The potential risks to the fetus or infant should be evaluated before initiating pharmacotherapy during pregnancy and breastfeeding. However, safety information and experiences during pregnancy and breastfeeding are often lacking because these populations are generally excluded from clinical drug development studies. Perinatal mental health is important. Based on quantitative and toxicological analyses, we focused on medications used in psychiatry and neurology during the perinatal period. As the placenta serves as a temporary but crucial organ for ensuring successful pregnancy and appropriate fetal growth, we assessed the effects of antiepileptic drugs (AEDs) on placental functions such as transport mechanisms, nutrient transport, and trophoblast differentiation. Several AEDs have been suggested to be transported to the placenta via carrier-mediated pathways in a series of studies. Valproic acid, known to pose several risks to the fetus, affects the gene expression of nutrient transporters and trophoblast differentiation. Furthermore, we established several quantitative methods, such as those for antianxiety and hypnotic drugs, to evaluate the safety of pharmacotherapy during breastfeeding using liquid chromatography-tandem mass spectrometry. The validated methods were applied to clinical samples donated by lactating women. In a series of studies, the importance of choosing a suitable method for sample preparation for each biological matrix has been highlighted. The results obtained from the clinical samples suggest the possibility of differences in transfer properties among drugs categorized in the same class. Furthermore, this research emphasizes the critical need to assess breast milk transfer in human studies because species differences have been suggested in some cases.

Boron neutron capture therapy (BNCT), a type of nuclear capture-based radiotherapy, has received extensive attention because of its strong anticancer effects, especially in head and neck cancers. This therapy was approved for clinical use in 2020 in Japan. This study demonstrated a technique that effectively uses the electrostatic interactions of negatively charged borane cage (polyhedral borane anion) of disodium mercaptoundecahydro-closo-dodecaborate (BSH) and positively charged cationic lipids or arginine-rich cell-penetrating peptides as carriers to enhance the efficiency of cellular uptake. Mixing of fluorescein isothiocyanate (FITC)-labeled BSH (FITC-BSH) with the cationic lipids led to increased cytosolic release and nuclear accumulation of FITC-BSH, resulting in superior cancer cell-killing activity following thermal neutron irradiation. This simple technique and our experimental results provide essential insights for the further development of BNCT.

Recombinant adeno-associated virus (AAV) vectors are widely used for manipulating gene expression. AAVrh.10 is a highly infectious AAV serotype for the central nervous system and various tissues. Owing to its potential use in research, we aimed to optimize the production strategy and develop a simple purification protocol for the AAVrh.10 vector. In this study, we explored a simple production and purification strategy for the AAVrh.10 vector via chloroform extraction and ultrafiltration. Initially, we evaluated the optimal conditions for AAVrh.10-CAG-GFP production using AAV-293 cells. AAVrh.10-CAG-GFP was successfully produced in a serum-free medium after plasmid transfection. Moreover, the culture medium contained a substantial amount of the virus. Therefore, both AAVrh.10-containing cell lysate and culture medium should be used to prepare the AAVrh.10 viral vector. To purify and concentrate AAVrh.10-CAG-GFP from the crude lysate and medium, we optimized the chloroform extraction and ultrafiltration strategies. Subsequently, purified AAVrh.10-CAG-GFP was used to infect HEK-293T cells. Overall, this study provides a simple and effective AAVrh.10 vector preparation strategy for basic and preclinical research.

Vancomycin (VCM) combined with piperacillin/tazobactam (PIPC/TAZ) is used as an empiric therapy in patients with severe infections, including sepsis. Recent research has found an increased incidence of acute kidney injury (AKI) in patients receiving combination therapy with these antibiotics. However, the pharmacological mechanism by which this combination worsens kidney function remains unclear. In this study, we investigated the direct cytotoxicity of VCM, PIPC, and TAZ on HK-2 cells and human renal proximal tubular epithelial cells (RPTEC). VCM, PIPC/TAZ, or PIPC significantly reduced cell viability in a concentration-dependent manner; the potency was in the order of VCM, PIPC/TAZ, and PIPC (IC₅₀ values were 1717, 2491, and 3020 μg/mL, respectively). The combined treatment with PIPC/TAZ or PIPC significantly enhanced the VCM-induced decrease in cell viability. Furthermore, PIPC/TAZ or PIPC increased lactate dehydrogenase leakage, indicating membrane cytotoxicity, whereas no such effect was observed with VCM or TAZ. VCM increased caspase-3/-7 activity, whereas PIPC did not. The VCM-induced increase in neutrophil gelatinase-associated lipocalin (NGAL) production was amplified by concomitant PIPC treatment. Synergistic effects were detected for both the cell viability and NGAL production, suggesting that the direct toxicity of PIPC to RPTEC was responsible for the increased AKI incidence in patients treated with VCM. Our results may contribute to a better understanding of how AKI is exacerbated, as well as provide tips for preventing AKI after VCM and PIPC/TAZ combined therapy.

