A multi-layered network model identifies Akt1 as a common modulator of neurodegeneration.

The accumulation of misfolded and aggregated proteins is a hallmark of neurodegenerative proteinopathies. Although multiple genetic loci have been associated with specific neurodegenerative diseases (NDs), molecular mechanisms that may have a broader relevance for most or all proteinopathies remain poorly resolved. In this study, we developed a multi-layered network expansion (MLnet) model to predict protein modifiers that are common to a group of diseases and, therefore, may have broader pathophysiological relevance for that group. When applied to the four NDs Alzheimer's disease (AD), Huntington's disease, and spinocerebellar ataxia types 1 and 3, we predicted multiple members of the insulin pathway, including PDK1, Akt1, InR, and sgg (GSK-3ß), as common modifiers. We validated these modifiers with the help of four Drosophila ND models. Further evaluation of Akt1 in human cell-based ND models revealed that activation of Akt1 signaling by the small molecule SC79 increased cell viability in all models. Moreover, treatment of AD model mice with SC79 enhanced their long-term memory and ameliorated dysregulated anxiety levels, which are commonly affected in AD patients. These findings validate MLnet as a valuable tool to uncover molecular pathways and proteins involved in the pathophysiology of entire disease groups and identify potential therapeutic targets that have relevance across disease boundaries. MLnet can be used for any group of diseases and is available as a web tool at http://ssbio.cau.ac.kr/software/mlnet.

The conserved microRNA miR-8-3p coordinates the expression of V-ATPase subunits to regulate ecdysone biosynthesis for Drosophila metamorphosis.

The steroid hormone ecdysone is the central regulator of insect metamorphosis, during which a growing, immature larva is remodeled, through pupal stages, to a reproductive adult. However, the underlying mechanisms of ecdysone-mediated metamorphosis remain to be fully elucidated. Here, we identified metamorphosis-associated microRNAs (miRNAs) and their potential targets by cross-linking immunoprecipitation coupled with deep sequencing of endogenous Argonaute 1 protein in Drosophila. Interestingly, miR-8-3p targeted five Vha genes encoding distinct subunits of vacuolar H+ -ATPase (V-ATPase), which has a vital role in the organellar acidification. The expression of ecdysone-responsive miR-8-3p is normally downregulated during Drosophila metamorphosis, but temporary overexpression of miR-8-3p in the whole body at the end of larval development led to defects in metamorphosis and survival, hallmarks of aberrant ecdysone signaling. In addition, miR-8-3p was expressed in the prothoracic gland (PG), which produces and releases ecdysone in response to prothoracicotropic hormone (PTTH). Notably, overexpression of miR-8-3p or knockdown of its Vha targets in the PG resulted in larger than normal, ecdysone-deficient larvae that failed to develop into the pupal stage but could be rescued by ecdysone feeding. Moreover, these animals showed defective PTTH signaling with a concomitant decrease in the expression of ecdysone biosynthetic genes. We also demonstrated that the regulatory network between the conserved miR-8-3p/miR-200 family and V-ATPase was functional in human cells. Consequently, our data indicate that the coordinated regulation of V-ATPase subunits by miR-8-3p is involved in Drosophila metamorphosis by controlling the ecdysone biosynthesis.

Ecdysone-responsive microRNA-252-5p controls the cell cycle by targeting Abi in Drosophila.

