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Currently, polypropylene (PP) is used in various products, thus leading to high daily exposure in humans. Thus, it is necessary to evaluate the toxicological effects, biodistribution, and accumulation of PP microplastics in the human body. In this study, administration of two particle sizes of PP microplastics (approximately 5 and 10–50 µm) did not lead to any significant changes in several toxicological evaluation parameters, including body weight and pathological examination, compared with the control group in ICR mice. Therefore, the approximate lethal dose and no-observed-adverse-effect level of PP microplastics in ICR mice were established as ≥2000 mg/kg. Furthermore, we manufactured cyanine 5.5 carboxylic acid (Cy5.5-COOH)-labeled fragmented PP microplastics to monitor real-time in vivo biodistribution. After oral administration of the Cy5.5-COOH-labeled microplastics to the mice, most of the PP microplastics were detected in the gastrointestinal tract and observed to be out of the body after 24 h in IVIS Spectrum CT. Therefore, this study provides a new insight into the short-term toxicity, distribution, and accumulation of PP microplastics in mammals.

Glutamate is an important excitatory neurotransmitter, and vesicular glutamate transporters (VGLUTs) are critical in regulating the extent of glutamate release. Because initial increase and late stage decrease in VGLUT expression were implicated in progressive Alzheimer’s disease (AD), VGLUTs can be a new imaging target for diagnosing AD. In this study, we have designed and synthesized a new multimodal gadolinium contrast agent conjugated with rose bengal to target VGLUTs (Gd-DO3A-RB). In addition to high kinetic and pH stability, magnetic resonance and fluorescence imaging using Gd-DO3A-RB showed high affinity to VGLUT1. Using the AD animal model (5XFAD), in vivo MRI with a Gd-DO3A-RB showed less MR signal enhancement in 5XFAD mouse brain than that of age-matched normal C57BL/6 mouse, suggesting a late stage decrease in VGLUT expression in AD animal model. Therefore, this new VGLUT targeting gadolinium contrast agent demonstrated a strong potential as a new diagnostic tool for AD with multimodal imaging capability.

This paper presents the results of a study of a voltage management technology called parameter determination of voltage regulator (PDVR) implemented in a 22.9 kV distribution network. Korea Electric Power Corporation has developed this PDVR technology for increasing the hosting capacity without causing any voltage problems in LV (220/380 V) networks. PDVR calculates and provides the operation settings of voltage regulators such as on load tap changer, pole mounted automatic voltage regulator, and distributed generation (DG). Subsequently, these settings are applied to the voltage regulators, which further optimize the 22.9 kV network voltage without the control of the distribution management system (DMS). A study of the operations and performance of PDVR through the analysis of test results is presented in this paper.

Changes in the proteome of different human tissues with advancing age are poorly characterized. Here, we studied the proteins present in primary skin fibroblasts collected from 82 healthy individuals across a wide age spectrum (22-89 years old) who participated in the GESTALT (Genetic and Epigenetic Signatures of Translational Aging Laboratory Testing) study of the National Institute on Aging, NIH. Proteins were extracted from lysed fibroblasts and subjected to liquid chromatography-mass spectrometry analysis, and the expression levels of 9341 proteins were analyzed using linear regression models. We identified key pathways associated with skin fibroblast aging, including autophagy, scavenging of reactive oxygen species (ROS), ribosome biogenesis, DNA replication, and DNA repair. Changes in these prominent pathways were corroborated using molecular and cell culture approaches. Our study establishes a framework of the global proteome governing skin fibroblast aging and points to possible biomarkers and therapeutic targets.