FR429, an ellagitannin isolated and purified from the whole herb Polygonum capitatum (P. capitatum), possesses a robust pharmacological profile, which is particularly noteworthy for its anti-inflammatory and anticancer properties. Despite these established effects, its potential in mitigating hepatic injury remains to be fully explored. The present investigation delineates the hepatoprotective efficacy of FR429 and unveils its underlying molecular mechanisms. Initially, of the tested compounds, 10 compounds (specifically, compounds 2, 4, 5, 6, 7, 8, 9, 12, 13, and 14) exhibited significant protective effects at a concentration of 10 μM, elevating HepG2 (human liver cancer cell) cell viability from 43.4 to 70% following carbon tetrachloride (CCl₄) exposure. Among them, compounds 2 (FR429, half-maximum effective concentration (EC₅₀) = 6.46 μM) and 6 (2ʺ-O-galloylquercitrin, EC₅₀ = 5.36 μM) demonstrated the highest cytoprotective activities. In the murine model, FR429 dramatically attenuated serum levels of alanine transaminase, aspartate transaminase, and alkaline phosphatase, indicative of its hepatoprotective potential. Histopathological evaluation further substantiated these findings, as FR429 noticeably mitigated CCl₄-induced hepatic lesions, involving necrosis, ballooning degeneration, and neutrophil infiltration. Transcriptomic analysis unveiled 178 differentially expressed genes in FR429-treated mice liver tissue, with significant alterations indicative of a hepatoprotective response. Mechanistic investigations revealed that FR429’s hepatoprotective effects involve modulation of the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway, evidenced by downregulation of toll-like receptor 2, phosphorylated PI3K, phosphorylated Akt, nuclear factor-kappa-B, interleukin-1 beta, and tumor necrosis factor-alpha expression. Furthermore, FR429 modulated the gene and protein expression levels of apoptotic markers (apoptotic protein (Bax) and B-lymphoblastoma-2 gene (Bcl2)), reinforcing its anti-hepatic damage efficacy. This study represents the first report establishing FR429 as an effective hepatoprotective compound, paving the way for further investigation into its therapeutic applications.

Although myocarditis is listed as a serious adverse reaction in the package inserts of mesalazine and some other anti-ulcerative colitis (UC) drugs currently in use, and regulatory authorities have issued related warnings, the detailed characteristics of anti-UC drug-induced myocarditis remain unknown. We aimed to investigate the association between UC drugs and myocarditis development as an adverse event and its characteristics using data from a spontaneous adverse drug reaction reporting database. We searched for adverse event signals of five drugs, mesalazine, sulfasalazine, azathioprine, mercaptopurine, and budesonide, listed in the treatment guidelines for UC, using VigiBase. The information component was calculated, and a signal was considered present when the lower limit of the 95% confidence interval of the information component exceeded zero. The total number of VigiBase and myocarditis (as a target adverse event) reports was 38459016 and 71571, respectively. No trend was identified based on age or sex. Analysis of the five UC drugs for severity in VigiBase showed that most patients recovered, with a low reported mortality rate. However, the time to onset of adverse drug reactions varied among the drugs. Mesalazine signals were detected regardless of age or sex. This finding suggests that myocarditis, an adverse event, may be a potential complication regardless of patient characteristics. Our results also indicate that UC itself may induce myocarditis. Our findings warrant multifaceted investigations, including basic and clinical studies, on the characteristics of each drug regarding the development of myocarditis as an adverse event.

