Kobayashi Pharmaceutical (Japan)

This study investigates how muscle viscoelasticity changes during static stretching by measuring the state of the muscle during stretching using continuous time-series data. We used a device that applied a force to the muscle during stretching and measured the reaction force. The device was attached to each participant, and time-series data of the reaction force (stress-relaxation data) during stretching were obtained. The spring-pot model, which uses fractional calculus, was selected as the viscoelastic model for the muscle, to which the stress relaxation data were fitted on a straight line on a double-logarithmic plot. The stress-relaxation data formed a broken line comprising two segments on the double-logarithmic plot, showing that viscoelasticity changed abruptly at a particular time during static stretching. Considering two viscoelastic states, before and after the change, the stress-relaxation curve was fitted through segmented regression to the double-logarithmic data with high accuracy (R2 = 0.99 and NRSME = 0.0018). We compared the parameters of the spring-pot model before and after the change in muscle viscoelasticity. By examining these continuous time-series data, we also investigated the time taken for the effects of stretching to become apparent. Furthermore, we measured the changes in muscle viscoelasticity during static stretching before and after a short-term exercise load of running on a treadmill.

Osteoarthritis, a degenerative joint disease, requires innovative therapies due to the limited ability of cartilage to regenerate. Since mesenchymal stem cells (MSCs) provide a cell source for chondrogenic cells, we hypothesize that chemicals capable of enhancing the chondrogenic potential of MSCs with transforming growth factor-beta (TGFβ) in vitro may similarly promote chondrogenesis in articular cartilage in vivo. Chemical compounds that enhance the TGFβ signaling for chondrogenesis were investigated utilizing mesenchymal stem cells derived from human induced pluripotent stem cells. The mechanisms of action underlying the identified compound were explored in vitro, and its therapeutic effects were validated in vivo using a mouse model of exercise-induced osteoarthritis. Hydroxycitric acid (HCA) emerged as the lead compound. In vitro, HCA effectively enhanced chondrogenesis by inhibiting ATP citrate lyase, inducing citrate and alpha-ketoglutarate (α-KG), while reducing cytosolic acetyl coenzyme A (Ac-CoA). This induction of α-KG promoted collagen prolyl-4-hydroxylase activity, boosting hydroxyproline production and matrix formation. The reduction of Ac-CoA attenuated the inhibitory effect of β-catenin on mitochondrial activity by diminishing its acetylation. In vivo, orally administered HCA accumulated in joint tissues of mice and histological examination demonstrated newly synthesized cartilage tissues in damaged area. Analysis of joint tissue extracts revealed a downregulation of Ac-CoA and an upregulation of citrate and α-KG, accompanied by a systemic increase in an anabolic biomarker. HCA demonstrates promise as an osteoarthritis therapy by enhancing chondrogenic differentiation. Its ability to modulate crucial metabolic pathways and facilitate cartilage repair suggests potential for clinical translation, addressing a critical need in the treatment of osteoarthritis.

This study searched for volatile organic compounds (VOCs）having antibacterial and antiviral efficacy. The antibacterial efficacy of volatilized components was evaluated and (2E,4E）-2,4-hexadienal, α- angelica lactone, 2-cyclohexen-1-one and 2-cyclopenten-1-one were found to inhibit the formation of colonies of Staphylococcus aureus and Escherichia coli. Evaluating the antimicrobial efficacy of the surfaces to which each VOC adhered to, these four compounds were revealed to have antibacterial efficacy (antibacterial activity value (A-value）against S. aureus; ≧2.63, A-value against Klebsiella pneumoniae; >5.07, A-value against E. coli; ≧2.17）. Furthermore, (2E,4E）-2,4-hexadienal and α-angelica lactone were found to have antiviral efficacy against Influenza A virus (H1N1）and Feline calicivirus on the cotton cloths to which it adheres to (antiviral activity value (R-value）against Influenza A virus; >2.94, R-value against Feline calicivirus; ≧2.31）. Using these components, it might be possible to develop antimicrobial products that exhibit antibacterial and antiviral efficacy.

