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Yoshinori Sato,1 Nami Hatayama,1 Tsuneyuki Ubagai,1 Shigeru Tansho-Nagakawa,1 Yasuo Ono,1,2 Yusuke Yoshino1 1Department of Microbiology and Immunology, Teikyo University School of Medicine, Itabashi-ku, Tokyo, 173-8605, Japan; 2Teikyo Heisei University, Faculty of Health and Medical Science, Toshima-ku, Tokyo, 170-8445, JapanCorrespondence: Yoshinori Sato, Department of Microbiology and Immunology, Teikyo University School of Medicine, 2-11-1 Kaga, Itabashi-ku, Tokyo, 173-8605, Japan, Tel +81-3-3964-1211, Fax +81-3-5375-5284, Email satoy@med.teikyo-u.ac.jpPurpose: To determine the ability of human neutrophils to kill multidrug-resistant Acinetobacter baumannii (MDRAB) in the presence of tigecycline (TGC).Methods: Clinical isolates of MDRAB were cultured with human neutrophils and H2O2 in the presence of TGC. The numbers of viable bacteria, catalase activity, gene expression at the K locus of the MDRAB, reactive oxygen species (ROS) production, and granule exocytosis in human neutrophils were determined.Results: There was a time-dependent increase in the numbers of MDRAB after co-culturing with human neutrophils, whereas there was a significant decrease in the MDRAB numbers when co-cultured with both, human neutrophils and TGC for 6 h. The presence or absence of TGC did not affect total ROS production or the expression of CD11b, CD15, and CD63 on human neutrophils occurred when co-cultured with MDRAB. TGC significantly suppressed catalase activity and gene expression at the K locus of MDRAB, and significantly reduced the thickness of the capsule. Additionally, the bacterial viability of TGC-treated MDRAB cultured with H2O2 was lower than that without H2O2 after 6 h of culture.Conclusion: TGC significantly suppressed the expression of catalase and the capsule in MDRAB without adverse effects on neutrophil function, allowing human neutrophils to kill MDRAB. TGC is an effective antibiotic for treating MDRAB infections.Keywords: multidrug-resistant Acinetobacter baumannii, MDRAB, catalase, capsule, tigecycline, TGC, human neutrophils

Background: Recently, the number of patients with metabolic dysfunction-associated steatotic liver disease (MASLD) and its more advanced condition, metabolic dysfunction-associated steatohepatitis (MASH), has been increasing. These patients are at a higher risk of cardiovascular events and thromboembolism. However, the direct impact of high-fat diet (HFD), a cause of MASLD, on liver coagulation function is not well understood. Previously, we demonstrated that a short-term, 4-day intake of a HFD exacerbates concanavalin A (Con A)-induced acute liver injury in mice by promoting coagulation and inflammation. This model demonstrates that the liver exposed to a short-term HFD is vulnerable even before disease onset. In this study, using this model, we elucidated the detailed mechanisms by which short-term HFD intake promotes coagulation, considering primary and secondary hemostasis., Methods: C57BL/6 mice normally fed a normal diet (ND) were subjected to a HFD for 4 days. Liver tissue and blood samples were collected before and 4 and 24 h after Con A administration. Histological analysis, flow cytometry for platelet analysis, and blood coagulation tests related to secondary hemostasis were performed., Results: Even with short-term consumption of a HFD alone, platelet and fibrinogen levels increased in the peripheral blood and liver. Additionally, when Con A was administered to mice on a short-term HFD, an increase in P-selectin expression was observed in the liver, with no upregulation in peripheral blood platelets. Furthermore, in mice subjected to a short-term HFD and treated with Con A, prolonged prothrombin time (PT) and activated partial thromboplastin time (APTT) were observed., Conclusions: Consuming a HFD in short-term can enhance primary and secondary hemostasis, thereby increasing the risk of thrombosis. These conditions are presumed to be a risk factor that exacerbates Con A-induced liver injury. The findings provide insight into early intervention strategies for chronic liver diseases, such as MASLD and MASH., Competing Interests: Declarations. Ethics approval and consent to participate: All procedures involving animals in this study were conducted in accordance with protocols approved by the Aichi Medical University Animal Care and Use Committee (approval number 2023–53). Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Rapid and accurate identification of bacterial species is essential for the effective treatment of infectious diseases and suppression of antibiotic-resistant strains. The unique autofluorescence properties of bacterial cells are exploited for rapid and cost-effective identification that is suitable for point-of-care applications. Fluorescence spectroscopy is combined with machine learning to improve the diagnostic accuracy. Good training data for machine learning can be obtained to achieve the same diagnostic accuracy for bacterial species as when each wavelength is measured in detail over a broad spectral width. Experiments were performed testing 14 bacterial strains. The excitation-emission matrix was analyzed, and Bayesian optimization was used to identify the most effective combinations of wavelengths. The results showed that fluorescence spectra using three specific excitation light regions or excitation spectra using two broad fluorescence detection regions could be used as supervised data to realize diagnostic accuracy comparable to that obtained with more complex instruments., (© 2024 Wiley‐VCH GmbH.)

We investigated the effects of Clostridioides difficile toxin B (TcdB), a major virulence factor in C. difficile infection (CDI), on human neutrophils. TcdB inhibits neutrophil migration via loss of polarity of F-actin polymerization in response to interleukin-8. TcdB facilitates CDI by allowing C. difficile to avert the host immune system., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

A 44-year-old woman with a subacute onset of an altered mental status, urinary retention, and fluctuating blood pressure was initially diagnosed with anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis, meeting the criteria of Graus et al. Cardiac arrest occurred, which required pacemaker placement. She subsequently showed profound flaccid limb paralysis, with magnetic resonance imaging demonstrating focal necrotic lesions localized in the anterior horn of the longitudinal segments of the spinal cord and in the pontine tegmentum. Enteroviruses or autoimmune encephalitis-associated autoantibodies were not detected. We herein report a case of acute flaccid myelitis with profound psychiatric symptoms and dysautonomia, resembling NMDAR encephalitis.

