Ohu University

Discrimination of cancer cells from normal growing cells is crucial to specifically target cancer cells. The transcription factor E2F1 is the principal target of the tumor suppressor pRB. E2F1 activated by growth stimulation activates cell cycle-related genes and facilitates cell proliferation. E2F1 activated by loss of pRB control, such as forced inactivation of pRB, activates tumor suppressor genes such as ARF and TAp73 and induces apoptosis. We show here that these genes are specifically activated by exogenously expressed E2F1 or forced inactivation of pRB but not by growth stimulation in epithelial cells. This observation indicates that E2F1 activity induced by forced inactivation of pRB contains distinct E2F1 activity that activates these tumor suppressor genes. Cancer cells survive with concomitant dysfunction of apoptosis-inducing pathways, suggesting the presence of distinct E2F1 activity specifically in cancer cells. We determined the presence of distinct E2F1 activity using E2F responsive elements of the ARF and TAp73 genes by reporter assay. All 33 cancer cell lines tested possessed distinct E2F1 activity, but five normal growing cell lines did not. These results indicate that distinct E2F1 activity is a unique characteristic of cancer cells that facilitates discrimination from normal growing cells to specifically target cancer cells.

Various methods have been investigated for constructing UAV routes using ad hoc network protocols implemented on wireless devices, such as residential smart meters. In our previous study, we proposed routereservation methods to prevent link conflicts when many UAVs operate simultaneously. To reduce route construction failures caused by reserved links, we introduced hierarchical link layer approaches. However, these methods did not sufficiently reduce route construction failures. In this study, we propose a method to dynamically configure the number of link layers based on real-time UAV traffic demand. This method utilizes the Erlang B formula from circuit-switching theory to determine the optimal number of layers. These methods aim to reduce route construction failures by optimizing the number of layers. We evaluate the effectiveness of the proposed methods through computer simulations.

In 2011, the Great East Japan Earthquake hit the Futaba District on the northeast coast of Japan, followed by a tsunami and a nuclear power plant accident. In this study, we investigated the impact of post-earthquake life on the onset of chronic kidney disease (CKD) among the residents of the Futaba District. Data on 17,859 residents of the Futaba District (7333 men, 10,526 women; mean age: 61.0 ± 10.2years; mean follow-up period: 3.42 ± 1.51years) who underwent health checkups and completed self-administered questionnaires in the Fukushima Mental Health and Lifestyle Survey were analyzed. These residents were confirmed to be CKD-free in 2012. Hence, they were assessed for the onset of CKD from 2013 to 2017. Univariate analysis results showed significant differences between residents with and without CKD. Differences in age, diabetes mellitus, body mass index (BMI), dyslipidemia, hypertension, hyperuricemia, Kessler 6 Psychological Distress Scale (K6) score, smoking habit, alcohol drinking history, exercise habit, history of job change, history of job loss, and evacuation experience were observed. Multivariate analysis was conducted to adjust for multiple factors, and age, BMI, dyslipidemia, hypertension, hyperuricemia, and K6 score were identified as significant promotional factors for CKD onset. Among the well-recognized risk factors, severe stress reflected by a high K6 score was established to be correlated with CKD onset among residents originally without CKD. Stress management may be another treatment strategy for treating CKD.

In this study, we evaluated the preparation conditions, stability, and durability of 2,2,6,6‐tetramethylpiperidine 1‐oxyl (TEMPO)‐modified glassy carbon (GC) electrodes prepared using electrolytic polymerization in an aqueous solvent, and examined the applicability of the electrodes as electrochemical sensors for pharmaceutical analysis. First, we synthesized a TEMPO‐substituted phenol (2‐PH‐TEMPO) and examined the conditions for its efficient immobilization on an electrode using the electrolytic polymerization method in an aqueous solvent. The TEMPO‐modified GC electrode showed high stability and durability in a low‐concentration substrate environment and effectively functioned as an electrolytic oxidation catalyst for ethanol, a compound with a hydroxyl group. In an analysis of pharmaceuticals, a high oxidative response was obtained for lidocaine, clarithromycin, and azithromycin, which have tertiary amines or multiple oxidizable hydroxyl groups, suggesting that the electrode can function as a highly sensitive electrochemical sensor. The use of this modified electrode enables quantification in the clinical concentration range of vancomycin, and it is expected to be applied to rapid and accurate pharmaceutical concentration measurements in therapeutic drug monitoring. These results suggest that TEMPO‐modified GC electrodes are useful as stable and sensitive electrochemical sensors in the fields of pharmaceutical analysis and biosensors, and are expected to contribute to the development of quantitative electrochemical methods.