Non-Darcy flows are a vital event leading to the inaccuracy in flow performance evaluation, particularly in the fractured wells in shale gas and tight oil deposits. The internal flow commonly indicates high flow rates at the early period of production, owing to the large geometric size of hydraulic fracture and high porosity in proppant packs. After that, the production flow rate decays steeply until the Darcy flow regime is attained. Therefore, accurate porous flow analyses from the Darcy to Forchheimer and then turbulent flow regimes are essential for optimal facility designs and economic productions in the unconventional deposits. The Comiti capillary model is recognised as the leading non-Darcy flow correlation for simple packed beds comprising mono-size grains. However, in actual hydraulic fractures, various types of proppants are used simultaneously and generally combined with numerous soil grains of different sizes and shapes. In this study, the Comiti model is modified by incorporating the effective diameters of the porous media and then examined for mixed complex multi-size packs. Subsequently, a novel type of non-Darcy flow equation is presented according to the logarithmic turbulent friction factor based on the physical variables determined only in the Darcy flow regime. The new equation presents accurate results for all the types of proppant packs under wide porosity and permeability ranges. The generalised non-Darcy flow correlation, which can be extensively employed from the Darcy to non-Darcy flow analyses, particularly beyond the Forchheimer regime, is presented for the accurate flow evaluation of the fractured reservoirs.

Background Epithelial–mesenchymal transition (EMT) is the most common cause of death in colorectal cancer (CRC). In this study, we investigated the functional roles of miRNA-17-5p in EMT of CRC cells. Methods In order to determine if miRNA-17-5p regulated EMT, the precursors and inhibitors of miR-17-5p were transduced into four CRC cells. To evaluate the regulatory mechanism, we performed argonaute 2 (Ago2) immunoprecipitation (IP) and luciferase assay. In addition, we used an intra-splenic injection mouse model of BALB/c nude mice to investigate the metastatic potential of miRNA-17-5p in vivo. Results The miRNA-17-5p expression was lower in primary CRC tissues with metastasis than in primary CRC tissues without metastasis in our RNA sequencing data of patient tissue. Real-time quantitative PCR revealed that miRNA-17-5p was inversely correlated with that of vimentin in five CRC cell lines. Over-expression of miRNA-17-5p decreased vimentin expression and inhibited cell migration and invasion in both LoVo and HT29 cells. However, inhibition of miRNA-17-5p showed the opposite effect. Ago2 IP and luciferase assay revealed that miRNA-17-5p directly bound to the 3′UTR of VIM mRNA. Furthermore, miRNA-17-5p inhibited the metastasis of CRC into liver in vivo. Conclusions Our results demonstrated that miRNA-17-5p regulates vimentin expression, thereby regulating metastasis of CRC.

We explored the influencing factors of the usage intention of a metaverse education application platform that directly influence the optimization of its service function, improve the usage intention, and realize the promotion and application of metaverse technology in the education domain. Based on the characteristics of the metaverse education application platform, we integrated the PPM (push–pull–mooring) model and the TAM (technology acceptance model) to construct the model of influencing factors of usage intention. Ultimately, 275 valid questionnaires were collected through expert demonstration, pre-investigation, formal investigation, and other processes. In addition, our paper used the SEM (structural equation model) and fsQCA (fuzzy-set qualitative comparative analysis) to analyze the influencing factors of user willingness and their configuration paths. The study found that personalized learning, contextualized teaching, perceived usefulness, perceived ease of use, social needs, and social impact play significant positive roles in the willingness to use the metaverse education platform. Meanwhile, the obtained findings show that the experience-led community-driven mode, personality-led community-driven mode, and social-led utility-driven mode serve as potential guidelines for usage intention enhancement.

Thiazole-containing π-conjugated moieties are important structural units in the development of new electronic and photochromic materials. We have developed a Pd-catalyzed syn-hydroarylation reaction of diaryl alkynes with thiazoles that provides access to thiazole-containing triarylethylenes. Pd(II) complexes derived from Pd(0) species and carboxylic acids facilitated C–H functionalization of the unsubstituted thiazole with high C5 selectivity. The catalytic system was also compatible with other azoles, such as oxazoles and a pyrazole, allowing the stereoselective syntheses of various trisubstituted olefins.