Staphylococcus pseudintermedius has been isolated from dogs, cats, and horses and is also known as an emergent zoonotic agent. We administered orbifloxacin, a fluoroquinolone, to treat bacterial infections of cutaneous wounds caused by excessive grooming of the skin in contact with the subcutaneous port of the subcutaneous ureteral bypass (SUB) system in a cat. However, after 80 days of treatment, a severe abscess was observed in the wound and fluoroquinolone-resistant S. pseudintermedius was isolated from the abscess. The isolate was identified as a novel sequence type (ST) 2660 and contained genes for leukocidins (lukS and lukF), exfoliative toxin (siet), and biofilm regulation (icaA and icaD). The isolate was resistant to macrolide, lincosamide, fluoroquinolone, and tetracycline classes. In addition, the isolate had strong biofilm-forming ability which significantly increased with culturing at 39 °C compared with that at 37 °C, suggesting that the isolate prefers a cats' body temperature as the optimal biofilm growth condition. Notably, the biofilms were increased in the presence of doxycycline with culturing at 39 °C. This study is the first report in Japan on the new sequence type of S. pseudintermedius isolated from a companion animal and clarifies the distinctive virulence of S. pseudintermedius., (© 2024. The Author(s).)

We established a compact machine perfusion system for whole blood perfusion of rat liver by making use of oxygenation filters as an artificial lung. Livers removed from rats were divided into Krebs-Henseleit (control), 50% blood (hemoglobin: 7 g/dL), and whole blood (hemoglobin: 14 g/dL) groups, then perfused (total perfusate volume: 25 ml) with a small oxygenation filter at 37 °C for 120 min. Blood or perfusate was collected over time, and blood gas and blood cell were measured. In addition, bile volume and portal venous pressure measurements were taken. In all groups, the partial pressure of oxygen was controlled to approximately 400 mmHg. Flow rates were maintained at approximately about 20-30 ml/min according to liver size. Portal venous pressure was normal in the 50% blood and whole blood groups, while lower than the reference value in the Krebs-Henseleit group. Twice as much bile was produced in the 50% blood and whole blood groups relative with the Krebs-Henseleit group. We observed no differences in hemoglobin and red blood cell levels. Lactate levels were normal in the 50% blood and whole blood groups, but were elevated in the Krebs-Henseleit group. Our compact perfusion system using oxygenation filters was able to maintain rat liver function by perfusing a small amount of extracorporeal blood. This system is simple and stable, and may contribute to the future development of machine perfusion systems., (© 2024. The Author(s), under exclusive licence to the Japanese Society for Artificial Organs.)

Cardiovascular diseases, particularly ischemic heart disease, area leading cause of morbidity and mortality worldwide. Myocardial infarction (MI) results in extensive cardiomyocyte loss, inflammation, extracellular matrix (ECM) degradation, fibrosis, and ultimately, adverse ventricular remodeling associated with impaired heart function. While heart transplantation is the only definitive treatment for end-stage heart failure, donor organ scarcity necessitates the development of alternative therapies. In such cases, methods to promote endogenous tissue regeneration by stimulating growth factor secretion and vascular formation alone are insufficient. Techniques for the creation and transplantation of viable tissues are therefore highly sought after. Approaches to cardiac regeneration range from stem cell injections to epicardial patches and interposition grafts. While numerous preclinical trials have demonstrated the positive effects of tissue transplantation on vasculogenesis and functional recovery, long-term graft survival in large animal models is rare. Adequate vascularization is essential for the survival of transplanted tissues, yet pre-formed microvasculature often fails to achieve sufficient engraftment. Recent studies report success in enhancing cell survival rates in vitro via tissue perfusion. However, the transition of these techniques to in vivo models remains challenging, especially in large animals. This review aims to highlight the evolution of cardiac patch and stem cell therapies for the treatment of cardiovascular disease, identify discrepancies between in vitro and in vivo studies, and discuss critical factors for establishing effective myocardial tissue regeneration in vivo.

Myelin oligodendrocyte glycoprotein antibody-associated disease (MOGAD) has highly heterogeneous clinical presentations, in which encephalitis is an important phenotype. Moreover, MOGAD has been reported to exhibit diverse imaging findings. However, there have been no previous reports of cases with perivascular radial gadolinium enhancement in periventricular regions, commonly reported in autoimmune glial fibrillary acidic protein (GFAP) astrocytopathy. In this paper, we present two cases of MOGAD with this MRI feature, both of which underwent brain biopsy for the lesions. Brain biopsies revealed perivenous demyelination and inflammation consistent with acute disseminated encephalomyelitis (ADEM), with pronounced axonal damage in Case 1 and minimal axonal involvement in Case 2. Case 1 exhibited more severe cerebral atrophy than Case 2, correlating with the extent of axonal damage. Through these cases, we highlight the heterogeneity of radiological manifestations of MOGAD, expanding the spectrum beyond previously defined MRI patterns. Furthermore, histopathological analysis revealed distinct axonal involvement as a potential prognostic marker of brain atrophy. These observations emphasize the importance of considering MOGAD in the differential diagnosis, even in cases with atypical imaging findings, and highlight the significance of brain biopsy in guiding both diagnosis and prognosis., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Shimizu, Beck, Murayama, Hoshi, Sumikura, Higashida, Fukasaka, Shimada, Nagashima, Fujioka, Hatayama, Okuno, Mochizuki and Sakaguchi.)

Rapid identification of bacterial species in patient samples is essential for the treatment of infectious diseases and the economics of health care. In this study, we investigated an algorithm to improve the accuracy of bacterial species identification with fluorescence spectroscopy based on autofluorescence from bacteria, and excitation wavelengths suitable for identification. The diagnostic accuracy of each algorithm for ten bacterial species was verified in a machine learning classifier algorithm. The three machine learning algorithms with the highest diagnostic accuracy, extra tree (ET), logistic regression (LR), and multilayer perceptron (MLP), were used to determine the number and wavelength of excitation wavelengths suitable for the diagnosis of bacterial species. The key excitation wavelengths for the diagnosis of bacterial species were 280 nm, 300 nm, 380 nm, and 480 nm, with 280 nm being the most important. The median diagnostic accuracy was equivalent to that of 200 excitation wavelengths when two excitation wavelengths were used for ET and LR, and three excitation wavelengths for MLP. These results demonstrate that there is an optimum wavelength range of excitation wavelengths required for spectroscopic measurement of bacterial autofluorescence for bacterial species identification, and that measurement of only a few wavelengths in this range is sufficient to achieve sufficient accuracy for diagnosis of bacterial species., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

