Dysbiosis leads to decreased intestinal barrier function, causing systemic inflammation and possibly the development of age-related cognitive decline. In this study, we investigated the effect of an Alaska pollack protein (APP) diet on cognitive function, gut microbiota composition, intestinal barrier function, and neuroinflammation in senescence-accelerated mouse prone8 (SAMP8) and senescence-resistant AKR/J (SAMR1) mice. The APP diet produced significant improvements across multiple parameters. It enhanced glucose tolerance in both strains and prevented short-term memory decline in SAMP8 mice. Microbiome analysis revealed that APP intake promoted beneficial bacteria growth, specifically increasing Lactobacillus in SAMR1 and butyrate-producing Lachnospiraceae in SAMP8. Notably, while APP diet increased butyrate-producing bacteria in SAMP8, short-chain fatty acids (SCFAs) analysis showed increased aetate but unchanged butyrate levels, suggesting complex metabolic interactions beyond simple bacterial abundance. Moreover, the APP diet significantly suppressed neuroinflammation in SAMP8, evidenced by decreased proinflammatory cytokine expression, microglia and astrocyte activation, and attenuated demyelination in the hippocampus. These findings suggest that APP intake prevents age-related short-term memory decline through beneficial gut microbiota modulation, and reduced neuroinflammation, supporting the role of the gut-brain axis in cognitive aging.

Proteolysis-targeting chimeras (PROTACs) are promising next-generation therapeutics for the degradation of disease-associated proteins. However, optimizing the physicochemical properties of PROTACs, particularly their poor cell membrane permeability, remains challenging. Traditionally, PROTAC linkers have been manually designed to improve cell membrane permeability. Although recent machine learning-based approaches have enabled the rational design of PROTAC linkers, no linker design methods that explicitly address cell membrane permeability have been reported. In this study, we developed PROTAC-TS, a linker generative model that combines a chemical language model and reinforcement learning to control cell membrane permeability. We first constructed a prediction model of cell membrane permeability, which achieved high prediction performance (R 2 = 0.710). By integrating this prediction model into the generative model, we successfully designed linkers of PROTACs with high predicted cell membrane permeability while considering PROTAC likeness. Our results highlight the potential of PROTAC-TS in accelerating PROTAC development with favorable cell membrane permeability.

The effects of salmon nasal cartilage-derived proteoglycan complexes (SNC-PG) on the gut microbiota were examined by assessing changes in microbial composition, concentrations of short-chain fatty acids (SCFA), epithelial cell morphology, distal ileal gene expression levels, and fecal mucin content. Mice were assigned to one of two groups and fed either the standard AIN93G diet (control group) or the same diet supplemented with 0.04 % (w/w) SNC-PG (SNC-PG group) for 12 weeks. The SNC-PG diet increased fecal mucin content, reduced expression of inflammatory cytokine genes in the distal ileal mucosa, and high levels of acetic and butyric acid in the cecal contents compared with the control group. The relative abundance of Desulfobacterota, a bacterium that uses sulfate as an energy source, was higher in the SNCPG group than in the control group. These findings suggest that SNC-PG intake may contribute to increased SCFA levels and fecal mucin content.

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Fish oil contains not only major fatty acids with double bonds at the n-3, n-6, n-7, and n-9 positions but also those with a double bond at the n-1 position, such as 6,9,12,15-hexadecatetraenoic acid (C16:4 n-1; HDTA). However, intracellular bioconversion and metabolic fate of n-1 polyunsaturated fatty acids (PUFA) remain unclear. Therefore, in this study, we aimed to assess the intracellular bioconversion and metabolic fate of HDTA and its metabolite, 8,11,14,17- octadecatetraenoic acid (C18:4 n-1; ODTA), using HepG2 cells. Based on the results of cell viability and cytotoxicity assays for HDTA and ODTA, the concentration of each fatty acid supplemented in the experiments was set at 10 μM. HepG2 cell culture with HDTA revealed C20:4 n-1 as a new HDTA metabolite, along with previously reported ODTA. Our findings suggest that the HDTA taken up by HepG2 cells undergoes elongation to form ODTA and C20:4 n-1. Following supplementation with HDTA, ODTA, and 5,8,11,14,17-eicosapentaenoic acid (C20:5 n-3; EPA), fatty acids disappeared from the culture medium within 24 h. Notably, the total relative level of HDTA and its metabolites, including ODTA and C20:4 n-1 in HDTA- and ODTA-supplemented cells were significantly lower than the total relative level of EPA and its metabolites, including 7,10,13,16,19-docosapentaenoic acid (C22:5 n-3), C24:6 n-3, and 4,7,10,13,16,19-docosahexaenoic acid (C22:6 n-3) in the EPA-supplemented cells. Except for a portion that was intracellularly elongated, most HDTA was taken up by HepG2 cells and may undergo rapid fatty acid β-oxidation. However, RNA-sequencing and real-time polymerase chain reaction analysis revealed no significant changes in fatty acid β-oxidation-related gene expression levels in HDTA-supplemented cells. Collectively, these results provide novel insights into the intracellular bioconversion mechanisms and metabolic fate of HDTA and ODTA in HepG2 cells, suggesting that the metabolic fate of n-1 PUFA is distinct from that of common PUFA.

