Segment Polarity Research Articles

MiR-145-5p mitigates dysregulated Wnt1/β-catenin signaling pathway in rheumatoid arthritis

Fibroblast-like synoviocytes (FLS) lining the arthritic synovial joint region have been implicated to be a key player in bone remodeling. The uncontrolled proliferation of this cell subtype is strictly regulated by various molecular elements including microRNAs (miRNAs). The Wnt1/β-catenin signaling pathway plays a crucial role in the survival of FLS cells. This study explores the underlying mechanism of miR-145-5p towards the Wnt1/β-catenin pathway. MiR-145-5p depicted a strong binding affinity towards frizzled class receptor 4 (FZD4) 3′ UTR, a key receptor complex essential for recognizing circulating Wnt1 molecules. Adjuvant induced arthritic fibroblast-like synoviocytes (AA-FLS) isolated from rats stimulated with Wnt1 (10 ng/ml) elicited active Wnt1/β-catenin signaling. Transfection of miR-145-5p mimic (50 pmol) to AA-FLS stimulated with Wnt1 elicited reduced expression levels of various factors of Wnt1/β-catenin signaling including low-density lipoprotein receptor-related protein 5 (LRP5), dishevelled segment polarity protein 1 (Dvl1) and β-catenin transcription factor. Moreover, pro-inflammatory cytokines (TNFα, IL-1β, IL-6 and IL-23) were regulated compared to the diseased groups. Furthermore, miR-145-5p counterbalanced the levels of receptor activator of nuclear factor kappa B ligand (RANKL) and osteoprotegerin (OPG) at the cellular level, essential for bone remodeling. Hence, we suggest that miR-145-5p regulates the survival/proliferation of FLS cells in RA disease condition through attenuation of Wnt1/β-catenin signaling.

DFT meets the segmented polarization consistent basis sets: Performances in the computation of molecular structures, rotational and vibrational spectroscopic properties

Quantum-chemical calculations assist the analysis of laboratory spectra, and often provide the only means to determine spectroscopic data that cannot be accessed experimentally. For the purpose, reliable predictions of structural and spectroscopic parameters are required. Although coupled cluster theory in conjunction with to large basis sets and composite schemes can reach impressive accuracies for structural, thermochemical and spectroscopic properties, it is still limited to small sized molecules. DFT represents the working option for medium to large molecular systems. In this context, systematic investigations are required aimed at characterizing the performances of the different DFT model chemistries. In this work, the accuracy of the popular hybrid B3LYP and the double hybrid B2PLYP functionals coupled to the segmented polarization consistent (aug-)pcs-n basis sets in the prediction of molecular structures and rotational- and vibrational spectroscopic parameters are investigated using a benchmark set of molecules of both atmospheric and astrochemical relevance. For comparison purposes, different flavors of Dunning’s triple-ζ basis sets and the SNSD basis set, are also employed. The convergence behavior of the pcs-n hierarchy with n = 1–4 is also addressed to some extent. The results indicate the B3LYP-D3 functional in conjunction with the aug-pcs-1 or SNSD basis sets as a cost-effective model chemistry for applications in the field of rotational and vibrational spectroscopies. Improved accuracy is obtained by coupling the B2PLYP-D3 functional with the aug-pcs-2 or aug-cc-pVTZ triple-ζ basis sets that show an accuracy around 0.003 Å and 0.3° for bond lengths and angles, 1% and 3% for rotational and quartic centrifugal distortion constants, respectively, 12 cm−1 for fundamental frequencies and 3 km mol−1 for IR intensities. The B2PLYP-D3/maug-cc-pVTZ-dH level keeps the same accuracy, with slightly larger deviations for intensities.

LncRNA testis-specific transcript, Y-linked 15 (TTTY15) promotes proliferation, migration and invasion of colorectal cancer cells via regulating miR-29a-3p/DVL3 axis.