The shoulder joint has a high degree of freedom and an extremely complex and unstable kinematic mechanism. Coordinated contraction of the rotator cuff muscles that stop around the humeral head and the deltoid muscles and the extensibility of soft tissues, such as the joint capsule, labrum, and ligaments, contribute to shoulder-joint stability. Understanding the mechanics of shoulder-joint movement, including soft-tissue characteristics, is important for disease prevention and the development of a device for disease treatment. This study aimed to create a musculoskeletal shoulder model to represent the realistic behavior of joint movement and soft-tissue deformation as a dynamic simulation using a rigid-body model for bones and a soft-body model for soft tissues via a spring-damper-mass system. To reproduce the muscle-contraction properties of organisms, we used a muscle-expansion representation and Hill's mechanical muscle model. Shoulder motion, including the movement of the center of rotation in joints, was reproduced, and the strain in the joint capsule during dynamic shoulder movement was quantified. Furthermore, we investigated narrowing of the acromiohumeral distance in several situations to induce tissue damage due to rotator cuff impingement at the anterior-subacromial border during shoulder abduction. Given that the model can analyze exercises under disease conditions, such as muscle and tendon injuries and impingement syndrome, the proposed model is expected to help elucidate disease mechanisms and develop treatment guidelines., (© 2024. The Author(s), under exclusive licence to Japanese Association of Anatomists.)

Multidrug-resistant Acinetobacter baumannii (MDRAB) is an important pathogen that causes nosocomial infections and is resistant to almost all antibiotics, including carbapenems. Cefiderocol is a novel siderophore cephalosporin active against a broad spectrum of gram-negative bacteria. However, the susceptibility of MDRAB to cefiderocol has not yet been reported in Japan. In this study, we measured the minimum inhibitory concentrations (MICs) of antibiotics including cefiderocol against MDRAB clinical isolates collected during a nosocomial outbreak between 2009 and 2010 at the Teikyo University Hospital in Japan. We found that all 10 MDRAB clinical isolates tested were susceptible to cefiderocol, with an MIC range of 0.12 to 1 μg/mL. All the isolates also exhibited resistance to ampicillin-sulbactam and an intermediate resistance to colistin, whereas nine of them were susceptible to tigecycline. DNA sequencing revealed that all strains harbored an OXA-51-like carbapenemase, a major cause of carbapenem resistance in A. baumannii in Japan. In conclusion, this study showed that the cefiderocol susceptibility of MDRAB clinical isolates in Japan was equivalent to that to colistin or tigecycline, and thus cefiderocol is a potential treatment option for MDRAB infections.

Background: Sprengel's deformity is a congenital abnormality of the shoulder girdle. Because scapular retraction, such as the Green procedure, is usually performed during childhood to improve esthetics and shoulder function, Sprengel's deformity is rarely found in older patients., Case Presentation: We presented a unique case of a Japanese female cadaver with Sprengel's deformity at the age of 80 years. Anatomical dissection and radiological imaging revealed musculoskeletal anomalies associated with Sprengel's deformity, including Klippel-Feil syndrome, presence of an omovertebral bone, and absence of the trapezius muscle. In addition, bilateral cervical ribs were in contact with the brachial plexus. These anomalies may lead to numbness, pain, and limited range of motion of the neck and upper girdle with aging., Conclusions: Because most adult patients with Sprengel's deformity experience neck pain and limited movement of the shoulder, the presented case is a rare case of neglected Sprengel's deformity in an 80-year-old cadaver., (© 2024. The Author(s).)

Peritoneal membrane dysfunction in peritoneal dialysis (PD) is primarily attributed to angiogenesis; however, the integrity of vascular endothelial cells can affect peritoneal permeability. Hyaluronan, a component of the endothelial glycocalyx, is reportedly involved in preventing proteinuria in the normal glomerulus. One hypothesis suggests that development of encapsulating peritoneal sclerosis (EPS) is triggered by protein leakage due to vascular endothelial injury. We therefore investigated the effect of hyaluronan in the glycocalyx on peritoneal permeability and disease conditions. After hyaluronidase-mediated degradation of hyaluronan on the endothelial cells of mice, macromolecules, including albumin and β2 microglobulin, leaked into the dialysate. However, peritoneal transport of small solute molecules was not affected. Pathologically, hyaluronan expression was diminished; however, expression of vascular endothelial cadherin and heparan sulfate, a core protein of the glycocalyx, was preserved. Hyaluronan expression on endothelial cells was studied using 254 human peritoneal membrane samples. Hyaluronan expression decreased in patients undergoing long-term PD treatment and EPS patients treated with conventional solutions. Furthermore, the extent of hyaluronan loss correlated with the severity of vasculopathy. Hyaluronan on endothelial cells is involved in the peritoneal transport of macromolecules. Treatment strategies that preserve hyaluronan in the glycocalyx could prevent the leakage of macromolecules and subsequent related complications., (© 2024. The Author(s).)

