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Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

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At least 865 records · Page 48
All sourcesNASA NTRSDOE OSTI
DOE OSTI2017

Identification and molecular cloning of novel transcripts of the human kallikrein-related peptidase 10 (KLK10) gene using next-generation sequencing

Tissue kallikrein and kallikrein-related peptidases (KLKs) form the largest group of serine proteases in the human genome, sharing many structural and functional characteristics. Multiple alternative transcripts have been reported for the most human KLK genes, while many of them are aberrantly expressed in various malignancies, thus possessing significant prognostic and/or diagnostic value. Alternative splicing of cancer-related genes is a common cellular mechanism accounting for cancer cell transcriptome complexity, as it affects cell cycle control, proliferation, apoptosis, invasion, and metastasis. In this study, we describe the identification and molecular cloning of eight novel transcripts of the human KLK10 gene using 3′ rapid amplification of cDNA ends (3′ RACE) and next-generation sequencing (NGS), as well as their expression analysis in a wide panel of cell lines, originating from several distinct cancerous and normal tissues. Bioinformatic analysis revealed that the novel KLK10 transcripts contain new alternative splicing events between already annotated exons as well as novel exons. In addition, investigation of their expression profile in a wide panel of cell lines was performed with nested RT-PCR using variant-specific pairs of primers. Since many KLK mRNA transcripts possess clinical value, these newly discovered alternatively spliced KLK10 transcripts appear as new potential biomarkers for diagnostic and/or prognostic purposes or as targets for therapeutic strategies. - Highlights: • NGS was used to identify novel transcripts of the human KLK10 gene. • 8 novel KLK10 transcripts were identified. • A novel 3′UTR was detected and characterized. • The expression profiles of all 8 novel KLK10 transcripts were identified.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2017

Vitamin D supplement ameliorates hippocampal metabolism in diabetic rats

Vitamin D deficiency induced by diabetes mellitus is tightly associated with neurodegenerative diseases, but the mechanism is still unknown. Endoplasmic reticulum stress (ER stress) is involved in hippocampal lesion and promote diabetic neuropathy, so we focus on the effects of 1, 25-dihydroxy vitamin D{sub 3} on ER stress in hippocampus of diabetic rats. Streptozotocin (STZ)-induced diabetic rats were administrated with different doses of vitamin D and divided into 3 groups: high, low, and blank, compared to wild-type rats which were received the same treatment. At the end of 12 weeks of treatment, the brains of the rats were analyzed by proton magnetic resonance spectroscopy ({sup 1}H-MRS). Rats were then weighed, tested for blood glucose, serum Ca, P, and vitamin D{sub 3}, and sacrificed for histopathological analysis of the hippocampus. Neuronal nitric oxide synthase (nNOS) and vitamin D receptor (VDR) expression were measured, as well as ER stress markers, including glucose-regulated protein78 (GRP78), protein kinase-like endoplasmic reticulum kinase (PERK) phosphorylation, eukaryotic initiation factor 2α(eIF-2α) phosphorylation, and CCAAT enhancer-binding protein homologous protein (CHOP). Our study showed that treated with appropriate concentration of active vitamin D could decrease the number of pathological pyramidal neurons, improve hippocampal nerve metabolism, and reduce the over-expression of nNOS, along with the relieved activation of ER stress in hippocampus of diabetic rats. These results suggest that 1,25-dihydroxy vitamin D{sub 3} treatment can ameliorate hyperglycemia-induced damage on hippocampal metabolism, possibly through alleviating the aberrant activation of ER stress.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2017

miR-137 attenuates Aβ-induced neurotoxicity through inactivation of NF-κB pathway by targeting TNFAIP1 in Neuro2a cells

