TMEM41B Is a Pan-flavivirus Host Factor.

Flaviviruses pose a constant threat to human health. These RNA viruses are transmitted by the bite of infected mosquitoes and ticks and regularly cause outbreaks. To identify host factors required for flavivirus infection, we performed full-genome loss of function CRISPR-Cas9 screens. Based on these results, we focused our efforts on characterizing the roles that TMEM41B and VMP1 play in the virus replication cycle. Our mechanistic studies on TMEM41B revealed that all members of the Flaviviridae family that we tested require TMEM41B. We tested 12 additional virus families and found that SARS-CoV-2 of the Coronaviridae also required TMEM41B for infection. Remarkably, single nucleotide polymorphisms present at nearly 20% in East Asian populations reduce flavivirus infection. Based on our mechanistic studies, we propose that TMEM41B is recruited to flavivirus RNA replication complexes to facilitate membrane curvature, which creates a protected environment for viral genome replication.

Cellular functions of eukaryotic RNA helicases and their links to human diseases.

RNA helicases are highly conserved proteins that use nucleoside triphosphates to bind or remodel RNA, RNA-protein complexes or both. RNA helicases are classified into the DEAD-box, DEAH/RHA, Ski2-like, Upf1-like and RIG-I families, and are the largest class of enzymes active in eukaryotic RNA metabolism - virtually all aspects of gene expression and its regulation involve RNA helicases. Mutation and dysregulation of these enzymes have been linked to a multitude of diseases, including cancer and neurological disorders. In this Review, we discuss the regulation and functional mechanisms of RNA helicases and their roles in eukaryotic RNA metabolism, including in transcription regulation, pre-mRNA splicing, ribosome assembly, translation and RNA decay. We highlight intriguing models that link helicase structure, mechanisms of function (such as local strand unwinding, translocation, winching, RNA clamping and displacing RNA-binding proteins) and biological roles, including emerging connections between RNA helicases and cellular condensates formed through liquid-liquid phase separation. We also discuss associations of RNA helicases with human diseases and recent efforts towards the design of small-molecule inhibitors of these pivotal regulators of eukaryotic gene expression.

Mapping RNA-protein interactions with subcellular resolution using colocalization CLIP.

RNA-binding proteins (RBPs) are essential for RNA metabolism and profoundly impact health and disease. The subcellular organization of RBP interaction networks with target RNAs remains largely unexplored. Here, we develop colocalization CLIP (coCLIP), a method that combines cross-linking and immunoprecipitation (CLIP) with proximity labeling, to explore in-depth the subcellular RNA interactions of the RBP human antigen R (HuR). Using this method, we uncover HuR's dynamic and location-specific interactions with RNA, revealing alterations in sequence preferences and interactions in the nucleus, cytosol, or stress granule (SG) compartments. We uncover HuR's unique binding preferences within SGs during arsenite stress, illuminating intricate interactions that conventional methodologies cannot capture. Overall, coCLIP provides a powerful method for revealing RBP-RNA interactions based on localization and lays the foundation for an advanced understanding of RBP models that incorporate subcellular location as a critical determinant of their functions.

The moderating effect of Korean preschoolers' receptive and expressive language skills on the link between Korean PA and English PA.

The purpose of the current study was to examine whether oral language skills moderate the effect of Korean phonological awareness (PA) on English PA for Korean preschoolers in the initial stage of learning English as a second language. The study participants comprised 81 five- to six-year-old Korean preschoolers attending Korean-medium preschools in South Korea. The findings demonstrated that Korean PA was significantly associated with English PA. In addition, Korean receptive and expressive language skills had moderating effects on the relationship between Korean PA and English PA, respectively. This study is discussed not only in terms of cross-language PA transfer in processing two phonologically and orthographically different languages but also in light of the importance of native language skills interacting with native PA in the second-language PA development of preschool children in the initial stage of learning a second language.

The neural correlates of attachment security in typically developing children.

