Neural substrates of marriage on self-parents processing and the association with a parents-oriented perspective shift in a collectivistic culture.

Relationship with parents is a special bond that shapes self-other representations and have an impact on adult-child's marriage, especially in the early stages of marriage. This study sought to investigate the neural mechanisms underlying self-parents processing as well as their relationship with marriage. Seventy-eight premarital Korean participants were scanned in functional MRI while evaluating traits of the self and parents. Then, 21 of them returned after being married to engage in the identical task three years later. The precuneus and temporoparietal junction were identified to activate stronger for parents than self at both marital statuses. The dorsal anterior cingulate cortex, posterior cingulate cortex, parietal operculum, and caudate activated more for self than parents before marriage, but their activities changed during marriage. The activation increase of the parietal operculum between marital statuses in the parents condition was negatively correlated with the level of marital dissatisfaction, and this association only appeared among participants with a child. Self-parents processing may recruit brain regions involved in autobiographical memory and self-other distinction, and marriage has an impact on the way individuals process rewards and multimodal sensory information during this processing. Marriage may lead to changes in brain function that affect the processing of emotions toward parents and a more parents-oriented perspective shift in collectivistic societies.

Phagocytosis-associated genes in Acanthamoeba castellanii feeding on Escherichia coli.

Acanthamoeba species are free-living amoebae those are widely distributed in the environment. They feed on various microorganisms, including bacteria, fungi, and algae. Although majority of the microbes phagocytosed by Acanthamoeba spp. are digested, some pathogenic bacteria thrive within them. Here, we identified the roles of 3 phagocytosis-associated genes (ACA1_077100, ACA1_175060, and AFD36229.1) in A. castellanii. These 3 genes were upregulated after the ingestion of Escherichia coli. However, after the ingestion of Legionella pneumophila, the expression of these 3 genes was not altered after the consumption of L. pneumophila. Furthermore, A. castellanii transfected with small interfering RNS (siRNA) targeting the 3 phagocytosis-associated genes failed to digest phagocytized E. coli. Silencing of ACA1_077100 disabled phagosome formation in the E. coli-ingesting A. castellanii. Alternatively, silencing of ACA1_175060 enabled phagosome formation; however, phagolysosome formation was inhibited. Moreover, suppression of AFD36229.1 expression prevented E. coli digestion and consequently led to the rupturing of A. castellanii. Our results demonstrated that the ACA1_077100, ACA1_175060, and AFD36229.1 genes of Acanthamoeba played crucial roles not only in the formation of phagosome and phagolysosome but also in the digestion of E. coli.

Identifying the function of genes involved in excreted vesicle formation in Acanthamoeba castellanii containing Legionella pneumophila.

BACKGROUND: Legionella spp. can survive and replicate inside host cells such as protozoa and macrophages. After enough growth, Legionella is released from the host cells as free legionellae or Legionella-filled vesicles. The vesicles support Legionella to survive for a long time in the environment and transmit to a new host. In this study, we identified the differentially expressed genes of Acanthamoeba infected by Legionella (ACA1_114460, ACA1_091500, and ACA1_362260) and examined their roles in the formation of the excreted vesicles and escape of Legionella from the Acanthamoeba. METHODS: After ingestion of Escherichia coli and Legionella pneumophila, expression levels of target genes in Acanthamoeba were measured by real-time polymerase chain reaction (PCR) analysis. The roles of target genes were investigated by transfection of small interfering RNA (siRNA). The formation of Legionella-containing excreted vesicles and the vesicular co-localization with the lysosomes were examined by Giemsa stain and LysoTracker stain. RESULTS: ACA1_114460, ACA1_091500, and ACA1_362260 were upregulated after ingestion of Legionella in Acanthamoeba. ACA1_114460- and ACA1_091500-silenced Acanthamoeba failed to form the Legionella-containing excreted vesicles. Legionella was released as free legionellae from the Acanthamoeba. When the ACA1_362260 of Acanthamoeba was silenced, Legionella-containing excreted vesicles were fused with the lysosome. CONCLUSIONS: These results indicated that ACA1_114460, ACA1_091500, and ACA1_362260 of Acanthamoeba played important roles in the formation of Legionella-containing excreted vesicles and inhibition of the lysosomal co-localization with the phagosome.

Evaluating the Diagnostic Potential of Chorismate Mutase Poly-Clonal Peptide Antibody for the Acanthamoeba Keratitis in an Animal Model.

