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Supplementary Figure 3 from Human ZNF312b Promotes the Progression of Gastric Cancer by Transcriptional Activation of the K-ras Gene

Supplementary Figure 2 from Human ZNF312b Promotes the Progression of Gastric Cancer by Transcriptional Activation of the K-ras Gene

Gastric cancer ranks second among the most common causes of cancer deaths worldwide. Recent studies reported target molecules that are candidates for new therapeutic interventions; however, their molecular mechanism has not been clearly defined. In this study, we found that ZNF312b plays a role in tumor progression and metastasis in gastric cancer via transcriptional activation of the K-ras oncogene. ZNF312b seems to be specifically overexpressed in gastric cancer tissues and cell lines. The overexpression of ZNF312b induces cancer-like phenotypes, including accelerated proliferation and increased tumor masses in nude mice, which are completely reversed by its knockdown in gastric cancer cell lines, implying direct involvement in gastric tumor progression. From analyses using deletion mutants of ZNF312b and K-ras promoter-driven luciferase reporters, we found that it translocates into the nucleus via the proline-rich domain of its COOH terminus to activate transcription of the K-ras gene, resulting in an enhancement of the extracellular signal-regulated kinase signaling pathway that governs cell proliferation. Taken together, these results suggest that ZNF312b contributes to the promotion of gastric cancer by triggering K-ras oncogene expression. The current study is the first to report that ZNF312b, a novel transcription factor, was associated with tumorigenicity of gastric cancer. This might be a valuable target that could provide new insight into the development of new therapeutic modalities for patients with gastric cancer. [Cancer Res 2009;69(7):3131–9]

Supplementary Figure 1 from Human ZNF312b Promotes the Progression of Gastric Cancer by Transcriptional Activation of the K-ras Gene

Supplementary Figure 4 from Human ZNF312b Promotes the Progression of Gastric Cancer by Transcriptional Activation of the K-ras Gene

Supplementary Methods, Figures Legends 1-5 from Human ZNF312b Promotes the Progression of Gastric Cancer by Transcriptional Activation of the K-ras Gene

Supplementary Figure 5 from Human ZNF312b Promotes the Progression of Gastric Cancer by Transcriptional Activation of the K-ras Gene

Supplementary Table 1 from Human ZNF312b Promotes the Progression of Gastric Cancer by Transcriptional Activation of the K-ras Gene

In the Concealed Information Test (CIT), differential responses between crime-relevant and crime-irrelevant items are indicative of concealed knowledge of a crime, and are used to classify an individual as either "guilty" or "innocent". However, when crime-relevant items are leaked before the test, an innocent examinee can exhibit enhanced responses to the crime-relevant items, thus causing such examinee to be wrongly classified as guilty. In an attempt to solve this problem, we examined the role of retroactive memory interference (RI) in differentiating informed innocents from guilty participants, using a P300-based CIT. Participants acquired crime-related knowledge either by committing a mock crime (guilty group) or reading a paper that described a mock crime (informed innocent group). Subsequently, the participants within each condition were randomly assigned to either an RI group, where they were exposed to new crime-related details before the CIT, or a control group. We found an interaction between guilty and RI groups: in the guilty group, there was a significant difference in P300 amplitude between the probe and irrelevant items, regardless of RI manipulation, whereas in the informed innocent group, a difference in P300 amplitude between the probe and irrelevant items was significant only in the control group, but not in the RI group. This led to an improved detection rate of the informed innocents (31% for the control group vs. 77% for the RI group). These results suggest that RI manipulation could be used to reduce the false positive outcomes of informed innocents without affecting the detection rate of guilty participants.

Many studies have consistently proven that repeatedly watching virtual reality (VR) content can reduce cybersickness. Moreover, the discomfort level decreases when the VR content includes an unusual orientation, such as an inverted scene. However, few studies have investigated the physiological changes during these experiences. The present study aimed to identify psychophysiological correlates, especially the neural processing, of cybersickness. Twenty participants experienced two types of VR orientation (upright and inverted), which were repeated three times. During the experience, we recorded the participants’ subjective levels of discomfort, brain waves, cardiac signals, and eye trajectories. We performed a heartbeat-evoked potential (HEP) analysis to elucidate the cortical activity of heartbeats while experiencing cybersickness. The results showed that the severity of cybersickness decreased as the participants repeatedly watched the VR content. The participants also reported less nausea when watching the inverted orientation. We only found a significant suppression at the fronto-central HEP amplitudes in the upright orientation for the physiological changes. This study provides a comprehensive understanding of bodily responses to varying degrees of cybersickness. In addition, the HEP results suggest that this approach might reflect the neural correlates of transient changes in heartbeats caused by cybersickness.