Most antidepressants, including selective serotonin reuptake inhibitors (SSRIs), initiate their drug actions by rapid elevation of serotonin, but they take several weeks to achieve therapeutic onset. This therapeutic delay suggests slow adaptive changes in multiple neuronal subtypes and their neural circuits over prolonged periods of drug treatment. Mossy cells are excitatory neurons in the dentate hilus that regulate dentate gyrus activity and function. Here we show that neuronal activity of hippocampal mossy cells is enhanced by chronic, but not acute, SSRI administration. Behavioral and neurogenic effects of chronic treatment with the SSRI, fluoxetine, are abolished by mossy cell-specific knockout of p11 or Smarca3 or by an inhibition of the p11/AnxA2/SMARCA3 heterohexamer, an SSRI-inducible protein complex. Furthermore, simple chemogenetic activation of mossy cells using Gq-DREADD is sufficient to elevate the proliferation and survival of the neural stem cells. Conversely, acute chemogenetic inhibition of mossy cells using Gi-DREADD impairs behavioral and neurogenic responses to chronic administration of SSRI. The present data establish that mossy cells play a crucial role in mediating the effects of chronic antidepressant medication. Our results indicate that compounds that target mossy cell activity would be attractive candidates for the development of new antidepressant medications.

Depression is a leading cause of disability. Current pharmacological treatment of depression is insufficient, and development of improved treatments especially for treatment-resistant depression is desired. Understanding the neurobiology of antidepressant actions may lead to development of improved therapeutic approaches. Here, we demonstrate that dopamine D1 receptors in the dentate gyrus act as a pivotal mediator of antidepressant actions in mice. Chronic administration of a selective serotonin reuptake inhibitor (SSRI), fluoxetine, increases D1 receptor expression in mature granule cells in the dentate gyrus. The increased D1 receptor signaling, in turn, contributes to the actions of chronic fluoxetine treatment, such as suppression of acute stress-evoked serotonin release, stimulation of adult neurogenesis and behavioral improvement. Importantly, under severely stressed conditions, chronic administration of a D1 receptor agonist in conjunction with fluoxetine restores the efficacy of fluoxetine actions on D1 receptor expression and behavioral responses. Thus, our results suggest that stimulation of D1 receptors in the dentate gyrus is a potential adjunctive approach to improve therapeutic efficacy of SSRI antidepressants.

Malignant characteristics of cancers, represented by rapid cell proliferation and high metastatic potential, are a major cause of high cancer-related mortality. As a multifunctional RNA-binding protein, heterogeneous nuclear ribonucleoprotein K (hnRNPK) is closely associated with cancer progression in various types of cancers. In this study, we sought to identify hnRNPK-regulated long intergenic non-coding RNAs (lincRNAs) that play a critical role in the regulation of cancer malignancy. We found that hnRNPK controlled malignant phenotypes including invasiveness, proliferation, and clonogenicity. RNA sequencing and functional studies revealed that LINC00263, a novel target of hnRNPK, is involved in the oncogenic functions of hnRNPK. Knockdown of LINC00263 mitigated the malignant capabilities. Conversely, increased malignant phenotypes were observed in LINC00263-overexpressing cells. Since LINC00263 was mainly localized in the cytosol and highly enriched in Argonaute 2-immunoprecipitation (Ago2-IP), we hypothesized that LINC00263 acts as a competitive endogenous RNA (ceRNA), and thus sought to identify LINC00263-associated microRNAs. Using small RNA sequencing followed by antisense oligonucleotide pull-down, miR-147a was selected for further study. We found that miR-147a negatively regulates LINC00263 via direct interaction, thus suppressing malignant capabilities. Moreover, knockdown of hnRNPK and LINC00263 upregulated miR-147a, indicating that LINC00263 serves as a ceRNA for miR-147a. By analyzing RNA sequencing data and miRNA target prediction, calpain 2 (CAPN2) was identified as a putative target of miR-147a. Ago2-IP and luciferase reporter assay revealed that miR-147a suppressed CAPN2 expression by directly binding to the 3′UTR of CAPN2 mRNA. In addition, we found that the weakened malignant capabilities following knockdown of hnRNPK or LINC00263 were restored by miR-147a inhibition or CAPN2 overexpression. Furthermore, our findings were validated in various other types of cancer cells including lung cancer, colorectal cancer, neuroblastoma, and melanoma. Collectively, we demonstrate that hnRNPK-regulated LINC00263 plays an important role in cancer malignancy by acting as a miR-147a decoy and thus upregulating CAPN2.