Background: Accumulation of β-amyloid (Aβ) and neuroinflammation are implicated in the pathogenesis and development of Alzheimer's disease (AD). Neuron-enriched miR-137 was aberrantly downregulated and may be associated with the pathogenesis of AD. However, the detailed function of miR-137 in AD pathogenesis and the molecular mechanism have not been elucidated. Methods: The expressions of miR-137 and tumor necrosis factor alpha (TNFα)-induced protein 1 (TNFAIP1) at mRNA and protein levels in primary mouse cortical neurons and Neuro2a (N2a) cells exposed to different concentrations of Aβ{sub 25-35} were examined by qRT-PCR and western blot. Luciferase reporter assay was used to confirm the potential target of miR-137. MTT assay, flow cytometry analysis, caspase-3 activity assay, Enzyme-linked immunosorbent assay (ELISA), and western blot were used to detect cell viability, apoptosis, caspase-3 activity, Nuclear factor-kappa B (NF-κB) activity and level, respectively. Results: Aβ{sub 25-35} downregulated miR-137 and upregulated TNFAIP1 in primary mouse cortical neurons and N2a cells. In addition, miR-137 was found to directly target TNFAIP1 and suppress its mRNA and protein levels. Moreover, miR-137 restoration and TNFAIP1 knockdown facilitate Aβ{sub 25-35}-induced cell toxicity, apoptosis, caspase-3 activity, and activated NF-κB in N2a cells, which was partially abolished by TNFAIP1 overexpression. Conclusion: miR-137 attenuated Aβ-induced neurotoxicity through inactivation of NF-κB pathway by targeting TNFAIP1 in N2a cells, shedding light on the molecular mechanism of miR-137 underlying Aβ-induced neurotoxicity. - Highlights: • Aβ{sub 25-35} downregulated miR-137 and upregulated TNFAIP1 in cortical neurons and N2a cells. • miR-137 suppressed TNFAIP1 expression through direct binding at the 3′UTR. • miR-137 overexpression contributed to Aβ{sub 25-35}-induced cell toxicity and apoptosis. • miR-137 overexpression on Aβ{sub 25-35}-induced neurotoxicity activated NF-κB pathway.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2017

Tacrolimus induces fibroblasts apoptosis and reduces epidural fibrosis by regulating miR-429 and its target of RhoE

Tacrolimus (FK506) has been demonstrated to reduce epidural fibrosis. However, the detailed mechanism of action has not been elucidated. Aberrant miR-429 is involved in many diseases. The aim of this study was to describe the exact mechanism of FK506 induced apoptosis in fibroblasts and the prevention of epidural fibrosis. FK506 induced fibroblast apoptosis was evaluated using CCK-8 assays, flow cytometry, and western blotting. The expression of miR-429 in fibroblasts treated with FK506 was determined by RT-qPCR. Additionally, luciferase activity assays were used to determine the target relationship between miR-429 and RhoE. Flow cytometry and western blot analysis were used to determine the effects of FK506 and miR-429 on fibroblast apoptosis. The effects of FK506 and RhoE on fibroblast apoptosis were determined by CCK-8 assay, flow cytometry, and western blotting. We also evaluate the effects of FK506 and miR-429 on epidural fibrosis in rats by using histological analysis and TUNEL-staining. The results revealed FK506 induces fibroblast apoptosis and significantly downregulates miR-429 expression in fibroblasts. Additionally, miR-429 downregulation caused the apoptosis of fibroblasts. The luciferase activity assay confirmed that RhoE is a direct target of miR-429 and RhoE promotes fibroblast apoptosis. The rat model demonstrated miR-429 inhibition promotes fibroblast apoptosis and epidural fibrosis, which is consistent with the results of FK506 treatment. Our study demonstrates that FK506 induces fibroblast apoptosis and reduces epidural fibrosis by regulating miR-429 expression and its target of RhoE. - Highlights: • FK506 could inhibit the proliferation and induce the apoptosis of human fibroblast. • FK506 could downregulate the expression of miR-429 and RhoE is a target gene of miR-429. • Inhibits the function of RohE could reduce the apoptosis of human fibroblast after treatment of FK506. • Inhibits the miR-429 function could inhibit proliferation and induce apoptosis of fibroblast and reduce epidural fibrosis.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2017

