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1.
Plant J ; 117(3): 747-765, 2024 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-37926922

RESUMEN

Brassinazole Resistant 1 (BZR1) and bri1 EMS Suppressor 1 (BES1) are key transcription factors that mediate brassinosteroid (BR)-responsive gene expression in Arabidopsis. The BZR1/BES1 family is composed of BZR1, BES1, and four BES1/BZR1 homologs (BEH1-BEH4). However, little is known about whether BEHs are regulated by BR signaling in the same way as BZR1 and BES1. We comparatively analyzed the functional characteristics of six BZR1/BES1 family members and their regulatory mechanisms in BR signaling using genetic and biochemical analyses. We also compared their subcellular localizations regulated by the phosphorylation status, interaction with GSK3-like kinases, and heterodimeric combination. We found that all BZR1/BES1 family members restored the phenotypic defects of bri1-5 by their overexpression. Unexpectedly, BEH2-overexpressing plants showed the most distinct phenotype with enhanced BR responses. RNA-Seq analysis indicated that overexpression of both BZR1 and BEH2 regulates BR-responsive gene expression, but BEH2 has a much greater proportion of BR-independent gene expression than BZR1. Unlike BZR1 and BES1, the BR-regulated subcellular translocation of the four BEHs was not tightly correlated with their phosphorylation status. Notably, BEH1 and BEH2 are predominantly localized in the nucleus, which induces the nuclear accumulation of other BZR1/BES1 family proteins through heterodimerization. Altogether, our comparative analyses suggest that BEH1 and BEH2 play an important role in the functional interaction between BZR1/BES1 family transcription factors.


Asunto(s)
Proteínas de Arabidopsis , Arabidopsis , Triazoles , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Brasinoesteroides/metabolismo , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Regulación de la Expresión Génica de las Plantas , Glucógeno Sintasa Quinasa 3/genética , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Factores de Transcripción/genética , Factores de Transcripción/metabolismo
2.
Small ; 20(29): e2311652, 2024 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-38361217

RESUMEN

Modern strides in energy storage underscore the significance of all-solid-state batteries (ASSBs) predicated on solid electrolytes and lithium (Li) metal anodes in response to the demand for safer batteries. Nonetheless, ASSBs are often beleaguered by non-uniform Li deposition during cycling, leading to compromised cell performance from internal short circuits and hindered charge transfer. In this study, the concept of "bottom deposition" is introduced to stabilize metal deposition based on the lithiophilic current collector and a protective layer composed of a polymeric binder and carbon black. The bottom deposition, wherein Li plating ensues between the protective layer and the current collector, circumvents internal short circuits and facilitates uniform volumetric changes of Li. The prepared functional binder for the protective layer presents outstanding mechanical robustness and adhesive properties, which can withstand the volume expansion caused by metal growth. Furthermore, its excellent ion transfer properties promote uniform Li bottom deposition even under a current density of 6 mA·cm-2. Also, scanning electron microscopy analysis reveals a consistent plating/stripping morphology of Li after cycling. Consequently, the proposed system exhibits enhanced electrochemical performance when assessed within the ASSB framework, operating under a configuration marked by a high Li utilization rate reliant on an ultrathin Li.

3.
Small ; : e2403147, 2024 Jul 11.
Artículo en Inglés | MEDLINE | ID: mdl-38989706

RESUMEN

All-solid-state batteries (ASSBs) possess the advantage of ensuring safety while simultaneously maximizing energy density, making them suitable for next-generation battery models. In particular, sulfide solid electrolytes (SSEs) are viewed as promising candidates for ASSB electrolytes due to their excellent ionic conductivity. However, a limitation exists in the form of interfacial side reactions occurring between the SSEs and cathode active materials (CAMs), as well as the generation of sulfide-based gases within the SSE. These issues lead to a reduction in the capacity of CAMs and an increase in internal resistance within the cell. To address these challenges, cathode composite materials incorporating zinc oxide (ZnO) are fabricated, effectively reducing various side reactions occurring in CAMs. Acting as a semiconductor, ZnO helps mitigate the rapid oxidation of the solid electrolyte facilitated by an electronic pathway, thereby minimizing side reactions, while maintaining electron pathways to the active material. Additionally, it absorbs sulfide-based gases, thus protecting the lithium ions within CAMs. In this study, the mass spectrometer is employed to observe gas generation phenomena within the ASSB cell. Furthermore, a clear elucidation of the side reactions occurring at the cathode and the causes of capacity reduction in ASSB are provided through density functional theory calculations.