Acanthamoeba spp. is the causative agent of Acanthamoeba keratitis (AK), a vision-threatening parasitic disease whose primary risk factor has been attributed to poor contact lens hygiene. Unfortunately, differential diagnosis of AK is challenging as the clinical manifestations for AK are similar to those of bacterial, fungal, or even viral keratitis. Since delayed AK diagnosis can incur permanent vision impairment, a rapid and sensitive diagnostic method is urgently needed. Here, the diagnostic potential of polyclonal antibodies targeting the chorismate mutase (CM) of Acanthamoeba spp. was evaluated in AK animal models. CM antibody specificity against Acanthamoeba trophozoites and cysts was confirmed by immunocytochemistry after co-culturing Acanthamoeba with Fusarium solani, Pseudomonas aeruginosa, and Staphylococcus aureus, and human corneal epithelial (HCE) cells. Enzyme-linked immunosorbent assay (ELISA) was performed using CM-specific immune sera raised in rabbits, which demonstrated that the antibodies specifically interacted with the Acanthamoeba trophozoites and cysts in a dose-dependent manner. To evaluate the diagnostic potential of the CM antibody, AK animal models were established by incubating contact lenses with an inoculum containing A. castellanii trophozoites and subsequently overlaying these lenses onto the corneas of BALB/c mice for 7 and 21 days. The CM antibody specifically detected Acanthamoeba antigens in the murine lacrimal and eyeball tissue lysates at both time points. Our findings underscore the importance of antibody-based AK diagnosis, which could enable early and differential AK diagnosis in clinical settings.

Predicting social anxiety in young adults with machine learning of resting-state brain functional radiomic features.

Social anxiety is a symptom widely prevalent among young adults, and when present in excess, can lead to maladaptive patterns of social behavior. Recent approaches that incorporate brain functional radiomic features and machine learning have shown potential for predicting certain phenotypes or disorders from functional magnetic resonance images. In this study, we aimed to predict the level of social anxiety in young adult participants by training machine learning models with resting-state brain functional radiomic features including the regional homogeneity, fractional amplitude of low-frequency fluctuation, fractional resting-state physiological fluctuation amplitude, and degree centrality. Among the machine learning models, the XGBoost model achieved the best performance with balanced accuracy of 77.7% and F1 score of 0.815. Analysis of input feature importance demonstrated that the orbitofrontal cortex and the degree centrality were most relevant to predicting the level of social anxiety among the input brain regions and the input type of radiomic features, respectively. These results suggest potential validity for predicting social anxiety with machine learning of the resting-state brain functional radiomic features and provide further understanding of the neural basis of the symptom.

The Applicability of Virtual Reality-Based Training for Controlling Anger in Aggressive Individuals.

Because a failure of anger control leads to emotional and social problems, appropriate anger management may be important for social well-being. Virtual reality (VR) may potentially be effectively utilized in anger management, and this study aimed to verify the applicability of the VR-based anger control training program. The data obtained by having 60 young male participants divided into 2 groups, the high aggression group and the low aggression group, based on their Aggression Questionnaire scores to execute this program were analyzed. The program consisted of "Anger Exposure Training" for provoking anger and facilitating anger control and "Mindfulness Training" for providing the meditation experience for controlling anger. The anger scores and comfort scores obtained from these tasks, respectively, were analyzed for differences between the groups and between the experimental conditions. The anger regulation and comfort enhancement rates were analyzed for correlations with psychological variables. In Anger Exposure Training, the anger scores in angry expression were reduced in managed expression of anger in both groups. In Mindfulness Training, meditation increased comfort score as well in both groups, and the comfort enhancement rates were negatively correlated with the levels of self-differentiation and open communication with mother only in the high aggression group. These results indicate that the VR environments can provide an effective means of trainings for managing anger. Therefore, further research on the effectiveness of the VR-based anger control training program is worthy conducting in individuals who express excessive aggression.

Enhanced monoterpene emission in transgenic orange mint (Mentha × piperita f. citrata) overexpressing a tobacco lipid transfer protein (NtLTP1).

MAIN CONCLUSION: Overexpression of the tobacco lipid transfer protein (NtLTP1) gene in transgenic orange mint resulted in enhanced accumulation of monoterpenes in the cavity of head cells of glandular trichomes, which resulted in enhanced emission of monoterpenes from transgenic orange mints. Plants in the genus Mentha (Lamiaceae) produce volatile oils that accumulate in peltate glandular trichomes in the aerial parts of plants. A lipid transfer protein (NtLTP1) in tobacco showed glandular trichome-specific expression and supported the secretion of diterpenoid lipids from head cells of glandular trichomes (Choi et al., Plant J 70:480-491,2012). Here, we constructed transgenic orange mint (Mentha × piperita f. citrata) overexpressing the tobacco NtLTP1 gene via Agrobacterium-mediated transformation. Transgenic lines of orange mint overexpressing NtLTP1 were confirmed by genomic PCR and RT-PCR. Immunoblotting analysis using an NtLTP1 polyclonal antibody showed clear dark spots at the position of the lipid exudates from tobacco glandular trichomes and the squeezed out lipids from the glandular trichomes of transgenic orange mint. Heads of glandular trichomes in transgenic plants overexpressing the NtLTP1 gene showed a larger diameter than those of the wild-type control. The enhanced size of trichome heads in transgenic orange mint was confirmed by scanning electron microscopy. Volatile components were extracted from wild-type and transgenic orange mint by solid-phase microextraction (SPME) and analyzed by headspace-gas chromatography-mass spectrometry (HS/GC/MS). Linalyl acetate was the most abundant component among the eleven identified monoterpenes in the volatile compounds extracted from both the wild-type and transgenic lines of orange mint. Overexpression of NtLTP1 in transgenic orange mint plants resulted in enhanced emission of volatile monoterpenoids compared with that of volatile monoterpenoids in the wild-type control plants.