Long non-coding RNA testis-specific transcript, Y-linked 15 (TTTY15) is oncogenic in prostate cancer, however its expression and function in colorectal cancer remain largely unknown. Paired colorectal cancer samples/normal tissues were collected, and the expression levels of TTTY15, miR-29a-3p and disheveled segment polarity protein 3 (DVL3) were examined by quantitative real-time polymerase chain reaction (qRT-PCR); TTTY15 shRNA and overexpression plasmids were transfected into HT29 and HCT-116 cell lines using lipofectamine reagent, respectively; the proliferation and colony formation were detected by CCK-8 assay and plate colony formation assay; qRT-PCR and Western blot were used to analyze the changes of miR-29a-3p and DVL3; dual-luciferase reporter gene assay was used to determine the regulatory relationships between miR-29a-3p and TTTY15, miR-29a-3p and DVL3. TTTY15 was significantly up-regulated in cancerous tissues of colorectal cancer samples, positively correlated with the expression of DVL3, while negatively correlated with the expression of miR-29a-3p. After TTTY15 shRNAs were transfected into colorectal cancer cells, the proliferation and metastasis of cancer cells were significantly inhibited, while TTTY15 overexpression had opposite biological effects. TTTY15 shRNA could reduce the expression of DVL3 on both mRNA and protein levels, and the luciferase activity of TTTY15 sequence was also inhibited by miR-29a-3p. DVL3 was also validated as a target gene of miR-29a-3p, and it could be repressed by miR-29a-3p mimics or TTTY15 shRNA. TTTY15 is abnormally upregulated in colorectal cancer tissues, and it can modulate the proliferation and metastasis of colorectal cancer cells. It functions as the ceRNA to regulate the expression of DVL3 by sponging miR-29a-3p.

Two pair-rule responsive enhancers regulate wingless transcription in the Drosophila blastoderm embryo.

While many developmentally relevant enhancers act in a modular fashion, there is growing evidence for nonadditive interactions between distinct cis-regulatory enhancers. We investigated if nonautonomous enhancer interactions underlie transcription regulation of the Drosophila segment polarity gene, wingless. We identified two wg enhancers active at the blastoderm stage: wg 3613u, located from -3.6 to -1.3 kb upstream of the wg transcription start site (TSS) and 3046d, located in intron two of the wg gene, from 3.0 to 4.6 kb downstream of the TSS. Genetic experiments confirm that Even Skipped (Eve), Fushi-tarazu (Ftz), Runt, Odd-paired (Opa), Odd-skipped (Odd), and Paired (Prd) contribute to spatially regulated wg expression. Interestingly, there are enhancer specific differences in response to the gain or loss of function of pair-rule gene activity. Although each element recapitulates aspects of wg expression, a composite reporter containing both enhancers more faithfully recapitulates wg regulation than would be predicted from the sum of their individual responses. These results suggest that the regulation of wg by pair-rule genes involves nonadditive interactions between distinct cis-regulatory enhancers.

Output feedback stabilizer design of Boolean networks based on network structure

In genetic regulatory networks, a stable configuration can represent the evolutionary behavior of cell death or unregulated growth in genes. We present analytical investigations on output feedback stabilizer design of Boolean networks (BNs) to achieve global stabilization via the semi-tensor product method. Based on network structure information describing coupling connections among nodes, an output feedback stabilizer is designed to achieve global stabilization. Compared with the traditional pinning control design, the output feedback stabilizer design is not based on the state transition matrix of BNs, which can efficiently determine pinning control nodes and reduce computational complexity. Our proposed method is efficient in that the calculation of the state transition matrix with dimension 2n × 2n is avoided; here n is the number of nodes in a BN. Finally, a signal transduction network and a D. melanogaster segmentation polarity gene network are presented to show the efficiency of the proposed method. Results are shown to be simple and concise, compared with traditional pinning control for BNs.

Death march of a segmented and trilobate bilaterian elucidates early animal evolution.

The origin of motility in bilaterian animals represents an evolutionary innovation that transformed the Earth system. This innovation probably occurred in the late Ediacaran period-as evidenced by an abundance of trace fossils (ichnofossils) dating to this time, which include trails, trackways and burrows1-3. However, with few exceptions4-8, the producers of most of the late Ediacaran ichnofossils are unknown, which has resulted in a disconnection between the body- and trace-fossil records. Here we describe the fossil of a bilaterian of the terminal Ediacaran period (dating to 551-539million years ago), which we name Yilingia spiciformis (gen. et sp. nov). This body fossil is preserved along with the trail that the animal produced during a death march. Yilingia is an elongate and segmented bilaterian with repetitive and trilobate body units, each of which consists of a central lobe and two posteriorly pointing lateral lobes, indicating body and segment polarity. Yilingia is possibly related to panarthropods or annelids, and sheds light on the origin of segmentation in bilaterians. As one of the few Ediacaran animals demonstrated to have produced long and continuous trails, Yilingia provides insights into the identity of the animals that were responsible for Ediacaran trace fossils.

Amyloid-like Assembly Activates a Phosphatase in the Developing Drosophila Embryo.