4.
Cell Tissue Res ; 395(1): 53-62, 2024 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-37985496

RESUMEN

Glomerular epithelial protein-1 (Glepp1), a R3 subtype family of receptor-type protein tyrosine phosphatases, plays important role in the activation of Src family kinases and regulates cellular processes such as cell proliferation, differentiation, and apoptosis. In this study, we firstly examined the functional evaluation of Glepp1 in tooth development and morphogenesis. The precise expression level and developmental function of Glepp1 were examined by RT-qPCR, in situ hybridization, and loss and gain of functional study using a range of in vitro organ cultivation methods. Expression of Glepp1 was detected in the developing tooth germs in cap and bell stage of tooth development. Knocking down Glepp1 at E13 for 2 days showed the altered expression levels of tooth development-related signaling molecules, including Bmps, Dspp, Fgf4, Lef1, and Shh. Moreover, transient knock down of Glepp1 revealed alterations in cellular physiology, examined by the localization patterns of Ki67 and E-cadherin. Similarly, knocking down of Glepp1 showed disrupted enamel rod and interrod formation in 3-week renal transplanted teeth. In addition, due to attrition of odontoblastic layers, the expression signals of Dspp and the localization of NESTIN were almost not detected after knock down of Glepp1; however, their expressions were increased after Glepp1 overexpression. Thus, our results suggested that Glepp1 plays modulating roles during odontogenesis by regulating the expression levels of signaling molecules and cellular events to achieve the proper structural formation of hard tissue matrices in mice molar development.


Asunto(s)
Proteínas Tirosina Fosfatasas Clase 3 Similares a Receptores , Diente , Animales , Ratones , Regulación del Desarrollo de la Expresión Génica , Morfogénesis , Odontogénesis , Proteínas Tirosina Fosfatasas/metabolismo , Proteínas Tirosina Fosfatasas Clase 3 Similares a Receptores/metabolismo , Transducción de Señal , Diente/metabolismo
5.
J Invertebr Pathol ; 206: 108166, 2024 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-39004164

RESUMEN

Peptidoglycan recognition proteins (PGRPs) and Toll-like receptors (TLRs) are highly conserved pattern recognition receptors (PRRs). Earthworms possess genes encoding TLRs that specifically respond to Gram-positive bacteria. In addition, several PGRPs have been recently identified, which are predicted to exhibit amidase activity but lack receptor function. In lophotrochozoans, a membrane-bound PRR responsible for detecting Gram-negative bacteria remains unidentified. This study reveals several novel transmembrane peptidoglycan recognition proteins (Ean-PGRPLs) in earthworms, whose mRNA expression increases in response to Gram-negative but not Gram-positive bacteria. This indicates that Ean-PGRPLs may serve as a PRR associated with intracellular signaling for Gram-negative bacteria.


Asunto(s)
Proteínas Portadoras , Oligoquetos , Animales , Oligoquetos/microbiología , Oligoquetos/metabolismo , Proteínas Portadoras/metabolismo , Proteínas Portadoras/genética , Bacterias Gramnegativas , Bacterias Grampositivas
6.
Biochem Biophys Res Commun ; 666: 21-28, 2023 07 23.
Artículo en Inglés | MEDLINE | ID: mdl-37167720

RESUMEN

This study aimed to develop a new symmetric-end antimicrobial peptide (AMP) with cell selectivity, antibiofilm, and anti-inflammatory activities. Two symmetric-end AMPs, Lf6-pP and Lf6-GG, were designed based on the sequence RRWQWRzzRWQWRR, which contains two symmetric repeat sequences connected by a ß-turn-promoting sequence (zz) that can be a rigid turn by D-Pro-Pro (pP) or a flexible turn by Gly-Gly (GG). Both Lf6-pP and Lf6-GG exhibited potent antibacterial activity without causing hemolysis, but Lf6-pP exhibited better cell selectivity, likely due to the more significant impact of the rigid pP turn. Compared to Lf6-GG, Lf6-pP demonstrated approximately three times higher antimicrobial activity against drug-resistant bacteria, had a low incidence of drug resistance, and maintained its activity in the presence of physiological salts and human serum. Additionally, Lf6-pP was more effective than Lf6-GG in inhibiting biofilm formation and eradicating mature biofilms. The BODIPY-cadaverine assay indicated that the potent anti-inflammatory activity of Lf6-pP may be attributed to its direct interaction with LPS, resulting in decreased TNF-α and IL-6 levels in LPS-stimulated macrophages. Mechanistic studies, including membrane depolarization, outer/inner membrane permeation, and membrane integrity change, demonstrated that Lf6-pP exerts its antibacterial action through an intracellular-target mechanism. Overall, we propose that Lf6-pP has potential as a novel antibacterial, antibiofilm, and anti-inflammatory agent against drug-resistant bacterial infections.