The steroid hormone ecdysone has a central role in the developmental transitions of insects through its control of responsive protein-coding and microRNA (miRNA) gene expression. However, the complete regulatory network controlling the expression of these genes remains to be elucidated. In this study, we performed cross-linking immunoprecipitation coupled with deep sequencing of endogenous Argonaute 1 (Ago1) protein, the core effector of the miRNA pathway, in Drosophila S2 cells. We found that regulatory interactions between miRNAs and their cognate targets were substantially altered by Ago1 in response to ecdysone signaling. Additionally, during the larva-to-adult metamorphosis, miR-252-5p was up-regulated via the canonical ecdysone-signaling pathway. Moreover, we provide evidence that miR-252-5p targets Abelson interacting protein ( Abi) to decrease the protein levels of cyclins A and B, controlling the cell cycle. Overall, our data suggest a potential role for the ecdysone/miR-252-5p/Abi regulatory axis partly in cell-cycle control during metamorphosis in Drosophila.-Lim, D.-H., Lee, S., Han, J. Y., Choi, M.-S., Hong, J.-S., Seong, Y., Kwon, Y.-S., Lee, Y. S. Ecdysone-responsive microR-252-5p controls the cell cycle by targeting Abi in Drosophila.

TrpA1 Regulates Defecation of Food-Borne Pathogens under the Control of the Duox Pathway.

Pathogen expulsion from the gut is an important defense strategy against infection, but little is known about how interaction between the intestinal microbiome and host immunity modulates defecation. In Drosophila melanogaster, dual oxidase (Duox) kills pathogenic microbes by generating the microbicidal reactive oxygen species (ROS), hypochlorous acid (HOCl) in response to bacterially excreted uracil. The physiological function of enzymatically generated HOCl in the gut is, however, unknown aside from its anti-microbial activity. Drosophila TRPA1 is an evolutionarily conserved receptor for reactive chemicals like HOCl, but a role for this molecule in mediating responses to gut microbial content has not been described. Here we identify a molecular mechanism through which bacteria-produced uracil facilitates pathogen-clearing defecation. Ingestion of uracil increases defecation frequency, requiring the Duox pathway and TrpA1. The TrpA1(A) transcript spliced with exon10b (TrpA1(A)10b) that is present in a subset of midgut enteroendocrine cells (EECs) is critical for uracil-dependent defecation. TRPA1(A)10b heterologously expressed in Xenopus oocytes is an excellent HOCl receptor characterized with elevated sensitivity and fast activation kinetics of macroscopic HOCl-evoked currents compared to those of the alternative TRPA1(A)10a isoform. Consistent with TrpA1's role in defecation, uracil-excreting Erwinia carotovora showed higher persistence in TrpA1-deficient guts. Taken together, our results propose that the uracil/Duox pathway promotes bacteria expulsion from the gut through the HOCl-sensitive receptor, TRPA1(A)10b, thereby minimizing the chances that bacteria adapt to survive host defense systems.

Character strengths as protective factors against depression and suicidality among male and female employees.

BACKGROUND: So far, studies on workplace mental health have only focused on work-related environmental risk factors, disregarding both protective and individual factors of employees. Therefore, we aimed to identify character strengths that act as protective factors against depressive moods and suicidality in Korean employees. METHODS: In total, 84 male and 151 female employees (aged 19-50 years) reported their sociodemographic characteristics; depressive symptoms, as measured by the Beck Depression Inventory-II; suicidality, as measured by the Korean version of the MINI International Neuropsychiatric Interview suicidality module; and character strengths, as measured by the 24 Character Strength Alphas on the Values in Action Survey-72. We conducted a hierarchical logistic regression, in which depressive mood and suicidality served as the categorical outcome variables. RESULTS: In females, scores on the "curiosity" (B = 1.107, Wald = 10.207, odds ratio = 3.026, p = .001) and "love" (B = .862, Wald = 5.767, odds ratio = 2.367, p = .016) sub scales of the 24 Character Strength Alphas on the Values in Action Survey-72 were statistically significant predictors of having depressive mood. Additionally, females' scores on "judgment" (B = - 1.405, Wald = 5.663, odds ratio = .245, p = .017) and "kindness" (B = - 1.456, Wald = 6.486, odds ratio = .233, p = .011) were protective factors against suicidality. In males, the "love" (B = 1.746, Wald = 4.279, odds ratio = 5.729, p = .039) score was a predictor of having depressive mood, while "teamwork" (B = - 2.204, Wald = 4.666, odds ratio = .110, p = .031) and "creativity" (B = - 1.384, Wald = 4.202, odds ratio = .251, p = .040) scores were protective factors against having depressive mood and suicidality, respectively. CONCLUSIONS: We suggest that focusing on "judgement" and "kindness" in female employees, and "teamwork" and "creativity" in male employees, and engaging in activities that use these strengths at the workplace can be protective factors against depression and suicidality. Future research should focus on developing interventions to promote these character strengths among employees at the workplace.