Prion-like proteins can assume distinct conformational and physical states in the same cell. Sequence analysis suggests that prion-like proteins are prevalent in various species; however, it remains unclear what functional space they occupy in multicellular organisms. Here, we report the identification of a prion-like protein, Herzog (CG5830), through a multimodal screen in Drosophila melanogaster. Herzog functions as a membrane-associated phosphatase and controls embryonic patterning, likely being involved in TGF-β/BMP and FGF/EGF signaling pathways. Remarkably, monomeric Herzog is enzymatically inactive and becomes active upon amyloid-like assembly. The prion-like domain of Herzog is necessary for both its assembly and membrane targeting. Removal of the prion-like domain impairs activity, while restoring assembly on the membrane using a heterologous prion-like domain and membrane-targeting motif can restore phosphatase activity. This study provides an example of a prion-like domain that allows an enzyme to gain essential functionality via amyloid-like assembly to control animal development.

Spatiotemporal Expression of Anticoagulation Factor Antistasin in Freshwater Leeches.

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.

Casiopeina II‑gly acts on lncRNA MALAT1 by miR‑17‑5p to inhibit FZD2 expression via the Wnt signaling pathway during the treatment of cervical carcinoma.

The present study investigated the underlying regulatory network involved in the differential expression of metastasis associated lung adenocarcinoma transcript 1 (MALAT1) long non-coding (lnc)RNA, microRNA-17-5p (miR-17-5p) and frizzled class receptor 2 (FZD2) mRNA under the influence of Casiopeina II-gly (Cas-II-gly) via the Wnt signaling pathway in cervical carcinoma (CC). The gene expression data were obtained from the Gene Expression Omnibus database (https://www.ncbi.nlm.nih.gov/geo/), and the differentially expressed genes were determined using R software. The R ClusterProfiler and enrichplot packages were applied for gene-set enrichment analysis based on the Gene Ontology biological process and Kyoto Encyclopedia of Genes and Genomes databases. TargetScan and the starBase database were used to predict the targeting associations between the miRNAs and lncRNAs/mRNAs. The MALAT1/miR-17-5p/mRNA FZD2 expression levels were measured via reverse transcription-quantitative polymerase chain reaction. The protein expression was monitored by western blot analysis. The target association among the lncRNA MALAT1, miR-17-5p and FZD2 was validated via a dual luciferase reporter assay. Cell viability and apoptosis were determined via MTT assays, EdU staining and flow cytometry. The results indicated that the expression levels of lncRNA MALAT1 and FZD2 mRNA were downregulated, while miR-17-5p expression was upregulated in HeLa and CaSki cells treated with increasing Cas-II-gly concentrations. The cell viability was decreased, and the apoptosis rate was increased in HeLa and CaSki cells following Cas-II-gly treatment. Furthermore, western blot analysis results demonstrated that Cas-II-gly and the MALAT1/miR-17-5p/FZD2 axis could affect the expression of proteins associated with the Wnt signaling pathway, including disheveled segment polarity protein, glycogen synthase kinase-3β and β-catenin, and via the MALAT1/miR-17-5p/FZD2/Wnt signaling pathway axis.

Loss of lncRNA-SNHG7 Promotes the Suppression of Hepatic Stellate Cell Activation via miR-378a-3p and DVL2

Small nuclear RNA host gene 7 (SNHG7), a novel long non-coding RNA (lncRNA), acts as an oncogene in cancers. However, whether SNHG7 is involved in hepatic stellate cell (HSC) activation during liver fibrosis is still unclear. In this study, upregulation of SNHG7 was found in vivo and in vitro during liver fibrosis. Silencing of SNHG7 led to the suppression of HSC activation, with a reduction in cell proliferation and collagen expression. SNHG7 knockdown also resulted in the suppression of liver fibrosis in vivo. Interestingly, miR-378a-3p was a target of SNHG7. SNHG7 and miR-378a-3p were co-located in the cytoplasm. Downregulation of miR-378a-3p blocked down the effects of loss of SNHG7 on HSC activation. Notably, SNHG7 could enhance Wnt/β-catenin pathway activation to contribute to liver fibrosis, with an increase in T cell factor (TCF) activity and a reduction in P-β-catenin level. It was found that miR-378a-mediated dishevelled segment polarity protein 2 (DVL2) was responsible for SNHG7-activated Wnt/β-catenin pathway. DVL2 was confirmed as a target of miR-378a-3p. SNHG7-induced HSC activation was almost blocked down by DVL2 knockdown. Accordingly, enhanced Wnt/β-catenin by SNHG7 was suppressed by loss of DVL2. Collectively, we demonstrate that SNHG7 reduces miR-378a-3p and attenuates its control on DVL2, leading to aberrant Wnt/β-catenin activity, which contributes to liver fibrosis progression.

