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Fungal keratitis (FK) is a sight-threatening infectious disease that can result in blindness if not appropriately treated. Although keratitis associated with filamentous fungi was rarely reported in the past, the incidence of fungal keratitis due to contact lens usage has increased. The clinical manifestations of fungal keratitis often resemble those of other corneal infections, potentially delaying accurate diagnosis and treatment. In this study, we developed a Fusarium -specific polyclonal peptide antibody targeting the cutinase of F. solani and evaluated its diagnostic potential for FK in an animal model. To assess the specificity of the cutinase antibody, we employed enzyme-linked immunosorbent assay (ELISA). The ELISA results demonstrated that the cutinase antibody specifically interacted with the cell lysates and conditioned media of F. solani . Additionally, an immunocytochemistry assay confirmed the specificity of the cutinase antibody for F. solani in samples co-cultured with human corneal epithelial (HCE) cells and other keratitis-causing agents. To validate our in vitro findings, FK animal models were established by infecting the corneas of BALB/c mice with F. solani . The cutinase antibody specifically detected Fusarium antigens in the tear-wash samples and eyeball lysates of FK mice. These results demonstrate that the cutinase antibody is highly specific to F. solani antigens, indicating its potential utility in developing an antibody-based diagnostic method for FK.

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.

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.

Individuals with generalized anxiety disorder (GAD) often face challenges with self-regulation and limited access to traditional therapy. Although acceptance and commitment therapy (ACT) has demonstrated both efficacy and effectiveness in promoting psychological flexibility, scalable solutions are necessary to address these barriers. This study introduces Anzeilax, an ACT-based digital therapeutic (DTx) that incorporates self-talk as a novel mechanism of action (MoA) to enhance psychological flexibility in the treatment of GAD. This study aimed to evaluate the efficacy of Anzeilax in reducing anxiety symptoms in individuals with GAD. A 10-week, parallel-group, superiority randomized controlled trial (RCT) was conducted with 96 participants diagnosed with GAD (Generalized Anxiety Disorder 7-item scale [GAD-7] scores≥10, age≥19 years). The participants were randomly assigned (1:1) to receive either Anzeilax alongside treatment as usual (TAU, treatment group; n=48, 50%) or TAU alone (control group; n=48). Only the outcome evaluators were blinded to the group assignment. The primary outcome was the change in the GAD-7 score from baseline to week 10. The secondary outcomes included the Beck Anxiety Inventory (BAI) for anxiety symptoms, the Penn State Worry Questionnaire (PSWQ) for pathological worry, and the Hospital Anxiety and Depression Scale (HADS) for anxiety (HADS-A) and depression (HADS-D) symptoms. All self-report outcomes were assessed at baseline and at weeks 5 (midintervention), 10 (postintervention), and 15 (follow-up). During the trial, 34 (71%) and 31 (65%) participants in the treatment group maintained at least 80% of the prescribed usage frequency at weeks 5 and 10, respectively. Based on the full analysis set (FAS), participants using Anzeilax demonstrated significant improvement in anxiety symptoms compared to the control group. Analysis of the primary outcome at 10 weeks postintervention compared to baseline exhibited a significant reduction in GAD-7 scores (adjusted mean difference -2.26, 95% CI -3.78 to -0.74, P=.002). Secondary outcomes at the same time point indicated consistent improvements, with significant group-by-time interactions observed in the GAD-7 (Cohen d=0.60, P=.008), BAI (Cohen d=0.50, P=.008), PSWQ (Cohen d=0.62, P=.002), and HADS-A (Cohen d=0.50, P=.01) scores. These improvements were sustained at the follow-up assessment (week 15). Although the differences in depressive symptoms between the two groups did not present statistical significance, notable improvements were observed in the treatment group. Anzeilax demonstrated clinically meaningful efficacy in reducing anxiety symptoms when combined with TAU. The results showed consistent improvements across multiple anxiety measures, with effects sustained through follow-up. The incorporation of context-sensitive self-talk within an ACT-based DTx framework offers a promising and accessible solution for treating individuals with GAD. ClinicalTrials.gov NCT06010654; https://clinicaltrials.gov/study/NCT06010654.

The increasing number of contact lens users correlates with a rise in the incidence of fungal keratitis. Fungal keratitis can lead to blindness if not treated promptly, and its early symptoms are similar to those of bacterial and amoebic keratitis, making rapid diagnosis challenging. This study aimed to assess the potential of using a peptide antibody against the fungal-specific protein ERG24, which encodes sterol C-14 reductase, to differentiate fungal keratitis from other forms of keratitis. The specificity of the ERG24 antibody was assessed through Western blot and enzyme-linked immunosorbent assay (ELISA). Immunocytochemistry (ICC) was performed by co-culturing two types of fungi, Acanthamoeba , and two bacterial strains with human corneal epithelial (HCE) cells. Additionally, to evaluate the diagnostic potential of the ERG24 antibody, animal models of fungal, amoebic, and bacterial keratitis were developed, and ELISA was conducted on tear and ocular lysates from these models. The results demonstrated that the antibody specifically reacted with Fusarium solani in Western blot, and both ELISA and ICC confirmed that the ERG24 antibody did not react with HCE cells, Acanthamoeba , or bacteria, but was specific to the two fungal species. In vivo experiments further showed that the ERG24 antibody significantly detected F. solani in tear-wash samples and eyeball lysates from the fungal keratitis model, without reacting with samples from amoebic and bacterial keratitis models. This study suggests that the ERG24 peptide antibody could provide valuable information for developing differential diagnostic methods for fungal keratitis compared to other forms of keratitis.

