Genetic predispositions can shape the gut microbiome, which in turn modulates host gene expression and impacts host physiology. The complex interplay between host genetics and the gut microbiome likely contributes to the development of neuropsychiatric disorders, yet the mechanisms behind these interactions remain largely unexplored. In this study, we investigated the gut microbiota in <i>Negr1</i> knockout (KO) mice, which exhibit anxiety- and depression-like behaviors, as NEGR1 (neuronal growth regulator 1) is a cell adhesion molecule linked to neuronal development and neuropsychiatric disorders. Our findings show significant early-life alterations in the gut microbiota composition of <i>Negr1</i> KO mice, most notably a marked reduction in <i>Akkermansia</i> spp. along with reduced dendritic arborization and spine density in the nucleus accumbens (NAc) and the dentate gyrus (DG) of the hippocampus. Remarkably, daily administration of an <i>Akkermansia</i> strain isolated from wild-type mice reversed the neuronal structural abnormalities and ameliorated anxiety- and depression-like behaviors in <i>Negr1</i> KO mice. Transcriptomic profiling revealed upregulation of mitochondrial genome-encoded genes in the NAc and hippocampus of <i>Negr1</i> KO mice, along with a predisposition toward a pro-inflammatory state in the colon of <i>Negr1</i> KO mice. The <i>Akkermansia</i> supplementation downregulated these mitochondrial genes in the NAc and hippocampus and upregulated genes involved in T cell activation and immune homeostasis in the colon. These findings demonstrate a novel gene-microbiome interaction in the pathophysiology of <i>Negr1</i> KO mice, positioning <i>Akkermansia</i> spp. as a key mediator that improves neuronal atrophy and modulates anxiety- and depression-like behaviors. Our study provides compelling evidence for bidirectional interactions between host genetics and the gut microbiome in modulating neuropsychiatric phenotypes, offering new insights for addressing genetically influenced mental disorders.

Commensal bacteria are critically involved in the establishment of tolerance against inflammatory challenges, the molecular mechanisms of which are just being uncovered. All kingdoms of life produce aminoacyl-tRNA synthetases (ARSs). Thus far, the non-translational roles of ARSs have largely been reported in eukaryotes. Here, we report that the threonyl-tRNA synthetase (AmTARS) of the gut-associated bacterium Akkermansia muciniphila is secreted and functions to monitor and modulate immune homeostasis. Secreted AmTARS triggers M2 macrophage polarization and orchestrates the production of anti-inflammatory IL-10 via its unique, evolutionary-acquired regions, which mediates specific interactions with TLR2. This interaction activates the MAPK and PI3K/AKT signaling pathways, which converge on CREB, leading to an efficient production of IL-10 and suppression of the central inflammatory mediator NF-κB. AmTARS restores IL-10-positive macrophages, increases IL-10 levels in the serum, and attenuates the pathological effects in colitis mice. Thus, commensal tRNA synthetases can act as intrinsic mediators that maintain homeostasis.

Ischaemic heart disease (IHD) is the leading cause of death worldwide. Although myocardial cell death plays a significant role in myocardial infarction (MI), its underlying mechanism remains to be elucidated. To understand the progression of MI and identify potential therapeutic targets, we performed tandem mass tag (TMT)-based quantitative proteomic analysis using an MI mouse model. Gene ontology (GO) analysis and gene set enrichment analysis (GSEA) revealed that the glutathione metabolic pathway and reactive oxygen species (ROS) pathway were significantly downregulated during MI. In particular, glutathione peroxidase 4 (GPX4), which protects cells from ferroptosis (an iron-dependent programme of regulated necrosis), was downregulated in the early and middle stages of MI. RNA-seq and qRT-PCR analyses suggested that GPX4 downregulation occurred at the transcriptional level. Depletion or inhibition of GPX4 using specific siRNA or the chemical inhibitor RSL3, respectively, resulted in the accumulation of lipid peroxide, leading to cell death by ferroptosis in H9c2 cardiomyoblasts. Although neonatal rat ventricular myocytes (NRVMs) were less sensitive to GPX4 inhibition than H9c2 cells, NRVMs rapidly underwent ferroptosis in response to GPX4 inhibition under cysteine deprivation. Our study suggests that downregulation of GPX4 during MI contributes to ferroptotic cell death in cardiomyocytes upon metabolic stress such as cysteine deprivation.