Fatigue is a prevalent and debilitating symptom of non-communicable diseases (NCDs); however, its biological basis are not well-defined. This exploratory study aimed to identify key biological drivers of fatigue by integrating metabolomic, microbiome, and genetic data from blood and saliva samples using a multi-omics approach. Metabolomic, microbiome, and single nucleotide polymorphisim analyses were conducted on saliva and blood samples from 52 patients with NCDs. Fatigue dimensions were assessed using the Multidimensional Fatigue Inventory and correlated with biological markers. LightGBM, a gradient boosting algorithm, was used for fatigue prediction, and model performance was evaluated using the F1-score, accuracy, and receiver operating characteristic area under the curve using leave-one-out cross-validation. Statistical analyses included correlation tests and multiple comparison adjustments (p < 0.05; false discovery rate <0.05). This study was approved by the Yokohama City University Hospital Ethics Committee (F230100022). Plasmalogen synthesis was significantly associated with physical fatigue in both blood and saliva samples. Additionally, homocysteine degradation and catecholamine biosynthesis in the blood were significantly associated with mental fatigue (Holm p < 0.05). Microbial imbalances, including reduced levels of Firmicutes negativicutes and Patescibacteria saccharimonadia, correlated with general and physical fatigue (r = − 0.379, p = 0.006). Genetic variants in genes, such as GPR180, NOTCH3, SVIL, HSD17B11, and PLXNA1, were linked to various fatigue dimensions (r range: −0.539–0.517, p < 0.05). Machine learning models based on blood and salivary biomarkers achieved an F1-score of approximately 0.7 in predicting fatigue dimensions. This study provides preliminary insights into the potential involvement of alterations in lipid metabolism, catecholamine biosynthesis disruptions, microbial imbalances, and specific genetic variants in fatigue in patients with NCDs. These findings lay the groundwork for personalized interventions, although further validation and model refinement across diverse populations are needed to enhance the prediction performance and clinical applicability.