Objective: Acute limb ischemia (ALI) is a rapid decrease in lower limb blood flow due to acute occlusion of peripheral arteries or bypass grafts. This study aimed to establish an ALI model using microsized gelatin beads and to investigate the pathophysiological conditions., Methods: Male Sprague-Dawley rats were anesthetized, and a low or high dose of microsized gelatin beads was administered into the left femoral artery on days 0 and 7. A control, that is, normal saline (NS) group in which NS was administered in the left femoral artery, a femoral artery cut (FAC) group in which the left femoral artery was cut, and a sciatic nerve cut (SNC) group in which the left sciatic nerve was cut were prepared. After 21 days, the temperature changes and the muscle weights in the lower limbs were measured. To assess nerve damage, the L1-6 sympathetic ganglia were immunostained with activating transcription factor 3 (ATF3) antibody., Results: In the Low-dose, High-dose, and FAC groups, a decrease in temperature was predominantly observed in the left limb. In the High-dose and SNC groups, the weight of the soleus muscle and extensor digitorum longus in the left limb decreased; however, no weight changes were observed in the Low-dose and FAC groups. Conversely, the weight of the gastrocnemius muscle significantly decreased in the Low-dose, High-dose, FAC, and SNC groups. In the High-dose and SNC groups, the number of ATF3-positive cells in the sympathetic ganglia significantly increased, and in the Low-dose, a small number of ATF3-positive cells were observed. However, ATF3-positive cells were rarely observed in the FAC and NS groups., Conclusion: We established an ALI rat model using microsized gelatin beads. The results of this study suggest that autonomic neuropathy in ALI is related to both muscle damage and peripheral neuropathy., Competing Interests: Declaration of conflicting interestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Objectives: This study aimed to determine the inhibitory and bactericidal effects of teicoplanin (TEC) on TEC-susceptible Staphylococcus haemolyticus isolated from a patient with cancer in whom infection persisted despite TEC therapy. We also focused on the biofilm-forming ability of the isolate in vitro., Methods: S. haemolyticus clinical isolate (strain 1369A) and its control strain, ATCC 29970 were cultured in Luria-Bertani (LB) broth with TEC. The inhibitory and bactericidal effects of TEC on planktonic, adherent, biofilm-dispersed, and biofilm-embedded cells of these strains were analyzed by using a biofilm formation/viability assay kit. The expression of biofilm-related genes was measured using quantitative real-time polymerase chain reaction (qRT-PCR). Biofilm formation was determined by using scanning electron microscopy (SEM)., Results: The clinical isolate of S. haemolyticus had enhanced ability to bacterial growth, adherence, aggregation, and biofilm formation, thus the inhibitory and bactericidal effects of TEC on planktonic, adherent, biofilm-dispersed, and biofilm-embedded cells of the isolate were attenuated. Additionally, TEC induced cell aggregation, biofilm formation, and some biofilm-related gene expression of the isolate., Conclusion: The clinical isolate of S. haemolyticus is resistant to TEC treatment due to cell aggregation and biofilm formation., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Yoshinori Sato reports financial support was provided by JSPS KAKENHI. Yoshinori Sato reports financial support was provided by ACRO Incubation Grants of Teikyo University., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Background: Endothelial cells are known to grow on the luminal surface of arteriovenous grafts (AVGs) used in hemodialysis. Although endothelial cells are important for preventing infection, a detailed growth of endothelial cells in AVGs is unknown. This study sought to create a simpler animal model of AVGs and to investigate how endothelial cells form on the luminal surface., Methods: Polyethylene grafts were placed between the cervical artery and vein of Wistar rats. The grafts were removed at 6 h, 24 h, 3 days, or 7 days after placement. The luminal surface was observed under optical and polarizing microscopy and stained with endothelial cell markers (LEL, CD31), the progenitor cell marker CD34, and the macrophage marker ED-1., Results: Microscopy demonstrated many diffuse vascular endothelial cells on the luminal surface of AVGs after placement. While there was no difference in the number of LEL-positive cells between the arterial side (AS) and venous side (VS) at 6 h or 7 days, there were significantly more of these cells on the VS at both 24 h and 3 days (p < 0.05). Analysis at 24 h showed some CD31-positive cells and few CD34-positive cells., Conclusions: This was the first study to use a simple rat model of AVG placement. Endothelial cell formation was initially more active on the VS than on the AS, but these cells subsequently increased in number across the luminal surface. Future clinical studies might contribute clinically by confirming whether AS versus VS puncture results in different infection rates., (© 2023. The Author(s), under exclusive licence to The Japanese Society of Nephrology.)

Arteries receive vascular branches (VBs) from peripheral nerves. VBs are thought to be involved in arterial constriction. Although the anterior tibial artery (ATA) receives VBs, information on their branching patterns and distribution areas remains limited. The aim of this study was to investigate the anatomical structures of the VBs reaching the ATA. Forty cadaver limbs were examined to assess the branching patterns and distribution areas of the VBs reaching the ATA. The VBs reaching the ATA ramified from the deep fibular nerve (DFN), and the ATA received two or three VBs in each limb. The following mean distances from the head of fibula to the points at which the VBs reached the ATA were measured: all the VBs, 1st VB, 2nd VB and 3rd VB. The measurements were 51.5 ± 23.2 mm, 33.3 ± 3.7 mm, 53.3 ± 18.6 mm, and 72.2 ± 24.5 mm, respectively. In all limbs, the DFN and the ATA converged after the DFN branched into the 1st VB. The 2nd VB in 38 of 40 limbs and the 3rd VB in 20 of 32 limbs were distributed in the ATA proximal to the convergence point of the ATA and the DFN. These findings revealed that all VBs reaching the ATA ramified from the DFN in all limbs. The ATA received two or three VBs, and all the 1st VBs distributed to the ATA proximal to the convergence point., (© 2022 American Association for Clinical Anatomists and the British Association for Clinical Anatomists.)

Objectives: High-fat diet (HFD) and high-carbohydrate diet (HCD) are strongly linked to nonalcoholic fatty liver disease, a hepatic manifestation of metabolic syndrome. The mechanism of pathologic progression from nonalcoholic fatty liver disease to nonalcoholic steatohepatitis, which is a more severe form associated with inflammation and fibrosis, remains poorly understood. Thus, the aim of this study was to investigate and compare the inflammatory and coagulative state of the liver in short-term HFD- or HCD-fed mice with acute liver injury induced by concanavalin A (Con A)., Methods: Histopathologic evaluation, real-time polymerase chain reaction, and immunohistochemical evaluation were performed on the liver of mice fed HFDs and HCDs for 4 d before and after Con A administration., Results: The liver of the HFD-fed mice had larger fibrinogen/fibrin depositions than those fed the HCD. HCD induced the expression of the proinflammatory cytokine tumor necrosis factor-α in the liver. Moreover, the expression of proinflammatory cytokines and chemokines was further enhanced after Con A stimulation in HCD (e.g., interleukin-1α, interleukin-6 at 1 h), with a strong tendency for inflammatory cell infiltration also found (24 h)., Conclusions: Short-term HCD and HFD increased susceptibility to liver injury. HCD tended to induce more intense inflammation, whereas HFD tended to induce more intense hypercoagulation, suggesting that HCD and HFD may have different mechanisms of pathologic progression to nonalcoholic steatohepatitis., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Purpose: To compare the decompressive effect around the optic nerve canal among 3 different decompression procedures (medial, balanced, and inferomedial) using 3D printed models., Methods: In this experimental study, based on data obtained from 9 patients (18 sides) with dysthyroid optic neuropathy, a preoperative control model and 3 plaster decompression models were created using a 3D printer (total, 72 sides of 36 models). A pressure sensor was placed at the optic foramen, and the orbital space was filled with silicone. The surface of the silicone was pushed down directly, and changes in pressure were recorded at 2-mm increments of pushing., Results: At 10 mm of pushing, there was significantly lower pressure in the medial (19,782.2 ± 4319.9 Pa, P = 0.001), balanced (19,448.3 ± 3767.4 Pa, P = 0.003), and inferomedial (15,855.8 ± 4000.7 Pa, P < 0.001) decompression models than in the control model (25,217.8 ± 6087.5 Pa). Overall, the statistical results for each 2-mm push were similar among the models up to 10 mm of pushing (P < 0.050). At each push, inferomedial decompression caused the greatest reduction in pressure (P < 0.050), whereas there was no significant difference in pressure between the medial and balanced decompression models (P > 0.050)., Conclusion: All 3 commonly performed decompression procedures significantly reduced retrobulbar pressure. Because inferomedial decompression models obtained the greatest reduction in pressure on the optic nerve canal, inferomedial decompression should be considered the most reliable procedure for rescuing vision in dysthyroid optic neuropathy., (© 2022. The Author(s).)