Proc Amer Assoc Cancer Res, Volume 47, 2006 2179 A promising target for tumor vasculature is phosphatidylserine (PS), an anionic phospholipid that resides exclusively on the inner leaflet of the plasma membrane under normal conditions. We have previously shown that PS becomes exposed on the surface of viable endothelial cells (EC) in solid tumors, likely in response to oxidative stresses that exist in the tumor microenvironment. To target PS on tumor vasculature, a murine monoclonal antibody (3G4) was developed. 3G4 specifically localizes to the vasculature of solid tumors. Treatment of mice with 3G4 inhibits growth of murine and human tumors and enhances the anti-tumor effects of standard radio- and chemo-therapeutic strategies. Therefore, 3G4 is a promising new therapeutic agent for treatment of solid tumor malignancies. We demonstrate here that the interaction between 3G4 and PS is dependent on antibody binding to the plasma glycoprotein beta2-glycoprotein I (β2GPI). When purified from a hybridoma converted to grow under serum-free conditions, 3G4 does not bind PS-coated ELISA plates unless serum is added to the binding reaction A similar phenomenon was described in the early 1990s, when “anti-cardiolipin antibodies” from patients with Anti-Phospholipid Syndrome (APS) were shown to require various plasma proteins (such as β2GPI or prothrombin) for binding to cardiolipin. Screening a panel of serum proteins for reactivity with 3G4 by ELISA and western blot identified β2GPI as the primary antigen for 3G4. β2GPI is a 50 kDa glycoprotein composed of five domains, including the positively charged domain V that binds anionic phospholipids. However, the interaction with anionic phospholipids is rather weak under physiological conditions. We demonstrate that 3G4 binds domain II of β2GPI and enhances the binding of β2GPI to EC induced to expose PS (PS-positive) 40- to 50-fold. We also show that binding of 3G4 to PS-positive EC is dependent on the lipid binding domain of β2GPI (domain V), since enzymatic cleavage of the lipid binding site abrogates both binding to PS and to EC. Finally, we demonstrate that the 3G4/β2GPI/PS binding interaction requires multivalent binding of 3G4 to β2GPI, since 3G4 Fab’ fragments do not bind PS-positive ECs. Together, the data suggest that 3G4 targets tumor vessels by increasing the binding affinity of β2GPI to PS exposed on tumor EC.

Genetic predispositions can shape the gut microbiome, which in turn modulates host gene expression and impacts host physiology. The complex interplay between host genetics and the gut microbiome likely contributes to the development of neuropsychiatric disorders, yet the mechanisms behind these interactions remain largely unexplored. In this study, we investigated the gut microbiota in <i>Negr1</i> knockout (KO) mice, which exhibit anxiety- and depression-like behaviors, as NEGR1 (neuronal growth regulator 1) is a cell adhesion molecule linked to neuronal development and neuropsychiatric disorders. Our findings show significant early-life alterations in the gut microbiota composition of <i>Negr1</i> KO mice, most notably a marked reduction in <i>Akkermansia</i> spp. along with reduced dendritic arborization and spine density in the nucleus accumbens (NAc) and the dentate gyrus (DG) of the hippocampus. Remarkably, daily administration of an <i>Akkermansia</i> strain isolated from wild-type mice reversed the neuronal structural abnormalities and ameliorated anxiety- and depression-like behaviors in <i>Negr1</i> KO mice. Transcriptomic profiling revealed upregulation of mitochondrial genome-encoded genes in the NAc and hippocampus of <i>Negr1</i> KO mice, along with a predisposition toward a pro-inflammatory state in the colon of <i>Negr1</i> KO mice. The <i>Akkermansia</i> supplementation downregulated these mitochondrial genes in the NAc and hippocampus and upregulated genes involved in T cell activation and immune homeostasis in the colon. These findings demonstrate a novel gene-microbiome interaction in the pathophysiology of <i>Negr1</i> KO mice, positioning <i>Akkermansia</i> spp. as a key mediator that improves neuronal atrophy and modulates anxiety- and depression-like behaviors. Our study provides compelling evidence for bidirectional interactions between host genetics and the gut microbiome in modulating neuropsychiatric phenotypes, offering new insights for addressing genetically influenced mental disorders.