Features of Resting-State Electroencephalogram Theta Coherence in Somatic Symptom Disorder Compared With Major Depressive Disorder: A Pilot Study.

OBJECTIVE: Somatic symptom disorder (SSD) often co-occurs with major depressive disorder (MDD). Both conditions share common psychobiological and biobehavioral characteristics, but little is known about differential patterns in brain function. In this study, we compared resting-state functional brain connectivity between SSD and MDD using quantitative electroencephalography. METHODS: Fifteen patients with SSD (SSD group), 15 patients with MDD (MDD group), and 15 healthy volunteers (healthy control [HC] group) participated in this study. Participants were assessed with quantitative electroencephalography using a 21-channel electroencephalogram system. Electroencephalogram coherence in the theta frequency range (3.5-7.5 Hz) was assessed between the following seven electrode pairs: Fp1 and Fp2, F7 and T3, F8 and T4, T5 and P3, P4 and T6, P3 and Pz, and Pz and P4. Differences in coherence between groups were analyzed using analysis of variance. RESULTS: Theta coherence between the F7 and T3 electrodes was lower in the SSD group than the MDD and HC groups (F(2,42) = 6.67, p = .0030). Theta coherence between the T5 and P3 electrodes was lower in the SSD and MDD groups than the HC group (F(2,42) = 5.65, p = .0067). Theta coherence between the Pz and P4 electrodes was lower in the SSD group than the MDD group (F(2,42) = 6.41, p = .0037). CONCLUSIONS: Both SSD and MDD patients commonly showed decreased functional connectivity within the left temporoparietal junction, which has neurophysiological implications for cognitive-attentional processing and social interaction. Frontostriatal circuit dysfunction affects processes that control perception and emotion, as well as misperception of somatosensory data in the parietal somatosensory area, and is more likely to be a neuropathology of SSD than MDD.

MicroRNA signatures associated with thioacetamide-induced liver fibrosis in mice.

Multiple etiologies of liver injury are associated with fibrosis in which the key event is the activation of hepatic stellate cells (HSCs). Although microRNAs (miRNAs) are reportedly involved in fibrogenesis, the complete array of miRNA signatures associated with the disease has yet to be elucidated. Here, deep sequencing analysis revealed that compared to controls, 80 miRNAs were upregulated and 21 miRNAs were downregulated significantly in the thioacetamide (TAA)-induced mouse fibrotic liver. Interestingly, 58 of the upregulated miRNAs were localized to an oncogenic miRNA megacluster upregulated in liver cancer. Differential expression of some of the TAA-responsive miRNAs was confirmed, and their human orthologs were similarly deregulated in TGF-ß1-activated HSCs. Moreover, a functional analysis of the experimentally validated high-confidence miRNA targets revealed significant enrichment for the GO terms and KEGG pathways involved in HSC activation and liver fibrogenesis. This is the first comprehensive report of miRNAs profiles during TAA-induced mouse liver fibrosis.

Comparison of QEEG Findings between Adolescents with Attention Deficit Hyperactivity Disorder (ADHD) without Comorbidity and ADHD Comorbid with Internet Gaming Disorder.