Asunto(s)
Péptidos Catiónicos Antimicrobianos , Péptidos Antimicrobianos , Humanos , Péptidos Catiónicos Antimicrobianos/farmacología , Lipopolisacáridos/farmacología , Antibacterianos/farmacología , Antiinflamatorios/farmacología , Biopelículas , Pruebas de Sensibilidad Microbiana
7.
J Invertebr Pathol ; 201: 108010, 2023 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-37865158

RESUMEN

Toll-like receptors (TLRs), an ancient and well-conserved group of pattern recognition receptors (PRRs), recognize conserved pathogen-associated molecular patterns. TLRs consist of three domains: the extracellular N-terminal domain, containing one or more leucine-rich repeats (LRRs), responsible for the recognizing and binding of antigens; the type-I transmembrane domain; and the intracellular domain known as the Toll/Interleukin-1 receptor (TIR) domain required for the downstream signaling pathway. We identified six new full-length complementary DNA (cDNA) sequences, Ean-TLR1/2/3/4/5/6. The deduced amino acid sequences indicate that Ean-TLRs consist of one signal peptide, one LRR N-terminal domain (Ean-TLR4/5), varying numbers of LRRs, one (Ean-TLR1/2/3/4/5) or two (Ean-TLR6) LRR C-terminal domains, one type-I transmembrane domain, and a TIR domain. In addition, a TIR domain alignment revealed that three conserved motifs, designated as Box 1, Box 2, and Box 3, contain essential amino acid residues for downstream signaling activity. Phylogenetic analysis of earthworm TLRs generated two separate evolutionary branches representing single (sccTLR) and multiple (mccTLR) cysteine cluster TLRs. Ean-TLR1/2/3/4 (sccTLR type) and Ean-TLR6 (mccTLR type) were clustered with corresponding types of previously reported earthworm TLRs as well as TLRs from Clitellata and Polychaete. As PRRs, earthworm TLRs should be capable of sensing a diverse range of pathogens. Except for Ean-TLR3, which was not responsive to any bacteria, earthworm TLR expression was significantly induced by Gram-positive but not Gram-negative bacteria. Moreover, it is likely that earthworms can differentiate between different species of Gram-positive bacteria via their TLR responses. The ligand specificity of earthworm TLRs suggests that their pathogenic ligand recognition is likely to be as specific and diverse as the mammalian TLR pathogen-sensing system.


Asunto(s)
Oligoquetos , Animales , Filogenia , Receptor Toll-Like 1/genética , Ligandos , Receptor Toll-Like 6/genética , Receptores Toll-Like/genética , Receptores Toll-Like/metabolismo , Receptores de Reconocimiento de Patrones/genética , Bacterias/metabolismo , Inmunidad Innata/genética , Mamíferos/metabolismo
8.
J Neurosci ; 39(50): 9989-10001, 2019 12 11.
Artículo en Inglés | MEDLINE | ID: mdl-31704784