This study investigated neural correlates of children's attachment security using functional magnetic resonance imaging. Fifty-one boys' attachment styles (age mean = 9.5 years, SD = 0.61) were assessed with the Separation Anxiety Test (SAT). We created an fMRI version of the SAT to activate children's attachment system in fMRI environment and contrasted two conditions in which children were instructed to infer the specific feeling of the boy in the picture or to identify objects or physical activities. In the final fMRI analysis (N = 21), attachment security could be detected at the neural level corresponding to the behavioural differences in the attachment interview. Securely attached children showed greater activation in the frontal, limbic and basal ganglia area which included the dorsolateral prefrontal cortex, amygdala, cingulate cortex and striatum, compared to other children who had lower quality of attachment. These regions have a key role in socio-emotional information processing and also represent a brain network related to approach and avoidance motivation in humans. Especially the striatum, strongly linked to reward processing underpinning social approach and avoidance motivation, showed the largest effects in these differences and also positively correlated with emotional openness scores in SAT. This suggests that the quality of attachment configures the approach and avoidance motivational system in our brain mediated by the striatum.

Reduced orbitofrontal cortical thickness in male adolescents with internet addiction.

BACKGROUND: The orbitofrontal cortex (OFC) has consistently been implicated in the pathology of both drug and behavioral addictions. However, no study to date has examined OFC thickness in internet addiction. In the current study, we investigated the existence of differences in cortical thickness of the OFC in adolescents with internet addiction. On the basis of recently proposed theoretical models of addiction, we predicted a reduction of thickness in the OFC of internet addicted individuals. FINDINGS: Participants were 15 male adolescents diagnosed as having internet addiction and 15 male healthy comparison subjects. Brain magnetic resonance images were acquired on a 3T MRI and group differences in cortical thickness were analyzed using FreeSurfer. Our results confirmed that male adolescents with internet addiction have significantly decreased cortical thickness in the right lateral OFC (p<0.05). CONCLUSION: This finding supports the view that the OFC alterations in adolescents with internet addiction reflect a shared neurobiological marker of addiction-related disorders in general.

Mapping RNA-Protein Interactions with Subcellular Resolution Using Colocalization CLIP.

RNA binding proteins (RBPs) are essential for RNA metabolism and profoundly impact health and disease. The subcellular organization of RBP interaction networks with target RNAs remains largely unexplored. Here, we develop colocalization CLIP, a method that combines CrossLinking and ImmunoPrecipitation (CLIP) with proximity labeling, to explore in-depth the subcellular RNA interactions of the well-studied RNA-binding protein HuR. Using this method, we uncover HuR's dynamic and location-specific interactions with RNA, revealing alterations in sequence preferences and interactions in the nucleus, cytosol, or stress granule compartments. We uncover HuR's unique binding preferences within stress granules during arsenite stress, illuminating intricate interactions that conventional methodologies cannot capture. Overall, coCLIP provides a powerful method for revealing RBP:RNA interactions based on localization and lays the foundation for an advanced understanding of RBP models that incorporate subcellular location as a critical determinant of their functions.

Two distinct binding modes provide the RNA-binding protein RbFox with extraordinary sequence specificity.

Specificity of RNA-binding proteins for target sequences varies considerably. Yet, it is not understood how certain few proteins achieve markedly higher sequence specificity than most others. Here we show that the RNA Recognition Motif of RbFox accomplishes extraordinary sequence specificity by employing functionally and structurally distinct binding modes. Affinity measurements of RbFox for all binding site variants reveal the existence of two distinct binding modes. The first exclusively accommodates cognate and closely related RNAs with high affinity. The second mode accommodates all other RNAs with reduced affinity by imposing large thermodynamic penalties on non-cognate sequences. NMR studies indicate marked structural differences between the two binding modes, including large conformational rearrangements distant from the RNA-binding site. Distinct binding modes by a single RNA-binding module explain extraordinary sequence selectivity and reveal an unknown layer of functional diversity, cross talk and regulation in RNA-protein interactions.

A selectable, plasmid-based system to generate CRISPR/Cas9 gene edited and knock-in mosquito cell lines.

