National Institute of Health Sciences

Based on a comparative study on the morphological characteristics of over sixty specimens, Capila pauripunetata tamdaoensis Devyatkin, 1996 syn. nov. is proposed to be a junior subjective synonym of C. pauripunetata Chou & Gu, 1994. Genitalia of both sexes of C. pauripunetata from Hainan are described. Information on the distribution of this species is discussed.

Fab has been extensively studied to elucidate the structural and physicochemical principles underlying antigen recognition. While most investigations have focused on IgG-derived Fabs, comparatively little is known about those from other isotypes. To investigate the role of the IgM constant domain Cμ1, we engineered a chimeric Fab (Cμ1Fab) in which the CH1 domain of adalimumab Fab (IgG1) was replaced by the human IgM Cμ1 domain. Cμ1Fab was expressed in CHO cells and its physicochemical properties were evaluated. SDS-PAGE analysis with or without PNGase F treatment demonstrated that the N166-N167-S168 motif was N-glycosylated. SPR analysis demonstrated that Cμ1Fab retained antigen-binding activity comparable to that of the wild-type Fab, and elimination of the glycosylation motif (N166A) did not alter binding affinity. Notably, N-linked glycosylation within the Cμ1 domain suppressed aggregation, whereas the N166A variant showed marked aggregation. DSC further revealed that the glycosylation affected thermal stability. The melting temperature (Tm) of the N166A variant was 60.5°C, substantially lower than that of the Cμ1Fab (Tm = 66.0°C), although Tm of the wild-type Fab was 75.2°C. Collectively, these findings establish the critical role of native Cμ1 N-linked glycosylation in Fab stability, offering new insights for antibody engineering.

Ziziphi Fructus (the fruit of Ziziphus jujuba Miller var. inermis Rehder; Japanese name "Taiso"), listed in the 18th edition of the Japanese Pharmacopoeia (JP18), is widely used in Kampo (Japanese traditional) medicine. However, a specific identification test for Ziziphi Fructus was not prescribed in JP18. Hence, in this study, we establish a TLC-based identification test for quality assurance of the crude drug Ziziphi Fructus. The HPLC experiments identified 3-O-cis-p-coumaroylalphitolic acid and 3-O-trans-p-coumaroylaliphitolic acid as suitable indicators. After developing the samples to a distance of 10 cm using ethyl acetate (EtOAc)/n-hexane/acetic acid (AcOH)/H2O (60 : 40 : 2 : 1) as the TLC-developing solvent and UV light at 254 nm, two spots with an Rf value of approximately 0.4-0.6 were detected. Additionally, based on the TLC-based experiments, alphitolic and maslinic acids emerged as potential indicator components. Treatment of samples with EtOAc/n-hexane/AcOH (60 : 40 : 1) serving as the developing solvent, followed by exposure to dilute sulfuric acid solution, heat, and UV light at 365 nm, revealed two clear spots with Rf values of approximately 0.2-0.3. These findings potentially serve as a basis for the development of an identification test to identify Ziziphi Fructus.

The increasing spread of toxin-producing Gambierdiscus dinoflagellates has led to cases of ciguatera poisoning in previously nonendemic regions. Ciguatoxins (CTXs) are potent, heat-stable compounds responsible for ciguatera, and their rapid and reliable analysis in seafood is essential to prevent illness. This study presents the development of the first lateral flow assay (LFA) for the detection of major Pacific CTXs congeners. The assay employs previously developed monoclonal antibodies in a sandwich format: specific antibodies against CTX1B and CTX3C are immobilized on a single test line, while the reporter antibody is a single antibody recognizing major Pacific CTX congeners that is conjugated to gold nanoparticles. The LFA provides one-step visual results in 30 min, making it suitable for resource-limited settings. It exhibits a visual limit of detection of 400 pg/mL and the assay is highly specific, showing no cross-reactivity with other marine toxins. It successfully detected Pacific CTXs spiked in noncontaminated fish extracts, as well as in naturally contaminated samples as previously confirmed by LC-MS/MS, with an estimated cutoff of approximately 0.1 μg/kg. Overall, this LFA demonstrates great potential as a rapid screening tool for outbreak investigations and research programs.

The tumor microenvironment (TME) not only influences cancer progression but also has been shown to have a significant effect on the prognosis. One of the major TME components is cancer-associated fibroblasts (CAFs), of which several subtypes have been identified in tumor tissues. Although single-cell RNA sequencing (scRNA-seq) technology is a powerful tool for investigating the proportions and composition of cells in tissues, large-scale datasets are needed to analyze small cell populations. Here, we constructed an integrated scRNA-seq dataset for non-small cell lung cancer (NSCLC) by compiling publicly available data and observed differences in TME heterogeneity and cellular composition between lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC). In particular, we identified significant differences in CAF subtypes between LUAD and LUSC. Inflammatory CAFs (iCAFs) were predominantly found in LUAD, whereas myofibroblastic CAFs (mCAFs) were more common in LUSC. A coculture analysis of lung fibroblasts confirmed that LUAD cells induced the iCAF phenotype and, in contrast, that LUSC cells promoted myofibroblastic differentiation. A correlation analysis with the prognosis identified mCAFs as poor prognostic factors in both LUAD and LUSC, but iCAFs were found to be a poor prognostic factor only in LUSC, with the opposite pattern observed in LUAD. This work highlights important considerations regarding the CAF subtype dominance in LUAD and LUSC, which may be related to the prognosis.

