Setsunan University

Tacrolimus, a topical calcineurin inhibitor, is used to treat atopic dermatitis (AD). AD is classically T helper type (Th) 2-driven, but Th17cells are implicated in chronic AD, yet its efficacy against Th17-dependent pathology remains unclear. We investigated the effects of tacrolimus using a murine model of Th17-mediated allergic skin inflammation. In CD4+ T cells from DO11.10/Rag2-/- mice, which express ovalbumin (OVA)-specific T cell receptor, tacrolimus differentially suppressed stimulation-induced cytokine expression by Th2 cells. At similar concentrations, tacrolimus suppressed Il21, but not Il17a or Il22, expression in Th17cells. Subcutaneous OVA challenge elicited ear thickening in BALB/c mice after adoptive transfer of Th2 or Th17cells, with a stronger response in Th17-transferred mice, but less than OVA-immunized controls. Topical tacrolimus reduced Th17-mediated ear swelling, corroborated by histopathology. In Th2-transferred mice, tacrolimus tended to reduce early skin thickening (day 3) but did not affect late responses (day 7). In Th17-transferred mice, tacrolimus significantly reduced allergen-specific T-cell accumulation in OVA-injected skin and tended to reduce Il21 expression, whereas Th2-cell accumulation and cytokine expression were unaffected. Tacrolimus exerts stronger inhibitory effects on Th17- than Th2-driven responses in this model, suggesting that suppression of Th17 pathways may contribute to its therapeutic benefit in AD.

BackgroundEdoxaban is the only direct-acting oral anticoagulant (DOAC) indicated for preventing venous thromboembolism (VTE) in patients undergoing arthroplasty and can be administered for a short period (1–2 weeks). The dosage of edoxaban is set according to renal function and the presence or absence of concomitant medications with P-glycoprotein inhibitory effects. However, a small number of patients still develop VTE after lower-extremity orthopedic surgery (LE-OS), despite edoxaban administration. This single-center, exploratory cohort study retrospectively investigated patients prescribed edoxaban after LE-OS in order to explore the potential of D-dimer as a signal of VTE.MethodsOf the 1,457 patients who received edoxaban after total hip arthroplasty or total knee arthroplasty between January 2016 and December 2020, 1,244 who cleared the exclusion criteria were divided into the non-VTE group (1,219 patients) and VTE group (25 patients). To reduce bias in VTE occurrence, patient age, sex, body weight, serum creatinine, edoxaban underdosing, previous thromboembolism, bed rest duration ≥ 48 h, and blood transfusion were matched using propensity score matching (PSM). We compared the routinely measured D-dimer values of blood samples collected on postoperative day 7 among non-VTE and VTE groups using the Mann–Whitney U test.ResultsAfter PSM, there were 23 patients in each of the non-VTE and VTE groups. The median (interquartile range) D-dimer value on postoperative day 7 was 6.2 (4.2–8.1) µg/mL in the non-VTE group and 9.6 (6.3–12.4) µg/mL in the VTE group (p = 0.005).ConclusionsThis retrospective exploratory study demonstrated that D-dimer values in patients receiving edoxaban after LE-OS may serve as a signal for VTE occurrence.

Approximately 5% to 10% of asthma patients are resistant to glucocorticoid therapy; however, the mechanisms by which this resistance develops remain unclear. The present study investigated whether and how mechanistic target of rapamycin (mTOR) is involved in the development of steroid resistance using in vitro and in vivo murine models. The interleukin (IL)-33/thymic stromal lymphopoietin (TSLP)/IL-7-induced growth of group 2 innate lymphoid cells (ILC2) in vitro was resistant to dexamethasone (DEX), but suppressed by everolimus, an mTOR inhibitor, in a concentration-dependent manner. The inhibition of ILC2 growth by the combination of DEX and everolimus was significantly stronger than that by everolimus monotherapy. The combination of the pan-class I phosphatidylinositide-3 kinase inhibitor, buparlisib and the pan-Akt inhibitor, capivasertib also attenuated the resistance of IL-33/TSLP/IL-7-exposed ILC2s to DEX. The expression of the antiapoptotic factor, B-cell lymphoma-extra large, induced by IL-33/TSLP/IL-7 in ILC2s was significantly reduced by everolimus. Additionally, everolimus effectively suppressed the IL-33/TSLP/IL-7-induced phosphorylation of glucocorticoid receptors (GR) at the Ser234 residue, which has been reported to cause GR dysfunction. In the steroid-resistant asthma mouse model in vivo, under treatment with everolimus, DEX inhibited the development of airway remodeling and increased the number of ILC2s in the lungs. The present results suggest that the phosphoinositide 3-kinase/protein kinase B/mTOR pathway plays an essential role in the development of steroid resistance in ILC2s and asthma pathogenesis through both the expression of B-cell lymphoma-extra large and phosphorylation of GR. Therefore, mTOR inhibitors have the potential to restore glucocorticoid sensitivity and, thus, exert steroid-sparing effects in severe asthma patients. SIGNIFICANCE STATEMENT: Approximately 5% to 10% of asthma patients exhibit resistance to steroid therapy, posing a major clinical challenge due to limited treatment options. This study demonstrates that activation of the phosphoinositide 3-kinase/protein kinase B/mechanistic target of rapamycin (mTOR) pathway induces steroid resistance in group 2 innate lymphoid cells. Targeting mTOR with everolimus restores steroid sensitivity, highlighting mTOR inhibition as a promising pharmacotherapy for steroid-resistant asthma.