Background: Integrins are cell surface proteins and major cell adhesion transmembrane receptors that play an important role in cell adhesion andmultifaceted roles in cell growth, survival, and cytoskeleton formation by transmitting stimuli from ligands to the cell. Integrin α6 acts as a receptor for laminin, an extracellular matrix, and is involved in growth, invasion, and resistance to therapy in many cancers. However, the role of integrin α6 and interaction with laminin in multiple myeloma (MM) have not yet been investigated. Therefore, we focused on the regulation of integrin α6 expression in MM cells. Additionally, we focused on the binding of integrin α6 to laminin and examined proliferation and signal transduction by laminin binding. Methods: ITGA6 expression was measured by RT-qPCR in CD138-positive plasma cells from 106 patients with MM, 13 patients with SMM, 72 patients with MGUS, 17 controls, and human myeloma cell lines (HMCL): KMS27, KMS11, KMS28BM, MM1S, KMS26, RPMI8226, and KMM1. The expression of integrin α6 on the HMCL cell surface was measured by flow cytometry. In HMCL, cell proliferation with laminin and molecules involved in signal transduction was evaluated using the CCK-8 assay and Western blotting. Results: ITGA6 mRNA expression was significantly higher in the MM group than in the control and MGUS groups (p <0.001). The median ITGA6 mRNA expression levels in the control, MGUS, SMM, and MM groups were 0.359, 2.033, 35.230, and 7.961, respectively. There were no differences in ITGA6 expression in MM samples according to ISS (p=0.772) and cytogenetic risk (p=0.371). The overall survival and progression free survival did not differ between high and low ITGA6 expressions (OS; 5.1 years vs not reached; p = 0.394, PFS; 2.1 vs 2.2 years; p = 0.523). To examine the regulation of integrin α6 expression by Myc, JQ1 and Myc inhibitors were added to the cells, and mean fluorescence intensity (MFI) decreased from 7.03 to 1.97, 6.15 to 2.54, and 5.71 to 2.31 in KMS27, KMS11, and KMS28BM, respectively, as JQ1 was added. MFI decreased from 4.23 to 3.08, 3.32 to 2.14, and 3.17 to 2.14 in KMS27, MM1S, and KMS28BM, respectively, as the Myc inhibitor was added. Since it was reported that integrin α6 expression is regulated by HIF-1α, a hypoxia-inducible factor, we examined whether integrin α6 expression is regulated by HIF-1α in HMCL. Comparing cells cultured under normal oxygen and 1% O2 conditions, the MFI of integrin α6 changed from 5.55 to 3.43, 1.43 to 1.51, 1.01 to 0.99, and 5.53 to 4.40 in KMS11, KMS26, RPMI8226, and KMM1, respectively. We examined changes in cell proliferation caused by the binding of integrin α6 to laminin. We used laminins 111, 211, 411, and 511, which are known ligands for integrin α6. Laminin 411 promoted the proliferation of KMS27 and KMS11 cells expressing integrin α6 compared to those without laminin (p=0.0207, p=0.0532). In HMCL with high integrin α6 expression, stimulation with laminin 411 increased the phosphorylation of Akt and Erk1/2. KMS27 increased phosphorylated Akt by 12-fold, phosphorylated Erk1/2 by 2-fold, and 6-fold; KMS11 increased phosphorylated Akt by 15-fold, phosphorylated Erk1/2 by 34-fold, and 11-fold. Conversely, in RPMI8226 with no integrin α6 expression, stimulation of laminin 411 resulted in a 0.6- and 1-fold increase in phosphorylated Akt and Erk1/2, respectively. Conclusion: RT-qPCR results suggest that integrin α6 may be involved in disease onset and progression. Cell surface expression of integrin α6 is regulated by Myc but not by HIF-1α. Stimulation of MM cells by laminin stimulated proliferation and caused phosphorylated Akt and Erk, suggesting that laminin is involved in MM proliferation via integrin α6. This study is a step toward developing integrin α6-targeted therapy.

BackgroundA rodent autism spectrum disorder (ASD) model based on prenatal exposure to valproic acid (VPA) is widely recognized as a prominent model. Social behavior in rodent ASD models has primarily been evaluated through a three-chamber approach test. However, in this study, we focused on social attention in the VPA model of ASD.MethodsIn male C57BL/6 J mice, attentional behaviors toward conspecifics were examined through reaching tasks around 9–11 weeks of age. On embryonic day 12.5, pregnant mice underwent a subcutaneous injection of 600 mg/kg VPA sodium salt dissolved in 0.9% saline solution (VPA group) or saline solution alone (Sal group) into their neck fat. Thirty-six mice—nine each in the VPA and saline groups, and 18 partners—underwent training in reaching behavior. Subsequently, we examined whether the VPA or Sal group demonstrated focused attention toward their partners during reaching tasks. A two-way analysis of variance (ANOVA) (condition [VPA/Sal] × situation [face-to-face (attention)/not paying attention (not attention)]) was conducted on the average success rate of the situation. Additionally, we measured the duration of sniffing behavior between pairs of mice in an open field twice in total at 4 and 8 weeks of age before reaching task. The pairs were constructed by pairing a VPA or Sal group mouse with its partner, with the objective of facilitating initial encounters between the mice. A one-way ANOVA was conducted on the average duration of sniffing behavior data from 4 weeks and a second one-way ANOVA on data from 8 weeks.ResultsThe analysis revealed a significant interaction between condition and situation in the reaching task [F (1, 28) = 6.75, p = 0.015, ηp2 = 0.19]. The simple main effect test exhibited that the “not paying attention” rate was significantly higher than that of the “face-to-face” in the VPA group (p < 0.01). The results revealed a not significant difference in the average duration of sniffing behavior at 4 weeks [F (3, 32) = 2.71, p = 0.06, n.s., ηp2 = 0.20], but significant difference at 8 weeks [F (3, 32) = 4.12, p < 0.05, ηp2 = 0.28]. Multiple comparisons using the Bonferroni method revealed significant differences in the sniffing duration at 8 weeks between from the partner toward the VPA mouse and from the partner toward the Sal mouse (p < 0.05).ConclusionThe VPA rodent model of ASD exhibited differences in social attention compared to the saline group. By focusing on social attention and exploring various ASD models, insights can be gained from the neural mechanisms underlying gaze abnormalities during social interaction in individuals with ASD.