Identification of novel ALK2 inhibitors and their effect on cancer cells

Bone morphogenetic protein 9 (BMP9), a member of the TGF-β superfamily, is considered a regulator of glucose homeostasis as well as a neuronal differentiation factor. BMP9 induces phosphorylation of Smad1/5 through activin receptor-like kinase 1 and 2 (ALK1 and ALK2). Recently, many studies have shown that BMP9 contributes to tumorigenesis, and aberrant ALK2 expression is involved in many diseases. To investigate the role of BMP9-ALK2 signaling in cancer cells, we used TF-1 cells that require granulocyte-macrophage colony-stimulating factor (GM-CSF) for cell proliferation. BMP9 promoted the proliferation of TF-1 cells in media lacking GM-CSF. TF-1 cells overexpressing ALK2 resulted in the autophosphorylation of Smad1/5, leading to consequent increase in cell growth. Through high-throughput screening (HTS), we found two ALK2-specific inhibitors, KRC203 and KRC360, with IC{sub 50} values of 0.9 nM and 0.3 nM. These compounds were more potent and specific for the inhibition of ALK2 when compared to LDN193189. In cell-based assays, these compounds effectively inhibited the proliferation and migration of cancer cells induced by ALK2 and BMP9. Therefore, we propose that our compounds are promising candidates for the treatment of cancer or diseases with abnormal ALK2 or BMP9 signaling. - Highlights: • BMP9 signaling promoted the proliferation of cancer cells via Smad1/5 pathway. • Overexpressing ALK2, a BMP9 receptors, promoted the proliferation of cancer cells. • KRC203 and KRC360 that we found through high-throughput screening showed high specific inhibition against ALK2. • Our compounds inhibited specificity BMP9 signaling and inhibited the migration of hepatocellular carcinoma. • Our compounds also effectively inhibited the proliferation of cancer cells by BMP9 and ALK2 signaling.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2017

Direct effects of mitochondrial dysfunction on poor bone health in Leigh syndrome

Mitochondrial diseases are the result of aberrant mitochondrial function caused by mutations in either nuclear or mitochondrial DNA. Poor bone health has recently been suggested as a symptom of mitochondrial diseases; however, a direct link between decreased mitochondrial function and poor bone health in mitochondrial disease has not been demonstrated. In this study, stem cells from human exfoliated deciduous teeth (SHED) were isolated from a child with Leigh syndrome (LS), a mitochondrial disease, and the effects of decreased mitochondrial function on poor bone health were analyzed. Compared with control SHED, LS SHED displayed decreased osteoblastic differentiation and calcium mineralization. The intracellular and mitochondrial calcium levels were lower in LS SHED than in control SHED. Furthermore, the mitochondrial activity of LS SHED was decreased compared with control SHED both with and without osteoblastic differentiation. Our results indicate that decreased osteoblast differentiation potential and osteoblast function contribute to poor bone health in mitochondrial diseases. - Highlights: • Stem cells from human exfoliated deciduous teeth (SHED) were isolated. • Osteoblast differentiation was decreased in Leigh syndrome (LS)-derived SHED. • Both intracellular and mitochondrial calcium levels were decreased in LS SHED. • Mitochondrial activity was also decreased in LS SHED. • Decreased mitochondrial activity directly contributes to poor bone health in LS.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2017

Sez6l2 regulates phosphorylation of ADD and neuritogenesis

Highlights: • Sez6l2 interacts with ADD and GluR1. • Extracellular domains of Sez6l2 and GluR1 are required for their interactions. • Sez6l2 may be one of the auxiliary subunits with AMPA receptors. • Sez6l2 may link GluR1 to ADD and be an up-regulator of ADD phosphorylation. Increasing evidence shows that immune-mediated mechanisms may contribute to the pathogenesis of central nervous system disorders including cerebellar ataxias, as indicated by the aberrant production of neuronal surface antibodies. We previously reported a patient with cerebellar ataxia associated with production of a new anti-neuronal antibody, anti-seizure-related 6 homolog like 2 (Sez6l2). Sez6l2 is a type 1 membrane protein that is highly expressed in the hippocampus and cerebellar cortex and mice lacking Sez6l2 protein family members develop ataxia. Here we used a proteomics-based approach to show that serum derived from this patient recognizes the extracellular domain of Sez6l2 and that Sez6l2 protein binds to both adducin (ADD) and glutamate receptor 1 (GluR1). Our results indicate that Sez6l2 is one of the auxiliary subunits of the AMPA receptor and acts as a scaffolding protein to link GluR1 to ADD. Furthermore, Sez6l2 overexpression upregulates ADD phosphorylation, whereas siRNA-mediated downregulation of Sez612 prevents ADD phosphorylation, suggesting that Sez6l2 modulates AMPA-ADD signal transduction.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2017