ABSTRACT Sugarcane (Saccharum spp. hybrids) is a globally important crop for food and bioenergy, but its production is increasingly threatened by drought driven by climate change. Drought causes complex interactions between root function and leaf photosynthesis, but their underlying mechanisms remain largely unstudied in sugarcane. To address this knowledge gap, we grew up to eight sugarcane cultivars in one field and two pot experiments inside a rain-out shelter. The relative growth rate of these eight cultivars under both well-watered and water-limited conditions was largely explained by net assimilation rate, which depended on bleeding sap rate caused by root pressure. Among the cultivars, Harunoogi – a first-generation backcross cultivar between a wild relative (S. spontaneum) and a commercial cultivar – exhibited improved drought avoidance through adequate root water supply, whereas the commercial cultivar NiTn18 showed the opposite. Throughout the drought and recovery, Harunoogi exhibited significantly higher leaf area, stomatal conductance, chlorophyll content, and maximum amplitude of the reaction center chlorophyll of PSI than cultivar NiTn18, which resulted in a higher net CO₂ assimilation rate and biomass. These results highlight the potential of improving both root function and photosynthesis as an effective approach to developing drought-tolerant sugarcane in the field.

Pigs have been utilized as the donor animal for xenotransplantation, a promising solution to the continued shortage of organ donor. However, porcine endogenous retrovirus (PERV), a gamma retrovirus present in all pig genomes, remains a major concern for the microbiological safety of donor pigs. Therefore, accurately assessing the risk of PERV infection is essential for the clinical application of xenotransplantation. To improve the evaluation of PERV infectivity, here we developed a new in vitro co-culture method for porcine and human cells, applying flowcytometric cell sorter for specific isolation of human cells. This method allows direct contact between porcine and human cells without special treatment that would affect PERV replication, which is better simulating the situation observed in in vivo xenotransplantation. Using this method, we identified several possible factors affecting PERV infectivity, including the period during which porcine and human cells are in contact, the number of porcine cells used, and the levels of PERV mRNA expression in porcine cells. Notably, we found that the PERV infection of human cells can occur within the first 24 h of contact if porcine cells are of a high-risk type for PERV infection, which is not confirmed by any types of conventional co-culture methods. Our result emphasized the importance of selecting appropriate donor pigs with lower risk of PERV infection. Our newly developed method may help the testing and selection of pigs with lower risk of PERV infection.

Microwave Rocket, a next-generation space transportation system, generates thrust by utilizing millimeter-wave energy beamed from the ground. The thruster operates cyclically, alternating between thrust generation—where air is compressed and exhausted via millimeter-wave-supported detonation—and an air-refilling process. Although reed valve systems have been studied for air intake, their effect on thrust performance has not been experimentally verified. This study experimentally evaluates the propulsion characteristics of a thruster equipped with a six-stage reed valve air intake system (36 valves in total, length 500 mm, radius 28 mm) under ground testing. A Gaussian beam with a frequency of 170 GHz, an output power of 550 kW, and a pulse width of 1.2 ms was repeatedly applied at pulse detonation engine (PDE) cycle frequencies ranging from 50 to 250 Hz, and the resulting thrust impulse was measured. Results show that approximately 60% of the first-cycle impulse was sustained up to 150 Hz, beyond which the partial filling effect led to a decrease in impulse. The time-averaged thrust, defined as the product of impulse and PDE cycle frequency, reached a peak of 8.3 kN per square meter at 206 Hz due to the trade-off between impulse degradation and frequency increase. Furthermore, modifications to the reed valve design, specifically reducing stiffness and fundamental frequency, led to up to 80% impulse recovery and enhanced thrust performance at lower PDE cycle frequencies. • Thrust performance of Microwave Rocket with reed valves evaluated experimentally. • Approximately 60% of first-cycle impulse maintained in subsequent cycles. • Transient characteristics of intake clarified by varying PDE cycle frequency. • Optimal PDE frequency for maximum thrust influenced by reed valve design.

The prevalence of allergic diseases has been increasing, and sensitization to allergens such as cedar pollen and house dust mites has become a social issue. Allergic inflammation is primarily driven by type 2 inflammation, which is mediated by interleukin (IL)-4, IL-5, and IL-13 produced by Th2 cells and group 2 innate lymphoid cells (ILC2s). Recent studies have suggested that extracellular vesicle (EV) also plays critical roles in the pathogenesis of allergic diseases. EVs are lipid bilayer-enclosed particles containing proteins, mRNA, and microRNA (miRNA), which function as carriers of cytokines, antigens, and miRNAs, thereby activating Th2 cells and ILC2s and contributing to the progression of various inflammatory diseases. In contrast, we demonstrated that EVs contributed to the regression of allergic disease: EVs derived from the serum of allergen immunotherapy-treated mice exhibited suppression of ILC2 activation. Given their dual roles in both promoting and suppressing immune responses, EVs are emerging as promising targets and tools for novel treatment strategies. Understanding the immunomodulatory mechanisms mediated by EVs will be a crucial step toward developing safer and more effective therapies for allergic diseases. This review provides an overview of the role of EVs in allergic inflammation and highlights our recent findings on how allergen immunotherapy influences the properties and functions of EVs, thereby contributing to the regulation of immune responses and alleviation of allergic symptoms.