Komatsu University

Although the SARS-CoV-2 pandemic has subsided with thewidespreadrollout of vaccines, emerging variants continue to pose significantpublic health challenges. While current diagnostic technologies cansensitively detect specific variants, there is an increasing needfor broad-spectrum detection methods that are less affected by viralmutations. In particular, the development of diagnostic techniquesthat are not only broadly applicable but also rapid and cost-effectiveis highly desirable for large-scale screening and long-term surveillance.In this study, the applicability of a noncompetitive fluorescencepolarization aptamer assay (NC-FPAA) was evaluated for detecting SARS-CoV-2,including both the wild-type strain and various variants. Two fluorescentlylabeled aptamers, a 77-mer (K1) and a 51-mer (M40), previously reportedto have broad affinity for spike proteins of different SARS-CoV-2variants, were used as probes. Detection experiments were conductedusing eight SARS-CoV-2 variants, and specificity was further assessedusing influenza viruses. The results demonstrate that NC-FPAA hasthe potential to serve as a diagnostic tool for the rapid, simple,and low-cost detection of multiple SARS-CoV-2 variants, thereby supportingbroad and effective viral surveillance.

The plasticity of blood mononuclear cells (MCs) and their role in vascular remodeling have been the focus of many studies; however, their in vitro differentiation efficiency remains poorly understood. Herein, we demonstrate that the inflammatory response accelerates the efficiency of MCs differentiation into endothelial-like cells through chemical cues in vitro. RT-PCR and RNA sequencing revealed that the differentiated cells exhibited upregulated pathways associated with vascular remodeling and regeneration. In contrast, MCs collected from normal blood showed a differentiation bias toward macrophages. Notably, under inflammatory conditions, primarily monocytes transitioned into the CD14++/CD16+/CD163+ subset, which contributed significantly to vascular remodeling. This transition was triggered by inflammation, as confirmed by in vitro cytokine treatment.

Rodent models are commonly used in preclinical studies on bioengineered conduits to induce nerve regeneration, but they often fail to predict human outcomes accurately. It is suspected that conventional evaluation methods, such as evoked electromyography and motor function assessments, are prone to errors and biases, which can lead to misleading conclusions. In this study, the limitations of conventional evaluation methods are highlighted by using muscle MRI to characterize pathophysiological changes in a rodent nerve injury model. Then, an alternative MRI-based evaluation method is proposed that quantified muscle cross-sectional area and T2 signal intensity, offering a non-invasive, repeatable evaluation with minimized errors and bias. Comparative analysis demonstrated the superior validity of the MRI-based evaluations. These findings suggest that MRI evaluation can enhance the validity of preclinical studies for developing nerve guide conduits and improve translational relevance to clinical outcomes.

We reconstructed in three dimensions all seminiferous tubules in the testes of 1- and 8-week-old musk shrews (Suncus murinus), a small experimental mammal belonging to the family Soricidae, order Eulipotyphla that preserves the primitive characteristics of mammalian species. Quantitative analyses revealed that the basic structure of seminiferous tubules at 1 week postpartum remained consistent into adulthood. A single dominant seminiferous tubule with extensive branching accounted for more than 90% of the length of all tubules, a feature distinct from that in mice and rats. Pachytene spermatocytes were observed in the seminiferous epithelia of most tubules at 1 week, which was markedly earlier than in mice and rats. At 8 weeks, a mosaic pattern of active and inactive areas of spermatogenesis was observed through the length of seminiferous tubules, a feature distinct from that in mice and rats. The percentage of active areas was significantly lower in tubule portions near the rete testis. The unique characteristics of seminiferous tubules in the musk shrew will provide a basis for investigating not only the phylogeny of testis functions but also the regulation of spermatogenesis and possibly the pathogenesis of azoospermia.