RESUMEN

Hearing loss is the biggest risk factor for tinnitus, and hearing-loss-related pathological changes in the auditory pathway have been hypothesized as the mechanism underlying tinnitus. However, due to the comorbidity of tinnitus and hearing loss, it has been difficult to differentiate between neural correlates of tinnitus and consequences of hearing loss. In this study, we dissociated tinnitus and hearing loss in FVB mice, which exhibit robust resistance to tinnitus following monaural noise-induced hearing loss. Furthermore, knock-down of glutamate decarboxylase 65 (GAD65) expression in auditory cortex (AI) by RNA interference gave rise to tinnitus in normal-hearing FVB mice. We found that tinnitus was significantly correlated with downregulation of GAD65 in the AI. By contrast, cortical map distortions, which have been hypothesized as a mechanism underlying tinnitus, were correlated with hearing loss but not tinnitus. Our findings suggest new strategies for the rehabilitation of tinnitus and other phantom sensation, such as phantom pain.SIGNIFICANCE STATEMENT Hearing loss is the biggest risk factor for tinnitus in humans. Most animal models of tinnitus also exhibit comorbid hearing loss, making it difficult to dissociate the mechanisms underlying tinnitus from mere consequences of hearing loss. Here we show that, although both C57BL/6 and FVB mice exhibited similar noise-induced hearing threshold increase, only C57BL/6, but not FVB, mice developed tinnitus following noise exposure. Although both strains showed frequency map reorganization following noise-induced hearing loss, only C57BL/6 mice had reduced glutamate decarboxylase 65 (GAD65) expression in the auditory cortex (AI). Knocking down GAD65 expression in the AI resulted in tinnitus in normal-hearing FVB mice. Our results suggest that reduced inhibitory neuronal function, but not sensory map reorganization, underlies noise-induced tinnitus.


Asunto(s)
Corteza Auditiva/metabolismo , Vías Auditivas/metabolismo , Regulación hacia Abajo , Glutamato Descarboxilasa/metabolismo , Pérdida Auditiva Provocada por Ruido/metabolismo , Plasticidad Neuronal/fisiología , Acúfeno/metabolismo , Animales , Corteza Auditiva/fisiopatología , Vías Auditivas/fisiopatología , Percepción Auditiva/fisiología , Mapeo Encefálico , Pérdida Auditiva Provocada por Ruido/fisiopatología , Masculino , Ratones , Acúfeno/fisiopatología
9.
Biochem Biophys Res Commun ; 521(1): 15-18, 2020 01 01.
Artículo en Inglés | MEDLINE | ID: mdl-31640857

RESUMEN

To investigate whether earthworm cellulases contribute to the innate immune system, the responsiveness of cellulase activity and mRNA expression to bacterial challenge was examined by zymography and RNA sequencing. A zymographic analysis revealed that the activity levels of earthworm cellulases were upregulated in response to either a bacterial (Bacillus subtilis or Escherichia coli) or LPS challenge. After the challenge, significant increases in cellulase 1 and cellulase 2 activity levels were observed within 8-16 and 16-24 h, respectively. In the coelomic fluid, both activities were significantly upregulated at 8 h post-injection with B. subtilis. Based on RNA sequencing, cellulase-related mRNAs encoding beta-1,4-endoglucanases were upregulated by 3-fold within 6 h after B. subtilis injection. Our results clearly demonstrated that earthworm cellulases are upregulated by bacterial challenge at the mRNA and protein levels. These results support the view that earthworm cellulases act as inducible humoral effectors of innate immunity against bacterial infection.


Asunto(s)
Bacillus subtilis/metabolismo , Celulasas/inmunología , Escherichia coli/metabolismo , Inmunidad Innata/inmunología , Oligoquetos/enzimología , ARN Mensajero/genética , Animales , Secuencia de Bases , Celulasas/metabolismo , Oligoquetos/metabolismo , Oligoquetos/microbiología , ARN Mensajero/metabolismo , Análisis de Secuencia de ARN , Regulación hacia Arriba
10.
Sensors (Basel) ; 20(20)2020 Oct 19.
Artículo en Inglés | MEDLINE | ID: mdl-33086561

RESUMEN

LiDAR-based Simultaneous Localization And Mapping (SLAM), which provides environmental information for autonomous vehicles by map building, is a major challenge for autonomous driving. In addition, the semantic information has been used for the LiDAR-based SLAM with the advent of deep neural network-based semantic segmentation algorithms. The semantic segmented point clouds provide a much greater range of functionality for autonomous vehicles than geometry alone, which can play an important role in the mapping step. However, due to the uncertainty of the semantic segmentation algorithms, the semantic segmented point clouds have limitations in being directly used for SLAM. In order to solve the limitations, this paper proposes a semantic segmentation-based LiDAR SLAM system considering the uncertainty of the semantic segmentation algorithms. The uncertainty is explicitly modeled by proposed probability models which are come from the data-driven approaches. Based on the probability models, this paper proposes semantic registration which calculates the transformation relationship of consecutive point clouds using semantic information with proposed probability models. Furthermore, the proposed probability models are used to determine the semantic class of the points when the multiple scans indicate different classes due to the uncertainty. The proposed framework is verified and evaluated by the KITTI dataset and outdoor environments. The experiment results show that the proposed semantic mapping framework reduces the errors of the mapping poses and eliminates the ambiguity of the semantic information of the generated semantic map.