Background: Tibialis anterior (TA) muscle is the largest dorsiflexor of the ankle joint and plays an important role during gait movement. However, descriptions of the TA attachment site are inconsistent even among major anatomy textbooks, and its origin, especially the attachment site for the tibia, has not been reported in detail. This study is the first experimental attempt to investigate the origin of the TA in detail, paying particular attention to the relationship with the shape of the tibia, including sex differences., Methods: Forty legs (20 males, 20 females) from twenty Japanese cadavers were examined. Gross anatomical examination of the TA's attachment site to the tibia and the tibia's shape was performed., Results: The location of the distal end of the TA's attachment on tibia was significantly more distal in males than in females (p < 0.01). The anterior border of the tibia had a gentle S-like curve, with a medially convex curve in the proximal region and a laterally convex curve in the distal region in frontal plane. The most protruding point of the distal curve of the anterior border located significantly more proximal in females than in males (p = 0.02)., Conclusions: There were sex differences in the distal end of the attachment site on tibia of the TA and the shape of the tibia. Consequently, the variations in the attachment site of TA were considered to provide for differences in function of TA. In males, the TA may enable advantageous power exertion, whereas in females it may work efficiently for dorsiflexion of ankle, respectively. Sex differences in TA's attachment site and the shape of the tibia may be involved in gait movement as well as frequency of lower leg disorders such as chronic exertional compartment syndrome., (© 2022. The Author(s).)

Formaldehyde has been traditionally used for embalming human cadavers for gross anatomy education and surgical skills training. However, exposure to formaldehyde negatively affects human health. This study aimed to assess the efficacy of reperfusing urea solution to a formalin-embalmed cadaver for surgical skills training and then investigate the cadaver's tissue elasticity alteration after being soaked into the urea solution. Twelve surgeons evaluated the similarity of tissue characteristics between the cadaver (embalmed by formalin solution and reperfused by urea solution) and a living human body. Furthermore, the tissue formaldehyde content and mechanical properties of the formalin-fixated femoral skin and artery specimens with or without soaking into urea solution were measured. Results showed that the tactile assessment, skin incision, vessel ligation and suture, and decollement were better and more useful in the cadaver reperfused by urea solution than in the cadaver merely fixated by formalin solution. In the urea-reperfused cadaver, the volatilized, or tissue formaldehyde levels declined. The stiffness and Young's modulus of the femoral skin and artery were also lower in the specimen than in the mere formalin-fixated specimen. In conclusion, reperfusion of urea solution to the formalin-fixated cadaver makes anatomical education and surgical skills training more efficient with fewer requirements for cadaver management., (© 2022. The Author(s), under exclusive licence to Japanese Association of Anatomists.)

Objective: Anorectal transplantation is a challenging procedure but a promising option for patients with weakened or completely absent anorectal function., Summary Background Data: We constructed a canine model of anorectal transplantation, evaluated the long-term outcomes, and controlled rejection and infection in allotransplantation., Methods: In the pudendal nerve function study, 6 dogs were randomly divided into 2 groups, transection and anastomosis, and were compared with a control using anorectal manometry, electromyography, and histological examination. In the anorectal transplantation model, 4 dogs were assigned to 4 groups: autotransplant, allotransplant with immunosuppression, allotransplant without immunosuppression, and normal control. Long-term function was evaluated by defecography, videography, and histological examination., Results: In the pudendal nerve function study, anorectal manometry indicated that the anastomosis group recovered partial function 6 months postoperatively. Microscopically, the pudendal nerve and the sphincter muscle regenerated in the anastomosis group. Anorectal transplantation was technically successful with a 3-stage operation: colostomy preparation, anorectal transplantation, and stoma closure. The dog who underwent allotransplantation and immunosuppression had 2 episodes of mild rejection, which were reversed with methylprednisolone and tacrolimus. The dog who underwent allotransplantation without immunosuppression had a severe acute rejection that resulted in graft necrosis. Successful dogs had full defecation control at the end of the study., Conclusions: We describe the critical role of the pudendal nerve in anorectal function and the first long-term success with anorectal transplantation in a canine model. This report is a proof-of-concept study for anorectal transplantation as a treatment for patients with an ostomy because of anorectal dysfunction., Competing Interests: The authors report no conflicts of interest., (Copyright © 2022 The Author(s). Published by Wolters Kluwer Health, Inc.)