P‐41: Border Fade Pattern for a Segmented Active Polarizer Dimmable Lens

Display units that utilize a dimmable smart lens may be utilized to reduce the required luminance compared to a fixed neutral density lens to provide a hidden display appearance. A lens configuration that utilizes an active polarizer in conjunction with a linear polarizer has previously been developed and reported. The use of a segmented active polarizer dimmable lens with a unique border fade pattern is explored.

Transcriptome Analysis of Larval Segment Formation and Secondary Loss in the Echiuran Worm Urechis unicinctus.

The larval segment formation and secondary loss in echiurans is a special phenomenon, which is considered to be one of the important characteristics in the evolutionary relationship between the Echiura and Annelida. To better understand the molecular mechanism of this phenomenon, we revealed the larval transcriptome profile of the echiuran worm Urechis unicinctus using RNA-Seq technology. Twelve cDNA libraries of U. unicinctus larvae, late-trochophore (LT), early-segmentation larva (ES), segmentation larva (SL), and worm-shaped larva (WL) were constructed. Totally 243,381 unigenes were assembled with an average length of 1125 bp and N50 of 1836 bp, and 149,488 unigenes (61.42%) were annotated. We obtained 70,517 differentially expressed genes (DEGs) by pairwise comparison of the larval transcriptome data at different developmental stages and clustered them into 20 gene expression profiles using STEM software. Based on the typical profiles during the larval segment formation and secondary loss, eight signaling pathways were enriched, and five of which, mTOR, PI3K-AKT, TGF-β, MAPK, and Dorso-ventral axis formation signaling pathway, were proposed for the first time to be involved in the segment formation. Furthermore, we identified 119 unigenes related to the segment formation of annelids, arthropods, and chordates, in which 101 genes were identified in Drosophila and annelids. The function of most segment polarity gene homologs (hedgehog, wingless, engrailed, etc.) was conserved in echiurans, annelids, and arthropods based on their expression profiles, while the gap and pair-rule gene homologs were not. Finally, we verified that strong positive signals of Hedgehog were indeed located on the boundary of larval segments using immunofluorescence. Data in this study provide molecular evidence for the understanding of larval segment development in echiurans and may serve as a blueprint for segmented ancestors in future research.

Dynamic expression of ZNF382 and its tumor-suppressor role in hepatitis B virus-related hepatocellular carcinogenesis

Hepatitis B virus (HBV) infection is the primary cause of hepatocellular carcinoma (HCC). Zinc-finger protein 382 (ZNF382), which belongs to zinc-finger protein family, has been documented to be downregulated in certain types of cancer. However, its role in HCC remains largely unknown. In this study, we demonstrated that ZNF382 expression was significantly elevated in HBV-infected liver cirrhosis tissues relative to HBV-negative normal liver tissues at protein levels, but not at mRNA levels, and was positively correlated with the levels of HBV DNA and hepatitis B virus X protein (HBx). Further studies revealed that ZNF382 was a target of miR-6867, and HBx promoted the translation of ZNF382 during HBV chronic infection through Erk-mediated miR-6867 inhibition. In addition, our data showed that ZNF382 was frequently downregulated by promoter methylation in HBV-related HCCs relative to HBV-infected liver cirrhosis tissues, and decreased expression of ZNF382 was strongly correlated with poor survival in early-stage HCC patients. Functional studies demonstrated that ZNF382 was a potent tumor suppressor in HCC cells through inhibiting cell proliferation, colony formation, migration, invasion, and tumorigenic potential in nude mice, and inducing cell apoptosis. Mechanistically, ZNF382 exerted its tumor-suppressor functions in HCC through transcriptionally repressing its downstream targets such as Fos proto-oncogene (FOS), Jun proto-oncogene (JUN), disheveled segment polarity protein 2 (DVL2), and frizzled class receptor 1 (FZD1), thereby impairing the activities of activating protein 1 (AP-1) and Wnt/β-catenin pathways and activating p53 signaling. Altogether, our data show that ZNF382 acts as a tumor suppressor, and is co-regulated by HBx and epigenetic mechanism in HBV-related hepatocellular carcinogenesis.