11.
Int J Mol Sci ; 21(7)2020 Mar 26.
Artículo en Inglés | MEDLINE | ID: mdl-32225111

RESUMEN

Echiura is one of the most intriguing major subgroups of phylum Annelida because, unlike most other annelids, echiuran adults lack metameric body segmentation. Urechis unicinctus lives in U-shape burrows of soft sediments. Little is known about the molecular mechanisms underlying the development of U. unicinctus. Herein, we overviewed the developmental process from zygote to juvenile U. unicinctus using immunohistochemistry and F-actin staining for the nervous and muscular systems, respectively. Through F-actin staining, we found that muscle fibers began to form in the trochophore phase and that muscles for feeding were produced first. Subsequently, in the segmentation larval stage, the transversal muscle was formed in the shape of a ring in an anterior-to-posterior direction with segment formation, as well as a ventromedian muscle for the formation of a ventral nerve cord. After that, many muscle fibers were produced along the entire body and formed the worm-shaped larva. Finally, we investigated the spatiotemporal expression of Uun_st-mhc, Uun_troponin I, Uun_calponin, and Uun_twist genes found in U. unicinctus. During embryonic development, the striated and smooth muscle genes were co-expressed in the same region. However, the adult body wall muscles showed differential gene expression of each muscle layer. The results of this study will provide the basis for the understanding of muscle differentiation in Echiura.


Asunto(s)
Anélidos/crecimiento & desarrollo , Desarrollo de Músculos , Actinas/genética , Actinas/metabolismo , Animales , Músculos/metabolismo , Sistema Nervioso/crecimiento & desarrollo , Sistema Nervioso/metabolismo , Transcriptoma
12.
Biomed Microdevices ; 21(4): 86, 2019 08 27.
Artículo en Inglés | MEDLINE | ID: mdl-31451957

RESUMEN

In this study, a novel film-based immunochromatographic microfluidic device (IMD) has been developed for malaria diagnosis. A microfluidic channel was patterned on a polyethylene terephthalate (PET) double-sided adhesive film using a plotting cutter and was assembled with a polycarbonate (PC) film. The PC film used for the probe immobilization layer was activated using oxygen plasma treatment to modify the film surface with avidin-biotin linker to immobilize a capture antibody. A fluorescent labeled Pan type mAb conjugate was prepared for signal indicator after undergoing a sandwich enzyme-linked immunosorbent assay (ELISA). Target antigens include Plasmodium falciparum (P. falciparum) lactate dehydrogenase (LDH) and Plasmodium vivax (P. vivax) LDH which were injected into the sample inlet. Target antigens combined with the conjugate and then flowed to the detection chamber where two test dots and a control dot (Ctrl) exist. In the presence of P. falciparum LDH, three detection dots including test dot 1 (T1), test dot 2 (T2) and Ctrl revealed fluorescence signals where P. falciparum mAb, Pan type pLDH mAb and goat anti-mouse IgG were immobilized, respectively. When P. vivax LDH was present, T2 and Ctrl dots showed fluorescence signals while no signal was detected with the negative control. P. falciparum LDH and P. vivax LDH were successfully detected on the IMD with a detection limit of 50 ng/mL and 100 ng/mL, respectively. The IMD provides a point-of-care diagnosis platform which is able to analyze pathogenic bacteria and viruses that can be applied in the field of clinical diagnosis and food safety testing.


Asunto(s)
Cromatografía de Afinidad/instrumentación , Dispositivos Laboratorio en un Chip , Malaria/diagnóstico , Especificidad de Anticuerpos , Ensayo de Inmunoadsorción Enzimática , L-Lactato Deshidrogenasa/metabolismo , Límite de Detección , Plasmodium falciparum/enzimología , Plasmodium falciparum/inmunología , Plasmodium falciparum/fisiología , Plasmodium vivax/enzimología , Plasmodium vivax/inmunología , Plasmodium vivax/fisiología
13.
Nature ; 493(7433): 526-31, 2013 Jan 24.
Artículo en Inglés | MEDLINE | ID: mdl-23254933

