In Vitro Tracking of Human Umbilical Vein Endothelial Cells Using Ultra-Sensitive Quantum Dot-Embedded Silica Nanoparticles.

The nanoscale spatiotemporal resolution of single-particle tracking (SPT) renders it a powerful method for exploring single-molecule dynamics in living cells or tissues, despite the disadvantages of using traditional organic fluorescence probes, such as the weak fluorescent signal against the strong cellular autofluorescence background coupled with a fast-photobleaching rate. Quantum dots (QDs), which enable tracking targets in multiple colors, have been proposed as an alternative to traditional organic fluorescence dyes; however, they are not ideally suitable for applying SPT due to their hydrophobicity, cytotoxicity, and blinking problems. This study reports an improved SPT method using silica-coated QD-embedded silica nanoparticles (QD2), which represent brighter fluorescence and are less toxic than single QDs. After treatment of QD2 in 10 µg/mL, the label was retained for 96 h with 83.76% of labeling efficiency, without impaired cell function such as angiogenesis. The improved stability of QD2 facilitates the visualization of in situ endothelial vessel formation without real-time staining. Cells retain QD2 fluorescence signal for 15 days at 4 °C without significant photobleaching, indicating that QD2 has overcome the limitations of SPT enabling long-term intracellular tracking. These results proved that QD2 could be used for SPT as a substitute for traditional organic fluorophores or single quantum dots, with its photostability, biocompatibility, and superior brightness.

Novel role for a bacterial nucleoid protein in translation of mRNAs with suboptimal ribosome-binding sites.

In Escherichia coli, the major nucleoid protein H-NS limits transcription by acting as a repressor or transcriptional silencer, presumably by its ability to close the looped chromosome domains in the nucleoid through DNA-protein-DNA bridging. Here, we demonstrate the direct involvement of H-NS as a positive factor stimulating translation of the malT mRNA. In vitro studies showed that H-NS facilitates a repositioning of the 30S preinitiation complex on the malT mRNA. H-NS stimulation of translation depended on the AU-rich -35 to -40 region of the mRNA. Several additional examples were found demonstrating a novel function for H-NS in translation of genes with suboptimal ribosome-binding sequences.

A new plant protein interacts with eIF3 and 60S to enhance virus-activated translation re-initiation.

The plant viral re-initiation factor transactivator viroplasmin (TAV) activates translation of polycistronic mRNA by a re-initiation mechanism involving translation initiation factor 3 (eIF3) and the 60S ribosomal subunit (60S). QJ;Here, we report a new plant factor-re-initiation supporting protein (RISP)-that enhances TAV function in re-initiation. RISP interacts physically with TAV in vitro and in vivo. Mutants defective in interaction are less active, or inactive, in transactivation and viral amplification. RISP alone can serve as a scaffold protein, which is able to interact with eIF3 subunits a/c and 60S, apparently through the C-terminus of ribosomal protein L24. RISP pre-bound to eIF3 binds 40S, suggesting that RISP enters the translational machinery at the 43S formation step. RISP, TAV and 60S co-localize in epidermal cells of infected plants, and eIF3-TAV-RISP-L24 complex formation can be shown in vitro. These results suggest that RISP and TAV bridge interactions between eIF3-bound 40S and L24 of 60S after translation termination to ensure 60S recruitment during repetitive initiation events on polycistronic mRNA; RISP can thus be considered as a new component of the cell translation machinery.

Early Osteogenic-Induced Adipose-Derived Stem Cells and Canine Bone Regeneration Potential Analyzed Using Biodegradable Scaffolds.

The complex process of bone regeneration is influenced by factors such as inflammatory responses, tissue interactions, and progenitor cells. Currently, multiple traumas can interfere with fracture healing, causing the prolonging or failure of healing. In these cases, bone grafting is the most effective treatment. However, there are several drawbacks, such as morbidity at the donor site and availability of suitable materials. Advantages have been provided in this field by a variety of stem cell types. Adipose-derived stem cells (ASCs) show promise. In the radiological examination of this study, it was confirmed that the C/S group showed faster regeneration than the other groups, and Micro-CT also showed that the degree of bone formation in the defect area was highest in the C/S group. Compared to the control group, the change in cortical bone area in the defect area decreased in the sham group (0.874), while it slightly increased in the C/S group (1.027). An increase in relative vascularity indicates a decrease in overall bone density, but a weak depression filled with fibrous tissue was observed outside the compact bone. It was confirmed that newly formed cortical bone showed a slight difference in bone density compared to surrounding normal bone tissue due to increased distribution of cortical bone. In this study, we investigated the effect of bone regeneration by ADMSCs measured by radiation and pathological effects. These data can ultimately be applied to humans with important clinical applications in various bone diseases, regenerative, and early stages of formative differentiation.