Introduction: TP53 is a tumor suppressor gene located on chromosome 17p 13.1 that plays a critical role in preventing and reducing the aggressiveness of many tumor types, including multiple myeloma (MM). 17p deletions and TP53 mutations are associated with poor clinical outcomes in patients with MM. Rho GTPase signaling is involved in cancer progression, dissemination, and chemoresistance. Among the three major Rho-GTPases (RhoA, Rac1, and Cdc42) we selected Rac1 because a previous study on lymphoma demonstrated an interaction between RaC1 and p53. In this study, we evaluated the role of Rac1 in MM, to inform novel therapy development. Materials and Methods: Bone marrow plasma cells obtained with informed consent from 114 MM patients, 70 MGUS patients, and 15 controls, and purified via anti-CD138 antibody and magnetic beads were included in this study. The study was approved by Gunma University's IRB, and followed the Declaration of Helsinki guidelines. Three human myeloma cell lines (HMCLs), KMS11, KMS26, and MM.1S, whose respective TP53 statuses were deficient, mutated, and wild type, were used. KMS26 and KMS11 cells expressing DOX-inducible wild type (WT) p53 (KMS26/Tet-on p53 and KMS11/Tet-on p53, respectively) and p53 MM.1S knockdown by shp53 were used. RAC1 mRNA levels were assessed by RT-qPCR, with ACTB serving as an endogenous control, and MM.1S as a reference sample. Protein expression levels of p53, p21, Mdm2, and Rac1 were determined via western blotting. To quantify cell proliferation or induce cell death, EdU and Annexin V assays were performed. RNA-seq was performed using an Illumina Next Seq 500. Results: KMS11/Tet-on p53 cell proliferation was not affected by p53 induction, while that of KMS26/Tet-on p53 cells was significantly reduced. Gene Ontology (GO) analysis using RNA-seq data demonstrated enhanced Rho-GTPase signaling in KMS11 compared to KMS26. RAC1 mRNA levels in purified BM plasma cells were significantly higher in patients with NDMM than in controls (p < 0.01). Rac1 inhibitor 1A-116 (50 μM) significantly reduced survival rates of both KMS11 and KMS26 cells at 72 h. This effect was more prominent in KMS11 cells than in KMS26 cells. 1A-116 also significantly reduced MM.1S cell survival at 72 h with p53 induction by MDM2 inhibitor Nutlin-3 (1 µM), or p53 knockdown by Shp53, although knockdown of WT p53 alone or Nutlin-3 alone did not affect survival. In KMS11/Tet-on p53, KMS26/Tet-on p53, and MM.1S cells, cotreatment with Nutlin-3 and 1A-116 did not increase p53, p21, or Mdm2 protein expression. In MM.1S cells with p53 knockdown, treatment with 1A-116 did not increase p53 or Mdm2 protein expression, but did increase that of p21. 1A-116 treatment significantly reduced EdU incorporation in all three HMCLs, indicating that Rac1 inhibition arrests the cell cycle, but the magnitude of this effect was attenuated in KMS11 cells. Apoptotic and dead cells, defined by annexin V-positivity and 7-AAD incorporation, were significantly increased after 1A-116 treatment in KMS11 and MM.1S cells. KMS11 and KMS26 cell survival at 72 h after treatment declined when CRBN modulators lenalidomide, pomalidomide, and iberdomide were combined with the Rac1 inhibitor 1A-116 (25µM) compared with CRBN modulator treatment alone. In contrast, Rac1 inhibitor showed no additive effect on cell survival after 24 h of bortezomib treatment in HMCLs. Median overall survival (OS) time of patients with high RAC1 mRNA expression (above median value) was significantly shortened (4.3 years vs not reached; p = 0.01), although progression free survival (PFS) was not significantly different (2.0 years vs 3.1 years; p = 0.21). In multivariate analysis, ASCT and RAC1 mRNA expression were independent prognostic factors for OS (hazard ratio [HR], 0.409; p = 0.04; RAC1 mRNA high: HR, 2.211; p = 0.02). Among patients who underwent ASCT, both OS and PFS were significantly lower in those with high RAC1 mRNA expression than in those with low RAC1 expression (OS: 5.1 years vs not reached, p = 0.02; PFS: 2.7 years vs not reached, p = 0.01). Conclusions: Rac1 affects HMCL survival regardless of p53 status and Rac1 associates with CRBN modulator sensitivity. High RAC1 mRNA expression in intramedullary plasma cells of patients with NDMM is associated with worse prognosis. Our research provides new insights for development of novel therapies targeting the Rac1 pathway to improve MM patient prognosis, including patients with p53 dysfunction.