Thermoplastic polyetherester elastomer (TPEE) based on poly(butylene terephthalate) (PBT) as a hard segment and poly(tetramethylene ether glycol terephthalate) (PTMGT) as a soft segment was synthesized with hard segment content (HSC) 35 wt%. The high molecular weight TPEE copolymer was successfully prepared by using MDI as a chain extender from the TPEE synthesized. The intrinsic viscosity of the TPEE copolymer containing 3 wt% of MDI increased from 1.56 dl/g to 2.13 dl/g and the melt index decreased from 54.2 g/min to 11.5 g/min. The breaking stress of TPEE-MDI monofilament increased with MDI content and the breaking strain decreased with increasing MDI content. However, it is noticeable that the breaking stress of TPEE-MDI 3 wt% copolymer monofilament showed 41 % enhanced breaking stress while still retained pretty high breaking strain, i.e. 825 %. The elastic recovery of TPEE-MDI 1 wt% monofilament was 80.7 % in the first cycle and 76.0 % in the final cycle, which were higher than that of TPEE both in the first and the final cycles. These results meant that TPEE-MDI copolymer fiber could have enhanced elastic recovery as well as enhanced breaking stress.

A number of inflammatory lung diseases, including chronic obstructive pulmonary disease, idiopathic pulmonary fibrosis, and pneumonia, are modulated by WNT/β-catenin signaling. However, the underlying molecular mechanisms remain unclear. Here, starting with a forward genetic screen in mouse, we identify the WNT coreceptor Related to receptor tyrosine kinase (RYK) acting in mesenchymal tissues as a cell survival and antiinflammatory modulator.Rykmutant mice exhibit lung hypoplasia and inflammation as well as alveolar simplification due to defective secondary septation, and deletion ofRykspecifically in mesenchymal cells also leads to these phenotypes. By analyzing the transcriptome of wild-type and mutant lungs, we observed the up-regulation of proapoptotic and inflammatory genes whose expression can be repressed by WNT/RYK signaling in vitro. Moreover, mesenchymalRykdeletion at postnatal and adult stages can also lead to lung inflammation, thus indicating a continued role for WNT/RYK signaling in homeostasis. Our results indicate that RYK signaling through β-catenin and Nuclear Factor kappa B (NF-κB) is part of a safeguard mechanism against mesenchymal cell death, excessive inflammatory cytokine production, and inflammatory cell recruitment and accumulation. Notably, RYK expression is down-regulated in the stromal cells of pneumonitis patient lungs. Altogether, our data reveal that RYK signaling plays critical roles as an antiinflammatory modulator during lung development and homeostasis and provide an animal model to further investigate the etiology of, and therapeutic approaches to, inflammatory lung diseases.

Goblet cell metaplasia and mucus hypersecretion are observed in many pulmonary diseases, including asthma, chronic obstructive pulmonary disease (COPD), and cystic fibrosis. However, the regulation of goblet cell differentiation remains unclear. Here, we identify a regulator of this process in an N -ethyl- N -nitrosourea (ENU) screen for modulators of postnatal lung development; Ryk mutant mice exhibit lung inflammation, goblet cell hyperplasia, and mucus hypersecretion. RYK functions as a WNT coreceptor, and, in the developing lung, we observed high RYK expression in airway epithelial cells and moderate expression in mesenchymal cells as well as in alveolar epithelial cells. From transcriptomic analyses and follow-up studies, we found decreased WNT/β-catenin signaling activity in the mutant lung epithelium. Epithelial-specific Ryk deletion causes goblet cell hyperplasia and mucus hypersecretion but not inflammation, while club cell-specific Ryk deletion in adult stages leads to goblet cell hyperplasia and mucus hypersecretion during regeneration. We also found that the airway epithelium of COPD patients often displays goblet cell metaplastic foci, as well as reduced RYK expression. Altogether, our findings reveal that RYK plays important roles in maintaining the balance between airway epithelial cell populations during development and repair, and that defects in RYK expression or function may contribute to the pathogenesis of human lung diseases.