Stability Analysis of Boolean Networks With Stochastic Function Perturbations

Using the semi-tensor product method, this paper studies the set stability of Boolean networks (BNs) with stochastic function perturbations. First, the definition of one-column function perturbation for BNs is defined, and two kinds of stochastic function perturbations are formulated. Second, by constructing a state transition matrix, a new criterion is proposed for the set stability of BNs with probabilistic function perturbation. Third, the set stability of BNs with Markov jump function perturbation is studied by calculating the state probability distribution. Finally, the obtained results are applied to D. melanogaster segmentation polarity gene network.

Exploring Observability of Attractor Cycles in Boolean Networks for Biomarker Detection.

Boolean Network (BN) is a simple and popular mathematical model that has attracted significant attention from systems biology due to its capacity to reveal genetic regulatory network behavior. In addition, observability, as an important network feature, plays a vital role in deciphering the underlying mechanisms driving a genetic regulatory network and has been widely investigated. Prior studies examined observability of BNs and other complex networks. That said, observability of attractor, which can serve as a biomarker for disease, has not been fully examined in the literature. In this study, we formulated a new definition for singleton or cyclic attractor observability in BNs and developed an effective methodology to resolve the captured problem. We also showed complexity is of O(Pmn), when the maximal period of cyclic attractor is P, the number of attractor is m and the number of genes is n. Importantly, we have confirmed our method can faithfully predict the expression pattern of segment polarity genes in Drosophila melanogaster and showed it can effectively and efficiently deal with the captured observability problem.

Identifying (un)controllable dynamical behavior in complex networks.

We present a technique applicable in any dynamical framework to identify control-robust subsets of an interacting system. These robust subsystems, which we call stable modules, are characterized by constraints on the variables that make up the subsystem. They are robust in the sense that if the defining constraints are satisfied at a given time, they remain satisfied for all later times, regardless of what happens in the rest of the system, and can only be broken if the constrained variables are externally manipulated. We identify stable modules as graph structures in an expanded network, which represents causal links between variable constraints. A stable module represents a system “decision point”, or trap subspace. Using the expanded network, small stable modules can be composed sequentially to form larger stable modules that describe dynamics on the system level. Collections of large, mutually exclusive stable modules describe the system’s repertoire of long-term behaviors. We implement this technique in a broad class of dynamical systems and illustrate its practical utility via examples and algorithmic analysis of two published biological network models. In the segment polarity gene network of Drosophila melanogaster, we obtain a state-space visualization that reproduces by novel means the four possible cell fates and predicts the outcome of cell transplant experiments. In the T-cell signaling network, we identify six signaling elements that determine the high-signal response and show that control of an element connected to them cannot disrupt this response.

Growth zone segmentation in the milkweed bug Oncopeltus fasciatus sheds light on the evolution of insect segmentation

BackgroundOne of the best studied developmental processes is the Drosophila segmentation cascade. However, this cascade is generally considered to be highly derived and unusual, with segments being patterned simultaneously, rather than the ancestral sequential segmentation mode. We present a detailed analysis of the segmentation cascade of the milkweed bug Oncopletus fasciatus, an insect with a more primitive segmentation mode, as a comparison to Drosophila, with the aim of reconstructing the evolution of insect segmentation modes.ResultsWe document the expression of 12 genes, representing different phases in the segmentation process. Using double staining we reconstruct the spatio-temporal relationships among these genes. We then show knock-down phenotypes of representative genes in order to uncover their roles and position in the cascade.ConclusionsWe conclude that sequential segmentation in the Oncopeltus germband includes three slightly overlapping phases: Primary pair-rule genes generate the first segmental gene expression in the anterior growth zone. This pattern is carried anteriorly by a series of secondary pair-rule genes, expressed in the transition between the growth zone and the segmented germband. Segment polarity genes are expressed in the segmented germband with conserved relationships. Unlike most holometabolous insects, this process generates a single-segment periodicity, and does not have a double-segment pattern at any stage. We suggest that the evolutionary transition to double-segment patterning lies in mutually exclusive expression patterns of secondary pair-rule genes. The fact that many aspects of the putative Oncopeltus segmentation network are similar to those of Drosophila, is consistent with a simple transition between sequential and simultaneous segmentation.

Transcriptome analysis of scyphozoan jellyfish Rhopilema esculentum from polyp to medusa identifies potential genes regulating strobilation.