Dietary factors, particularly proteins, have been extensively explored to combat cognitive impairment. We have previously reported that dietary fish (Alaska Pollock) protein (APP) is more effective than casein (CAS) or fish oil in maintaining short-term memory in senescence-accelerated mice prone 10 (SAMP10). To examine the specificity of the protective effect of APP intakes against short-term memory decline, we assessed the impact of various dietary animal proteins, including APP, CAS, chicken breast protein (CP), and whey protein (WP), against age-related cognitive function in SAMP10 mice. After feeding the experimental diets for 5 months, memory was assessed using the Y-maze. The APP group exhibited a significant increase in spontaneous alternation behavior as an indicator of working memory when group compared with groups fed with other protein source. Additionally, the APP group displayed significantly higher neurofilament heavy chain positivity than the CAS and CP groups, as evidenced immunohistochemical analysis. Gut microbiota analysis indicated that dietary APP significantly enhanced the relative abundance of Lactobacillus, which positively correlated with spontaneous alternation behavior. Collectively, these findings suggest that dietary APP is more effective than CAS, CP, or WP in preventing age-related short-term memory decline and morphological abnormalities in the hippocampal axons of SAMP10 mice. Moreover, APP-mediated improvements in cognitive deficits may be associated with changes in microbiota diversity. PRACTICAL APPLICATION: This research suggests that dietary fish protein from Alaska Pollock may be more efficient in prevention short-term memory decline in mice, compared to other animal proteins. This finding has practical implications for nutritional optimization, developing the new health food products, and elucidating the relationship between the impact of specific proteins on gut microbiota and prevention of age-related cognitive decline.

Several studies show that genetic and environmental factors contribute to the onset and progression of neurodevelopmental disorders. Maternal immune activation (MIA) during gestation is considered one of the major environmental factors driving this process. The kynurenine pathway (KP) is a major route of the essential amino acid L-tryptophan (Trp) catabolism in mammalian cells. Activation of the KP following neuro-inflammation can generate various endogenous neuroactive metabolites that may impact brain functions and behaviors. Additionally, neurotoxic metabolites and excitotoxicity cause long-term changes in the trophic support, glutamatergic system, and synaptic function following KP activation. Therefore, investigating the role of KP metabolites during neurodevelopment will likely promote further understanding of additional pathophysiology of neurodevelopmental disorders, including autism spectrum disorder (ASD). In this review, we describe the changes in KP metabolism in the brain during pregnancy and represent how maternal inflammation and genetic factors influence the KP during development. We overview the patients with ASD clinical data and animal models designed to verify the role of perinatal KP elevation in long-lasting biochemical, neuropathological, and behavioral deficits later in life. Our review will help shed light on new therapeutic strategies and interventions targeting the KP for neurodevelopmental disorders.