We report a comprehensive Cu ${L}_{3}$-edge resonant x-ray scattering (RXS) study of two- and three-dimensional (2D and 3D) incommensurate charge correlations in single crystals of the underdoped high-temperature superconductor ${\mathrm{YBa}}_{2}{\mathrm{Cu}}_{3}{\mathrm{O}}_{6.67}$ under uniaxial compression up to 1% along the two inequivalent $\mathrm{Cu}─\mathrm{O}─\mathrm{Cu}$ bond directions ($a$ and $b$) in the ${\mathrm{CuO}}_{2}$ planes. We confirm the strong in-plane anisotropy of the 2D charge correlations and observe their symmetric response to pressure: pressure along $a$ enhances correlations along $b$, and vice versa. Our results imply that the underlying order parameter is uniaxial. In contrast, 3D long-range charge order is only observed along $b$ in response to compression along $a$. Spectroscopic RXS measurements show that the 3D charge order resides exclusively in the ${\mathrm{CuO}}_{2}$ planes and may thus be generic to the cuprates. We discuss implications of these results for models of electronic nematicity and for the interplay between charge order and superconductivity.

α-RuCl3 is a major candidate for the realization of the Kitaev quantum spin liquid, but its zigzag antiferromagnetic order at low temperatures indicates deviations from the Kitaev model. We have quantified the spin Hamiltonian of α-RuCl3 by a resonant inelastic x-ray scattering study at the Ru L3 absorption edge. In the paramagnetic state, the quasi-elastic intensity of magnetic excitations has a broad maximum around the zone center without any local maxima at the zigzag magnetic Bragg wavevectors. This finding implies that the zigzag order is fragile and readily destabilized by competing ferromagnetic correlations. The classical ground state of the experimentally determined Hamiltonian is actually ferromagnetic. The zigzag state is stabilized by quantum fluctuations, leaving ferromagnetism - along with the Kitaev spin liquid - as energetically proximate metastable states. The three closely competing states and their collective excitations hold the key to the theoretical understanding of the unusual properties of α-RuCl3 in magnetic fields., European Research Council, Alexander von Humbolft Foundation, Czech Science Foundation, Projekt DEAL

Coloboma, heart defect, atresia choanae, retarded growth and development, genital hypoplasia, ear anomalies/deafness (CHARGE) syndrome is a congenital disorder affecting multiple organs and mainly caused by mutations in CHD7, a gene encoding a chromatin-remodeling protein. Immunodeficiency and reduced T cells have been noted in CHARGE syndrome. However, the mechanisms underlying T lymphopenia are largely unexplored. Herein, we observed dramatic decrease of T cells in both chd7knockdown and knockout zebrafish embryos. Unexpectedly, hematopoietic stem and progenitor cells and, particularly, lymphoid progenitor cells were increased peripherally in nonthymic areas in chd7-deficient embryos, unlikely to contribute to the T-cell decrease. Further analysis demonstrated that both the organogenesis and homing function of the thymus were seriously impaired. Chd7 might regulate thymus organogenesis through modulating the development of both neural crest cell-derived mesenchyme and pharyngeal endoderm-derived thymic epithelial cells. The expression of foxn1, a central regulator of thymic epithelium, was remarkably down-regulated in the pharyngeal region in chd7-deficient embryos. Moreover, the T-cell reduction in chd7-deficient embryos was partially rescued by overexpressing foxn1, suggesting that restoring thymic epithelium may be a potential therapeutic strategy for treating immunodeficiency in CHARGE syndrome. Collectively, the results indicated that chd7 was critical for thymic development and T-lymphopenia in CHARGE syndrome may be mainly attributed to the defects of thymic organogenesis. The current finding may benefit the diagnosis and therapy of T lymphopenia and immunodeficiency in CHARGE syndrome.

A novel chemically bonded phosphate ceramic (CBPC) binder was developed for the simultaneous treatment of the top five most toxic heavy metals (Hg, Pb, As, Cr, and Cd). Various CBPC binders were synthesized and tested, and their toxicity characteristic leaching procedure (TCLP) values were obtained. A magnesium/calcium-potassium phosphate ceramic binder with FeCl2 (M/C-KP-FeCl2) simultaneously stabilized multiple heavy metals. The TCLP value of the final product for industrial waste (IW) treatment using the M/C-KP-FeCl2 technology was well below the Universal Treatment Standard (UTS). Additionally, the compressive strength of the final product was below the US Nuclear Regulatory Commission Standard.