Aedes (Ae.) aegypti and Ae. albopictus mosquitoes transmit arthropod-borne diseases around the globe, causing ~ 700,000 deaths each year. Genetic mutants are valuable tools to interrogate both fundamental vector biology and mosquito host factors important for viral infection. However, very few genetic mutants have been described in mosquitoes in comparison to model organisms. The relative ease of applying CRISPR/Cas9-based gene editing has transformed genome engineering and has rapidly increased the number of available gene mutants in mosquitoes. Yet, in vivo studies may not be practical for screening large sets of mutants or possible for laboratories that lack insectaries. Thus, it would be useful to adapt CRISPR/Cas9 systems to common mosquito cell lines. In this study, we generated and characterized a mosquito optimized, plasmid-based CRISPR/Cas9 system for use in U4.4 (Ae. albopictus) and Aag2 (Ae. aegypti) cell lines. We demonstrated highly efficient editing of the AGO1 locus and isolated U4.4 and Aag2 cell lines with reduced AGO1 expression. Further, we used homology-directed repair to establish knock-in Aag2 cell lines with a 3xFLAG-tag at the N-terminus of endogenous AGO1. These experimentally verified plasmids are versatile, cost-effective, and efficiently edit immune competent mosquito cell lines that are widely used in arbovirus studies.

Attachment security and striatal functional connectivity in typically developing children.

Attachment security is formed through interactions with a main caregiver during the first three years of life and reflects inter-individual differences in mental representations for the relationship. The striatum is known to be a key structure to initiate attachment behaviours and maintain attachment relationships as well as to modulate reward-related processing as part of the approach module in current neurobiological models of human attachment. Although findings have suggested critical roles of the striatum in inter-individual differences in attachment, most studies were based on a wide variety of tasks and very few have investigated these associations in intrinsic brain connectivity in typically developing children. In the present study, using resting-state functional magnetic resonance imaging, we examined the striatal functional connectivity according to children's attachment security in 68 nine-year-olds (Secure attachment = 39, Insecure attachment = 29, mean age/SD = 9.62/0.69). Children with secure attachment demonstrated increased functional connectivity in the tempro-limbic region, compared to children with insecure attachment. In addition, the child-reported attachment security scores were negatively associated with the caudate-prefrontal connectivity, but positively with the putamen-visual area connectivity. These data demonstrate that inter-individual differences in attachment can be captured in striatal functional connectivity organization in the typical brain.

Argonaute-CLIP delineates versatile, functional RNAi networks in Aedes aegypti, a major vector of human viruses.

Argonaute (AGO) proteins bind small RNAs to silence complementary RNA transcripts, and they are central to RNA interference (RNAi). RNAi is critical for regulation of gene expression and antiviral defense in Aedes aegypti mosquitoes, which transmit Zika, chikungunya, dengue, and yellow fever viruses. In mosquitoes, AGO1 mediates miRNA interactions, while AGO2 mediates siRNA interactions. We applied AGO-crosslinking immunoprecipitation (AGO-CLIP) for both AGO1 and AGO2, and we developed a universal software package for CLIP analysis (CLIPflexR), identifying 230 small RNAs and 5,447 small RNA targets that comprise a comprehensive RNAi network map in mosquitoes. RNAi network maps predicted expression levels of small RNA targets in specific tissues. Additionally, this resource identified unexpected, context-dependent AGO2 target preferences, including endogenous viral elements and 3'UTRs. Finally, contrary to current thinking, mosquito AGO2 repressed imperfect targets. These findings expand our understanding of small RNA networks and have broad implications for the study of antiviral RNAi.

Sleep habits and disturbances in Malaysian children with epilepsy.

AIMS: To compare sleep habits and disturbances between Malaysian children with epilepsy and their siblings (age range 4-18 years) and to determine the factors associated with greater sleep disturbance. METHODS: The Sleep Disturbance Scale for Children (SDSC) questionnaire was completed by the primary caregiver for 92 epileptic children (mean age 11.1 years, 50 male, 42 females) and their healthy siblings (mean age 11.1 years, 47 males, 45 females). Details of sleep arrangements and illness severity were obtained. Multiple regression analysis was used to determine factors associated with high Total SDSC scores in epileptic patients. RESULTS: Compared with their siblings, epileptic children had significantly higher total SDSC score (difference between means 8.7, 95% confidence interval (CI) 6.4-11.1) and subscale scores in disorders of initiating and maintaining sleep (3.9, 95% CI 2.8-5.2), sleep-wake transition disorders (2.1, 95% CI 1.3-2.9), sleep-disordered breathing (0.7, 95% CI 0.3-1.1) and disorders of excessive sleepiness (1.5, 95% CI 0.6-2.4). Epileptic children had a higher prevalence of co-sleeping (73.7% vs 31.5%) and on more nights per week (difference between means 3, 95% CI 2.0-3.9) than their siblings. Higher Epilepsy Illness Severity scores were associated with higher total SDSC scores (P= 0.02). CONCLUSION: Co-sleeping was highly prevalent in children with epilepsy, who also had more sleep disturbances (especially problems with initiating and maintaining sleep and sleep-wake transition disorders) than their siblings. Epilepsy severity contributed to the sleep disturbances. Evaluation of sleep problems should form part of the comprehensive care of children with severe epilepsy.