Cisplatin is a platinum-based drug that is widely used to treat various types of cancer. However, cisplatin is known to cause severe adverse effects, such as nephrotoxicity and ototoxicity. Clock genes, such as Bmal1 and Clock, regulate cisplatin-related homeostasis genes, such as Oct2 and Mate1. Although these clock genes may be involved in cisplatin-induced nephrotoxicity, their associations with other clock genes remain unclear. The aim of the present study was to investigate whether seven clock genes (Ciart, cryptochrome 1 (Cry1), Cry2, Npas2, Per1, Per2, and Per3) regulate cisplatin-induced renal toxicity in a renal cortex tubule cell line (MuRTE61). Cisplatin treatment decreases MuRTE61 cell viability in a dose-dependent manner. Cry2 expression levels increased after treatment with cisplatin for 24 h. Notably, Cry2 overexpression alleviated cisplatin-induced suppression of cell proliferation, apoptosis, and platinum content in MuRTE61 cells. Moreover, Cry2 overexpression upregulated the efflux-related transporters (Atp7a and Mrp2). These results suggest that Cry2 protects against cisplatin toxicity by reducing Pt accumulation and increasing the expression of Atp7a and Mrp2.

The optimal pharmacokinetics (PK) of orally administered nanoparticles (NPs) varies depending on their application (e.g., drug delivery, adsorbent, and adjuvant). Therefore, engineering NPs to achieve optimal PK is essential for the development of drug designs. Some studies have demonstrated that individual NP factors change the intestinal absorption of NPs; however, no technology has been established to control the biodistribution of orally administered NPs. In this study, a database about the influence of NP characteristics on biodistribution after oral administration was provided. A library of N-isopropylacrylamide polymer NPs with various characteristics that could influence the biodistribution after oral administration, such as size, flexibility, hydrophobicity, surface charges, and surface chemistries, were prepared. NPs with various sizes were synthesized by tuning the surfactant concentration only during synthesis, whereas NPs with different flexibility, hydrophobicity, surface charge, and surface chemistry were synthesized by feeding the corresponding functional monomer. The total amount of NPs accumulated in the organs decreased with increasing NP size, rigidity, hydrophobicity, electric potential (whether positive or negative), and polyethylene glycol modification. The results indicated that the absorption of orally administered NPs can be controlled by optimizing the characteristics of NP such as size, flexibility, hydrophobicity, surface charge, and surface chemistry. The results of this study will provide useful information to design NP formulations.

Selenium (Se) is an essential micronutrient for animals. Various chemical forms of Se exist in nature, each with distinct physiological, nutritional, and toxicological properties. In this study, we aimed to determine whether dimethyldiselenide (DMDSe, a monomethylated Se (MMSe) compound) and dimethylselenide (DMSe, a dimethylated Se compound), known gut bacterial metabolites, could serve as Se sources in rats. DMDSe could be utilized for selenoprotein biosynthesis and was metabolized into urinary selenometabolites. By contrast, DMSe was not utilized for selenoprotein biosynthesis but was further methylated to trimethylselenonium ion (TMSe), one of the urinary Se metabolites. Our findings indicate that dimethylated Se is not readily available as an Se source in rats, unlike MMSe. Selenoprotein biosynthesis requires selenide, an unmethylated form of Se, in the metabolic pathway. Our observations support the hypothesis that demethylation occurs on MMSe as a reversible methylation step but not on dimethylated Se. This suggests that the second methylation step is crucial for inactivating Se and plays a significant role in metabolism to maintain Se homeostasis in animals. Gut microbiota, which can synthesize both DMDSe and DMSe, may contribute to host Se metabolism through methylation processes.

Surveillance of antimicrobial consumption (AMC) is important for controlling antimicrobial resistance (AMR). In recent years, the landscape of infectious diseases has changed due to factors such as the introduction of the National Action Plan (NAP) on AMR and the coronavirus disease 2019 (COVID-19) pandemic. However, their impact on the consumption of broad-spectrum antimicrobial and anti-methicillin-resistant Staphylococcus aureus (MRSA) agents remains unexplored. This study aimed to clarify trends in the consumption of these agents up to 2021, considering the spread of NAP and the COVID-19 pandemic. We used sales data from IQVIA Japan, which were analyzed using an interrupted time-series analysis, with April 2016 (introduction of NAP) and April 2020 (first declaration of a state of emergency) as key change points. The oral broad-spectrum antimicrobial agents consumption decreased, and the spread of the NAP (p-value: 8.15 * 10⁻³, 95% confidence intervals (95% CI): −7.70 * 10⁻³ to −2.06 * 10⁻³) and behavioral restrictions for the COVID-19 pandemic (p value: 1.60 * 10⁻⁸, 95% CI: −0.35 to −0.17) were significantly related to this change. Conversely, there was no notable change in the consumption of anti-MRSA agents from 2013 to 2021. Thus, the introduction of NAP and the COVID-19 pandemic may have been more effective in decreasing the consumption of oral broad-spectrum antimicrobial agents. Since antibiotics are used to treat infections across multiple anatomical therapeutic chemical classifications, continuous evaluation based on treatment purposes is important.

