is determining the differing pathophysiological mechanisms of preterm birth in twins, including the role of the myometrium and premature uterine contraction. We performed RNA sequencing (RNA-Seq) of human myometrium from singleton and twin pregnancies at term (> 37+0 weeks) and preterm (< 37+0 weeks), collected during pre-labour Caesarean Section. RNA-Seq libraries were prepared from polyA-selected RNA and sequenced on the Illumina HiSeq 4000 platform. Differentially expressed genes (DEGs), GO (Gene Ontology) and KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway enrichment were conducted using R software. Significance was determined with a false discovery rate-adjusted P value of <0.05. Only 3 DEGs were identified between gestational age-matched singleton and twin myometrium and only 1 DEG identified between singleton term and twin preterm tissues. Comparison of singleton preterm myometrium with twin term myometrium however, revealed 75 down-regulated and 24 up-regulated genes in twin myometrium. This included genes associated with inflammation and immune response, T cell maturation and differentiation and steroid biosynthesis. GO and KEGG enrichment analyses for biologically relevant processes and functions also revealed several terms related to inflammation and immune response, as well as cytokine-cytokine receptor interaction and chemokine receptor signalling. Data indicate that little or no differences exist in the transcriptome of singleton and twin myometrium when matched for gestational age. The significant up- and down-regulation of genes identified between preterm singleton and twin myometrium at term may point to transcriptome changes associated with the chronic levels of uterine stretch in twin pregnancy or genes associated with the myometrium transitioning to labour onset.

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An Axonal Blueprint: Generating Neuronal Polarity with Light-Inducible Proteins.

In this issue of Cell Chemical Biology, Woo et al. (2019) show that activation of a photoinducible form of the TrkB protein in a single neurite induces multiple aspects of axonal differentiation. This study exemplifies the ability of optical methods to relate protein functions to complex phenotypes in living cells.

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Quantitative PCR Assays Developed for and for Phomopsis Stem Canker Diagnosis and Germplasm Screening in Sunflower ().

Phomopsis stem canker of sunflower is caused by two fungal pathogens, and , in the United States. In this study, two quantitative PCR (qPCR) assays were developed to detect and quantify and in sunflower. The two assays differentiated the two fungi from each other, other species of the genus , and pathogens, and they have high efficiency (>90%). The qPCR assays detected the two pathogens on plant samples exhibiting Phomopsis stem canker symptoms sampled from commercial sunflower fields in Minnesota, Nebraska, North Dakota, and South Dakota. Furthermore, the assays were used to screen cultivated sunflower accessions for resistance to and . The disease severity index (DSI) of the accessions significantly correlated ( < 0.0001) with the amount of pathogen DNA from the qPCR assays. The qPCR assays identified PI664232 and PI561918 to be significantly less susceptible ( ≤ 0.05) to and , respectively, when compared with the susceptible check cultivar HA 288, and this was in agreement with the DSI. These results suggest that the qPCR assays for and can be used as a reliable tool to diagnose Phomopsis stem canker and screen sunflower germplasm for disease resistance.

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A lab-on-chip ultrasonic platform for real-time and nondestructive assessment of extracellular matrix stiffness.

Extracellular matrix (ECM) mechanical stiffness and its dynamic change is one of the main cues that directly affects the differentiation and proliferation of normal cells as well as the progression of disease processes such as fibrosis and cancer. Recent advancements in biomaterials have enabled a wide range of polymer matrices that could mimic the ECM of different tissues for a wide range of in vitro basic research and drug discovery. However, most of the technologies utilized to quantify the stiffness of such ECM are either destructive or expensive, and therefore are unsuitable for the in situ, long-term monitoring of variations in ECM stiffness for on-chip cell culture applications. This work demonstrates a novel noninvasive on-chip platform for characterization of ECM stiffness in vitro, by monitoring ultrasonic wave attenuation through the targeted material. The device is composed of a pair of millimeter scale ultrasonic transmitter and receiver transducers with the test medium placed in between them. The transmitter generates an ultrasonic wave that propagates through the material, triggers the piezoelectric receiver and generates a corresponding electrical signal. The characterization reveals a linear (r2 = 0.86) decrease in the output voltage of the piezoelectric receiver with an average sensitivity of -15.86 μV kPa-1 by increasing the stiffnesses of hydrogels (from 4.3 kPa to 308 kPa made with various dry-weight concentrations of agarose and gelatin). The ultrasonic stiffness sensing is also demonstrated to successfully monitor dynamic changes in a simulated in vitro tissue by gradually changing the polymerization density of an agarose gel, as a proof-of-concept towards future use for 3D cell culture and drug screening. In situ long-term ultrasonic signal stability and thermal assessment of the device demonstrates its high robust performance even after two days of continuous operation, with negligible (<0.5 °C) heating of the hydrogel in contact with the piezoelectric transducers. In vitro biocompatibility assessment of the device with mammary fibroblasts further assures that the materials used in the platform did not produce a toxic response and cells remained viable under the applied ultrasound signals in the device.

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