A Cell-Penetrant Peptide Disrupting the Transcription Factor CP2c Complexes Induces Cancer-Specific Synthetic Lethality.

Despite advances in precision oncology, cancer remains a global public health issue. In this report, proof-of-principle evidence is presented that a cell-penetrable peptide (ACP52C) dissociates transcription factor CP2c complexes and induces apoptosis in most CP2c oncogene-addicted cancer cells through transcription activity-independent mechanisms. CP2cs dissociated from complexes directly interact with and degrade YY1, leading to apoptosis via the MDM2-p53 pathway. The liberated CP2cs also inhibit TDP2, causing intrinsic genome-wide DNA strand breaks and subsequent catastrophic DNA damage responses. These two mechanisms are independent of cancer driver mutations but are hindered by high MDM2 p60 expression. However, resistance to ACP52C mediated by MDM2 p60 can be sensitized by CASP2 inhibition. Additionally, derivatives of ACP52C conjugated with fatty acid alone or with a CASP2 inhibiting peptide show improved pharmacokinetics and reduced cancer burden, even in ACP52C-resistant cancers. This study enhances the understanding of ACP52C-induced cancer-specific apoptosis induction and supports the use of ACP52C in anticancer drug development.

ARP-T1-associated Bazex-Dupré-Christol syndrome is an inherited basal cell cancer with ciliary defects characteristic of ciliopathies.

Actin-Related Protein-Testis1 (ARP-T1)/ACTRT1 gene mutations cause the Bazex-Dupré-Christol Syndrome (BDCS) characterized by follicular atrophoderma, hypotrichosis, and basal cell cancer. Here, we report an ARP-T1 interactome (PXD016557) that includes proteins involved in ciliogenesis, endosomal recycling, and septin ring formation. In agreement, ARP-T1 localizes to the midbody during cytokinesis and the basal body of primary cilia in interphase. Tissue samples from ARP-T1-associated BDCS patients have reduced ciliary length. The severity of the shortened cilia significantly correlates with the ARP-T1 levels, which was further validated by ACTRT1 knockdown in culture cells. Thus, we propose that ARP-T1 participates in the regulation of cilia length and that ARP-T1-associated BDCS is a case of skin cancer with ciliopathy characteristics.

Medroxyprogesterone Reverses Tolerable Dose Metformin-Induced Inhibition of Invasion via Matrix Metallopeptidase-9 and Transforming Growth Factor-ß1 in KLE Endometrial Cancer Cells.

This study was performed to evaluate the anticancer effects of tolerable doses of metformin with or without medroxyprogesterone (MPA) in endometrial cancer cells. Cell viability, cell invasion, and levels of matrix metallopeptidase (MMP) and transforming growth factor (TGF)-ß1 were analyzed using three human endometrial adenocarcinoma cell lines (Ishikawa, KLE, and uterine serous papillary cancer (USPC)) after treatment with different dose combinations of MPA and metformin. Combining metformin (0, 100, 1000 µM) and 10 µM MPA induced significantly decreased cell viability in a time- and dose-dependent manner in Ishikawa cells, but not in KLE and USPC cells. In KLE cells, metformin treatment alone significantly inhibited cell invasion in a dose-dependent manner. The inhibitory effect of metformin was reversed when 10 µM MPA was combined, which was significantly inhibited again after treatment of MMP-2/9 inhibitor and/or TGF-ß inhibitor. Changes of MMP-9 and TGF-ß1 according to combinations of MPA and metformin were similar to those of invasion in KLE cells. In conclusion, the anticancer effects of tolerable doses of metformin varied according to cell type and combinations with MPA. Anti-invasive effect of metformin in KLE cells was completely reversed by the addition of MPA; this might be associated with MMP-9 and TGF-ß1.

Differentiation of equine induced pluripotent stem cells into mesenchymal lineage for therapeutic use.