Exposure to cigarette smoke disturbs adipokines secretion causing intercellular damage and insulin resistance in high fructose diet-induced metabolic disorder mice

Highlights: • Cigarette smoke and fructose excess intake induce disruption to mechanisms involved in ROS generation and ER stress. • Damage of cellular organelles induced by cigarette smoke and high fructose feeding affects inflammatory response and alters secretion of adipokines. • Cigarette smoking and fructose excess intake are associated with metabolic syndrome. A large amount of fructose intake along with smoking is associated with increased incidence of diseases linked to metabolic syndrome. More research is necessary to understand the complex mechanism that ultimately results in metabolic syndrome and the effect, if any, of high fructose dietary intake and smoking on individual health. In this study, we investigated changes in ER-Golgi network and disturbance to secretion of adipokines induced by cigarette smoking (CS) and excess fructose intake and their contribution to the disruption of metabolic homeostasis. We used high fructose-induced metabolic disorder mice model by feeding them with high fructose diet for 8 weeks. For CS exposure experiment, these mice were exposed to CS for 28 days according to OECD guideline 412. Our results clearly showed that the immune system was suppressed and ER stress was induced in mice with exposure to CS and fed with high fructose. Furthermore, their concentrations of adipokines including leptin and adiponectin were aberrant. Such alteration in secretion of adipokines could cause insulin resistance which may lead to the development of type 2 diabetes.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Dystonia-4 (DYT4)-associated TUBB4A mutants exhibit disorganized microtubule networks and inhibit neuronal process growth

Highlights: • Dystonia-4-associated TUBB4A mutants (R2G) exhibit disorganized tubulin networks. • Dystonia-4-associated TUBB4A mutants (A271T) exhibit disorganized tubulin networks. • Dystonia-4-associated mutants have diminished ability to interact with alpha-type tubulins. • Dystonia-4-associated mutants show inhibited neurite growth in a neuronal cell line. Dystonia-1 (DYT1) is an autosomal dominant early-onset torsion form of dystonia, a neurological disease affecting movement. DYT1 is the prototypic hereditary dystonia and is caused by the mutation of the tor1a gene. The gene product has chaperone functions important for the control of protein folding and stability. Dystonia-4 (DYT4) is another autosomal dominant dystonia that is characterized by onset in the second to third decade of progressive laryngeal dysphonia. DYT4 is associated with the mutation of the tubb4a gene, although it remains to be understood how disease-associated mutation affects biochemical as well as cell biological properties of the gene product as the microtubule component (a tubulin beta subunit). Herein we demonstrate that DYT4-associated TUBB4A missense mutants (Arg2-to-Gly or Ala271-to-Thr) form disorganized tubulin networks in cells. Transfected mutants are indeed expressed in cytoplasmic regions, as observed in wild-type transfectants. However, mutant proteins do not exhibit typical radial tubulin networks. Rather, they have diminished ability to interact with tubulin alpha subunits. Processes do not form in sufficient amounts in cells of the N1E-115 neuronal cell line expressing each of these mutants as compared to parental cells. Together, DYT4-associated TUBB4A mutants themselves form aberrant tubulin networks and inhibit neuronal process growth, possibly explaining progress through the pathological states at cellular levels.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Impaired autophagy promotes bile acid-induced hepatic injury and accumulation of ubiquitinated proteins