RESUMEN

Current genomic perspectives on animal diversity neglect two prominent phyla, the molluscs and annelids, that together account for nearly one-third of known marine species and are important both ecologically and as experimental systems in classical embryology. Here we describe the draft genomes of the owl limpet (Lottia gigantea), a marine polychaete (Capitella teleta) and a freshwater leech (Helobdella robusta), and compare them with other animal genomes to investigate the origin and diversification of bilaterians from a genomic perspective. We find that the genome organization, gene structure and functional content of these species are more similar to those of some invertebrate deuterostome genomes (for example, amphioxus and sea urchin) than those of other protostomes that have been sequenced to date (flies, nematodes and flatworms). The conservation of these genomic features enables us to expand the inventory of genes present in the last common bilaterian ancestor, establish the tripartite diversification of bilaterians using multiple genomic characteristics and identify ancient conserved long- and short-range genetic linkages across metazoans. Superimposed on this broadly conserved pan-bilaterian background we find examples of lineage-specific genome evolution, including varying rates of rearrangement, intron gain and loss, expansions and contractions of gene families, and the evolution of clade-specific genes that produce the unique content of each genome.


Asunto(s)
Tipificación del Cuerpo/genética , Evolución Molecular , Genoma/genética , Sanguijuelas/genética , Moluscos/genética , Filogenia , Poliquetos/genética , Animales , Secuencia Conservada/genética , Genes Homeobox/genética , Ligamiento Genético , Especiación Genética , Humanos , Mutación INDEL/genética , Intrones/genética , Sanguijuelas/anatomía & histología , Moluscos/anatomía & histología , Familia de Multigenes/genética , Poliquetos/anatomía & histología , Sintenía/genética
14.
Int J Mol Sci ; 20(16)2019 Aug 16.
Artículo en Inglés | MEDLINE | ID: mdl-31426335

RESUMEN

Antistasin, which was originally discovered in the salivary glands of the Mexican leech Haementeria officinalis, was newly isolated from Helobdella austinensis. To confirm the temporal expression of antistasin during embryogenesis, we carried out semi-quantitative RT-PCR. Hau-antistasin1 was uniquely expressed at stage 4 of the cleavage and was strongly expressed in the late stages of organogenesis, as were other antistasin members. In order to confirm the spatial expression of antistasin, we performed fluorescence in situ hybridization in the late stages of organogenesis. The expression of each antistasin in the proboscis showed a similar pattern and varied in expression in the body. In addition, the spatial expression of antistasin orthologs in different leeches showed the possibility of different function across leech species. Hau-antistasin1 was expressed in the same region as hedgehog, which is a known mediator of signal transduction pathway. Hau-antistasin1 is probably a downstream target of Hedgehog signaling, involved in segment polarity signal pathway.


Asunto(s)
Anticoagulantes/análisis , Hormonas de Invertebrados/análisis , Sanguijuelas/química , Animales , Anticoagulantes/metabolismo , Regulación del Desarrollo de la Expresión Génica , Proteínas Hedgehog/análisis , Proteínas Hedgehog/metabolismo , Hormonas de Invertebrados/genética , Hormonas de Invertebrados/metabolismo , Sanguijuelas/embriología , Sanguijuelas/genética , Sanguijuelas/metabolismo , Filogenia , Transducción de Señal
15.
J Exp Zool B Mol Dev Evol ; 330(6-7): 341-350, 2018 09.
Artículo en Inglés | MEDLINE | ID: mdl-30280505

RESUMEN

The Forkhead box (Fox) gene family is an evolutionarily ancient gene family named after the Drosophila melanogaster forkhead gene (fkh). Fox genes are highly conserved transcription factors critical for embryogenesis and carcinogenesis. In the current study, we report a whole-genome survey of Fox genes and their expression patterns in the leech Helobdella austienesis. Phylogenetic analysis suggests that some Fox genes of leeches are correlated with other Lophotrochozoa and vertebrate Fox genes. Here we have performed semiquantitative reverse transcription polymerase chain reaction and whole-mount in situ hybridization of Fox genes throughout the embryonic development of H. austinensis. We found that each one of the leech Fox genes (FoxA1, FoxA3, FoxC, FoxL2, FoxO1, and FoxO2) is expressed in a specific set of cells or tissue type. From Stages 9-11, Hau-FoxA1 was expressed in the foregut of the anterior region, and Hau-FoxL2 was expressed in mesodermal muscle fiber. Hau-FoxA3 was temporally expressed in the ventral neuroectoderm. At Stage 11, Hau-FoxC was expressed in the foregut. Hau-FoxO genes have a ubiquitous expression. Our results provide more insight on the evolutionary linkage and role of the Fox gene function in Bilateria.