本研究の目的は，日本クリティカルケア看護学会せん妄ケア委員会より2020年に公開された「せん妄ケアリスト」を導入した臨床現場において，導入の目的や方法，導入による効果や課題をどのように看護師が認識しているかを明らかにすることであった．せん妄ケアリストを導入した臨床現場の看護師計8名を対象に半構造化面接を実施し，質的帰納的に分析した．分析の結果，18カテゴリーが抽出され，せん妄ケアリストの導入目的として【せん妄ケアの妥当性の検討】，導入方法として【臨床課題に応じた内容を部分的・段階的に導入】することなどが認識されていた．そして，せん妄ケアリストの導入によって，【日常生活支援が具体化・促進された】こと，【看護ケアの価値の再認識と促進につながった】ことが認識されており，日常生活を支援するという看護ケアの促進に繋がることが期待されていると考える．

High-fat diet (HFD)-induced obesity represents a significant challenge to male reproductive health, affecting approximately 13% of the global adult population. This comprehensive review synthesizes current evidence regarding mulberry (Morus alba L.) fruit extract's therapeutic potential for HFD-induced male reproductive dysfunction. Through comprehensive analysis of the peer-reviewed literature from multiple databases (PubMed, Web of Science, Scopus, and Google Scholar; 2005-2024), we evaluated mulberry extract's effects on testicular morphology, spermatogenesis, sperm parameters, and the underlying molecular mechanisms. Mechanistic studies reveal that standardized mulberry extract mediates protective effects through multiple pathways: enhanced antioxidant enzyme activities (SOD: +45%, Catalase: +38%, GPx: +35%), reduced inflammatory markers (TNF-α: -64%, IL-6: -58%), and modulated NF-κB signaling (-42.3%). These effects are facilitated by mulberry's rich phytochemical profile, particularly anthocyanins (2.92-5.35 mg/g dry weight) and polyphenols (4.23-6.38 mg/g). The extract demonstrates particular efficacy in preserving seminiferous tubule integrity and maintaining blood-testis barrier function, with treated groups maintaining up to 85% of normal tubular architecture compared to HFD controls. Key molecular mechanisms include AMPK/SIRT1 pathway activation (2.3-fold increase), enhanced mitochondrial function (67% increase in mtDNA copy number), and epigenetic regulation of metabolic pathways. Temporal analysis indicates optimal therapeutic effects after 28 days of treatment, with initial improvements observable within 14 days. While current evidence is promising, limitations include predominant reliance on rodent models and lack of standardized extraction protocols. Future research priorities include well-designed human clinical trials, standardization of preparation methods, and investigation of potential synergistic effects with other therapeutic agents. This comprehensive review indicates that mulberry extract is a promising therapeutic candidate for obesity-related male infertility, warranting further clinical investigation.

Ground reaction force (GRF) analysis is critical to evaluate gait and running performances. However, installing force plates on the ground surface is costly, and their transportation is challenging owing to weight constraints and restricted measurement areas. This study proposes the method by Extended Kalman filter (EKF) to estimate the three-axis force components in the GRF, utilizing vertical force data from insole sensors, angular velocity, and acceleration data from inertial measurement units (IMUs). Shear forces were calculated using the acceleration in global coordinates, with the equivalent mass calculated by the vertical force and acceleration. The force components were then compensated for sensitivity and interference using a matrix. Moreover, the method was extended to include the center of pressure and unit position vector. The experimental protocol comprised two phases: calibration and measurement, conducted on force plates installed on the ground surface. The error compensation for force components was based on actual gait motion data and the results were compared with the force plates' outputs. The findings confirmed the effectiveness of this method. This method will be expanded to include motion measurements during long-distance walking and outdoor environments.