Internet gaming disorder (IGD) is often comorbid with attention deficit hyperactivity disorder (ADHD). In this study, we compared the neurobiological differences between ADHD comorbid with IGD (ADHD+IGD group) and ADHD without comorbidity (ADHD-only group) by analyzing quantitative electroencephalogram (QEEG) findings. We recruited 16 male ADHD+IGD, 15 male ADHD-only adolescent patients, and 15 male healthy controls (HC group). Participants were assessed using Young's Internet Addiction Scale and ADHD Rating Scale. Relative power and inter- and intra-hemispheric coherences of brain waves were measured using a digital electroencephalography (EEG) system. Compared to the ADHD-only group, the ADHD+IGD group showed lower relative delta power and greater relative beta power in temporal regions. The relative theta power in frontal regions were higher in ADHD-only group compared to HC group. Inter-hemispheric coherence values for the theta band between F3-F4 and C3-C4 electrodes were higher in ADHD-only group compared to HC group. Intra-hemispheric coherence values for the delta, theta, alpha, and beta bands between P4-O2 electrodes and intra-hemispheric coherence values for the theta band between Fz-Cz and T4-T6 electrodes were higher in ADHD+IGD group compared to ADHD-only group. Adolescents who show greater vulnerability to ADHD seem to continuously play Internet games to unconsciously enhance attentional ability. In turn, relative beta power in attention deficit in ADHD+IGD group may become similar to that in HC group. Repetitive activation of brain reward and working memory systems during continuous gaming may result in an increase in neuronal connectivity within the parieto-occipital and temporal regions for the ADHD+IGD group.

Comparison of Electroencephalography (EEG) Coherence between Major Depressive Disorder (MDD) without Comorbidity and MDD Comorbid with Internet Gaming Disorder.

Internet gaming disorder (IGD) has many comorbid psychiatric problems including major depressive disorder (MDD). In the present study, we compared the neurobiological differences between MDD without comorbidity (MDD-only) and MDD comorbid with IGD (MDD+IGD) by analyzing the quantitative electroencephalogram (QEEG) findings. We recruited 14 male MDD+IGD (mean age, 20.0 ± 5.9 years) and 15 male MDD-only (mean age, 20.3 ± 5.5 years) patients. The electroencephalography (EEG) coherences were measured using a 21-channel digital EEG system and computed to assess synchrony in the frequency ranges of alpha (7.5-12.5 Hz) and beta (12.5-35.0 Hz) between the following 12 electrode site pairs: inter-hemispheric (Fp1-Fp2, F7-F8, T3-T4, and P3-P4) and intra-hemispheric (F7-T3, F8-T4, C3-P3, C4-P4, T5-O1, T6-O2, P3-O1, and P4-O2) pairs. Differences in inter- and intra-hemispheric coherence values for the frequency bands between groups were analyzed using the independent t-test. Inter-hemispheric coherence value for the alpha band between Fp1-Fp2 electrodes was significantly lower in MDD+IGD than MDD-only patients. Intra-hemispheric coherence value for the alpha band between P3-O1 electrodes was higher in MDD+IGD than MDD-only patients. Intra-hemispheric coherence values for the beta band between F8-T4, T6-O2, and P4-O2 electrodes were higher in MDD+IGD than MDD-only patients. There appears to be an association between decreased inter-hemispheric connectivity in the frontal region and vulnerability to attention problems in the MDD+IGD group. Increased intra-hemisphere connectivity in the fronto-temporo-parieto-occipital areas may result from excessive online gaming.

The comparison of temperament and character between patients with internet gaming disorder and those with alcohol dependence.

BACKGROUND: The differences in prevalence, natural history, and disease progression between Internet gaming disorder (IGD) and substance use disorder contribute to the controversy over IGD as a diagnosis under substance-related and addictive disorders. AIMS: The purpose of the current study was to assess the temperament and character of subjects with IGD in comparison with those with alcohol dependence (AD). METHODS: Temperament and character were assessed using Cloningernt temperament and character inventory (TCI). The severity of IGD or AD, depressed mood, anxiety, attention and impulsiveness were assessed using each of the six scales. RESULTS: Among patients with AD, after controlling for other variables, the severity of AD was positively correlated with harm avoidance (HA) score and depressed mood. Among patients with IGD, after controlling for other variables, the severity of IGD was positively correlated with novelty seeking (NS) score, impulsiveness and attention. CONCLUSIONS: There were significant differences in temperament and character between the IGD and AD groups as measured using the TCI. These results suggest that IGD and AD need to be categorized separately in a diagnostic classification system and benefit from different treatment approaches.