TMEM41B is a pan-flavivirus host factor.

Flaviviruses pose a constant threat to human health. These RNA viruses are transmitted by the bite of infected mosquitoes and ticks and regularly cause outbreaks. To identify host factors required for flavivirus infection we performed full-genome loss of function CRISPR-Cas9 screens. Based on these results we focused our efforts on characterizing the roles that TMEM41B and VMP1 play in the virus replication cycle. Our mechanistic studies on TMEM41B revealed that all members of the Flaviviridae family that we tested require TMEM41B. We tested 12 additional virus families and found that SARS-CoV-2 of the Coronaviridae also required TMEM41B for infection. Remarkably, single nucleotide polymorphisms (SNPs) present at nearly twenty percent in East Asian populations reduce flavivirus infection. Based on our mechanistic studies we hypothesize that TMEM41B is recruited to flavivirus RNA replication complexes to facilitate membrane curvature, which creates a protected environment for viral genome replication. HIGHLIGHTS: TMEM41B and VMP1 are required for both autophagy and flavivirus infection, however, autophagy is not required for flavivirus infection.TMEM41B associates with viral proteins and likely facilitates membrane remodeling to establish viral RNA replication complexes.TMEM41B single nucleotide polymorphisms (SNPs) present at nearly twenty percent in East Asian populations reduce flavivirus infection.TMEM41B-deficient cells display an exaggerated innate immune response upon high multiplicity flavivirus infection.

Gaming-addicted teens identify more with their cyber-self than their own self: Neural evidence.

According to existing neuroimaging studies of social cognition, individuals use knowledge about themselves to infer the mental states of others and to mentalize in a different way when the other is perceived to be similar versus dissimilar to oneself. In this study, we examined whether adolescent boys make mental state inferences for their online game characters and whether adolescents who were diagnosed as addicted to the internet game perceived their personal game character to be similar to themselves. Twelve internet-addicted adolescents and fifteen adolescents without addiction reported whether short phrases described themselves, a well-known historical person, or their own game character while undergoing a functional magnetic resonance imaging (fMRI). Different patterns of activity emerged for adolescents with internet game addiction compared to healthy adolescents when they were thinking about themselves, another person, and their game characters. Specifically, when addicted adolescents were thinking about their own game characters, more global and significant medial prefrontal (MPFC) and anterior cingulate (ACC) activations were observed, than even when compared to thinking about themselves. The ACC activation was correlated with the symptom severity. The activation patterns demonstrated that addicted adolescents were most attached to their game characters and equated their game characters to human.

A selective involvement of putamen functional connectivity in youth with internet gaming disorder.

Brain cortico-striatal circuits have consistently been implicated in the pathology of addiction related disorders. We applied a reliable seed-based analysis of the resting-state brain activity to comprehensively delineate the subdivisions of striatal functional connectivity implicated in internet gaming disorder. Among twelve right-handed male adolescents with internet gaming disorder and 11 right-handed and gender-matched healthy controls, we examined group differences in the functional connectivity of dorsal and ventral subdivisions of the caudate nucleus and putamen, as well as the association of these connectivity indices with behavioral measures of internet use. Adolescents with internet gaming disorder showed significantly reduced dorsal putamen functional connectivity with the posterior insula-parietal operculum. More time spent playing online games predicted significantly greater functional connectivity between the dorsal putamen and bilateral primary somatosensory cortices in adolescents with internet gaming disorder, and significantly lower functional connectivity between the dorsal putamen and bilateral sensorimotor cortices in healthy controls. The dorsal putamen functional connectivity was significantly and specifically different in adolescents with internet gaming disorder. The findings suggest a possible biomarker of internet gaming disorder.

[The effect of self-foot reflexology on the relief of premenstrual syndrome and dysmenorrhea in high school girls].