Depression can be associated with chronic systemic inflammation, and production of peripheral proinflammatory cytokines and upregulation of the kynurenine pathway have been implicated in pathogenesis of depression. However, the mechanistic bases for these comorbidities are not yet well understood. As tryptophan 2,3-dioxygenase (TDO) and indoleamine 2,3-dioxygenase (IDO), which convert tryptophan to kynurenine, are rate-limiting enzymes of the kynurenine pathway, we screened TDO or IDO inhibitors for effects on the production of proinflammatory cytokines in a mouse macrophage cell line. The TDO inhibitor 680C91 attenuated LPS-induced pro-inflammatory cytokines including IL-1β and IL-6. Surprisingly, this effect was TDO-independent, as it occurred even in peritoneal macrophages from TDO knockout mice. Instead, the anti-inflammatory effects of 680C91 were mediated through the suppression of signal transducer and activator of transcription (STAT) signaling. Furthermore, 680C91 suppressed production of proinflammatory cytokines and STAT signaling in an animal model of inflammatory bowel disease. Specifically, 680C91 effectively attenuated acute phase colon cytokine responses in male mice subjected to dextran sulfate sodium (DSS)-induced colitis. Interestingly, this treatment also prevented the development of anxiodepressive-like neurobehaviors in DSS-treated mice during the recovery phase. The ability of 680C91 to prevent anxiodepressive-like behavior in response to chemically-induced colitis appeared to be due to rescue of attenuated dopamine responses in the nucleus accumbens. Thus, inhibition of STAT-mediated, but TDO-independent proinflammatory cytokines in macrophages can prevent inflammation-associated anxiety and depression. Identification of molecular mechanisms involved may facilitate the development of new treatments for gastrointestinal-neuropsychiatric comorbidity.

骨粗鬆症は特に閉経後の女性に多い骨の疾患である.これまで骨粗鬆症の治療薬は骨量の増加効果に主眼をおいて開発されているが,治療のエンドポイントである骨折リスクの軽減には骨量と骨質を改善し,骨代謝バランスを正常にする有効性の高い治療薬の開発が重要である.近年,様々な漢方製剤の骨維持の有効性が臨床では示されているが,その作用機序については明らかにされていない.本研究では,閉経後の更年期障害の治療で汎用される漢方製剤の中で,温経湯(UKT)に着目し,破骨細胞分化誘導因子であるRANKL刺激による破骨細胞分化への影響を調べ,UKTの作用メカニズムについて明らかにした.我々がスクリーニングした様々な漢方製剤の中で,UKTが最も強い破骨細胞分化阻害効果を示した.UKTは破骨細胞の初期分化に不可欠な転写因子NFATc1を阻害した.またNFATc1の上流シグナルであるNF-κBの核移行を阻害した一方で,NFATc1を阻害するBlimp1-Bcl6シグナルを活性化し,破骨細胞の分化成熟を抑制することを明らかにした.さらに我々は活性型Caspase-3によって,UKTが単核破骨細胞のアポトーシスを誘導することを示した.本研究はUKTがBlimp1-Bcl6およびNF-κBシグナル伝達経路を介して,RANKL誘導による破骨細胞分化を抑制し,単核破骨細胞の細胞死を誘導することを初めて明らかにした研究であり,UKTが閉経後骨粗鬆症の効果的な治療薬となりうる可能性を示した.本稿では,我々の研究成果について概説する.

Physiological hypoxia is critical for placental mammalian development. However, the underlying mechanisms by which hypoxia regulates embryonic development remain unclear. We discovered that the expression of glycolytic genes partially depends on hypoxia in neuroepithelial cells of E8.25 mouse embryos. Consistent with this finding, inhibiting glycolysis during the early phase of neural tube closure (E8.0-8.5) resulted in a neural tube closure defect. In contrast, inhibiting the electron transport chain did not affect neural tube formation. Furthermore, inhibiting glycolysis affected cell proliferation, but not differentiation and survival. Inhibiting glycolysis repressed the phosphorylation of myosin light chain 2, and consequent neural plate folding. Our findings revealed that anaerobic glycolysis regulates neuroepithelial cell proliferation and apical constriction during the early phase of neural tube closure.

MicroRNAs (miRNAs and miRs) are small (19–25 base pairs) non-coding RNAs with the ability to modulate gene expression. Previously, we showed that the miR-34 family is downregulated in multiple myeloma (MM) as the cancer progressed. In this study, we aimed to clarify the mechanism of miRNA dysregulation in MM. We focused particularly on the interaction between MYC and the TP53-miR34 axis because there is a discrepancy between increased TP53 and decreased miR-34 expressions in MM. Using the nutlin-3 or Tet-on systems, we caused wild-type (WT) p53 protein accumulation in human MM cell lines (HMCLs) and observed upregulated miR-34 expression. Next, we found that treatment with an Myc inhibitor alone did not affect miR-34 expression levels, but when it was coupled with p53 accumulation, miR-34 expression increased. In contrast, forced MYC activation by the MYC-ER system reduced nutlin-3-induced miR-34 expression. We also observed that TP53 and MYC were negatively correlated with mature miR-34 expressions in the plasma cells of patients with MM. Our results suggest that MYC participates in the suppression of p53-dependent miRNA expressions. Because miRNA expression suppresses tumors, its inhibition leads to MM development and malignant transformation.