We have carried out Raman spectroscopy experiments to investigate two-magnon excitations in epitaxial thin films of the quasi-two-dimensional antiferromagnetic Mott insulator $\mathrm{S}{\mathrm{r}}_{2}\mathrm{Ir}{\mathrm{O}}_{4}$ under in-plane misfit strain. With in-plane biaxial compression, the energy of the two-magnon peak increases and the peak remains observable over a wider temperature range above the N\'eel temperature, indicating a strain-induced enhancement of the superexchange interactions between ${J}_{\mathrm{eff}}=1/2$ pseudospins. From density functional theory calculations, we have found an increase of the nearest-neighbor hopping parameter and exchange interaction with increasing biaxial compressive strain, in agreement with the experimental observations. Our experimental and theoretical results provide perspectives for systematic, theory-guided strain control of the primary exchange interactions in $5d$ transition-metal oxides.

// Jae-Hye Lee 1, 3, * , Hyun-Soo Cho 1, 3, * , Jeong-Ju Lee 1 , Soo Young Jun 1, 3 , Jun-Ho Ahn 1 , Ju-Sik Min 1 , Ji-Yong Yoon 1 , Min-Hyuk Choi 1, 3 , Su-Jin Jeon 1, 3 , Jung Hwa Lim 2 , Cho-Rok Jung 2 , Dae-Soo Kim 1, 3 , Hyun-Taek Kim 4 , Valentina M. Factor 5 , Yun-Han Lee 6 , Snorri S. Thorgeirsson 7 , Cheol-Hee Kim 4 , Nam-Soon Kim 1, 3 1 Genome Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon 305-333, Republic of Korea 2 Gene Therapy Research Unit, Korea Research Institute of Bioscience and Biotechnology, Daejeon 305-333, Republic of Korea 3 Department of Functional Genomics, Korea University of Science and Technology, Daejeon 305-333, Republic of Korea 4 Department of Biology, Chungnam National University, Daejeon 305-764, Republic of Korea 5 Laboratory of Molecular Pharmacology, Center for Cancer Research, NCI, NIH, Bethesda, MD 20892-5068, USA 6 Department of Molecular Medicine, Keimyung University School of Medicine, Daegu 704-701, Republic of Korea 7 Laboratory of Human Carcinogenesis, Center for Cancer Research, NCI, NIH, Bethesda, MD 20892-4255, USA * Both authors are shared co-first authorship Correspondence to: Nam-Soon Kim, email: nskim37@kribb.re.kr Keywords: liver cancer, metastasis, PGCP, Wnt/β-catenin Received: July 25, 2016 Accepted: October 14, 2016 Published: October 28, 2016 ABSTRACT Tumor metastasis is the leading cause of cancer death. In the metastatic process, EMT is a unique phenotypic change that plays an important role in cell invasion and changes in cell morphology. Despite the clinical significance, the mechanism underlying tumor metastasis is still poorly understood. Here we report a novel mechanism by which secreted plasma glutamate carboxypeptidase(PGCP) negatively involves Wnt/β-catenin signaling by DKK4 regulation in liver cancer metastasis. Pathway analysis of the RNA sequencing data showed that PGCP knockdown in liver cancer cell lines enriched the functions of cell migration, motility and mesenchymal cell differentiation. Depletion of PGCP promoted cell migration and invasion via activation of Wnt/β-catenin signaling pathway components such as phospho-LRP6 and β-catenin. Also, addition of DKK4 antagonized the Wnt/β-catenin signaling cascade in a thyroxine (T4)-dependent manner. In an in vivo study, metastatic nodules were observed in the lungs of the mice after injection of shPGCP stable cell lines. Our findings suggest that PGCP negatively associates with Wnt/β-catenin signaling during metastasis. Targeting this regulation may represent a novel and effective therapeutic option for liver cancer by preventing metastatic activity of primary tumor cells.