Highlights: • CDCA, CA and bile duct ligation caused defective autophagic clearance. • CDCA suppressed Beclin-1 expression, which exhibits a higher cytotoxicity than CA. • Pharmacological or genetic inhibition of autophagy enhanced bile acid-induced cell death. • Reduced Beclin-1 expression may lead to higher cytotoxicity of CDCA. Chronic exposure to hydrophobic bile acids such as chenodeoxycholic acid (CDCA) and cholic acid (CA) in the liver during cholestasis causes hepatotoxicity and inflammatory response. However, the detailed mechanisms regarding the role of autophagy in cholestatic hepatotoxicity remain largely unknown. Here we determined autophagic clearance in livers of bile duct-ligated mice, in which bile acids accumulate, and in human hepatoma HepG2 cells treated with CDCA and CA. The accumulation of bile acids caused defective autophagic clearance, shown by the accumulation of insoluble p62 and ubiquitinated proteins and cell death accompanied by caspase-3 processing. Hepatocytes exposed to bile acids also showed the accumulation of autophagosomes via suppressed autophagy flux. Treatment of CDCA markedly suppressed Beclin-1 expression, which exhibits a higher cytotoxicity than CA. Moreover, pharmacological or genetic inhibition of autophagy enhanced bile acid-induced cell death. Finally, in vivo, bile duct ligation led to aberrant accumulation of p62 and ubiquitinated proteins in the liver. Our data demonstrate that inhibited autophagy is an essential component of liver injury during cholestasis.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

AMOT is required for YAP function in high glucose induced liver malignancy

Highlights: • AMOT is identified to be O-GlcNAcylated. • High glucose stimulates expression and O-GlcNAcylation of AMOT. • AMOT is a stimulator of YAP in high glucose level, whereas a suppressor of YAP in normal glucose level. • High expression and nuclear accumulation of AMOT and YAP are found in high glucose mice. AMOT has been identified as a YAP interactor. However, how AMOT regulates YAP remains unclear and controversy. Here, we identified that besides YAP, AMOT was another Hippo signaling core factor which could be O-GlcNAcylated. Moreover, high glucose (HG) was able to enhance the expression and O-GlcNAcylation of AMOT. We also found that HG stimulated nuclear accumulation, transcription activity, interaction with transcription factor and transcription of target genes of YAP via AMOT, while AMOT acted as a suppressor of YAP in normal glucose level. Finally, we observed the upregulation and nuclear accumulation of AMOT and YAP in Streptozocin (STZ) induced high glucose mice. Collectively, we have uncovered that AMOT acts as a YAP stimulator in high glucose level. Targeting the aberrantly regulated core factors in Hippo pathway might be a more effective therapeutic approach for liver cancer associated with possibly diabetes.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

CCN3 secretion is regulated by palmitoylation via ZDHHC22

Highlights: • The CCN3 thrombospondin type-1 (TSP1) domain is required for its secretion. • The TSP1 domain is also involved in CNN3 intracellular function. • Residue C241 is important for CNN3 secretion, but not its intracellular function. • Palmitoylation at C241 is necessary for CCN3 secretion. • Zinc finger DHHC-type containing 22 interacts with CCN3. Normal extracellular secretion of nephroblastoma overexpressed (NOV, also known as CCN3) is important for the adhesion, migration, and differentiation of cells. In previous studies, we have shown that the intracellular accumulation of CCN3 inhibits the growth of prominent neurons. Increased intracellular CCN3 can be induced through various processes, such as transcription, detoxification, and posttranslational modification. In general, posttranslational modifications are very important for protein secretion. However, it is unclear whether posttranslational modification is necessary for CCN3 secretion. In this study, we have conducted mutational analysis of CCN3 to demonstrate that its thrombospondin type-1 (TSP1) domain is important for CCN3 secretion and intracellular function. Point mutation analysis confirmed that CCN3 secretion was inhibited by cysteine (C)241 mutation, and overexpression of CCN3-C241A inhibited neuronal axonal growth in vivo. Furthermore, we demonstrated that palmitoylation is important for the extracellular secretion of CCN3 and that zinc finger DHHC-type containing 22 (ZDHHC22), a palmityoltransferase, can interact with CCN3. Taken together, our results suggest that palmitoylation by ZDHHC22 at C241 in the CCN3 TSP1 domain may be required for the secretion of CCN3. Aberrant palmitoylation induces intracellular accumulation of CCN3, inhibiting neuronal axon growth.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

BMI1 activates WNT signaling in colon cancer by negatively regulating the WNT antagonist IDAX