Antimicrobial peptides (AMPs) have attracted attention as next-generation antimicrobial drugs. Designing AMPs while considering multiple properties, such as antimicrobial activities and toxicity, requires numerous trials and errors by chemists. In this study, we propose MODAN, a machine learning-assisted AMP design framework based on multi-objective Bayesian optimisation. The primary advantage of MODAN is its ability to handle various non-proteinogenic amino acids, which have recently shown the potential of the activity enhancement, and this flexibility has not been achieved by previous works. In addition, multi-objective Bayesian optimisation enables simultaneous improvement of antimicrobial activity and toxicity. We have succeeded in designing peptides that have potent antimicrobial and low haemolytic activities within two rounds of MODAN recommendation and experimentation, based on a strategy that chemists do not usually consider.

Tissue inhibitors of metalloproteinases (TIMPs) are endogenous matrix metalloproteinase inhibitors. TIMP1 is produced by cancer cells and has pleiotropic activities. However, its role and source in multiple myeloma (MM) are unclear. Here, we evaluated TIMP1 protein and mRNA levels in bone marrow (BM) plasma cells and assessed the effects of TIMP1 expression on fibroblast invasive capacity using three-dimensional spheroid cell invasion assays. TIMP1 mRNA and protein levels were elevated when patients progressed from monoclonal gammopathy of undetermined significance or smouldering myeloma to MM. Furthermore, TIMP1 levels decreased at complete response and TIMP1 protein levels increased with higher international staging. TIMP1 mRNA levels were markedly higher in extramedullary plasmacytoma and MM with t(4;14). Overall survival and post-progression survival were significantly lower in MM patients with high TIMP1 protein. Recombinant TIMP1 did not directly affect MM cells but enhanced the invasive capacity of fibroblasts; this effect was suppressed by treatment with anti-TIMP1 antibodies. Fibroblasts supported myeloma cell invasion and expansion in extracellular matrix. Overall, these results suggested that MM-derived TIMP1 induces the invasive phenotype in fibroblasts and is involved in disease progression. Further studies are required to elucidate the specific roles of TIMP1 in MM and facilitate the development of novel therapies targeting the TIMP1 pathway.

Septic encephalopathy (SE) is characterized by symptoms such as coma, delirium, and cognitive dysfunction, and effective therapeutic interventions for SE remain elusive. In this study, we aimed to investigate the potential alleviating effects of vagal nerve stimulation (VNS) on SE-associated signs. To evaluate our hypothesis, we utilized a mouse model of SE induced by intraperitoneal injection of lipopolysaccharide (0.3 mg per mouse) and administered noninvasive, high-frequency ultrasound VNS. To assess the efficacy of ultrasound VNS, we measured inflammation-related molecules, including the α7 nicotinic acetylcholine receptor (α7nAChR) expression in peritoneal macrophages and plasma interleukin 1β (IL-1β) levels. Consistent with our hypothesis, SE mice exhibited reduced α7nAChR expression in macrophages and elevated IL-1β levels in the blood. Remarkably, VNS in SE mice restored α7nAChR expression and IL-1β levels to those observed in control mice. Furthermore, we evaluated the effects of VNS on survival rate, body temperature, and locomotor activity. SE mice subjected to VNS demonstrated a modest, yet significant, improvement in survival rate, recovery from hypothermia, and increased locomotor activity. To investigate the impact on the brain, we examined the hippocampus of SE mice. In control mice, VNS increased the expression of c-fos, a marker of neuronal electrical excitability, in the hippocampus. In SE mice, VNS led to the restoration of aberrant firing patterns in hippocampal neurons. Additionally, proteomic analysis of hippocampal tissue in SE mice revealed abnormal increases in two proteins, tissue factor (TF) and acyl-CoA dehydrogenase family member 9 (ACAD9), which returned to control levels following VNS. Collectively, our findings support the value of exploring the beneficial effects of ultrasound VNS on SE.