Mineralocorticoid receptor (MR) blockers reduce cardiovascular complications as MRs play a crucial role in cardiovascular regulation. Diabetic cardiovascular complications are caused by vascular endothelial dysfunction. This study used a type 2 diabetic mouse model (DM) to investigate whether esaxerenone (ESAX), an MR blocker, ameliorates vascular endothelial dysfunction. ESAX (3 mg/kg/d) was administered via diet to KK-Ay mice or C57BL/6J mice, a nondiabetic control (Control), for 8 weeks, and metabolic parameters and blood pressure were measured. Vascular responses of the aortic segments were analyzed with acetylcholine, sodium nitroprusside, UK14304, or phenylephrine (PE). The other aortas were used for Western blot analysis. DM mice exhibited higher plasma glucose, insulin, metabolic parameters, and blood pressure levels than those of the Control mice. Parameters that did not include blood pressure were unaltered by DM or ESAX-administered DM (DM + ESAX). However, DM impaired UK14304-induced endothelial-dependent relaxation and nitric oxide production and elevated PE-induced contraction. ESAX administration ameliorated endothelial dysfunction and improved the protein kinase B (Akt) phosphorylation under α₂-agonist UK14304 stimulation in the aorta from DM mice compared with that of the Control mice. However, ESAX did not recover the increased G protein-coupled receptor kinase 2 (GRK2) expression and activity in the DM aorta. Furthermore, the DM-induced phosphorylation of serum and glucocorticoid-regulated kinase 1 (SGK1) was inhibited by ESAX. Overall, ESAX attenuates the development of DM-induced endothelial dysfunction by reducing SGK1 activity and enhancing Akt activity without affecting the GRK2 pathway. These results suggest that the vascular protective effects of ESAX could be employed for diabetic vascular complications.

Oral iron preparations for iron deficiency anemia have major side effects, such as nausea and vomiting, which are gastrointestinal symptoms widely known to occur with gastrointestinal motility disorders. However, it is unclear whether these symptoms are associated with gastrointestinal motility. This study aimed to explore the correlation between oral iron preparations that cause nausea and vomiting with gastrointestinal motility. Sodium ferrous citrate (SFC), a common ingredient in iron preparations, was used in this study. Gastrointestinal motility in mice was measured using the ¹³C-octanoic acid breath test to determine gastric emptying and colonic transport capacities using the bead expulsion test. SFC significantly delayed gastric emptying. However, it did not affect the colonic transport capacity. Treatment with the antiemetic palonosetron, a 5-hydroxytryptamine 3 (5-HT₃) receptor inhibitor, abolished the gastric evacuation retardation effect of SFC. However, the additive in the SFC formulation, palonosetron alone, did not affect the gastric emptying capacity. These results suggest that iron preparations cause vagal nausea and vomiting in the upper gastrointestinal tract due to chemical stimulation of the gastrointestinal blood with a concomitant reduction in gastric emptying capacity. Knowledge of the association of delayed gastric emptying with the onset of iron-induced nausea is useful for understanding iron-induced adverse effects.

Microglia-mediated neuroinflammation plays a critical role in the onset and progression of Alzheimer’s disease. In a previous study, we synthesized 6-hydroxy-3′-propyl-[1,1′-biphenyl]-3-propanoic acid (6OHA) based on the structure of magnaldehyde B, a natural compound that our group identified as a retinoid X receptor (RXR) agonist. However, its potential effects on inflammation in microglial cells remain unexplored. In this study, we specifically focused on the early-phase inflammatory responses to lipopolysaccharide (LPS) and evaluated the inhibitory effects of 6OHA on BV-2 microglial cells following 2 h of LPS exposure. Similar to the existing RXR agonist bexarotene (Bex), 6OHA treatment (0.1 and 1 μM) resulted in a dose-dependent decrease in the mRNA levels of proinflammatory mediators, including interleukin-1β (Il1b), Il6, and inducible nitric oxide synthase. However, these effects on proinflammatory mediators were effectively abolished by the RXR antagonist UVI3003. Additionally, 6OHA promoted M2 microglia polarization after 24 h of treatment, as evidenced by the increased mRNA levels of the M2 marker genes arginase-1 (Arg1), C-C motif chemokine ligand 6 (Ccl6), Ccl17, and Ccl22. Notably, 6OHA induced a distinct set of M2 microglial markers compared with IL-4, a known M2 microglial inducer. Furthermore, the transcription of Arg1, a key M2 marker gene, is regulated by retinoic acid receptor/RXR heterodimers and the IL-4 signaling pathway. Collectively, 6OHA suppressed the early inflammatory responses to LPS and promoted M2 microglial polarization through a mechanism distinct from that of IL-4. Therefore, RXR agonists, including 6OHA and Bex, may exhibit dual anti-inflammatory effects and serve as novel modulators of neuroinflammation.