Strobilation is a unique asexual reproduction mode of scyphozoan jellyfish, through which benthic polyp develops into pelagic medusa. It is an orderly metamorphosis process triggered by environmental signals. However, the knowledges of molecular mechanisms under the drastic morphological and physiological changes are still limited. In this study, the transcriptomes from polyps to juvenile medusae at different stages were characterized by RNA-seq in scyphozoan jellyfish Rhopilema esculentum. Among 96,076 de novo assembled unigenes, 7090 differentially expressed genes (DEGs) were identified during the developmental stages. The co-expression pattern analysis of DEGs yielded 15 clusters with different expression patterns. Among them, a cluster with 388 unigenes was related to strobila. In this specific cluster, the GO terms related to "sequence-specific DNA binding transcription factor activity" and "sequence-specific DNA binding" were significantly enriched. Transcription factors, including segmentation protein even-skipped-like, segmentation polarity protein engrailed-like, homeobox proteins Otx-like, Twist-like and Cnox2-Pc-like, as well as genes such as RxR-like and Dmrtf-like, were identified to be potentially involved in strobilation. Their expression patterns and the other 11 TFs/genes involved in strobilation were confirmed with qRT-PCR methods. The present study pointed out the role of transcription factors in strobilation and produced a list of novel candidate genes for further studies. It could provide valuable information for understanding the molecular mechanisms of jellyfish strobilation.

Reliable quantification of trace diarrhetic shellfish poisoning toxins in high-lipid bivalves by UHPLC-ESI-MS/MS in time segment polarity switching mode: Comparison of three extraction methods

Analysis of marine biotoxins in high-lipid bivalves has several limitations due to the coexistence of lipid matrices, low biotoxin concentrations, and the different physiochemical properties of the various biotoxins. In this study, a sensitive and reliable method to simultaneously determine six diarrhetic shellfish poisoning (DSP) toxins (five acidic and one neutral biotoxins) in oysters and mussels was developed based on UHPLC-MS/MS-multiple ion reaction monitoring (MRM) using time segment polarity switching. For effective sample pretreatment, three protocols including syringe filtration, Strata™-X solid phase extraction (SPE), and freezing lipid filtration combined with SPE (FLF + SPE) were evaluated, and FLF + SPE method was shown to be the most effective with high lipid removal efficiency. Aging effects of the UHPLC mobile phase on MS detection sensitivity and retention time shift of DSP toxins were carefully examined. The developed method provided good recoveries ranging from 81.0 to 119.6%, showing no significant matrix effect and assay was linear with coefficients of determination above R2 > 0.99 for all analytes. Validation tests using a certified reference material (CRM) and spiking DSP toxins (OA and DTX-1 at 1.07 μg/g, and DTX-2 at 0.86 μg/g) showed this method to be acceptable, with a relative standard deviation of <0.17% and recovery ranging from 87.29 to 109.97%. Finally, the established method was successfully applied to quantify DSP toxins in 40 oyster and mussel samples collected from Korean fishery markets, ensuring sea food safety.

Effects of FSS on the expression and localization of the core proteins in two Wnt signaling pathways, and their association with ciliogenesis.

Fluid shear stress (FSS) may alter ciliary structures and ciliogenesis, and it has been reported that the Wnt signaling pathway may regulate cilia assembly and disas-sembly. The present study aimed to investigate the effects of FSS on primary cilia, the Wnt/β-catenin and Wnt/PCP signaling pathways, and the association among them. In the present study, human umbilical vein endothelial cells were subjected to FSS of differing velocities for various periods of time using a shear stress device. Subsequently, immunofluorescence and quantitative polymerase chain reaction were used to detect the expression and localization of the following core proteins: β-catenin in the Wnt/β-catenin signaling pathway; and dishevelled segment polarity protein 2 (Dvl2), fuzzy planar cell polarity protein (Fuz) and VANGL planar cell polarity protein 2 (Vangl2) in the Wnt/planar cell polarity (PCP) signaling pathway. Furthermore, the colocalization of Dvl2 with the basal body was analyzed under low FSS and laminar FSS. The results demonstrated that low FSS promoted the expression of Dvl2 and its colocalization with the basal body. Although Fuz expression was decreased with increasing duration of FSS, no visible alterations were detected in its localization, it was ubiquitously localized in the ciliated region. Conversely, the expression of Vangl2 was increased by laminar FSS, and β-catenin was translocated into the nucleus at the early stage of low FSS. These findings suggested that Dvl2 may participate in low FSS-induced ciliogenesis and β-catenin may participate at the early stage, whereas Vangl2 may be associated with laminar FSS-induced cilia disassembly.