Impaired alveolar formation and maintenance are features of many pulmonary diseases that are associated with significant morbidity and mortality. In a forward genetic screen for modulators of mouse lung development, we identified the non-muscle myosin II heavy chain gene, Myh10. Myh10 mutant pups exhibit cyanosis and respiratory distress, and die shortly after birth from differentiation defects in alveolar epithelium and mesenchyme. From omics analyses and follow up studies, we find decreased Thrombospondin expression accompanied with increased matrix metalloproteinase activity in both mutant lungs and cultured mutant fibroblasts, as well as disrupted extracellular matrix (ECM) remodeling. Loss of Myh10 specifically in mesenchymal cells results in ECM deposition defects and alveolar simplification. Notably, MYH10 expression is downregulated in the lung of emphysema patients. Altogether, our findings reveal critical roles for Myh10 in alveologenesis at least in part via the regulation of ECM remodeling, which may contribute to the pathogenesis of emphysema., Abnormal alveolar development and homeostasis are common features of pulmonary disease. Here the authors show that Myh10 expression is reduced in emphysema patients, and that Myh10 loss of function impairs alveolar formation and lung morphogenesis via upregulation of matrix metalloproteinase activity and altered matrix remodeling.

Secreted Wnts play crucial roles in synaptogenesis and synapse maintenance, but endogenous factors promoting synapse elimination in central neurons remain unknown. Here we show that proline-rich 7 (PRR7) induces specific removal of excitatory synapses and acts as a Wnt inhibitor. Remarkably, transmembrane protein PRR7 is activity-dependently released by neurons via exosomes. Exosomal PRR7 is uptaken by neurons through membrane fusion and eliminates excitatory synapses in neighboring neurons. Conversely, PRR7 knockdown in sparse neurons greatly increases excitatory synapse numbers in all surrounding neurons. These non-cell autonomous effects of PRR7 are effectively negated by augmentation or blockade of Wnt signaling. PRR7 exerts its effect by blocking the exosomal secretion of Wnts, activation of GSK3β, and promoting proteasomal degradation of PSD proteins. These data uncover a proximity-dependent, reciprocal mechanism for the regulation of excitatory synapse numbers in local neurons and demonstrate the significance of exosomes in inter-neuronal signaling in the vertebrate brain., Wnts are important for synapse formation and maintenance. Here, the authors show that proline-rich 7 (PRR7) is a Wnt inhibitor that is secreted via exosomes to regulate excitatory synapse numbers.

Tubulogenesis is essential for the formation and function of internal organs. One such organ is the trachea, which allows gas exchange between the external environment and the lungs. However, the cellular and molecular mechanisms underlying tracheal tube development remain poorly understood. Here, we show that the potassium channel KCNJ13 is a critical modulator of tracheal tubulogenesis. We identify Kcnj13 in an ethylnitrosourea forward genetic screen for regulators of mouse respiratory organ development. Kcnj13 mutants exhibit a shorter trachea as well as defective smooth muscle (SM) cell alignment and polarity. KCNJ13 is essential to maintain ion homeostasis in tracheal SM cells, which is required for actin polymerization. This process appears to be mediated, at least in part, through activation of the actin regulator AKT, as pharmacological increase of AKT phosphorylation ameliorates the Kcnj13-mutant trachea phenotypes. These results provide insight into the role of ion homeostasis in cytoskeletal organization during tubulogenesis., Tubulogenesis is required for the formation of many internal structures including the trachea. Here, the authors show that the potassium channel KCNJ13 regulates tracheal tube formation, with shorter tracheas forming in mutant mice due in part to changes in actin organization in tracheal smooth muscle cells.

Epithelial tubes, comprised of polarised epithelial cells around a lumen, are crucial for organ function. However, the molecular mechanisms underlying tube formation remain largely unknown. Here, we report on the function of fibrillin (FBN)2, an extracellular matrix (ECM) glycoprotein, as a critical regulator of tracheal tube formation.We performed a large-scale forward genetic screen in mouse to identify regulators of respiratory organ development and disease. We identified Fbn2 mutants which exhibit shorter and narrowed tracheas as well as defects in tracheal smooth muscle cell alignment and polarity.We found that FBN2 is essential for elastic fibre formation and Fibronectin accumulation around tracheal smooth muscle cells. These processes appear to be regulated at least in part through inhibition of p38-mediated upregulation of matrix metalloproteinases (MMPs), as pharmacological decrease of p38 phosphorylation or MMP activity partially attenuated the Fbn2 mutant tracheal phenotypes. Analysis of human tracheal tissues indicates that a decrease in ECM proteins, including FBN2 and Fibronectin, is associated with tracheomalacia.Our findings provide novel insights into the role of ECM homeostasis in mesenchymal cell polarisation during tracheal tubulogenesis.