Shwachman-Diamond syndrome (SDS; OMIM #260400) is caused by variants in SBDS (Shwachman-Bodian-Diamond syndrome gene), which encodes a protein that plays an important role in ribosome assembly. Recent reports suggest that recessive variants in EFL1 are also responsible for SDS. However, the precise genetic mechanism that leads to EFL1-induced SDS remains incompletely understood. Here we present 3 unrelated Korean SDS patients who carry biallelic pathogenic variants in EFL1 with biased allele frequencies, resulting from a bone marrow–specific somatic uniparental disomy in chromosome 15. The recombination events generated cells that were homozygous for the relatively milder variant, allowing for the evasion of catastrophic physiologic consequences. However, the milder EFL1 variant was still solely able to impair 80S ribosome assembly and induce SDS features in cell line and animal models. The loss of EFL1 resulted in a pronounced inhibition of terminal oligopyrimidine element–containing ribosomal protein transcript 80S assembly. Therefore, we propose a more accurate pathogenesis mechanism of EFL1 dysfunction that eventually leads to aberrant translational control and ribosomopathy.

Background: Malignant characteristics of cancers, represented by rapid cell proliferation and high metastatic potential, are a major cause of high cancer-related mortality. As a multifunctional RNA-binding protein, heterogeneous nuclear ribonucleoprotein K (hnRNPK) is closely associated with cancer progression in various types of cancers. In this study, we sought to identify hnRNPK-regulated long intergenic non-coding RNAs (lincRNAs) that play a critical role in the regulation of cancer malignancy.Methods: Malignant phenotypes including metastatic and proliferative potential were examined using Transwell invasion assay, cell counting, and colony forming assay. To search for hnRNPK-regulated lincRNAs, RNA sequencing was performed. Using small RNA sequencing followed by antisense oligonucleotide pull-down, lincRNA-associated miRNAs were selected. The association between miRNA and target mRNA was determined by Argonaute 2-immunoprecipitation and luciferase reporter assay. Results: We found that hnRNPK controlled malignant phenotypes including invasiveness, proliferation, and clonogenicity. RNA sequencing and functional studies revealed that LINC00263, a novel target of hnRNPK, is involved in the oncogenic functions of hnRNPK. Knockdown of LINC00263 mitigated the malignant capabilities. Conversely, increased malignant phenotypes were observed in LINC00263-overexpressing cells. We also found that miR-147a negatively regulates LINC00263 via direct interaction, thus suppressing malignant capabilities. Moreover, knockdown of hnRNPK and LINC00263 upregulated miR-147a, indicating that LINC00263 serves as a ceRNA for miR-147a. By analyzing RNA sequencing data and miRNA target prediction, calpain 2 (CAPN2) was identified as a putative target of miR-147a. Ago2-IP and luciferase reporter assay revealed that miR-147a suppressed CAPN2 expression by directly binding to the 3'UTR of CAPN2 mRNA. Additionally, we found that the weakened malignant capabilities following knockdown of hnRNPK or LINC00263 were restored by miR-147a inhibition or CAPN2 overexpression.Conclusions: Our findings demonstrate that hnRNPK-regulated LINC00263 plays an important role in cancer malignancy by acting as a miR-147a decoy and thus upregulating CAPN2.

Prospective shale plays require a combination of good reservoir and completion qualities. Total organic carbon (TOC) is an important reservoir quality and brittleness is the most critical condition for completion quality. We analyzed seismically-derived brittleness and TOC to investigate the prospectivity of the Horn River Group shale (the Muskwa, Otter Park, Evie shales) of a shale-gas field in the western Horn River Basin, British Columbia, Canada. We used the λρ-μρ brittleness template, constructed from the mineralogy-based brittleness index (MBI) and elastic logs from two wells, to convert the λρ and μρ volumes from prestack seismic inversion to the volume for the brittleness petrotypes (most brittle, intermediate, and least brittle). The probability maps of the most brittle petrotype for the three shales were generated from Bayesian classification, based on the λρ-μρ template. The relationship between TOC and P-wave and S-wave velocity ratio ( V P / V S ) at the wells allowed the conversion of the V P / V S volume from prestack inversion to the TOC volume, which in turn was used to construct the TOC maps for the three shales. Increased TOC is correlated with high brittleness, contrasting with the commonly-held understanding. Therefore, the prospectivity of the shales in the study area can be represented by high brittleness and increased TOC. We propose a shale prospectivity index (SPI), computed by the arithmetic average of the normalized probability of the most brittle petrotype and the normalized TOC. The higher SPI corresponds to higher production rates in the Muskwa and Evie shales. The areas of the highest SPI have not been fully tested. The future drilling should be focused on these areas to increase the economic viability of the field.