Trafficking of LAG-3 to the surface on activated T cells via its cytoplasmic domain and protein kinase C signaling.

Lymphocyte activation gene-3 (LAG-3; CD223), a structural homolog of CD4, binds to MHC class II molecules. Recent research indicated that signaling mediated by LAG-3 inhibits T cell proliferation, and LAG-3 serves as a key surface molecule for the function of regulatory T cells. Previous reports demonstrated that the majority of LAG-3 is retained in the intracellular compartments and is rapidly translocated to the cell surface upon stimulation. However, the mechanism by which LAG-3 translocates to the cell surface was unclear. In this study, we examined the trafficking of human LAG-3 under unstimulated as well as stimulated conditions of T cells. Under the unstimulated condition, the majority of LAG-3 did not reach the cell surface, but rather degraded within the lysosomal compartments. After stimulation, the majority of LAG-3 translocated to the cell surface without degradation in the lysosomal compartments. Results indicated that the cytoplasmic domain without Glu-Pro repetitive sequence is critical for the translocation of LAG-3 from lysosomal compartments to the cell surface. Moreover, protein kinase C signaling leads to the translocation of LAG-3 to the cell surface. However, two potential serine phosphorylation sites from the LAG-3 cytoplasmic domain are not involved in the translocation of LAG-3. These results clearly indicate that LAG-3 trafficking from lysosomal compartments to the cell surface is dependent on the cytoplasmic domain through protein kinase C signaling in activated T cells.

Effectiveness of atomoxetine and methylphenidate for problematic online gaming in adolescents with attention deficit hyperactivity disorder.

OBJECTIVE: There is a high prevalence of problematic online gaming in adolescents with attention deficit hyperactivity disorder (ADHD). In the current study, we compared the effectiveness of atomoxetine (ATM) and methylphenidate (MPH) on problematic online gaming in adolescents with ADHD. METHODS: We recruited 86 adolescents diagnosed with ADHD together with Internet gaming disorder. These participants were divided into two treatment groups: 44 participants were treated with MPH for 12 weeks, and 42 participants were treated with ATM for 12 weeks. RESULTS: During the 3-month study period, the MPH group showed greater improvement in Korean ADHD rating scale scores than the ATM group. The ATM group showed greater improvement in Child Depression Inventory scores than the MPH group. However, Young Internet Addiction Scale and Behavioral Inhibition & Activation Scales score changes did not differ significantly between the MPH and ATM groups. In both groups, changes in Young Internet Addiction Scale scores were positively correlated with the changes in Behavioral Inhibition & Activation Scales scores. CONCLUSIONS: Both MPH and ATM reduced the severity of Internet gaming disorder symptoms, and this reduction was correlated with impulsivity reduction, which also resulted from both ADHD medications. These findings suggest impulsivity plays a critical role in the development of problematic online gaming.

Global identification of target recognition and cleavage by the Microprocessor in human ES cells.

The Microprocessor plays an essential role in canonical miRNA biogenesis by facilitating cleavage of stem-loop structures in primary transcripts to yield pre-miRNAs. Although miRNA biogenesis has been extensively studied through biochemical and molecular genetic approaches, it has yet to be addressed to what extent the current miRNA biogenesis models hold true in intact cells. To address the issues of in vivo recognition and cleavage by the Microprocessor, we investigate RNAs that are associated with DGCR8 and Drosha by using immunoprecipitation coupled with next-generation sequencing. Here, we present global protein-RNA interactions with unprecedented sensitivity and specificity. Our data indicate that precursors of canonical miRNAs and miRNA-like hairpins are the major substrates of the Microprocessor. As a result of specific enrichment of nascent cleavage products, we are able to pinpoint the Microprocessor-mediated cleavage sites per se at single-nucleotide resolution. Unexpectedly, a 2-nt 3' overhang invariably exists at the ends of cleaved bases instead of nascent pre-miRNAs. Besides canonical miRNA precursors, we find that two novel miRNA-like structures embedded in mRNAs are cleaved to yield pre-miRNA-like hairpins, uncoupled from miRNA maturation. Our data provide a framework for in vivo Microprocessor-mediated cleavage and a foundation for experimental and computational studies on miRNA biogenesis in living cells.