In previous work, we established an equine induced pluripotent stem cell line (E-iPSCs) from equine adipose-derived stem cells (ASCs) using a lentiviral vector encoding four transcription factors: Oct4, Sox2, Klf4, and c-Myc. In the current study, we attempted to differentiate these established E-iPSCs into mesenchymal stem cells (MSCs) by serial passaging using MSC-defined media for stem cell expansion. Differentiation of the MSCs was confirmed by analyzing expression levels of the MSC surface markers CD44 and CD29, and the pluripotency markers Nanog and Oct4. Results indicated that the E-iPSC-derived MSCs (E-iPSC-MSCs) retained the characteristics of MSCs, including the ability to differentiate into chondrogenic, osteogenic, or myogenic lineages. E-iPSC-MSCs were rendered suitable for therapeutic use by inhibiting immune rejection through exposure to transforming growth factor beta 2 (TGF-ß2) in culture, which down-regulated the expression of major histocompatibility complex class I (MHC class I) proteins that cause immune rejection if they are incompatible with the MHC antigen of the recipient. We reported 16 cases of E-iPSC-MSC transplantations into injured horses with generally positive effects, such as reduced lameness and fraction lines. Our findings indicate that E-iPSC-MSCs can demonstrate MSC characteristics and be safely and practically used in the treatment of musculoskeletal injuries in horses.

Four plant Dicers mediate viral small RNA biogenesis and DNA virus induced silencing.

Like other eukaryotes, plants use DICER-LIKE (DCL) proteins as the central enzymes of RNA silencing, which regulates gene expression and mediates defense against viruses. But why do plants like Arabidopsis express four DCLs, a diversity unmatched by other kingdoms? Here we show that two nuclear DNA viruses (geminivirus CaLCuV and pararetrovirus CaMV) and a cytoplasmic RNA tobamovirus ORMV are differentially targeted by subsets of DCLs. DNA virus-derived small interfering RNAs (siRNAs) of specific size classes (21, 22 and 24 nt) are produced by all four DCLs, including DCL1, known to process microRNA precursors. Specifically, DCL1 generates 21 nt siRNAs from the CaMV leader region. In contrast, RNA virus infection is mainly affected by DCL4. While the four DCLs are partially redundant for CaLCuV-induced mRNA degradation, DCL4 in conjunction with RDR6 and HEN1 specifically facilitates extensive virus-induced silencing in new growth. Additionally, we show that CaMV infection impairs processing of endogenous RDR6-derived double-stranded RNA, while ORMV prevents HEN1-mediated methylation of small RNA duplexes, suggesting two novel viral strategies of silencing suppression. Our work highlights the complexity of virus interaction with host silencing pathways and suggests that DCL multiplicity helps mediate plant responses to diverse viral infections.

[The development of an integrated suicide-violence prevention program for adolescents].

PURPOSE: The purpose of this study was to develop an integrated suicide-violence prevention program for adolescents. Another purpose was to evaluate the effects of the integrated suicide-violence prevention program on self-esteem, parent-child communication, aggression, and suicidal ideation in adolescents. METHODS: The study employed a quasi-experimental design. Participants for the study were high school students, 24 in the experimental group and 25 in the control group. Data was analyzed by using the SPSS/WIN. 11.5 program with chi2 test, t-test, and 2-way ANOVA. RESULTS: Participants in the integrated suicide-violence prevention program reported increased self-esteem scores, which was significantly different from those in the control group. Participants in the integrated suicide-violence prevention program reported decreased aggression and suicidal ideation scores, which was significantly different from those in the control group. CONCLUSION: The integrated suicide-violence prevention program was effective in improving self-esteem and decreasing aggression and suicidal ideation for adolescents. Therefore, this approach is recommended as the integrated suicide-violence prevention strategy for adolescents.

Establishment of stably expandable induced myogenic stem cells by four transcription factors.

Life-long regeneration of healthy muscle by cell transplantation is an ideal therapy for patients with degenerative muscle diseases. Yet, obtaining muscle stem cells from patients is very limited due to their exhaustion in disease condition. Thus, development of a method to obtain healthy myogenic stem cells is required. Here, we showed that the four transcription factors, Six1, Eya1, Esrrb, and Pax3, converts fibroblasts into induced myogenic stem cells (iMSCs). The iMSCs showed effective differentiation into multinucleated myotubes and also higher proliferation capacity than muscle derived stem cells both in vitro and in vivo. The iMSCs do not lose their proliferation capacity though the passaging number is increased. We further isolated CD106-negative and α7-integrin-positive iMSCs (sort-iMSCs) showing higher myogenic differentiation capacity than iMSCs. Moreover, genome-wide transcriptomic analysis of iMSCs and sort-iMSCs, followed by network analysis, revealed the genes and signaling pathways associated with enhanced proliferation and differentiation capacity of iMSCs and sort-iMSCs, respectively. The stably expandable iMSCs provide a new source for drug screening and muscle regenerative therapy for muscle wasting disease.

[Validation of a path model on adolescents' suicidal ideation and violent behavior].