Although the methods for medical education continue to evolve due to the development of medicines, the cadaver dissection course still plays a fundamental role. The cadaver dissection course allows students to learn to handle instruments correctly while actively exploring three-dimensional anatomy. However, dissection comes with the risk of accidental injury. In recent years, the number of classes offered for the cadaver dissection course has decreased while the amount of knowledge required in clinical medicine has increased. Simulation-based education (SBE) has been proven to be an effective educational method that enhances the development of practical skills by integrating learners' knowledge and skills. This study aimed to investigate the effect of SBE as a preparatory education course when taken prior to a medical student's enrollment in the cadaver dissection course. In the present study, an SBE assuming practical cadaver dissection course was performed in the Clinical Simulation Center. The frequency of injury rates per 1000 h of cadaver dissection course was significantly less in 2017 and 2018 compared to that in 2016. Two years after the implementation of the SBE, average student self-efficacy scores and written examination scores significantly increased, whereas self-contentment scores were relatively unchanged. The results showed that the implementation of SBE decreased the incidence of injuries and improved students' overall self-efficacy scores and increased acquisition of knowledge evident on written examination score. Therefore, SBE as a preparatory education course may effectively promote the combined development of dissection skills and anatomical knowledge in the subsequent fundamental cadaver dissection course., (© 2021 American Association for Anatomy.)

Vascular endothelial cells are covered with glycocalyx comprising heparan sulfate, hyaluronan, chondroitin sulfate, and associated proteins. Glomerular endothelial glycocalyx is involved in protecting against induction of proteinuria and structural damage, but the specific components in glycocalyx that represent therapeutic targets remain unclear. Anti-vascular endothelial growth factor (VEGF) therapy is associated with an increased risk of glomerular endothelial injury. This study investigated whether hyaluronan could provide a therapeutic target to protect against proteinuria. We conducted ex vivo and in vivo experiments to explore the effects of degrading glomerular hyaluronan by administering hyaluronidase and of supplementation with hyaluronan. We investigated hyaluronan expression using biotin-labeled hyaluronan-binding protein (HABP) in human kidney specimens or serum hyaluronan in endothelial injuries under inhibition of VEGF signaling. We directly demonstrated hyaluronan in glomerular endothelial layers using HABP staining. Ex vivo and in vivo experiments showed the development of proteinuria after digestion of hyaluronan in glomerular capillaries. Supplementation with hyaluronan after hyaluronidase treatment suppressed proteinuria. Mice in the in vivo study developed albuminuria after intraperitoneal injection of hyaluronidase with decreased glomerular hyaluronan and increased serum hyaluronan. In human kidneys with endothelial cell dysfunction and proteinuria due to inhibition of VEGF, glomerular expression of hyaluronan was reduced even in normal-appearing glomeruli. Serum hyaluronan levels were elevated in patients with pre-eclampsia with VEGF signaling inhibition. Our data suggest that hyaluronan itself plays crucial roles in preventing proteinuria and preserving the integrity of endothelial cells. Hyaluronan could provide a therapeutic target for preventing glomerular endothelial glycocalyx damage, including VEGF signaling inhibition., (© 2021 The Authors. Physiological Reports published by Wiley Periodicals LLC on behalf of The Physiological Society and the American Physiological Society.)

Infections of CTX-M extended-spectrum β-lactamase-producing Enterobacterales are a severe threat in clinical settings. CTX-M genes on plasmids have been transferred to many Enterobacterales species, and these species have spread, leading to the global problem of antimicrobial resistance. Here, we developed a lateral flow immunoassay (LFIA) based on an anti-CTX-M rabbit monoclonal antibody. This antibody detected CTX-M variants from the CTX-M-9, CTX-M-2, and CTX-M-1 groups expressed in clinical isolates. The LFIA showed 100% sensitivity and specificity with clinical isolates on agar plates, and its limit of detection was 0.8 ng/mL recombinant CTX-M-14. The rabbit monoclonal antibody did not cross-react with bacteria producing other class A β-lactamases, including SHV. In conclusion, we developed a highly sensitive and specific LFIA capable of detecting CTX-M enzyme production in Enterobacterales. We anticipate that our LFIA will become a point-of-care test enabling rapid detection of CTX-M in hospital and community settings as well as a rapid environmental test., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Experimental autoimmune orchitis (EAO) may be used as a model to investigate immunological infertility in men. Murine EAO is induced via immunization with auto-immunogenic antigens (AIAgs) from testicular germ cells (TGCs). CD4 + T cells play a crucial role in EAO induction. However, whether AIAgs induce an immune response remains unclear. We aimed to identify self-antigens that induce EAO by screening a phage display library of random TGC peptides using IgG from EAO-induced A/J mice. Twenty TGC-specific AIAgs were detected, and G protein-coupled receptor kinase 2 interacting protein-1 (GIT1) and heat shock protein A4L (HSPA4L) were identified as candidate AIAgs that induce EAO. Immunization with GIT1 or HSPA4L, emulsified in complete Freund's adjuvant, resulted in 66 % or 100 % incidence of EAO, respectively, indicating that HSPA4L is a most potent AIAg that induces EAO in mice. These findings may expectedly help improve the diagnostic procedures and treatment of immunological infertility in men., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The occurrence of a third head of the biceps femoris is very rare. We encountered the case of a 90-year-old Japanese male cadaver with a third head of the biceps femoris in the posterior aspect of the thigh during dissection at Aichi Medical University in 2016. It originated from the proximal part of the femur and fused with the muscle belly between the long and short heads of the biceps femoris. Additionally, three muscle tendons were connected to the gluteus maximus. To the best of our knowledge, this is the first report on the third head of the biceps femoris demonstrating two origins, i.e., the proximal part of the femur and the insertion tendon of the gluteus maximus. Moreover, the third head, as well as the short head, of the biceps femoris was innervated by the muscular branch of the common peroneal nerve. Based on the origin and innervation, it can be believed that the third head of the biceps femoris is analogous to its short head and is related to the tenuissimus, a phylogenetic remnant. Therefore, we concluded that this third head is an intermediate muscle type of the tenuissimus and short head of the biceps femoris.