In recent developments of shale reservoirs, it is important to estimate the permeabilities of hydraulic fractures accompanying the Non- Darcy effects and geometric changes. Accordingly, a new permeability estimation method that considers the varying geometric features under different flow regions is demanded. To this end, the present study introduces the generalized Darcy’s friction flow relation, especially for examining the friction factor-Reynolds number (f · Re) relationship of porous flow, which is originally used in general internal friction flow analyses. Moreover, simple hydraulic fractures comprising structured microbeads are simulated via computational fluid dynamics during fracture aperture variations under different flow conditions from laminar to turbulent. Frictional flow features, e.g., the preservation characteristics of f · Re values, are examined under different geometry and flow conditions, and the transient flow characteristics are investigated using streamline analyses. Consequently, it is verified that the f · Re values vary slightly in proportion to the geometric changes caused by aperture reduction in each medium. Even though the variations in the f · Re values are much smaller than the permeability variations, it seems to be contrary to our expectation. Otherwise, the almost linear-variation aspects of f · Re values were observed in both directional flow cases. The linear-variation aspect of f · Re values is expected to be useful in the permeability-variation estimations in porous media with changing basic geometric factors, such as hydraulic fracture closing. Moreover, it is demonstrated that regardless of aperture reduction in the same type of medium, each porous flow has a very similar power-law relation between f and Re values when the flow velocity changes from the laminar to the turbulent condition. This aspect can be effectively used for obtaining permeability estimations of the varied media, particularly under different flow conditions.

Although SRSF3 (Serine/arginine-rich splicing factor 3) plays a significant role in various biological processes, many of its functions still remain unclear. More particularly, little is known about SRSF3's involvement in the regulation of miRNA. In this report, we found that invasive and migratory abilities were inhibited in SRSF3-silenced U2OS and HeLa cells. We also found that a knockdown of SRSF3 results in a decreased expression level of REST (RE1-silencing transcription factor). The silencing of REST increased the expression of primary miR-132/212 as well as their mature forms. In particular, miR-132-3p and miR-212-3p possess an identical seed sequences and a common target gene. Overexpression of miR-132-3p and miR-212-3p suppressed the expression of YAP1 (Yes-associated protein 1) by directly binding to the 3՚UTR of its mRNA. CCND1 (Cyclin D1), which acts downstream of YAP1, was downregulated in both miR-132-3p and miR-212-3p-overexpressed cells, in correlation with diminished YAP1 levels. Taken together, our results reveal that SRSF3 controls the expression of the miR-132/212 cluster through regulating REST expression, and that the REST-elicited alteration of miRNA expression is implicated in enabling the migratory and invasive abilities of cancer cells.

Polo-like kinase 1 (PLK1) is a critical regulator of cell cycle progression and apoptosis. However, its regulation remains poorly understood. In the present study, we investigated the molecular mechanism underlying the post-transcriptional regulation of PLK1. We observed that heterogeneous nuclear ribonucleoprotein K (hnRNPK) and PLK1 were positively associated in several different cancers and high expression levels of them correlated with poor prognosis in patients with cancer. Knockdown of hnRNPK resulted in reduced expression of PLK1, whereas conversely, PLK1 expression was increased in hnRNPK-overexpressing cells. We found that hnRNPK regulated PLK1 expression through KH1- and KH2-dependent interactions with the 3′UTR of PLK1 mRNA. In addition, microRNA-149-3p (miR-149-3p) and miR-193b-5p suppressed PLK1 expression by targeting the 3′UTR of PLK1 mRNA. MicroRNA-elicited enrichment of PLK1 mRNA in Ago2 immunoprecipitation was altered by the presence or absence of hnRNPK. Furthermore, the deletion of the cytosine (C)-rich sequences of the 3′UTR of PLK1 mRNA abolished the decreased PLK1 expression observed via hnRNPK silencing and administration of miRNAs, a finding that suggests that hnRNPK shares this C-rich motif with miR-149-3p and miR-193b-5p. We also found that downregulation of PLK1 by either silencing hnRNPK or overexpression of miR-149-3p and miR-193b-5p decreased clonogenicity and induced apoptosis. Our findings from this study demonstrate that hnRNPK regulates PLK1 expression by competing with the PLK1-targeting miRNAs, miR-149-3p and miR-193b-5p.