Significance Emotion-related responses, such as fear and anxiety, are important behavioral phenomena in most animal species, as well as in humans. However, the underlying mechanisms of fear and anxiety in animals and in humans are still largely unknown, and anxiety disorders continue to represent a large unmet medical need in the human clinic. Animal models may speed up discovery of these mechanisms and may also lead to betterment of human health. Herein, we report the identification of a chemokine-like gene family, samdori (sam), and present functional characterization of sam2. We observed increased anxiety-related responses in both zebrafish and mouse knockout models. Taken together, these results support a crucial and evolutionarily conserved role of sam2 in regulating anxiety-like behavior., Emotional responses, such as fear and anxiety, are fundamentally important behavioral phenomena with strong fitness components in most animal species. Anxiety-related disorders continue to represent a major unmet medical need in our society, mostly because we still do not fully understand the mechanisms of these diseases. Animal models may speed up discovery of these mechanisms. The zebrafish is a highly promising model organism in this field. Here, we report the identification of a chemokine-like gene family, samdori (sam), and present functional characterization of one of its members, sam2. We show exclusive mRNA expression of sam2 in the CNS, predominantly in the dorsal habenula, telencephalon, and hypothalamus. We found knockout (KO) zebrafish to exhibit altered anxiety-related responses in the tank, scototaxis and shoaling assays, and increased crh mRNA expression in their hypothalamus compared with wild-type fish. To investigate generalizability of our findings to mammals, we developed a Sam2 KO mouse and compared it to wild-type littermates. Consistent with zebrafish findings, homozygous KO mice exhibited signs of elevated anxiety. We also found bath application of purified SAM2 protein to increase inhibitory postsynaptic transmission onto CRH neurons of the paraventricular nucleus. Finally, we identified a human homolog of SAM2, and were able to refine a candidate gene region encompassing SAM2, among 21 annotated genes, which is associated with intellectual disability and autism spectrum disorder in the 12q14.1 deletion syndrome. Taken together, these results suggest a crucial and evolutionarily conserved role of sam2 in regulating mechanisms associated with anxiety.

Cilia are microtubule-based structures that project into the extracellular space. Ciliary defects are associated with several human diseases, including polycystic kidney disease, primary ciliary dyskinesia, left-right axis patterning, hydrocephalus and retinal degeneration. However, the genetic and cellular biological control of ciliogenesis remains poorly understood. The IFT46 is one of the highly conserved intraflagellar transport complex B proteins. In zebrafish, ift46 is expressed in various ciliated tissues such as Kupffer׳s vesicle, pronephric ducts, ears and spinal cord. We show that ift46 is localized to the basal body. Knockdown of ift46 gene results in multiple phenotypes associated with various ciliopathies including kidney cysts, pericardial edema and ventral axis curvature. In ift46 morphants, cilia in kidney and spinal canal are shortened and abnormal. Similar ciliary defects are observed in otic vesicles, lateral line hair cells, olfactory pits, but not in Kupffer׳s vesicle. To explore the functions of Ift46 during mouse development, we have generated Ift46 knock-out mice. The Ift46 mutants have developmental defects in brain, neural tube and heart. In particular Ift46(-/-) homozygotes displays randomization of the embryo heart looping, which is a hallmark of defective left-right (L/R) axis patterning. Taken together, our results demonstrated that IFT46 has an essential role in vertebrate ciliary development.