Nuclear accidents, such as the Fukushima nuclear accident, have highlighted the necessity for accident‐tolerant fuel (ATF) cladding. Previous studies focused on coating the outside of Zr alloy currently used in nuclear reactors with an oxidation‐resistant material in a vacuum environment. This limits the coating to the inside of the cladding and does not tend to achieve a uniform coating on meter scale cladding. In this study, a room temperature and non‐vacuum‐based swaging–drawing process was demonstrated as an alternative cladding manufacturing process. It enables both the inner and outer sides of the 2‐m‐long Zr alloy cladding to be uniformly covered with a 100‐ μ m‐thick corrosion‐resistant material (316‐L stainless steel; SS316L), thereby minimizing its high‐temperature oxidation and avoiding failures. After the swaging–drawing process, there was a gap of less than 1  µ m between outer SS316L and Zr alloy and a gap of about 12  µ m between inner Zr alloy and SS316L. The high‐temperature oxidation properties of the resulting triplex Gachon ATF cladding tube (G‐tube) were evaluated up to 1,200°C in an atmospheric environment. Following heat treatment at 1,200°C, the control cladding completely oxidized and ruptured, potentially causing leakage of radioactive material during application. In contrast, only 15% of the G‐tube cladding manufactured by the swaging–drawing process was oxidized despite a gap, and the Zr alloy of the G‐tube changed phase from α ‐Zr to α ‐Zr (O) and prior β ‐Zr. The cladding microstructure, oxide layer, and oxidation mechanism were analyzed through microscopy, X‐ray diffraction, and thermogravimetric analysis. As a result, it was confirmed that SS316L completely prevented oxygen diffusion into the bulk Zr alloy. In addition, there was no elemental diffusion between SS316L and the Zr alloy. These results demonstrate the feasibility of using room temperature, nonvacuum environment‐based swaging–drawing process to fabricate structurally stable ATF cladding at extremely high temperatures.

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. Graphical Abstract

One’s own voice undergoes unique processing that distinguishes it from others’ voices, and thus listening to it may have a special neural basis for self-talk as an emotion regulation strategy. This study aimed to elucidate how neural effects of one’s own voice differ from those of others’ voices on the implementation of emotion regulation strategies. Twenty-one healthy adults were scanned using fMRI while listening to sentences synthesized in their own or others’ voices for self-affirmation and cognitive defusion, which were based on mental commitments to strengthen one’s positive aspects and imagining metaphoric actions to shake off negative aspects, respectively. The interaction effect between voice identity and strategy was observed in the superior temporal sulcus, middle temporal gyrus, and parahippocampal cortex, and activity in these regions showed that the uniqueness of one’s own voice is reflected more strongly for cognitive defusion than for self-affirmation. This interaction was also seen in the precuneus, suggesting intertwining of self-referential processing and episodic memory retrieval in self-affirmation with one’s own voice. These results imply that unique effects of one’s own voice may be expressed differently due to the degree of engagement of neural sharpening-related regions and self-referential networks depending on the type of emotion regulation.

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.

Virtual reality (VR) technologies can induce realistic emotions in controlled experimental settings, offering unprecedented opportunities to study how the human brain processes emotions under real-world conditions. The integration of VR experiences with electroencephalography (EEG) provides a promising potential for gaining novel insights into individual emotional states. However, the complex network dynamics underlying human emotions during VR experiences remain largely unexplored. To address this gap, we leveraged graph-theoretical approaches to investigate functional brain networks derived from EEG signals recorded during immersive VR experiments. We assessed key topological properties of functional brain networks across multiple frequency bands (delta, theta, alpha, beta, gamma, and high gamma) and compared network characteristics between different emotional states (negative, neutral, and positive). Furthermore, we evaluated whether these graph-based features could accurately distinguish between positive and negative emotions using machine learning approaches. Our findings revealed distinct network patterns associated with different emotional states. During negative emotional experiences, we observed two key neural signatures: increased high gamma band activity in the left central region and decreased theta band activity in the occipital region. Conversely, positive emotions were characterized by reduced activity across most frequency bands in the left frontal region. Our machine learning model achieved an average classification accuracy of 79% in differentiating positive and negative emotions using network features that combined graph-theoretical measures and connectivity weights across all frequency bands, with the high gamma band demonstrating particular importance for emotion processing. This study advances our understanding of how brain networks dynamically reorganize during VR-induced emotional experiences and establishes the potential of graph-based EEG features for robust emotion recognition, paving the way for personalized VR applications.