// Hye Ree Kim 1 , Chang Hoon Shin 1 , Hong Lee 1 , Kyung Hee Choi 1 , Do-Hyun Nam 1, 2 , Takbum Ohn 3 , Hyeon Ho Kim 1, 4 1 Department of Health Sciences and Technology, Samsung Advanced Institute for Health Sciences and Technology, Sungkyunkwan University, Seoul, Republic of Korea 2 Department of Neurosurgery, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Republic of Korea 3 Department of Cellular and Molecular Medicine, College of Medicine, Chosun University, Gwangju, Republic of Korea 4 Research Institute for Future Medicine, Samsung Medical Center, Seoul, Republic of Korea Correspondence to: Hyeon Ho Kim, email: hyeonhkim@skku.edu Takbum Ohn, email: tohn@chosun.ac.kr Keywords: SRSF3, FADS1, miR-1908-5p, NF-κB, NKIRAS2 Received: August 24, 2016 Accepted: November 22, 2016 Published: December 26, 2016 ABSTRACT Serine/arginine (SR)-rich proteins that contain RS domains and SR repeats have diverse cellular functions including transcription, polyadenylation, translation, and RNA export. The splicing factor SRSF3, also termed SRp20, is the smallest member of the SR protein family and is a known proto-oncogene. Although it is implicated in the malignant phenotypes of various cancer cells, the molecular mechanism underlying SRSF3-mediated cancer progression is still obscure. We investigated here the oncogenic functions of SRSF3 in osteosarcoma U2OS cells. Knockdown of SRSF3 inhibited proliferation, clonogenicity, and metastatic potential including migration and invasion. It also decreased the level of miR-1908 independent of its host gene FADS1 . Although FADS1 was not associated with SRSF3-mediated malignant properties, overexpression of miR-1908-5p increased cell proliferation, migration, and invasion, suggesting that miR-1908-5p is responsible for the oncogenic functions of SRSF3. Knockdown of SRSF3 decreased the expression of miR-1908-5p by inhibiting transactivation of NF-κB. We observed that miR-1908-5p downregulated NF-κB inhibitor interacting Ras-like 2 ( NKIRAS2 ), a negative regulator of the NF-κB pathway by directly binding to the 3’UTR of NKIRAS2 mRNA. Consistent with overexpression of miR-1908-5p, knockdown of NKIRAS2 diminished the expression level of IκB-β and provoked translocation of NF-κB into the nucleus where it transcriptionally activates its target genes including miR-1908-5p expression, thus elevating the proliferation and metastatic potential. Taken together, our results demonstrate that SRSF3 confers the malignant characteristics on cancer cells via the SRSF3/miR-1908-5p/NKIRAS2 axis.

Glioblastomas (GBM) are very difficult to treat and their aggressiveness is one of the main reasons for this as well as for the frequent recurrences. MicroRNAs post-transcriptionally regulate their target genes through interaction between their seed sequence and 3'UTR of the target mRNAs. We previously reported that miR-296-3p is regulated by neurofibromatosis 2 (NF2) and enhances the invasiveness of GBM cells via SOCS2/STAT3. In this study, we investigated whether miR-296-5p, which originates from the same precursor miRNA as miR-296-3p, can increase the invasiveness of GBM cells. It was observed that miR-296-5p potentiated the invasion of various GBM cells including LN229, T98G, and U87MG. Through bioinformatics approaches, two genes were identified as miR-296-5p targets: caspase-8 (CASP8) and nerve growth factor receptor (NGFR). From results obtained from Ago2 immunoprecipitation and luciferase assays, we found that miR-296-5p downregulates CASP8 and NGFR through direct interaction between seed sequence of the miRNA and 3'UTR of the target mRNA. Knockdown of CASP8 or NGFR also increased the invasive ability of GBM cells, indicating that CASP8 and NGFR are involved in potentiation of invasiveness by miR-296-5p. Consistent with our findings, CASP8 was downregulated in brain metastatic lung cancer cells, which have a high level of miR-296-5p, compared to parental cells, suggesting that miR-296-5p may be generally associated with the acquisition of invasiveness. Collectively, our results implicate miR-296-5p as a potential cause of invasiveness in cancer and suggest it as a promising therapeutic target for GBM.