The high-pressure gas (HPG) method with carbon monoxide (CO) and oxygen (O 2 ) mixture maintains the preserved rat heart function. The metabolites of rat hearts preserved using the HPG method (HPG group) and cold storage (CS) method (CS group) by immersion in a stock solution for 24 h were assessed to confirm CO and O 2 effects. Lactic acid was significantly lower and citric acid was significantly higher in the HPG group than in the CS group. Moreover, adenosine triphosphate (ATP) levels as well as some pentose phosphate pathway (PPP) metabolites and reduced nicotinamide adenine dinucleotide phosphate (NADPH) were significantly higher in the HPG group than in the CS group. Additionally, reduced glutathione (GSH), which protects cells from oxidative stress, was also significantly higher in the HPG group than in the CS group. These results indicated that each gas, CO and O 2 , induced the shift from anaerobic to aerobic metabolism, maintaining the energy of ischemic preserved organs, shifting the glucose utilization from glycolysis toward PPP, and reducing oxidative stress. Both CO and O 2 in the HPG method have important effects on the ATP supply and decrease oxidative stress for preventing ischemic injury. The HPG method may be useful for clinical application.

The liposome-protamine-DNA complex (LPD) is an effective cationic carrier of various nucleic acid constructs such as plasmid DNA and small interfering RNA (siRNA). Hyaluronic acid coated on LPD (LPDH) reduces cytotoxicity and maintains the silencing effect of LPD-encapsulated siRNA. Herein, we aim to develop LPD- or LPDH-containing spray-freeze-dried particles (SFDPs) for therapeutic delivery of siRNA to the lungs. LPD- or LPDH-containing SFDPs (LPD- or LPDH-SFDPs) were synthesized and their structure and function as gene carriers were evaluated using physical and biological methods. The particle size of LPDH, but not of LPD, was constant after re-dispersal from the SFDPs and the amount of siRNA encapsulated in LPDH was larger than that in LPD after re-dispersal from the SFDPs. The in vitro pulmonary inhalation properties of LPDH-SFDPs and LPD-SFDPs were almost the same. The cytotoxicity of LPDH-SFDPs in human umbilical vein endothelial cells (HUVEC) was greatly decreased compared with that of LPD-SFDPs. In addition, Bcl-2 siRNA in LPDH-SFDPs had a significant gene silencing effect in human lung cancer cells (A549), whereas Bcl-2 siRNA in LPD-SFDPs had little effect. These results indicate that compared with LPD, LPDH is more useful for developing SFDPs for siRNA pulmonary inhalation., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The sagittal band of the finger is an aponeurosis-like structure surrounding the metacarpophalangeal joint. Sagittal band rupture causes extensor tendon dislocation from the dorsal side of the metacarpophalangeal joint. The thumb has two extensor tendons: extensor pollicis longus and extensor pollicis brevis tendons. Multiple studies have reported variations of extensor pollicis brevis tendon insertion. However, it remains unclear how the thumb sagittal band envelopes the extensor pollicis longus and extensor pollicis brevis tendons. This study investigated the anatomical relationship between the sagittal band and the two extensor tendons of the thumb. One hundred hands (47 right, 53 left) from 54 cadavers were examined to assess the detailed structure of the thumb sagittal band and extensor pollicis longus and extensor pollicis brevis tendons. We found that the thumb sagittal band enveloped both the tendons either separately (type I) or collectively (type II). Thirty-four cases (34.0%) were type I and 66 cases (66.0%) were type II. The extensor pollicis longus and extensor pollicis brevis tendons enveloped in the type I thumb sagittal band were inserted on different sites, respectively, whereas those tendons enveloped in the type II thumb sagittal band were inserted on the same sites. This study demonstrated that differences in the type of thumb sagittal band are closely associated with variations in extensor pollicis brevis tendon insertion. We predicted that these differences contribute to the sliding distance between the extensor pollicis brevis and extensor pollicis longus tendons and affect the pathophysiology of extensor tendon dislocation.

Cutaneous nerves have vascular branches (VBs) that reach the arteries and are thought to be involved in arterial constriction. We aimed to examine the anatomical and histological relationship between the VBs of a cutaneous nerve in the foot and the acupuncture point LR3 (Taichong), which is a depression between the base of the first and second metatarsal bones on the dorsum of the foot and is a source point of the foot. We examined 40 cadaver feet to assess the distribution areas of the VBs of the medial branch of the deep peroneal nerve (MBDPN). MBDPNs were distally followed to identify the point where the VBs reached the arteries. The distance between the point and LR3 was measured. Sympathetic fibers in the VBs were histologically observed using tyrosine hydroxylase (TH) immunostaining. The VBs of the MBDPNs reaching the dorsal pedis arteries were observed in all specimens (100%). The mean distance between LR3 and the point where the VBs of the MBDPN reached the arteries was 3.2 ± 2.6 mm. Among the VBs, 70% were distributed proximal to LR3. Moreover, TH-positive fibers were present in the VBs. These findings revealed that a part of the MBDPN distributed the dorsal pedis artery and contained sympathetic fibers. We also found that the distribution area of the VBs was close to LR3. Our study provides anatomical evidence that LR3 is a specific area and its stimulation would be useful for treating peripheral circulatory failure., Competing Interests: The authors declare that there are no conflicts of interest regarding the publication of this paper., (Copyright © 2020 Kanae Umemoto et al.)

The fibers of the abductor pollicis longus (APL) and extensor pollicis brevis (EPB) muscles intersect the distal radius. This anatomical structure puts pressure on the dorsal surface of the distal radius when various wrist positions are adopted. An increase in this pressure is associated with the risk of intersection syndrome and with immobilization after Colles' fracture. However, the relationship between the pressure on the distal radius and various wrist positions remains unclear. This study was established to provide quantitative data on the mechanical effect of the pressure exerted by the APL and EPB. Ten cadaveric wrist models containing a force sensor were prepared and used to record pressure levels at various wrist positions, such as pronation, supination, flexion and dorsiflexion, and radial and ulnar deviation. A three-dimensional simulation model comprising four bones, one muscle, one tendon, and one tendon sheath was constructed and analyzed in detail using the finite element method. The contribution of the APL and EPB to the pressure exerted on the distal radius was quantified by dissecting muscles while measuring pressure. The position (pronation and ulnar deviation without flexion/dorsiflexion) associated with a strong force being exerted on the distal radius was determined by measuring and analyzing the mechanical effect. We concluded that this position increases the risk of intersection syndrome but provides effective immobilization after Colles' fracture. The cadaveric and computational method presented herein is the first to identify the anatomical relationship between the pressure on the distal radius and various wrist positions.