Miles–Carpenter syndrome (MCS) was described in 1991 as an XLID syndrome with fingertip arches and contractures and mapped to proximal Xq. Patients had microcephaly, short stature, mild spasticity, thoracic scoliosis, hyperextendable MCP joints, rocker-bottom feet, hyperextended elbows and knees. A mutation, p.L66H, in ZC4H2, was identified in a XLID re-sequencing project. Additional screening of linked families and next generation sequencing of XLID families identified three ZC4H2 mutations: p.R18K, p.R213W and p.V75in15aa. The families shared some relevant clinical features. In silico modeling of the mutant proteins indicated all alterations would destabilize the protein. Knockout mutations in zc4h2 were created in zebrafish and homozygous mutant larvae exhibited abnormal swimming, increased twitching, defective eye movement and pectoral fin contractures. Because several of the behavioral defects were consistent with hyperactivity, we examined the underlying neuronal defects and found that sensory neurons and motoneurons appeared normal. However, we observed a striking reduction in GABAergic interneurons. Analysis of cell-type-specific markers showed a specific loss of V2 interneurons in the brain and spinal cord, likely arising from mis-specification of neural progenitors. Injected human wt ZC4H2 rescued the mutant phenotype. Mutant zebrafish injected with human p.L66H or p.R213W mRNA failed to be rescued, while the p.R18K mRNA was able to rescue the interneuron defect. Our findings clearly support ZC4H2 as a novel XLID gene with a required function in interneuron development. Loss of function of ZC4H2 thus likely results in altered connectivity of many brain and spinal circuits.

Wnt signaling controls critical developmental processes including tissue/body patterning. Here we report the identification of a novel regulator of Wnt signaling, OTTOGI (OTG), isolated from a large-scale expression screening of human cDNAs in zebrafish embryos. Overexpression of OTG in zebrafish embryos caused dorso-anteriorized phenotype, inhibited the expression of Wnt target genes, and prevented nuclear accumulation of β-catenin. Conversely, knockdown of zebrafish otg using specific antisense morpholino promoted nuclear accumulation of β-catenin and caused ventralization. However, OTG failed to rescue headless-like phenotype induced by inhibition of GSK-3β activity, suggesting that OTG acts upstream of GSK-3β. OTG bound specifically to Frizzled8 (Fz8) receptor and caused retention of Fz8 in the endoplasmic reticulum possibly by preventing N-linked glycosylation of Fz8. Taken together, our data indicate that OTG functions as a novel negative regulator of Wnt signaling during development by the modulation of cell surface expression of Fz receptor.

WDR11 has been implicated in congenital hypogonadotropic hypogonadism (CHH) and Kallmann syndrome (KS), human developmental genetic disorders defined by delayed puberty and infertility. However, WDR11’s role in development is poorly understood. Here we report that WDR11 modulates the Hedgehog (Hh) signalling pathway and is essential for ciliogenesis. Disruption of WDR11 expression in mouse and zebrafish results in phenotypic characteristics associated with defective Hh signalling, accompanied by dysgenesis of ciliated tissues.Wdr11null mice also exhibit early onset obesity. We found that WDR11 shuttles from the cilium to the nucleus in response to Hh signalling. WDR11 was also observed to regulate the proteolytic processing of GLI3 and cooperate with EMX1 transcription factor to induce the expression of downstream Hh pathway genes and gonadotrophin releasing hormone production. The CHH/KS-associated human mutations result in loss-of-function of WDR11. Treatment with the Hh agonist purmorphamine partially rescued the WDR11-haploinsufficiency phenotypes. Our study reveals a novel class of ciliopathy caused by WDR11 mutations and suggests that CHH/KS may be a part of the human ciliopathy spectrum.

Zebrafish regenerate damaged myocardial tissue very effectively. Hence, insights into the molecular networks underlying zebrafish heart regeneration might help develop alternative strategies to restore human cardiac performance. While TGF-β signaling has been implicated in zebrafish cardiac regeneration, the role of its individual ligands remains unclear. Here, we report the opposing expression response during zebrafish heart regeneration of two genes, mstnb and inhbaa, which encode TGF-β family ligands. Using gain-of-function (GOF) and loss-of-function (LOF) approaches, we show that these ligands mediate inverse effects on cardiac regeneration and specifically on cardiomyocyte (CM) proliferation. Notably, we find that Inhbaa functions as a CM mitogen and that its overexpression leads to accelerated cardiac recovery and scar clearance after injury. In contrast, mstnb GOF and inhbaa LOF both lead to unresolved scarring after cardiac injury. We further show that Mstnb and Inhbaa inversely control Smad2 and Smad3 transcription factor activities through alternate Activin type 2 receptors., Zebrafish can regenerate damaged myocardial tissue but it is unclear how this is regulated. Here, the authors show that two TGF-β family members, Mstnb and Inhbaa, have opposite effects in regeneration, with mstnb overexpression or inhbaa loss-of-function causing cardiac scarring after injury.