Emerging studies suggest that microRNAs (miRNAs) play multiple roles in cancer malignancy, including proliferation and acquisition of metastatic potential. Differentially expressed miRNAs responsible for the malignancy of lung cancer were searched by miRNA microarray using a previously established brain metastatic lung cancer model. Twenty-five miRNAs were down-regulated in brain metastatic lung cancer cells. Among those, miR-193b-3p and -5p were chosen for further studies. Their function in metastatic potential and proliferation was examined using Transwell invasion, wound healing, and colony forming assays. The underlying mechanism of tumor-suppressor miR-193b-3p and -5p was explored using reverse transcriptase quantitative polymerase chain reaction (RT-qPCR), Western blot, Argonaute 2-RNA immunoprecipitation (Ago2-RIP), and reporter assays. Both strands of miR-193b were down-regulated in brain metastatic lung cancer cells and in tissues from lung cancer patients. Overexpression of miR-193b-3p and -5p inhibited invasive and migratory activities and diminished clonogenic ability. Conversely, inhibition of miR-193b-3p or -5p increased the metastatic potential and colony forming ability. Cyclin D1 (CCND1), Ajuba LIM Protein (AJUBA), and heart development protein with EGF like domains 1 (HEG1) were identified as common target genes of miR-193b-3p and -5p. A reporter assay and an Ago2-RIP experiment showed that both miRNAs directly bind to the 3′ untranslated region (3′UTR) of the target mRNA. Knockdown of target gene reduced the proliferative and metastatic potential of primary and metastatic lung cancer cells. Our results demonstrate miR-193b is a dual-strand tumor suppressor and a novel therapeutic target for lung cancer.

Melia azedarach L., called chinaberry, is native to Southeast Asia and Australia. The trees are commonly planted as ornamentals in the southern part of Korea. In October 2010, a leaf spot disease was observed on trees for the first time in Wando, Korea. Further surveys conducted from 2010 to 2012 showed that the disease occurs on trees in Jeju, Seogwipo, and Tongyeong cities as well as Wando county with nearly 100% incidence. Leaf spots were circular to semicircular, later becoming angular, small, pale brown in the center with a dark brown margin, and later becoming milky white. Leaf spots sometimes coalesced to blight the entire leaf and were capable of rapidly defoliating whole trees in late September. Fruiting was amphigenous, but mostly hypogenous. Stromata were substomatal, globular, dark brown, and 25 to 70 μm in diameter. Conidiophores were densely fasciculate, pale olivaceous to pale brown, substraight to mildly curved, not geniculate, 10 to 30 μm long, 2.5 to 4.5 μm wide, and aseptate or uniseptate. Conidia were pale olivaceous, generally darker than conidiophores, cylindric to obclavate, substraight in shorter ones, curved to mildly sinuous in longer ones, obconically truncate at the base, obtuse at the apex, 2- to 14-septate, 16 to 120 × 3 to 5 μm, guttulate, and had inconspicuous hila. Morphological characteristics of the fungus were consistent with the previous descriptions of Pseudocercospora subsessilis (Syd. & P. Syd.) Deighton (2). Voucher specimens (n = 6) were deposited in the Korea University Herbarium (KUS). An isolate from KUS-F25395 was deposited in the Korean Agricultural Culture Collection (KACC45688). The complete internal transcribed spacer (ITS) region of rDNA was amplified with the primers ITS1/ITS4 (3) and sequenced. The resulting sequence of 517 bp was deposited in GenBank (Accession No. JX993904). A BLAST search in GenBank revealed that the sequence shows >99% similarity (1 bp substitution) with a sequence of P. subsessilis ex M. azedarach from Cuba (GU269815). For pathogenicity tests, hyphal suspensions were prepared by grinding 3-week-old colonies grown on potato dextrose agar with distilled water using a mortar and pestle. Five 3-year-old chinaberry trees were inoculated with hyphal suspensions using a fine haired paint brush. Three healthy trees of the same age, serving as controls, were sprayed with sterile water. The plants were covered with plastic bags to maintain 100% relative humidity for 24 h and then transferred to a greenhouse. Typical symptoms of necrotic spots that appeared on the inoculated leaves 10 days after inoculation were identical to the ones observed in the field. P. subsessilis was reisolated from symptomatic leaf tissues, confirming Koch's postulates. No symptoms were observed on control plants. The disease has been reported in several Asian countries as well as in Cuba and the United States (1). To our knowledge, this is the first report of leaf spot on chinaberry caused by P. subsessilis in Korea. The observed high incidence and severity suggest that this disease can be a limiting factor in utilizing this tree species as ornamentals in public areas. References: (1) D. F. Farr and A. Y. Rossman. Fungal Databases. Syst. Mycol. Microbiol. Lab., Online publication, ARS, USDA, Retrieved October 22, 2012. (2) Y. L. Guo and W. H. Hsieh. The genus Pseudocercospora in China. International Academic Publishers, Beijing, China, 1995. (3) T. J. White et al. PCR Protocols. Academic Press, San Diego, CA, 1990.