Loss-of-function mutations in the GIRDIN/CCDC88A gene cause developmental epileptic encephalopathy (DEE) in humans. However, its pathogenesis is largely unknown. Global knockout mice of the corresponding orthologous gene (gKOs) have a preweaning lethal phenotype with growth failure, preventing longitudinal analysis. We aimed to overcome this lethality and elucidate DEE pathogenesis. We developed a novel lifelong feeding regimen (NLFR), which consists of providing mash food from postnatal day 14 (P14) until weaning (P28), followed by agar-bound food exclusively after weaning. Videography, electroencephalography (EEG), and histological analyses were performed. Conditional Girdin/Ccdc88a knockout mice (cKOs) of variable lineages (Nestin, Emx1, or Nkx2-1) were generated to identify the region responsible for epilepsy. Under the NLFR, gKOs survived beyond 1 year and displayed fully penetrant, robust epileptic phenotypes, including early-onset (P22.3 in average) generalized tonic-clonic seizures (GTCSs) (averaging eight per day), which were completely synchronized with fast rhythms on EEG, frequent interictal electroencephalographic spikes (averaging 430 per hour), and progressive deformation of visceral organs. In addition, gKOs had absence seizures, which were not always time-locked to frequent spike waves on EEG. The frequent GTCSs and interictal spikes in gKOs were suppressed by known antiepileptic drugs. Histologically, bilateral hippocampi in gKOs exhibited congenital cornu-ammonis splitting, granule cell dispersion, and astrogliosis. Furthermore, analysis of conditional knockouts using multiple Cre-deleters identified a defect in the delivery of interneuron precursors from the medial ganglionic eminence into the hippocampal primordium during embryogenesis as a major cause of epileptogenesis. These findings give rise to a new approach of lifelong caregiving to overcome the problem of preweaning lethality in animal models. We propose a useful model for studying DEE with hippocampal sclerosis and interneuronopathy. gKOs with NLFR combine the contradictory properties of robust epileptic phenotypes and long-term survivability, which can be used to investigate spontaneous epileptic wave propagation and therapeutic intervention in hippocampal sclerosis.

Although the stereoscopic effect in 3D virtual reality (VR) space has been studied, its influence on motor performance, specifically how stereoscopic objects affect behavioral outcomes like reaching, remains unclear. Previous research has primarily focused on perceptual aspects rather than on how stereoscopic visual input impacts motor control at the behavioral level. Thus, we investigated the effects of stereoscopic objects in a VR environment on reaching performance, examining whether the stereoscopic effect of objects is a significant aspect enough to affect performance at the behavioral level. While doing so, we investigated young and older adults separately, as age is a critical factor influencing motor performance. Fourteen young and 23 older participants performed a reaching task in the VR space. The target objects were 2D and 3D, deviating from the initial position by 10 and 20 cm. The movement attributed to feedforward control was analyzed using end-point error and smoothness of movement. Our results revealed that older participants had significantly worse predictive control than young participants in the 3D long task, suggesting that the positions of 3D objects were more difficult to predict than those of 2D objects. Young participants showed a significant difference in smoothness between 2D and 3D objects, which was not observed in older participants. This may reflect the floor effect in older participants. Under the short-distance condition, neither group showed a significant difference, suggesting the ceiling effect by distance. We confirmed that the effect of stereoscopic objects was not drastic but it did hamper the reaching performance.

Intrahepatic cholangiocarcinoma (iCCA) exhibits different blood imaging features and prognosis depending on histology. To clarity histopathological growth patterns (HGPs) and vascularization processes of iCCA, we collected 145 surgical specimens and histologically classified them into large bile duct (LBD) (20 cases), small bile duct (SBD) (54), cholangiolocarcinoma (CLC) (35), combined SBD–CLC (cSBD–CLC) (26), and ductal plate malformation (DPM) (10) (sub)types. According to the invasive pattern at the interface between tumor and adjacent background liver, HGPs were classified into desmoplastic, pushing, and replacing HGPs. Desmoplastic HGP predominated in LBD type (55.5%), while replacing HGP was common in CLC (82.9%) and cSBD–CLC (84.6%) subtypes. Desmoplastic HGP reflected angiogenesis, while replacing HGP showed vessel co-option in addition to angiogenesis. By evaluating microvessel density (MVD) using vascular markers, ELTD1 identified vessel co-option and angiogenesis, and ELTD1-positive MVD at invasive margin in replacing HGP was significantly higher than those in desmoplastic and pushing HGPs. REDD1, an angiogenesis-related marker, demonstrated preferably higher MVD in the tumor center than in other areas. iCCA (sub)types and HGPs were closely related to vessel co-option and immune-related factors (lymphatic vessels, lymphocytes, and neutrophils). In conclusion, HGPs and vascular mechanisms characterize iCCA (sub)types and vessel co-option linked to the immune microenvironment.