PURPOSE: This study examined the fitness of a path model on the relationship among stress, self-esteem, aggression, depression, suicidal ideation, and violent behavior for adolescents. METHODS: The subjects consisted of 1,177 adolescents. Data was collected through self-report questionnaires. The data was analyzed by the SPSS and AMOS programs. RESULTS: Stress, self-esteem, aggression, and depression showed a direct effect on suicidal ideation for adolescents, while stress, self-esteem, and aggression showed an indirect effect on suicidal ideation for adolescents. Stress, self-esteem, aggression, and suicidal ideation showed a direct effect on violent behavior for adolescents, while stress, self-esteem, aggression, and depression showed an indirect effect on violent behavior for adolescents. The modified path model of adolescent's suicidal ideation and violent behavior was proven correct. CONCLUSION: These results suggest that adolescent's suicidal ideation and violent behavior can be decreased by reducing stress, aggression, and depression and increasing self-esteem. Based on the outcomes of this study, it is necessary to design an intervention program that emphasizes reducing stress, aggression, and depression and increasing self-esteem in order to decrease adolescents' suicide ideation and violence.

[The study on predictors of depression for Korean female adolescents].

PURPOSE: The purposes of this study were 1) to compare the contribution of demographic-behavioral variables and psychological variables in explaining the variance of depression, 2) identify the most important predictors of depression for Korean female adolescents. METHOD: The participants were 840 female adolescents. Data was collected through self-report questionnaires, which were constructed to include demographicbehavioral factors, self-esteem, hostility, hopelessness, and depression. Data was analyzed using the SPSS program. RESULT: Female adolescents' demographicbehavioral variables explained 17% of the variance in depression, and perceived physical health status, history of physical abuse, smoking, satisfaction of body weight, parental alcohol abuse, parental divorce, and history of suicidal attempt were the significant predictors of depression for female adolescents. Psychological variables explained 50% of the variance in depression, and self-esteem, hostility, and hopelessness were the significant predictors of depression for female adolescents. The significant predictors of depression among female adolescents' demographicbehavioral variables and psychological variables were self-esteem, hostility, hopelessness, perceived physical health status, parental alcohol problem, and history of physical abuse, explaining 52% of the variance in depression. CONCLUSION: In order to reduce depression in female adolescents, it is necessary to design an intervention program that emphasizes improving self-esteem while reducing hostility and hopelessness.

[Factors on internet game addiction among adolescents].

PURPOSE: The purpose of this study was to explore factors related to internet game addiction for adolescents. METHOD: This study was a cross-sectional survey, and data was collected through self-report questionnaires. Data was analyzed using the SPSS program. RESULTS: In logistic regression analysis, the risk of being addicted to internet games was 2.22 times higher in males than females. Adolescents with low and middle academic performance also had a higher risk(2.08 times and 2.54 times) to become addicted to internet games. For the location of the computer, the risk of becoming addicted to internet games were .01 times lower in the living room or brother or sisters' room than in their own room. The risk of becoming addicted to internet games was 1.18 times higher in the higher usage time of internet games. The risk of becoming addicted to internet games was .49 times lower in the more accepting and autonomic parents' rearing attitude and .02 times lower in the high self-efficacy group than the low group. CONCLUSION: The result of this study suggests that there are noticeable relationships between internet game addiction and gender, academic performance, location of computer, usage time of internet games, parents' rearing attitude, and self efficacy.

Functional nanosome for enhanced mitochondria-targeted gene delivery and expression.

Mitochondria dysfunction plays a role in many human diseases. Therapeutic techniques for these disorders require novel delivery systems that can specifically target and penetrate mitochondria. In this study, we report a novel nanosome composed of dequalinium-DOTAP-DOPE (1,2 dioleoyl-3-trimethylammonium-propane-1,2-dioleoyl-sn-glycero-3-phosphoethanolamine) (DQA80s) as a potential mitochondria-targeting delivery vector. The functional DQAsome, DQA80s, showed enhanced transfection efficiency compared to a vector DQAsomes in HeLa cells and dermal fibroblasts. In addition, DQA80s/pDNA complexes exhibited rapid escape from the endosome into the cytosol. We observed the delivery of pDNA to mitochondria in living cells using flow cytometry, confocal microscopy, and TME imaging. More specifically, we confirmed our results by co-localization of hmtZsGreen constructs to mitochondria when delivered via DQAsomes and DQA80s in living cells. The mitochondria-targeting DQAsomes and DQA80s induced mitochondrial dysfunction through depolarization of mitochondrial membrane potential. Our data demonstrate that DQA80s show promise for use as a mitochondria-targeted carrier system for treatment of mitochondria diseases in vivo.