Genetically modified rice produces ginsenoside aglycone (protopanaxadiol).

MAIN CONCLUSION: Protopanaxadiol is dammarane-type tetracyclic triterpene sapogenin found in ginseng and has a high medicinal values. We successfully constructed transgenic rice producing protopanaxadiol by introducing the ginseng PgDDS and CYP716A47 genes in this crop plant. Protopanaxadiol (PPD), an aglycone of ginsenosides, possesses pleiotropic anticarcinogenesis activities in many cancers. Here, we constructed transgenic rice overexpressing the Panax ginseng dammarenediol-II synthase gene (PgDDS) and protopanaxadiol synthase gene (CYP716A47) driven by a rice endosperm-specific α-globulin promoter. Among more than 50 independent lines, five transgenic lines were selected. The introduction of the genes in the T1 generation of the transgenic lines was confirmed by genomic PCR. The expression of the introduced genes in T2 seeds was confirmed by qPCR. Methanol extracts of transgenic rice grains were analyzed by LC/MS to detect the production of PPD and dammarenediol-II (DD). The production of both PPD and DD was identified not only by comparing the retention times but also mass fraction patterns of authentic PPD and DD standards. The mean concentrations of PPD and DD in rice grains were 16.4 and 4.5 µg/g dry weight, respectively. The invention of genetically engineered rice grains producing PPD and DD can be applied to rice breeding to reinforce new medicinal values.

Cloning and Characterization of Oxidosqualene Cyclases Involved in Taraxasterol, Taraxerol and Bauerenol Triterpene Biosynthesis in Taraxacum coreanum.

Triterpenes, consisting of six isoprene units, are one of the largest classes of natural compounds in plants. The genus Taraxacum is in the family Asteraceae and is widely distributed in the Northern Hemisphere. Various triterpenes, especially taraxerol and taraxasterol, are present in Taraxacum plants. Triterpene biosynthesis occurs through the action of oxidosqualene cyclase (OSC), which generates various types of triterpenes from 2,3-oxidosqualene after the rearrangement of the triterpene skeleton. However, no functional characterization of the OSC genes involved in triterpene biosynthesis, except for a lupeol synthase in Taraxacum officinale, has been performed. Taraxacum coreanum, or Korean dandelion, grows in Korea and China. Putative OSC genes in T. coreanum plants were isolated by transcriptome analysis, and four of these (TcOSC1, TcOSC2, TcOSC3 and TcOSC4) were functionally characterized by heterologous expression in yeast. Both TcOSC1 and TcOSC2 were closely related to dammarenediol-II synthases. TcOSC3 and TcOSC4 were strongly grouped with ß-amyrin synthases. Functional analysis revealed that TcOSC1 produced several triterpenes, including taraxasterol; Ψ-taraxasterol; α-, ß- and δ-amyrin; and dammarenediol-II. TcOSC2 catalyzed the production of bauerenol and another unknown triterpene, TcOSC3 catalyzed the production of ß-amyrin. TcOSC4 catalyzed the production of taraxerol. Moreover, we identified taraxasterol, ψ-taraxasterol, taraxerol, lupeol, δ-amyrin, α-amyrin, ß-amyrin and bauerenol in the roots and leaves of T. coreanum. Our results suggest that TcOSC1, TcOSC2, TcOSC3 and TcOSC4 are key triterpene biosynthetic enzymes in T. coreanum. These enzymes are novel triterpene synthases involved in the production of taraxasterol, bauerenol and taraxerol.

ß-Amyrin synthase (EsBAS) and ß-amyrin 28-oxidase (CYP716A244) in oleanane-type triterpene saponin biosynthesis in Eleutherococcus senticosus.

Siberian ginseng (Eleutherococcus senticosus) is a woody medical shrub belonging to the Araliaceae family. E. senticosus contains various types of saponins, including oleanane, noroleanane, lupane, and 3,4-secolupane types, depending on the aglycone structure. Oleanane-type triterpenes are the major saponin components in E. senticosus. Two enzymes (ß-amyrin synthase and ß-amyrin 28-oxidase) are essential for oleanane-type saponin biosynthesis from 2,3-oxidosqualene. In the present study, two full-length cDNAs encoding EsBAS and CYP716A244 were isolated based on transcriptomics analysis of plant leaves. Both ß-amyrin synthase (EsBAS) and ß-amyrin 28-oxidase (CYP716A244), isolated from E. senticosus, were functionally characterised. ß-amyrin production was confirmed by heterologous expression of the EsBAS gene in yeast and tobacco. Oleanolic acid production was confirmed by co-expression of both EsBAS and CYP716A244 in engineered yeast and transgenic tobacco.