Utilization of Western and Traditional Korean Medicine for Children and Adolescents with Mental Disorders: a Nationwide Population-based Study from 2010 to 2012.

When in need of medical treatment, Korean citizens have a choice of practitioners of western medicine (WM) or Traditional Korean Medicine (TKM). However, the two branches frequently conflict with one another, particularly with regard to mental disorders. This study was designed to compare the utilization of WM and TKM, focusing on child/adolescent patients with mental disorders. We analyzed F-code (Mental and behavioral disorders) claims from the Korean Health Insurance Review and Assessment Service, including data from 0-18-year-old patients from 2010 to 2012. Slightly more men than women utilized WM, while TKM use was almost evenly balanced. WM claims increased with advancing age, whereas utilization of TKM was common for the 0-6 age group. In WM and TKM, the total number of claims relying on the National Health Insurance Service (NHIS) was 331,154 (92.78%) and 73,282 (97.85%), respectively, and the number of claims relying on medical aid was 25,753 (7.22%) and 1,610 (2.15%), respectively. The most frequent F-coded claim in WM was F90 (Hyperkinetic disorders), with 64,088 claims (17.96%), and that in TKM was F45 (Somatoform disorders), with 28,852 claims (38.52%). The prevalence of a single disorder without comorbidities was 168,764 (47.29%) in WM and 52,615 (70.25%) in TKM. From these data, we conclude that WM takes prevalence over TKM in cases of attention deficit/hyperactivity disorder (ADHD), as well as in psychological problems such as depression and anxiety. On the other hand, patients utilizing TKM more commonly present with physical health problems including somatoform problems, sleep, and eating disorders.

Comparative study of the effects of bupropion and escitalopram on Internet gaming disorder.

AIM: We compared the efficacy of bupropion and escitalopram treatments in Internet gaming disorder (IGD) patients. METHODS: We recruited 119 adolescents and adults with IGD. We treated these participants for 6 weeks in three groups as follows: 44 participants were treated with bupropion SR (bupropion group), 42 participants were treated with escitalopram (escitalopram group), and 33 patients without any medication were observed in the community (observation group). At baseline and at the 6-week follow-up visit, all subjects were evaluated using the Clinical Global Impression-Severity Scale, the Young Internet Addiction Scale, the Beck Depression Inventory, the ADHD Rating Scale, and the Behavioral Inhibition and Activation Scales. RESULTS: Both the escitalopram group and the bupropion group showed improvement on all clinical symptom scales after 6 weeks of treatment compared to the observation group. Additionally, the bupropion group showed greater improvement on scores for the Clinical Global Impression-Severity Scale, the Young Internet Addiction Scale, the ADHD Rating Scale, and the Behavioral Inhibition Scale than the escitalopram group. CONCLUSION: Both bupropion and escitalopram were effective in treating and managing IGD symptoms. Moreover, bupropion appeared to be more effective than escitalopram in improving attention and impulsivity in IGD patients. In addition, attention and impulsivity seem to be important for the management of IGD.

Changes in cellular microRNA expression induced by porcine circovirus type 2-encoded proteins.