Background: Infertility and gonadal dysfunction are well known side-effects by cancer treatment in males. In particularly, chemotherapy and radiotherapy induced testicular damage, resulting in prolonged azoospermia. However, information regarding therapeutics to treat spermatogenesis disturbance after cancer treatment is scarce. Recently, we demonstrated that Goshajinkigan, a traditional Japanese medicine, can completely rescue severe busulfan-induced aspermatogenesis in mice. In this study, we aimed to detect the effects of Goshajinkigan on aspermatogenesis after irradiation., Methods: This is animal research about the effects of traditional Japanese medicine on infertility after cancer treatment. C57BL/6 J male mice received total body irradiation (TBI: a single dose of 6Gy) at 4 weeks of age and after 60 days were reared a Goshajinkigan (TJ107)-containing or TJ107-free control diet from day 60 to day 120. Then, two untreated females were mated with a single male from each experimental group. On day 60, 120 and 150, respectively, the sets of testes and epididymis of the mice in each group after deep anesthetization were removed for histological and cytological examinations., Results: Histological and histopathological data showed that 6Gy TBI treatment decreased the fertility rate (4/10) in the control diet group; in contrast, in the TJ107-diet group, the fertility rate was 10/10 (p < 0.05 vs. 6Gy group). Supplementation with TJ107 was found to rescue the disrupted inter-Sertoli tight junctions via the normalization of claudin11, occludin, and ZO-1 expression and reduce serum anti-germ cell autoantibodies., Conclusions: These findings show the therapeutic effect on TBI-induced aspermatogenesis and the recovering disrupted gonadal functions by supplementation with TJ107.

(1) Background: Heme oxygenase-1 (HO-1) degrades heme and generates carbon monoxide (CO), producing various anti-inflammatory, anti-oxidative, and anti-apoptotic effects. This study aimed to confirm the effects of CO on the ischemia-reperfusion injury (IRI) of donor lungs using a high-pressure gas (HPG) preservation method. (2) Methods: Donor rat and canine lungs were preserved in a chamber filled with CO (1.5 atm) and oxygen (O 2 ; 2 atm) and were ventilated with either CO and O 2 mixture (CO/O 2 group) or air (air group) immediately before storage. Rat lungs were subjected to heterotopic cervical transplantation and evaluated after reperfusion, whereas canine lungs were subjected to allogeneic transplantation and evaluated. (3) Results: Alveolar hemorrhage in the CO/O 2 group was significantly milder than that in the air group. mRNA expression levels of HO-1 remained unchanged in both the groups; however, inflammatory mediator levels were significantly lower in the CO/O 2 group than in the air group. The oxygenation of graft lungs was comparable between the two groups, but lactic acid level tended to be higher in the air group. (4) Conclusions: The HO-1/CO system in the HPG preservation method is effective in suppressing IRI and preserving donor lungs.

We maintained the function of an extracted rat heart after 24-48 h preservation in a high-pressure gaseous mixture of carbon monoxide (CO) and oxygen (O 2 ). Here, we assessed the effects of different partial pressures of hyperbaric CO and O 2 for 24-48 h at 4 °C on rat heart preservation and compared conditions including immersion in University of Wisconsin solution. Preserved hearts were transplanted into recipient rats via heterotopic cervical heart transplantation for in vivo evaluation and perfused using the Langendorff system for ex vivo evaluation. The survival rate of transplanted hearts was 100% at postoperative day 7 in the CO + O 2 (PCO:PO 2  = 1.5:2.0 atm) group but only 33% in the CO + O 2 (PCO:PO 2  = 2.0:1.5 atm) group. Langendorff system and histopathological analysis revealed that the left ventricular pressure of preserved hearts in the CO + O 2 (PCO:PO 2  = 1.5:2.0 atm) group was better than the CO + O 2 (PCO:PO 2  = 2.0:1.5 atm). We demonstrate that exposure of rat hearts to hyperbaric CO and O 2 is superior to the immersion method and that partial pressure of hyperbaric CO and O 2 is crucial to preservation.

Formaldehyde (FA) is frequently used to embalm human cadavers that are employed to teach gross anatomy to medical and dental students. However, exposure to FA is harmful to both students and educators. The aim of this study was to reduce the FA levels in the anatomy dissection hall by spraying an FA scavenger solution. We measured the changes in FA levels after administering FA scavenger solutions to liquid, wet paper towels, organs, and cadavers containing FA. Among L-cysteine, N-ethyl urea, and urea, the latter was found to have the strongest scavenging power towards the FA in the liquid. The molar concentration of urea that most efficiently reduced the levels of volatilized FA from the wet paper towels was the same as that of the FA. After spraying the urea solution, the volatilized FA levels immediately decreased, reaching their minimum at 60 min, and remained low even after 240 min. Spraying the urea solution onto the organs reduced the levels of FA volatilized from the surfaces of organs but not those from the insides of the organs. In the dissection hall used for the gross anatomy course at Tokyo Medical University, the FA levels were significantly decreased after spraying the urea solution onto the cadavers. Moreover, dissection could be performed without the cadavers putrefying during the 4-month course. These results indicate that various institutes could use urea solution spray to effectively reduce the FA levels in the dissection hall and thus ensure the safety of students and educators.

Cold ischemia times ranging from <6 h to as long as 24 h are generally quoted as the limits for attempting the replantation of amputated extremities. In this study, we aimed to assess the effect of hyperbaric carbon monoxide (CO) and oxygen (O 2 ) on rat limb preservation. Donor rat limbs were preserved in a chamber filled with hyperbaric CO and O 2 for 3 days (CO + O 2 3 days) or 7 days (CO + O 2 7 days). Positive and negative control groups were created by using non-preserved limbs (NP) and limbs wrapped in saline-moistened gauze for 3 days (SMG 3 days), respectively. The survival rate of transplanted limbs at postoperative day 90 was 88% in the NP and 86% in the CO + O 2 3 days. The corresponding survival rate was 50% in the CO + O 2 7 days at postoperative day 90 but was 0% in the SMG 3 days at postoperative day 3. Muscle mass decreased in the CO + O 2 3 days and CO + O 2 7 days compared with the NP, but sciatic-tibial nerve conduction velocities did not differ. These results indicate that amputated extremities preservation with hyperbaric CO and O 2 could extend the time limits of preservation, maintaining their viability for replantation.