Commensal bacteria are critically involved in the establishment of tolerance against inflammatory challenges, the molecular mechanisms of which are just being uncovered. All kingdoms of life produce aminoacyl-tRNA synthetases (ARSs). Thus far, the non-translational roles of ARSs have largely been reported in eukaryotes. Here, we report that the threonyl-tRNA synthetase (AmTARS) of the gut-associated bacterium Akkermansia muciniphila is secreted and functions to monitor and modulate immune homeostasis. Secreted AmTARS triggers M2 macrophage polarization and orchestrates the production of anti-inflammatory IL-10 via its unique, evolutionary-acquired regions, which mediates specific interactions with TLR2. This interaction activates the MAPK and PI3K/AKT signaling pathways, which converge on CREB, leading to an efficient production of IL-10 and suppression of the central inflammatory mediator NF-κB. AmTARS restores IL-10-positive macrophages, increases IL-10 levels in the serum, and attenuates the pathological effects in colitis mice. Thus, commensal tRNA synthetases can act as intrinsic mediators that maintain homeostasis.

Ischaemic heart disease (IHD) is the leading cause of death worldwide. Although myocardial cell death plays a significant role in myocardial infarction (MI), its underlying mechanism remains to be elucidated. To understand the progression of MI and identify potential therapeutic targets, we performed tandem mass tag (TMT)-based quantitative proteomic analysis using an MI mouse model. Gene ontology (GO) analysis and gene set enrichment analysis (GSEA) revealed that the glutathione metabolic pathway and reactive oxygen species (ROS) pathway were significantly downregulated during MI. In particular, glutathione peroxidase 4 (GPX4), which protects cells from ferroptosis (an iron-dependent programme of regulated necrosis), was downregulated in the early and middle stages of MI. RNA-seq and qRT-PCR analyses suggested that GPX4 downregulation occurred at the transcriptional level. Depletion or inhibition of GPX4 using specific siRNA or the chemical inhibitor RSL3, respectively, resulted in the accumulation of lipid peroxide, leading to cell death by ferroptosis in H9c2 cardiomyoblasts. Although neonatal rat ventricular myocytes (NRVMs) were less sensitive to GPX4 inhibition than H9c2 cells, NRVMs rapidly underwent ferroptosis in response to GPX4 inhibition under cysteine deprivation. Our study suggests that downregulation of GPX4 during MI contributes to ferroptotic cell death in cardiomyocytes upon metabolic stress such as cysteine deprivation.

Proc Amer Assoc Cancer Res, Volume 47, 2006 2179 A promising target for tumor vasculature is phosphatidylserine (PS), an anionic phospholipid that resides exclusively on the inner leaflet of the plasma membrane under normal conditions. We have previously shown that PS becomes exposed on the surface of viable endothelial cells (EC) in solid tumors, likely in response to oxidative stresses that exist in the tumor microenvironment. To target PS on tumor vasculature, a murine monoclonal antibody (3G4) was developed. 3G4 specifically localizes to the vasculature of solid tumors. Treatment of mice with 3G4 inhibits growth of murine and human tumors and enhances the anti-tumor effects of standard radio- and chemo-therapeutic strategies. Therefore, 3G4 is a promising new therapeutic agent for treatment of solid tumor malignancies. We demonstrate here that the interaction between 3G4 and PS is dependent on antibody binding to the plasma glycoprotein beta2-glycoprotein I (β2GPI). When purified from a hybridoma converted to grow under serum-free conditions, 3G4 does not bind PS-coated ELISA plates unless serum is added to the binding reaction A similar phenomenon was described in the early 1990s, when “anti-cardiolipin antibodies” from patients with Anti-Phospholipid Syndrome (APS) were shown to require various plasma proteins (such as β2GPI or prothrombin) for binding to cardiolipin. Screening a panel of serum proteins for reactivity with 3G4 by ELISA and western blot identified β2GPI as the primary antigen for 3G4. β2GPI is a 50 kDa glycoprotein composed of five domains, including the positively charged domain V that binds anionic phospholipids. However, the interaction with anionic phospholipids is rather weak under physiological conditions. We demonstrate that 3G4 binds domain II of β2GPI and enhances the binding of β2GPI to EC induced to expose PS (PS-positive) 40- to 50-fold. We also show that binding of 3G4 to PS-positive EC is dependent on the lipid binding domain of β2GPI (domain V), since enzymatic cleavage of the lipid binding site abrogates both binding to PS and to EC. Finally, we demonstrate that the 3G4/β2GPI/PS binding interaction requires multivalent binding of 3G4 to β2GPI, since 3G4 Fab’ fragments do not bind PS-positive ECs. Together, the data suggest that 3G4 targets tumor vessels by increasing the binding affinity of β2GPI to PS exposed on tumor EC.