Trimeric intracellular cation channel A (TRIC-A) provides counter-ion support for sarcoplasmic reticulum (SR) Ca2+ release, yet its physiological role in the intact heart under stress remains poorly defined. Here, we demonstrate that TRIC-A is essential for maintaining balanced SR Ca2+ release, mitochondrial integrity, and cardiac resilience during β-adrenergic stimulation. Tric-a-/- cardiomyocytes exhibited Ca2+ transients evoked by electrical stimuli and exaggerated isoproterenol (ISO)-evoked Ca2+ release, consistent with SR Ca2+ overload. These defects were accompanied by selective upregulation of protein kinase A (PKA)-dependent phosphorylation of ryanodine receptor 2 (RyR2) (S2808) and phospholamban (PLB) (S16). Acute ISO challenge induced mitochondrial swelling, cristae disruption, and Evans Blue Dye uptake, and elevated circulating troponin T in Tric-a-/- hearts, hallmarks of necrosis-like cell death. Mitochondrial Ca2+ uptake inhibition with Ru360 markedly reduced membrane injury, establishing mitochondrial Ca2+ overload as the proximal trigger of cardiac cell death. With sustained β-adrenergic stimulation by ISO, Tric-a-/- hearts developed extensive interstitial and perivascular fibrosis without exaggerated hypertrophy. Cardiac fibroblasts lacked TRIC-A expression and displayed normal Ca2+ signaling and activation, indicating that fibrosis arises secondarily from cardiomyocyte injury rather than fibroblast-intrinsic abnormalities. These findings identify TRIC-A as a critical regulator of SR-mitochondrial Ca2+ coupling and a key molecular safeguard that protects the heart from catecholamine-induced injury and maladaptive remodeling.

Acrylamide (AA) is a neurotoxic and potentially carcinogenic compound whose detection in processed foods and food additives has raised significant public health concerns, underscoring the urgent need for sensitive yet straightforward analytical methods. Conventional analytical approaches based on GC or HPLC have limited applicability in case of AA due to its high polarity and low molecular weight. Therefore, various derivatization strategies have been explored; however, challenges remain in terms of selectivity. Herein, we report a novel HPLC-FLD method for AA analysis using 5-(dimethylamino)-N-(2-sulfanylethyl)naphthalene-1-sulfonamide (DNS-SH) as a fluorescent derivatization reagent. DNS-SH was selected to provide moderate hydrophobicity for reversed-phase retention, long-wavelength fluorescence distinct from that of food components, and a reactive thiol group for the selective derivatization of AA. The AA-DNS derivative was structurally confirmed by LC-HRMS, 1D-NMR, and 2D-NMR. The developed HPLC-FLD method exhibited excellent linearity (r² > 0.9999), reproducibility (RSD < 2.76 %), and a low LOQ of 0.1 µg/mL. Its applicability was validated using an “Amino Acid-Monosaccharide Reaction Product,” where AA was detected at 0.1 µg/g in spiked samples. This approach provides a reliable tool for monitoring AA in food additives and potentially broader food matrices.

Abstract Commercially available disposable products such as cell culture utensils and catheters do not necessarily possess sufficient quality for in vitro fertilization (IVF), from the perspective of pyrogen contamination. We aimed to comprehensively analyze the pyrogen contamination status, including bacterial endotoxins, of the products used for IVF in assisted reproductive technology (ART) and improve their cleanliness using a new sterilization technology. Pyrogen contamination levels were evaluated using a direct human cell-based pyrogen test that is not affected by the recovery ratio, unlike bacterial endotoxin tests. Pyrogen inactivation tests were performed using low-temperature ozone/hydrogen peroxide gas treatment. The residual hydrogen peroxide was colorimetrically quantified, and the effectiveness of its removal by drying treatment was evaluated using germ cell viability as an indicator. Significant amounts of pyrogen, from 0.014 to 1.110 EU/product, were detected in seven of the twenty products. Pyrogen contamination levels were reduced below the detection limit by ozone/hydrogen peroxide gas sterilization. Hydrogen peroxide remained on the surface of the GPS dish but was reduced to a level that did not affect human sperm viability and embryo development after drying at 80°C for 24 h following sterilization. These products may carry a potential risk of reducing ART success rates, and pyrogen contamination levels may exceed the previously reported allowable level of 0.01–0.02 EU/mL in human IVF during actual use. Our study suggests that the manufacturing of products free from pyrogens and without adverse effects on germ cells is possible using ozone/hydrogen peroxide gas sterilization and subsequent drying technologies.

Background: Plantar fasciitis leading cause of heel pain, frequently affects individuals engaged in prolonged standing, such as salespersons. The interplay between intrinsic foot muscle (IFM) strength and ankle foot dorsiflexion is critical in maintaining plantar fascia integrity and reducing mechanical overload. Objective: A review on the role of IFM strength and ankle dorsiflexion in plantar fascia health, emphasizing occupational implications for salespersons. Methods: A narrative review after studying databases including PubMed, Scopus and Google Scholar (2020–2025). Studies examining the relationship between IFM function, ankle dorsiflexion, and plantar fasciopathy were analyzed. Results: Limited dorsiflexion and weak intrinsic musculature increase plantar fascia strain via altered arch mechanics and early heel-off during gait. IFM-strengthening exercises and dorsiflexion mobility programs significantly improve symptoms, posture, and foot function. Salespersons exhibit elevated plantar fasciitis prevalence due to prolonged standing and cumulative load. Conclusion: Optimal plantar fascia health relies on adequate IFM strength and ankle mobility. Integrating strengthening, stretching, and ergonomic interventions offers preventive and therapeutic benefits for salespersons.