Porcine circovirus type 2 (PCV2) is the primary causative agent of postweaning multisystemic wasting syndrome, which leads to serious economic losses in the pig industry worldwide. While the molecular basis of PCV2 replication and pathogenicity remains elusive, it is increasingly apparent that the microRNA (miRNA) pathway plays a key role in controlling virus-host interactions, in addition to a wide range of cellular processes. Here, we employed Solexa deep sequencing technology to determine which cellular miRNAs were differentially regulated after expression of each of three PCV2-encoded open reading frames (ORFs) in porcine kidney epithelial (PK15) cells. We identified 51 ORF1-regulated miRNAs, 74 ORF2-regulated miRNAs, and 32 ORF3-regulated miRNAs that differed in abundance compared to the control. Gene ontology analysis of the putative targets of these miRNAs identified transcriptional regulation as the most significantly enriched biological process, while KEGG pathway analysis revealed significant enrichment for several pathways including MAPK signaling, which is activated during PCV2 infection. Among the potential target genes of ORF-regulated miRNAs, two genes encoding proteins that are known to interact with PCV2-encoded proteins, zinc finger protein 265 (ZNF265) and regulator of G protein signaling 16 (RGS16), were selected for further analysis. We provide evidence that ZNF265 and RGS16 are direct targets of miR-139-5p and let-7e, respectively, which are both down-regulated by ORF2. Our data will initiate further studies to elucidate the roles of ORF-regulated cellular miRNAs in PCV2-host interactions.

Identification of Oil-Loving Cupriavidus necator BM3-1 for Polyhydroxyalkanoate Production and Assessing Contribution of Exopolysaccharide for Vegetable Oil Utilization.

Polyhydroxyalkanoates (PHA) have received attention owing to their biodegradability and biocompatibility, with studies exploring PHA-producing bacterial strains. As vegetable oil provides carbon and monomer precursors for poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (P(3HB-co-3HHx)), oil-utilizing strains may facilitate PHA production. Herein, Cupriavidus necator BM3-1, which produces 11.1 g/L of PHB with 5% vegetable oil, was selected among various novel Cupriavidus necator strains. This strain exhibited higher preference for vegetable oils over sugars, with soybean oil and tryptone determined to be optimal sources for PHA production. BM3-1 produced 33.9 g/L of exopolysaccharides (EPS), which was three-fold higher than the amount produced by H16 (10.1 g/L). EPS exhibited 59.7% of emulsification activity (EI24), higher than that of SDS and of EPS from H16 with soybean oil. To evaluate P(3HB-co-3HHx) production from soybean oil, BM3-1 was engineered with P(3HB-co-3HHx) biosynthetic genes (phaCRa, phaARe, and phaJPa). BM3-1/pPhaCJ produced 3.5 mol% of 3HHx and 37.1 g/L PHA. BM3-1/pCB81 (phaCAJ) produced 32.8 g/L PHA, including 5.9 mol% 3HHx. Physical and thermal analyses revealed that P(3HB-co-5.9 mol% 3HHx) was better than PHB. Collectively, we identified a novel strain with high vegetable oil utilization capacity for the production of EPS, with the option to engineer the strain for P(3HB-co-3HHx).

Microarray analysis of Drosophila dicer-2 mutants reveals potential regulation of mitochondrial metabolism by endogenous siRNAs.