Highlights: • BMI1 is upregulated and activates WNT signaling in colon cancer. • IDAX is a direct target of BMI1 and is downregulated in colon cancer. • IDAX plays as a tumor suppressor in colon cancer through inhibiting WNT signaling. Aberrant activation of Wnt signaling plays a critical role in the development of colon cancer. BMI, a component of the polycomb repressive complex (PRC1), is upregulated in various types of cancer and contributes to epigenetic silencing of tumor suppressors. In this study, we showed that BMI1 is upregulated in colon cancer tissues and cell lines. Overexpression of BMI1 in primary epithelial colon cells promotes cellular growth and activates WNT pathway, while BMI1 silencing in colon cancer cells represses these effects. We also found that BMI1 binds to the promoter of IDAX, a Wnt antagonist, and decreases its transcription. Expression of IDAX is downregulated in colon cancer tissues and cell lines and negatively correlated with BMI1 in colon cancer tissues. Furthermore, Silencing of IDAX counteracts the effects of BMI1 suppression, while its overexpression reverses oncogenic effects of BMI1. Together, these findings indicate that BMI1-mediated IDAX epigenetic suppression is crucial for enhancement of colon carcinogenesis, suggesting that BMI1∖IDAX axis as a potential novel diagnostic and therapeutic target of colon cancer.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

SMAD7 methylation as a novel marker in atherosclerosis

Highlights: • MALDI-TOF MS is an advantageous way to analyze DNA methylation. • SMAD7 promoter is hyper-methylated in human atherosclerotic plaques and peripheral blood of atherosclerosis patients. • SMAD7 methylation levels positively correlate with homocysteine levels and carotid plaque scores. • Methylated SMAD7 may be a novel predicted marker and therapeutics target for atherosclerosis. Atherosclerosis is a complicated process comprising inflammation, accumulation of collagen matrix and aberrant DNA methylation. SMAD7 is known to play an important role in fibrosis and inflammation. In recent years, increasing research has concentrated on the connection between DNA methylation and atherosclerosis. The current study was designed to investigate methylation status of some specific gene with a focus on SMAD7 in atherosclerosis and elucidate their relationship. We found that SMAD7 expression was decreased and its promoter region was markedly methylated in atherosclerotic plaques when compared with normal artery walls. Using MALDI-TOF MS, increased DNA methylation levels of SMAD7 promoter at CpG unit 5.8.15.16 were found in peripheral blood of atherosclerosis patients relative to matched normal controls, respectively. Correlation analysis revealed that mean DNA methylation levels of SMAD7 promoter of CpG unit 5.8.15.16 were positively associated with homocysteine levels (r = 0.724, p p SMAD7 promoter is hyper-methylated both in human atherosclerotic plaques and atherosclerosis patients, which is positively associated with homocysteine levels and carotid plaque scores. Thus, methylated SMAD7 may be a novel predicted marker and therapeutics target for atherosclerosis.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Adamts18 deficiency increases arterial thrombus formation associated with vascular defects in mice

Highlights: • We validated ADAMTS18 modulates thrombosis using Adamts18 KO mice. • We validated ADAMTS18 affects carotid remodeling in mice. • We validated ADAMTS18 exerts little effects on platelet functions. • The increased thrombosis in Adamts18 KO mice is associated with the vascular defects. ADAMTS18 is a member of a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTSs) that are known for their crucial role in development, angiogenesis, inflammation and coagulation. It was previously reported that ADAMTS18 cleaved by thrombin induced platelet fragmentation, through which thrombus were dissolved. However, it remains unclear whether this represents a dominant physiologic mechanism controlling thrombus growth in vivo. Here, we used an established Adamts18 knockout (KO) mouse model to determine its function in thrombus formation. ADAMTS18 deficiency accelerated FeCl{sub 3}-induced carotid artery thrombosis and aggravated postischemic cerebral infarction in mice. However, this accelerated thrombus phenotype in Adamts18 KO mice was not due to the lack of ADAMTS18-mediated-platelet fragmentation. Moreover, Adamts18 deficiency exerted little effects on mouse platelet functions. The underlying molecular mechanisms could be attributed in part to the abnormal vascular remodeling, including deficiency of carotid body (glomus) and aberrant carotid basal lamina. These results indicate a novel function of ADAMTS18 in vascular remodeling and associated thrombus formation.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Nonmuscle myosin IIA and IIB differentially contribute to intrinsic and directed migration of human embryonic lung fibroblasts