PURPOSE: This study was aimed to identify the effect of self-foot reflexology on the relief of premenstrual syndrome and dysmenorrhea in high school girls. METHOD: Study subjects was 236 women residing in the community, teachers and nurses who were older than 45 were recruited. Data was collected with self administered questionnaires from July 1st to August 31st, 2003 and analysed using SPSS/WIN 10.0 with Xtest, t-test, and stepwise multiple logistic regression at a significant level of =.05. RESULT: The breast cancer screening rate was 57.2%, and repeat screening rate was 15.3%. With the multiple logistic regression analysis, factors associated with mammography screening were age and perceived barriers of action, and factors related to the repeat mammography screening were education level and other cancer screening experience. CONCLUSION: Based on the results, we recommend the development of an intervention program to decrease the perceived barrier of action, to regard mammography as an essential test in regular check-up, and to give active advertisement and education to the public to improve the rates of breast cancer screening and repeat screening.

[The effects of a stretching exercise program in elderly women].

PURPOSE: This study was aimed to identify the effects of a ten-week stretching exercise program on physiological, psychological functions, and activities of daily living(ADL) among elderly women. METHOD: Using a quasi-experimental design, the experimental group received a ten-week stretching exercise program 3 times a week from March to May in 2002. They were divided into 22 persons in the experimental groups in 2 halls where the program was incorporated, and 22 persons in the control group in 2 halls, where the program was not incorporated. Data was analyzed with descriptive statistics, the chi-square test, Fisher's Exact test, and ANCOVA was considered significant as a 2-tailed test. RESULT: There was a significant improvement in diastolic blood pressure(P=0.023), total cholesterol (P=0.019), triglycerides (P=0.002), spine ROM(P=0.000), trunk and hip-joint ROM(P=0.000), percent of body fat(P=0.039) as physiological functions, depression(P=0.041) as a psychological function, and activities of daily living(P=0.001) in the experimental group compared to the control group. CONCLUSION: A stretching exercise program showed good effects on improving physiological functions, psychological functions and activities of daily living among the elderly women in a city. Therefore, we recommend this program be utilized as a health promoting program for the elderly in the community.

[The development of educational CD-program for obesity prevention and management for primary school students].

PURPOSE: The study is designed to develop an educational CD-Program for prevention and control of obesity among primary school students. METHOD: The study is conducted from June 15, 2000 to April 15, 2002. Based on the course of program development suggested by Dick and Cray (1990), the study followed the planning, development, education and evaluation of a program. RESULT: The developed CD-Program consists 2 parts each for lower and higher grades of primary school students. The introduction part of the first trial for lower grade students uses quiz to encourage their motivations, the body proceeds with motion pictures and animations to trigger their interests. The introduction part of the second trial for the lower grades consists of remembering the exhibition lecture. The first trial for higher grades of primary school students builds on the contents of the low grades. Its body part, how to determine obesity and calculate ones own obesity, puts ones own weight and height in by the mouse. For the second trial of the higher grades, the body consists of life-style, diet, and regiments. CONCLUSION: The merits of this CD-Program are that to be possible an interaction between teachers and students.

Decreased thalamic volumes in adolescent inhalant users from Korea and Australia.

OBJECTIVES: Research investigating the impact of inhalant misuse on brain structure suggests abnormalities in subcortical regions. We investigated the association between inhalant misuse and subcortical brain volumes in adolescents. METHODS: Based on a collaborative dataset from South Korea (inhalant users: N = 15, mean age = 16.7, SD = 1.1; controls: N = 15, mean age = 15.4, SD = 1.2) and Australia (inhalant users: N = 7, mean age = 18.2, SD = 1.4; controls: N = 7, mean age = 18.9, SD = 2.6), the volumes of caudate nucleus, putamen, pallidum, amygdala, hippocampus, and thalamus were estimated in adolescent inhalant users and healthy adolescents using FreeSurfer. RESULTS: The results revealed a significantly decreased right thalamic volume in adolescent inhalant users (P = 0.042), along with a trend-level decrease in left thalamic volume (P = 0.061). A negative correlation (r = -0.544; P = 0.036) between thalamic volume and severity of inhalant use (i.e., reduced volumes associated with greater use) was identified among Korean participants. CONCLUSIONS: These findings suggest that compared with other subcortical structures, the thalamus is particularly sensitive to damage following chronic inhalant exposure during adolescence.