Mutations in ACTRT1 and its enhancer RNA elements lead to aberrant activation of Hedgehog signaling in inherited and sporadic basal cell carcinomas.

Basal cell carcinoma (BCC), the most common human cancer, results from aberrant activation of the Hedgehog signaling pathway. Although most cases of BCC are sporadic, some forms are inherited, such as Bazex-Dupré-Christol syndrome (BDCS)-a cancer-prone genodermatosis with an X-linked, dominant inheritance pattern. We have identified mutations in the ACTRT1 gene, which encodes actin-related protein T1 (ARP-T1), in two of the six families with BDCS that were examined in this study. High-throughput sequencing in the four remaining families identified germline mutations in noncoding sequences surrounding ACTRT1. These mutations were located in transcribed sequences encoding enhancer RNAs (eRNAs) and were shown to impair enhancer activity and ACTRT1 expression. ARP-T1 was found to directly bind to the GLI1 promoter, thus inhibiting GLI1 expression, and loss of ARP-T1 led to activation of the Hedgehog pathway in individuals with BDCS. Moreover, exogenous expression of ACTRT1 reduced the in vitro and in vivo proliferation rates of cell lines with aberrant activation of the Hedgehog signaling pathway. In summary, our study identifies a disease mechanism in BCC involving mutations in regulatory noncoding elements and uncovers the tumor-suppressor properties of ACTRT1.

Predictors of suicidal ideation among high school students by gender in South Korea.

The purpose of this study was to examine the evidence to determine if there are gender differences in suicidal ideation of adolescents. This study examined the main effect of risk factors from 5 domains and protective factors from 1 domain in relation to suicidal ideation by gender and identified the most important predictors of suicidal ideation for males (N = 654) and females (N = 658). This study was a cross-sectional survey, and data were collected through self-report questionnaires. In the univariate analysis, especially, risk factors from behavioral variables and psychosocial-environmental variables appeared to be gender skewed. For males, all behavioral variables were predictive of suicidal ideation. For the females, unlike the males, Wang-tta or victim of bullying behavior and sexual orientation as behavioral variables were predictive of suicidal ideation. For males, parental divorce and parental alcohol abuse as psychosocial-environmental variables were predictive of suicidal ideation. For the females, again unlike for the males, all the psychosocial-environmental variables were not predictive of suicidal ideation. The most important predictors of suicidal ideation for males as a result of the multivariate analysis were history of suicidal attempt, depression, hostility, smoking, parental alcohol abuse, communication with friends, and self-esteem. The most important predictors of suicidal ideation for females as a result of the multivariate analysis were depression, hostility, sexual orientation, and self-esteem. These results would indicate that an effective suicide screening and prevention program for adolescents should consider gender differences.

[Satisfaction with life and it's predictors of Korean adolescents].

PURPOSE: This study was done to investigate the satisfaction with life in adolescents, and to identify factors affecting satisfaction with life for them. METHOD: The participants were 1,057 adolescents. Data was collected through self-report questionnaires which were constructed to include satisfaction with life, self-esteem, hostility and hopelessness. The data was analyzed using the SPSS program. RESULT: Satisfaction with life for adolescents was significantly different according to school type, grade, scholastic achievement, religion, monthly income of family, conversation with parents and conversation with friends. Satisfaction with life in adolescents correlated with self-esteem, hostility and hopelessness. Significant predictors influencing satisfaction with life in adolescents were self-esteem, hopelessness, school type, conversation with parents, monthly income of family and religion, and these predictors accounted for 37.8% of the variance in satisfaction with life. CONCLUSION: The above findings indicate that satisfaction with life in adolescents is influenced by self-esteem, hostility and hopelessness. Therefore when nursing interventions are developed to improve satisfaction with life in adolescents, these factors need to be considered.

[The effects of a recovery education program on rehabilitation motivation, symptoms, and function for schizophrenic patients].

PURPOSE: This study investigated the effects of a recovery education program on rehabilitation motivation, symptoms, and function for schizophrenic patients. METHOD: The study employed a quasi-experimental design. Participants for the study were 27 patients with schizophrenia, 14 in the experimental group and the other 13 in the control group. Data were analyzed by using the SPSS/WIN 11.5 program with Fisher's exact test, t-test, and Repeated measures ANOVA. RESULTS: After a 7 week intervention, participants in the recovery education program group reported increased rehabilitation motivation and function scores, which was significantly different from those in the control group. CONCLUSION: A recovery education program was effective improving rehabilitation motivation and function for schizophrenic patients. Therefore, this program is recommended as a rehabilitation strategy for schizophrenic patients.