Shwachman-Diamond syndrome (SDS; OMIM: #260400) is caused by variants inSBDS(Shwachman-Bodian-Diamond syndrome gene), which encodes a protein that plays an important role in ribosome assembly. Recent reports suggest that recessive variants inEFL1are also responsible for SDS. However, the precise genetic mechanism that leads toEFL1-induced SDS remains incompletely understood. Here we present three unrelated Korean SDS patients that carry biallelic pathogenic variants inEFL1with biased allele frequencies, resulting from a bone marrow-specific somatic uniparental disomy (UPD) in chromosome 15. The recombination events generated cells that were homozygous for the relatively milder variant, allowing for the evasion of catastrophic physiological consequences. Still, the milderEFL1variant was solely able to impair 80S ribosome assembly and induce SDS features in cell line, zebrafish, and mouse models. The loss ofEFL1resulted in a pronounced inhibition of terminal oligo-pyrimidine element-containing ribosomal protein transcript 80S assembly. Therefore, we propose a more accurate pathogenesis mechanism of EFL1 dysfunction that eventually leads to aberrant translational control and ribosomopathy.

It is important to accurately estimate performances of a hydraulic fractured well, because it will be utilized to evaluate various completion parameters and furthermore to establish a future development plan. Shale gas reservoirs with fracture networks have high initial production rates but show drastic production decline as reservoirs are depleted by production. One of the reasons behind this phenomenon is an increased effective stress during production resulting in fracture closures. Gas mainly flows through hydraulic and natural fracture networks, so the fracture closures cause permeability reduction in the flow areas. In typical hydraulic fracturing operations, proppants are injected with fracturing fluid and placed in the fracture networks. Proppants play a crucial role to keep an induced hydraulic fracture open and retain a well productivity. However, only small portion of the fracture networks are filled with proppants (propped fracture) and the rest exist without proppants (unpropped fracture). Therefore, fracture closures of these regions are quite different. In this article, we have investigated to identify the combined effect of fracture closure and proppant placement on production estimation of a shale gas well. A numerical model has been developed to mimic well performances in Horn River Basin, BC, Canada. We have used pressure-dependent correlations based on experiments to consider fracture permeability alteration with changing reservoir pressure. Proppant placements are described using a fracture propagation model and this enables to classify a whole reservoir into sub regions such as propped, unpropped fracture, and matrix. By comparing with different cases, this article shows that reasonable results on gas production estimation are accomplished when considering fracture closure and proppant placement effects together.