RNA interference is a eukaryotic regulatory mechanism by which small non-coding RNAs typically mediate specific silencing of their cognate genes. In Drosophila, the RNase III enzyme Dicer-2 (Dcr-2) is essential for biogenesis of endogenous small interfering RNAs (endo-siRNAs), which have been implicated in regulation of endogenous protein-coding genes. Although much is known about microRNA-based regulatory networks, the biological functions of endo-siRNAs in animals remain poorly understood. We performed gene expression profiling on Drosophila dcr-2 null mutant pupae to investigate transcriptional effects caused by a severe defect in endo-siRNA production, and found 306 up-regulated and 357 down-regulated genes with at least a twofold change in expression compared with the wild type. Most of these up-regulated and down-regulated genes were associated with energy metabolism and development, respectively. Importantly, mRNA sequences of 39% of the up-regulated genes were perfectly complementary to the sequences of previously reported endo-siRNAs, suggesting they may be direct targets of endo-siRNAs. We confirmed up-regulation of five selected genes matching endo-siRNAs and concomitant down-regulation of the corresponding endo-siRNAs in dcr-2 mutant pupae. Most of the potential endo-siRNA target genes were associated with energy metabolism, including the citric acid cycle and oxidative phosphorylation in mitochondria, implying that these are major metabolic processes directly affected by endo-siRNAs in Drosophila. Consistent with this finding, dcr-2 null mutant pupae had lower ATP content compared with controls, indicating that mitochondrial energy production is impaired in these mutants. Our data support a potential role for the endo-siRNA pathway in energy homeostasis through regulation of mitochondrial metabolism.

Effects of polymorphisms in the porcine microRNA MIR206 / MIR133B cluster on muscle fiber and meat quality traits.

MicroRNAs are a class of small non-coding RNA molecules that repress gene expression primarily at the post-transcriptional level. Genetic variations in microRNA genes may contribute to phenotypic differences by altering the expression of microRNAs and their targets. Here, we identified 12 single nucleotide polymorphisms (SNPs) in the genomic region of the porcine MIR206 / MIR133B cluster, 10 and 2 of which were associated with MIR206 and MIR133B respectively. All 12 SNPs were located within primary microRNAs. Allele frequency determination in different pig breeds (Berkshire, n = 153; Landrace, n = 125; Yorkshire, n = 173) and association studies of muscle fiber characteristics, lean meat production and meat quality traits were performed on the MIR206 and MIR133B SNPs. The MIR206 SNPs were associated with the percentage of type IIa and IIb fibers for muscle fiber area composition, meat quality traits including drip loss and lightness, and backfat thickness, a parameter of lean meat production. In addition, we found significant association of the MIR133B SNPs with total muscle fiber number, loin eye area, and muscle pH. Furthermore, these SNPs significantly affected the levels of mature MIR206 and MIR133B , respectively, primarily by regulating the processing of primary microRNAs into precursor microRNAs. Interestingly, altered MIR206 levels correlated with phenotypic variability among genotypes of the MIR206 SNP. Our data suggest that polymorphisms in the porcine MIR206 / MIR133B cluster are a genetic factor affecting muscle and meat quality traits.

Home visitation program for detecting, evaluating and treating socially withdrawn youth in Korea.

AIM: The problems of youth social withdrawal (or hikikomori) became a hot-button social issue in Japan in the 1990s. Unfortunately, current nosology in the DSM-IV may not adequately capture the concept of socially withdrawn youth (SWY) or hikikomori. This study aimed to investigate core SWY issues, evaluate SWY's psychopathologies, and approach them therapeutically through a home visitation program. METHODS: Participants were 65 youth referred by community mental health centers and psychiatric clinics around Seoul and Kyongki-Do province. Among them, only 41 participants (31 male, 10 female, mean age 15 ± 3.6 years) fit our SWY criteria. In addition, 248 middle and high school students in Seoul were recruited as a baseline control group. Caseworkers interviewed the SWY participants and their parents in their homes, using our structured interview manual and a number of psychiatric scales. Caseworkers also approached the participants therapeutically. RESULTS: Participants' Depression Inventory, Trait Anxiety Inventory, Social Anxiety Scale, and Internet Addiction Scale scores were significantly higher than those of baseline controls. Participants' mean number of psychotherapeutic sessions was 2.8, and the mean number of parental interview sessions was 3.4. After the therapeutic sessions, Global Assessment Functioning scores and social activities had improved somewhat in 68.3% of participants. CONCLUSION: These findings suggest that SWY is a complex phenomenon, so an individual psychopathologic process is very important for treatment. The most difficult problem in SWY treatment was therapeutic access. Hence, the home visit approach with a structured manual may be a good gateway for solving this problem.