To investigate the main symptoms of female sexual dysfunction (FSD) and lower urinary tract symptoms associated with vulvovaginal atrophy (VVA) symptoms as the core symptoms of genitourinary syndrome of menopause. We extracted the data of 4134 Japanese women aged 40-79 years who participated in the GENitourinary syndrome of menopause in JApanese women (GENJA) study. All participants responded to web-based questionnaires assessing their health situation, including the Vulvovaginal Symptoms Questionnaire, the Female Sexual Function Index (FSFI), and the Core Lower Urinary Tract Symptom Score. Multivariable regression and multivariable logistic regression analyses were applied to analyze the association between VVA symptoms and FSD, and between VVA symptoms and lower urinary tract symptoms. Multivariable regression analysis revealed that VVA symptoms were associated with lower scores for arousal, lubrication, orgasm, satisfaction, and pain domains in the FSFI in sexually active women (p < 0.01). Regression coefficients were higher for lubrication and pain domains than for the other domains. Multivariable logistic regression analysis revealed that women reporting VVA symptoms were more likely to have increased daytime urinary frequency, nocturia, urgency, slow stream, straining to void, feeling of incomplete emptying, bladder pain, and feeling a bulge/lump from or in the vagina (p < 0.05). Adjusted odds ratios were particularly high for straining to void, feeling of incomplete emptying, and bladder pain. Vulvovaginal atrophy symptoms were significantly associated with decreased lubrication and dyspareunia in FSD, and urinary symptoms of straining to void, feeling of incomplete emptying, and bladder pain.

紅麹菌（Monascus）を米に生育させた紅麹米は，世界中で食品として利用されている．近年は天然色素としての着色料だけではなく，血圧低下作用やコレステロール低下効果など機能性食品としての利用も高まっている．紅麹菌はヒトの健康に良い効果をもたらすモナコリンKやアザフィロン色素を生産する一方で，健康を害すると考えられるカビ毒シトリニンも作る．したがって，安全に食品として利用できるカビ毒シトリニンをつくらない紅麹菌の産業利用が望まれる．我々は，紅麹3菌種について次世代シークエンサーを用いたゲノム解析とLCMSを用いた二次代謝物解析を行った．その結果，ゲノムレベルと代謝物レベルでカビ毒シトリニンを生産しない紅麹菌株としてMonascus pilosus NBRC 4520とMonascus ruber NBRC 4483を見出した．また，Monascus ruberについてはシトリニンを生産することが報告されているが，Monascus pilosusについてはゲノムレベルでシトリニンの生合成遺伝子が保存されていないことが複数の菌株で報告されている．したがって，食品として利用するための紅麹菌種として安全性が最も高いのはMonascus pilosusであると考えられる．これらの研究知見が，安全な紅麹菌の研究を通じて食品への利用，世界中のヒトの健康の維持増進へ貢献する端緒となれば幸いである．

Red yeast rice fermented by Monascus contains secondary metabolites such as monacolin K and azaphilone pigments. To date, no information on the localization of these secondary metabolites in red yeast rice has been reported. In this study, the localization of monacolin K, monascin, and rubropunctamine, produced by Monascus pilosus, was revealed through the use of matrix-assisted laser desorption/ionization–mass spectrometry imaging (MALDI-MSI). Monacolin K was visualized as being distributed throughout the rice during the fermentation process. By contrast, monascin and rubropunctamine were found to be localized near the surface of the rice. In addition, high-resolution imaging revealed that monacolin K was localized in the ascocarp of Monascus. This is the first such study to reveal the localization of secondary metabolites in red yeast rice during fermentation using MALDI-MSI. These findings will enhance our understanding of metabolite production in fungi.

The skin loses its moisture with advancing age, causing cosmetic issues such as wrinkles. In addition, the loss of moisture leads to hypersensitivity to external stimuli such as UV light. Transcutaneous supplementation with hyaluronic acid (HA) is an effective and safe method of recovering the moisturizing function and elasticity of the skin. However, the transcutaneous delivery of HA remains challenging owing to the barrier function of the stratum corneum (SC) layer. To penetrate the SC barrier, we used a reverse micelle formulation that does not require high energy consumption processes for preparation. We aimed to enhance the skin permeability of HA by incorporating glyceryl monooleate—a skin permeation enhancer—into the formulation. A fluorescently-labeled HA-loaded reverse micelle formulation showed significantly enhanced permeation across Yucatan micro pig skin. Fourier transform infra-red spectroscopy of the surface of the skin treated with the reverse micelle formulation showed blue shifts of the CH2 symmetric/asymmetric stretching peaks, indicating a reduction in the barrier function of the SC. Further study revealed that HA was released from the reverse micelles at the hydrophobic/hydrophilic interface between the SC and the living epidermis. The results demonstrated that our reverse micellar system is an easy-to-prepare formulation for the effective transcutaneous delivery of HA.