Clusterin, a protein associated with multiple functions, is expressed in a wide variety of mammalian tissues. Although clusterin is known to be involved in neurodegenerative diseases, ageing, and tumorigenesis, a detailed analysis of the consequences of gain- or loss-of-function approaches has yet to be performed to understand the underlying mechanisms of clusterin functions. Since clusterin levels change in neurological diseases, it is likely that clusterin contributes to cell death and degeneration in general. Zebrafish was investigated as a model system to study human diseases. During development, zebrafish clusterin was expressed in the notochord and nervous system. Embryonic overexpression of clusterin by mRNA microinjection did not affect axis formation, whereas its knock-down by anti-sense morpholino treatment resulted in neuronal cell death. To analyze the function of clusterin in neurodegeneration, a transgenic zebrafish was investigated, in which nitroreductase expression is regulated under the control of a neuron-specific huC promoter which is active between the stages of early neuronal precursors and mature neurons. Nitroreductase turns metronidazole into a cytotoxic agent that induces cell death within 12 h. After metronidazole treatment, transgenic zebrafish showed neuron-specific cell death. Interestingly, we also observed a dramatic induction of clusterin expression in the brain and spinal cord in these fish, suggesting a direct or indirect role of clusterin in neuronal cell death and thus, more generally, in neurodegeneration.

Both water quantity and quality are impacted by climate change. In addition, rapid urbanization has also brought an immeasurable loss of life and property resulting from floods. Hence, there is a need to predict changes in rainfall events to effectively design stormwater infrastructure to protect urban areas from disaster. This study develops a framework for predicting future short duration rainfall intensity and examining the effects of climate change on urban runoff in the Gunja Drainage Basin. Non-stationarities in rainfall records are first analysed using trend analysis to extrapolate future climate change scenarios. The US Environmental Protection Agency Storm Water Management Model (SWMM) was used for single event simulation of runoff quantity from the study area. For the 1-hour and 24-hour durations, statistically significant upward trends were observed. Although the 10-minute duration was only nearly significant at the 90% level, the steepest slope was observed for this short duration. Moreover, it was observed that the simulated peak discharge from SWMM increases as the short duration rainfall intensity increases. The proposed framework is thought to provide a means to review the current design of stormwater infrastructures to determine their capacity, along with consideration of climate change impact.

In light of a recent development in a coolant leakage detection method for pressurized heavy water reactors (PHWRs), which utilizes a novel leak detector based on laser absorption spectroscopy, we report the successful application of the said detection method for the detection and location of leaks in the fuel channel closure plug of the PHWRs. This paper contains the results of real-world performance tests performed on operational commercial PHWRs, and shows that the newly developed laser leak detector was fully capable of finding leak locations as well as detecting leaks with a sensitivity of less than ∼1 g/h. Based on these findings, we believe the laser leak detector can be a reliable apparatus for the early detection of heavy water leakage at fuel channel closure plugs during the normal operation of PHWRs.

본 연구에서는 사건관련전위(event-related potentials: ERPs)를 사용하여 외향적인 사람들과 내향적인 사람들이 경계과제를 수행할 때 스트레스 유무에 따라 발생하는 뇌신경 활동의 차이를 밝히고자 하였다. 마흔 세 명(외향성 21, 내향성 22)의 피험자들을 각각 외향성 집단과 내향성 집단으로 나누어서 사회적 스트레스 요인이 없는 상황과 사회적 스트레스 요인이 있는 상황에서 경계과제를 수행하는 동안 사건관련전위를 측정하였다. 행동반응에서는 두 집단 모두 사회적 스트레스 요인이 있는 경우에 정확도가 증가하였다. 사건관련전위에서는 첫 번째, 내향성 집단의 경우에 사회적 스트레스 요인 유무에 관계없이 N2-P2 최고점간 진폭과 CNV(contingent negative variation) 평균진폭이 외향성 집단 보다 유의미하게 크게 나타났다. 두 번째, 외향성 집단의 경우에 사회적 스트레스 요인이 있는 경우에만 전두엽 부위(Fz)에서 N2-P2 최고점간 진폭이 유의미하게 증가하였다. 이러한 결과는 동일한 사회적 스트레스 요인이 외향적-내향적 성격에 따라 상이한 뇌신경활동에 의해서 매개될 수 있다는 것을 보여주며, 내향성 집단이 외향성 집단 보다 더 높은 각성 수준과 주의 수준을 유지하고 있음을 시사한다.