Elastic fibers, which contribute to the flexibility of tissues such as the skin, alveoli, and arteries, have a long half-life and are not regenerated once formed during the fetal stage. Consequently, the degradation of elastic fibers due to aging or inflammation can significantly impact tissue function. In the dermis, degeneration of elastic fibers is characterized by degradation in photoaging, driven by UV radiation, and structural abnormalities of elastic fibers in intrinsic aging. However, the mechanisms driving the abnormalities associated with intrinsic aging remain incomplete. This study aimed to identify the factors involved in the elastic fiber abnormalities associated with intrinsic aging of the dermis. Through a comprehensive analysis of gene expression, this study focused on microfibril-associated protein 5 (MFAP5) as a candidate gene responsible for the elastic fiber abnormalities associated with intrinsic aging. Immunofluorescence staining revealed that aged fibroblasts highly expressed MFAP5 and strongly localized it to aggregated elastic fibers. Furthermore, the elimination of MFAP5 expression suppressed elastic fiber aggregation. The exogenous addition of MFAP5 induced thickening and disorganization of elastic fibers, effects that were not observed with the overexpression of MFAP5 in young fibroblasts, which merely express MFAP5. Moreover, MFAP5 inhibited the interaction between latent transforming growth factor β binding protein 4 and fibulin-5, which are crucial for elastic fiber formation. These results suggest that excess MFAP5 expression associated with aging causes abnormalities in elastic fibers. Understanding the role of MFAP5 in elastic fiber abnormalities highlights its potential as a therapeutic target for mitigating intrinsic dermal aging and improving skin elasticity.

One of the side effects of calcineurin inhibitors, such as tacrolimus (FK506) and cyclosporin A (CsA), is intestinal mucosal damage leading to ulceration and bleeding. However, it remains to be elucidated whether these side effects are direct effects of calcineurin inhibitors on intestinal epithelial cells (IECs). To determine whether IECs are directly damaged by calcineurin inhibitors, we analyzed the effects of calcineurin inhibitors on the intestinal barrier in Caco-2 cells, a human intestinal cell line. Treatment of Caco-2 with calcineurin inhibitors such as FK506, CsA, and deltamethrin inhibited expression of zonula occludens-1, a tight junction protein, and increased permeability of Lucifer Yellow. These effects were observed in confluent cells, but not clear in subconfluent cells. Our findings suggest that calcineurin inhibitors can directly damage IECs.

Thyroid hormone receptors (TRs) predominantly consist of three isoforms: TRβ1, TRβ2, and TRα1. Mutations in TRs that reduce or eliminate their ligand-dependent functions lead to a condition known as resistance to thyroid hormone (RTH), which is a genetic disorder caused in an autosomal-dominant manner. Recently, resmetirom, a selective TRβ agonist, has showen significant promise and was approved by the United States Food and Drug Administration as the first drug for the treatment of metabolic dysfunction-associated steatohepatitis and liver fibrosis. In this study, we performed a series of assays to determine how different mutations retain the activity of resmetirom in presumed clinical practice to screen patients based on previously published 15 studies. We generated and examined the transcriptional activation of 57 mutant TRβs, to evaluate the therapeutic potential of resmetirom in treating these mutations. Resmetirom functions as a partial agonist for TRβ and has been found to recruit and interact with all cofactors, albeit significantly less effectively than T3. Consequently, our results suggest that some residues in helices 1, 3, 5, 6, 7, 10, and 11 of TRβ, are still susceptible to resmetirom binding. However, mutants of helix 12, which forms part of the dimerization interface used to bind transcriptional coactivators, are non-responsive to resmetirom. While certain mutants do not respond to lower concentrations of resmetirom, some can be rescued when higher concentrations are applied. In clinical applications, resmetirom can contribute to RTH without considering the syndrome of inappropriate secretion of thyroid-stimulating hormones by the negatively regulated gene.