Transgenic mice with a gamma 2b transgene were produced to investigate whether gamma 2b can replace mu in the development of B lymphocytes. Transgenic gamma 2b is present on the surface of B cells. Young transgenic mice have a dramatic decrease in B cell numbers, however, older mice have almost normal B cell numbers. Strikingly, all gamma 2b-expressing B cells in the spleen also express mu. The same is true for mice with a hybrid transgene in which the mu transmembrane and intracytoplasmic sequences replace those of gamma 2b (gamma 2b-mumem). The B cell defect is not due to toxicity of gamma 2b since crosses between gamma 2b transgenic and mu transgenic mice have normal numbers of B cells. Presence of the gamma 2b transgene strongly enhances the feedback inhibition of endogenous heavy chain gene rearrangement. Light chain genes are expressed normally, and the early expression of transgenic light chains does not improve B cell maturation. When the endogenous mu locus is inactivated, B cells do not develop at all in gamma 2b transgenic mice. The data suggest that gamma 2b cannot replace mu in promoting the developmental maturation of B cells, but that it can cause feedback inhibition of heavy chain gene rearrangement. Thus, the signals for heavy chain feedback and B cell maturation appear to be different.

Few studies have examined how Artificial Intelligence-based computer-aided diagnosis (AI-CAD) may miss breast cancers on mammograms. Our goal was to investigate the false-negative results of AI-CAD in women newly diagnosed with breast cancer and to evaluate the standalone diagnostic value of diffusion-weighted imaging (DWI) in detecting breast cancers missed by AI-CAD.

In this paper, we propose a method to improve the accuracy of speech emotion recognition (SER) by using vision transformer (ViT) to attend to the correlation of frequency (y-axis) with time (x-axis) in spectrogram and transferring positional information between ViT through knowledge transfer. The proposed method has the following originality i) We use vertically segmented patches of log-Mel spectrogram to analyze the correlation of frequencies over time. This type of patch allows us to correlate the most relevant frequencies for a particular emotion with the time they were uttered. ii) We propose the use of image coordinate encoding, an absolute positional encoding suitable for ViT. By normalizing the x, y coordinates of the image to −1 to 1 and concatenating them to the image, we can effectively provide valid absolute positional information for ViT. iii) Through feature map matching, the locality and location information of the teacher network is effectively transmitted to the student network. Teacher network is a ViT that contains locality of convolutional stem and absolute position information through image coordinate encoding, and student network is a structure that lacks positional encoding in the basic ViT structure. In feature map matching stage, we train through the mean absolute error (L1 loss) to minimize the difference between the feature maps of the two networks. To validate the proposed method, three emotion datasets (SAVEE, EmoDB, and CREMA-D) consisting of speech were converted into log-Mel spectrograms for comparison experiments. As a result of the experiment, the proposed method achieved 99.47%, 99.76%, and 95.24% accuracy, significantly exceeding the state-of-the-art methods, with much less computational complexity on three emotion datasets.

Rho guanine nucleotide exchange factors (Rho GEFs) activate Rho GTPases, which act as molecular switches regulating various essential cellular functions. This study investigated the role of ARHGEF5, a Rho GEF known for its involvement in cell migration and invasion processes, in the context of brain development. We found that ARHGEF5 is essential for dendrite development during the early stages of neuronal growth. We also discovered that ARHGEF5 binds to Drebrin E, which is vital for coordinating actin and microtubule dynamics, and facilitates the interaction between Drebrin E and Cyclin-dependent kinase 5, which phosphorylates Drebrin E. Notably, ARHGEF5 deficiency resulted in a decrease in acetylated α-tubulin levels, and the expression of an α-tubulin acetylation mimetic mutant (K40Q) rescued the defects in dendrite development and neuronal migration, suggesting ARHGEF5's role in modulating microtubule stability. Additionally, ARHGEF5 was shown to influence Golgi positioning in the leading processes of migrating cortical neurons during brain development. Our study suggests that ARHGEF5 plays a crucial role in integrating cytoskeletal dynamics with neuronal morphogenesis and migration processes during brain development.