Asunto(s)
Factores de Transcripción Forkhead/metabolismo , Sanguijuelas/embriología , Sanguijuelas/metabolismo , Animales , Ectodermo/embriología , Ectodermo/metabolismo , Embrión no Mamífero/metabolismo , Factores de Transcripción Forkhead/genética , Regulación del Desarrollo de la Expresión Génica , Mesodermo/embriología , Mesodermo/metabolismo , Filogenia , Secuenciación del Exoma
16.
J Nanosci Nanotechnol ; 18(3): 1599-1605, 2018 Mar 01.
Artículo en Inglés | MEDLINE | ID: mdl-29448635

RESUMEN

In this paper, whole-bacteria SELEX (WB-SELEX) strategy was adopted to isolate specific aptamers against Vibrio parahaemolyticus. Round selection for V. parahaemolyticus was conducted 11 rounds, including two negative selection rounds. It was determined through real-time PCR amplification and post-SELEX experiment. The selected aptmers had high binding property and specificity to V. parahaemolyticus. Of 28 aptamers tested, VPCA-apta#1 had the highest binding affinity compared to other aptamer candidates obtained. To detect V. parahaemolyticus, aptamer based SPR biosensor platform was constructed and pathogenic bacteria sensing was conducted in two steps. The first step was to construct 5'-biotinylated VPCA-apta#1 binding probe. The second step was to incubate V. parahaemolyticus and test microbes in functionalized SA sensor chip in parallel. Our platform showed significant activity for detecting and discriminating V. parahaemolyticus from other enteric species such as Escherichia coli, Listeria monocytogenes, Sigella sonnei, and Vibrio fischeri. This is the first report on the use of whole-SELEX to isolate DNA aptamers specific for V. parahaemolyticus. We demonstrated the feasibility of using aptamer platform for the detection of V. parahaemolyticus in various food supplies. It might be used in multiple points of care for diagnosing Vibriosis.


Asunto(s)
Aptámeros de Nucleótidos , Técnicas Biosensibles , Técnica SELEX de Producción de Aptámeros , Resonancia por Plasmón de Superficie , Vibrio parahaemolyticus , Reacción en Cadena de la Polimerasa
17.
Environ Toxicol ; 33(1): 52-62, 2018 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-29044990

RESUMEN

Throughout life, the human eye is continuously exposed to sunlight and artificial lighting. Ambient light exposure can lead to visual impairment and transient or permanent blindness. To mimic benign light stress conditions, Mus musculus eyes were exposed to low-energy UVB radiation, ensuring no severe morphological changes in the retinal structure post-exposure. We performed RNA-seq analysis to reveal the early transcriptional changes and key molecular pathways involved before the activation of the canonical cell death pathway. RNA-seq analysis identified 537 genes that were differentially modulated, out of which 126 were clearly up regulated (>2-fold, P < .01) and 51 were significantly down regulated (<2-fold, P < .01) in response to UVB irradiation in the mouse retina. Gene ontology analysis revealed that UVB exposure affected pathways for cellular stress and signaling (eg, Creb3, Ddrgk1, Grin1, Map7, Uqcc2, Uqcrb), regulation of chromatin and gene expression (eg, Chd5, Jarid2, Kat6a, Smarcc2, Sumo1, Zfp84), transcription factors (eg, Asxl2, Atf7, Per1, Phox2a, Rxra), RNA processing, and neuronal genes (eg, B4gal2, Drd1, Grm5, Rnf40, Rnps1, Usp39, Wbp4). The differentially expressed genes from the RNA-seq analysis were validated by quantitative PCR. Both analyses yielded similar gene expression patterns. The genes and pathways identified here improve the understanding of early transcriptional responses to UVB irradiation. They may also help in elucidating the genes responsible for the inherent susceptibility of humans to UVB-induced retinal diseases.