Highlights: • Normal human fibroblasts show intrinsic and directed migration for a long duration. • NMIIA controls protrusion formation in the anterior region to steer migration. • NMIIB prevents protrusions in the posterior region to maintain front-rear polarity. • NMIIA and NMIIB differentially contribute to intrinsic and directed migration. Nonmuscle myosin II (NMII) plays an essential role in directional cell migration. In this study, we investigated the roles of NMII isoforms (NMIIA and NMIIB) in the migration of human embryonic lung fibroblasts, which exhibit directionally persistent migration in an intrinsic manner. NMIIA-knockdown (KD) cells migrated unsteadily, but their direction of migration was approximately maintained. By contrast, NMIIB-KD cells occasionally reversed their direction of migration. Lamellipodium-like protrusions formed in the posterior region of NMIIB-KD cells prior to reversal of the migration direction. Moreover, NMIIB KD led to elongation of the posterior region in migrating cells, probably due to the lack of load-bearing stress fibers in this area. These results suggest that NMIIA plays a role in steering migration by maintaining stable protrusions in the anterior region, whereas NMIIB plays a role in maintenance of front-rear polarity by preventing aberrant protrusion formation in the posterior region. These distinct functions of NMIIA and NMIIB might promote intrinsic and directed migration of normal human fibroblasts.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Membrane glycerolipid equilibrium under endoplasmic reticulum stress in Arabidopsis thaliana

Highlights: • Membrane glycerolipid biosynthesis under ER stress was investigated in Arabidopsis. • Gene expression of enzymes for PC and PE biosynthesis was changed during ER stress. • Lipid content was mostly unchanged despite significant changes at the transcriptional level. • The membrane glycerolipid profile may be maintained at post-transcriptional levels upon ER stress in Arabidopsis thaliana. Endoplasmic reticulum (ER) is an indispensable organelle for secretory protein synthesis as well as metabolism of phospholipids and their derivatives in eukaryotic cells. Various external and internal factors may cause an accumulation of aberrant proteins in the ER, which causes ER stress and activates cellular ER stress responses to cope with the stress. In animal research, molecular mechanisms for protein quality control upon ER stress are well documented; however, how cells maintain lipid homeostasis under ER stress is an emerging issue. The ratio of phosphatidylcholine (PC) to phosphatidylethanolamine (PE), two major phospholipid classes, is important under ER stress in animal cells. However, in seed plants, no study has reported on the changes in membrane lipid content under ER stress, although a number of physiologically important environmental stresses, such as heat and salinity, induce ER stress. Here, we investigated membrane glycerolipid metabolism under ER stress in Arabidopsis. ER stress transcriptionally affected PC and PE biosynthesis pathways differentially, with no significant changes in membrane glycerolipid content. Our results suggest that higher plants maintain membrane lipid equilibrium during active transcription of phospholipid biosynthetic genes under ER stress.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Circular RNA expression profiles in placental villi from women with gestational diabetes mellitus

Highlights: • More than 40,000 circRNAs are sequenced in the placental villi. • The expression profile of circRNAs in GDM is different from that in controls. • The potential effects of many differential circRNAs are involve in the development of GDM. • Some differential circRNAs have binding sites of GDM-related miRNAs. Circular RNAs (circRNAs) have recently been shown to exert their effects on multiple pathological processes by acting as microRNA (miRNA) sponges. However, the roles of circRNAs in gestational diabetes mellitus (GDM) are largely unknown. This study aimed to identify the circRNAs involved in GDM and predict their potential biological functions. We first performed next-generation sequencing (NGS) to generate unbiased placental villi circRNA expression profiles of GDM and normal controls. In total, 48,270 circRNAs from the placental villi of the two groups were sequenced. Of these, 227 circRNAs were significantly up-regulated and 255 circRNAs were significantly down-regulated. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) biological pathway analyses demonstrated that glycometabolism and lipometabolism processes, which are important in GDM development, were significantly enriched. Further analysis showed that most of the circRNAs harbored miRNA binding sites, and some were associated with GDM. These results showed that circRNAs are aberrantly expressed in the placental villi of GDM patients and play potential roles in the development of GDM.

60 APPLIED LIFE SCIENCES↗