Asunto(s)
Retina , Transcriptoma , Rayos Ultravioleta , Animales , Ratones , Proteína de Unión a Elemento de Respuesta al AMP Cíclico/genética , Proteína de Unión a Elemento de Respuesta al AMP Cíclico/metabolismo , Regulación hacia Abajo/efectos de la radiación , Perfilación de la Expresión Génica , Ratones Endogámicos C57BL , Proteínas Asociadas a Microtúbulos/genética , Proteínas Asociadas a Microtúbulos/metabolismo , Proteínas del Tejido Nervioso/genética , Proteínas del Tejido Nervioso/metabolismo , Análisis de Componente Principal , Reacción en Cadena en Tiempo Real de la Polimerasa , Receptores de N-Metil-D-Aspartato/genética , Receptores de N-Metil-D-Aspartato/metabolismo , Retina/citología , Retina/metabolismo , Retina/efectos de la radiación , ARN/química , ARN/aislamiento & purificación , ARN/metabolismo , Análisis de Secuencia de ARN , Transducción de Señal/efectos de la radiación , Transcriptoma/efectos de la radiación , Regulación hacia Arriba/efectos de la radiación
18.
Sensors (Basel) ; 18(12)2018 Nov 28.
Artículo en Inglés | MEDLINE | ID: mdl-30487399

RESUMEN

A High-Definition map (HD map) is a precise and detailed map composed of various landmark feature layers. The HD map is a core technology that facilitates the essential functions of intelligent vehicles. Recently, it has come to be required for the HD map to continuously add new feature layers in order to increase the performances of intelligent vehicles in more complicated environments. However, it is difficult to generate a new feature layer for the HD map, because the conventional method of generating the HD map based on several professional mapping cars has high costs in terms of time and money due to the need to re-drive on all of the public roads. In order to reduce these costs, we propose a crowd-sourced mapping process of the new feature layer for the HD map. This process is composed of two steps. First, new features in the environments are acquired from multiple intelligent vehicles. The acquired new features build each new feature layer in each intelligent vehicle using the HD map-based GraphSLAM approach, and these new feature layers are conveyed to a map cloud through a mobile network system. Next, the crowd-sourced new feature layers are integrated into a new feature layer in a map cloud. In the simulation, the performance of the crowd-sourced process is then analyzed and evaluated. Experiments in real driving environments confirm the results of the simulation.

19.
Sensors (Basel) ; 18(12)2018 Nov 23.
Artículo en Inglés | MEDLINE | ID: mdl-30477194

RESUMEN

Nowadays, many intelligent vehicles are equipped with various sensors to recognize their surrounding environment and to measure the motion or position of the vehicle. In addition, the number of intelligent vehicles equipped with a mobile Internet modem is increasing. Based on the sensors and Internet connection, the intelligent vehicles are able to share the sensor information with other vehicles via a cloud service. The sensor information sharing via the cloud service promises to improve the safe and efficient operation of the multiple intelligent vehicles. This paper presents a cloud update framework of occupancy grid maps for multiple intelligent vehicles in a large-scale environment. An evidential theory is applied to create the occupancy grid maps to address sensor disturbance such as measurement noise, occlusion and dynamic objects. Multiple vehicles equipped with LiDARs, motion sensors, and a low-cost GPS receiver create the evidential occupancy grid map (EOGM) for their passing trajectory based on GraphSLAM. A geodetic quad-tree tile system is applied to manage the EOGM, which provides a common tiling format to cover the large-scale environment. The created EOGM tiles are uploaded to EOGM cloud and merged with old EOGM tiles in the cloud using Dempster combination of evidential theory. Experiments were performed to evaluate the multiple EOGM mapping and the cloud update framework for large-scale road environment.

20.
Dev Genes Evol ; 227(4): 245-252, 2017 07.
Artículo en Inglés | MEDLINE | ID: mdl-28699036

RESUMEN

Genes of the twist family encode bHLH transcription factors known to be involved in the regulation and differentiation of early mesoderm. Here, we report our characterization of Hau-twist, a twist homolog from the leech Helobdella austinensis, a tractable lophotrochozoan representative. Hau-twist was expressed in segmental founder cells of the mesodermal lineage, in subsets of cells within the mesodermal lineage of the germinal plate, in circumferential muscle fibers of a provisional integument during segmentation and organogenesis stages and on the ventral side of the developing proboscis. Thus, consistent with other systems, our results suggest that twist gene of the leech Helobdella might function in mesoderm differentiation.


Asunto(s)
Sanguijuelas/crecimiento & desarrollo , Sanguijuelas/metabolismo , Proteína 1 Relacionada con Twist/metabolismo , Animales , Embrión no Mamífero/metabolismo , Sanguijuelas/citología , Mesodermo/citología , Mesodermo/metabolismo , Filogenia
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