ObjectiveTo clarify how the preoperatively determined position of the Cupid's bow peak on the lateral lip element affects postoperative nasolabial formation in patients with unilateral cleft lip. We hypothesize that setting the proposed peak of the Cupid's bow close to the nostril floor causes the cleft-side nasal alar base to droop while positioning it near the oral commissure shortens the red lip, causing an elevated cleft-side oral commissure.DesignRetrospective study.SettingSingle institution.PatientsTwenty-seven patients with non-syndromic complete unilateral cleft lip and palate (UCLP) who received primary lip plasty at our department.InterventionsPrimary lip repair for patients with UCLP.Main outcome measuresWe tested our hypothesis using three-dimensional facial images taken before and 1 year after primary lip repair. Bilateral measurements included lip height (distance between the nasal alar base and the Cupid's bow peak), red lip length, and the vertical height of the nasal alar base and oral commissure.ResultsCorrelation analysis showed a significant relationship between preoperative cleft-side lip height and the postoperative position of the cleft-side nasal alar base (P = .0024, Pearson's correlation analysis). Preoperative short red lip remained short afterwards (P = .000036, Pearson's correlation analysis), causing a lip cant with an elevated cleft-side oral commissure (P = .012, Pearson's correlation analysis).ConclusionsPostoperative nasolabial formation is influenced by the preoperatively proposed position of the Cupid's bow peak. This study provides a comprehensive principle explaining the relationship between the location of the proposed Cupid's bow peak on the lateral lip elements and postoperative nasolabial formation in primary lip repair in patients with unilateral cleft lip.

Abstract Purpose Topical local anesthetics have become increasingly important with the recent development of laser therapy. Lidocaine, the most commonly used anesthetic in clinical practice, not only has analgesic effects but also possesses properties such as suppression of neonatal seizures and antiarrhythmic, anti-inflammatory, antibacterial, and antifungal effects. This study aimed to improve the solubility and permeability of lidocaine (Lid) by preparing solid dispersions of Lid and ascorbic acid 2-glucoside (AG) and evaluating their physicochemical properties, solubility, and permeability. Methods Here, we evaluated the physicochemical properties and solubility of solid dispersions of Lid and AG prepared by freeze-drying (freeze-dried [FD] Lid/SSA = 1/1). Results For Differential scanning calorimetry measurements revealed the disappearance of the endothermic peaks associated with the melting of Lid and AG in the freeze-dried formulation (FD) (Lid/AG = 1/1), with the emergence of a new glass transition point. Powder X-ray diffraction measurements confirmed the absence of the characteristic peaks of both Lid and AG in the FD (Lid/AG = 1/1), showing a halo pattern. Near-infrared measurements indicated peak shifts and broadening of the -CH and -NH groups of Lid and the -OH and -CH groups of AG in the FD (Lid/AG = 1/1), suggesting the involvement of complex formation. Solubility tests revealed that the solubility of FD (Lid/AG = 1/1) was approximately 220 mg/mL, which is approximately 50 times higher than that of Lid (approximately 4.0 mg/mL). Nuclear Overhauser effect spectroscopy nuclear magnetic resonance (NMR) measurements revealed cross-peaks between the -CH groups of Lid and the -OH groups of AG, suggesting intermolecular interactions via hydrogen bonding. Diffusion-ordered spectroscopy NMR measurements showed that the diffusion coefficients of Lid and AG in the FD (Lid/AG = 1/1) converged, indicating that the formation of a complex with AG altered the dispersion behavior of Lid in the solvent. In the silicon membrane permeability test, the cumulative amount permeated after 24 h was approximately 700 µg/cm 2 for Lid, compared to approximately 4.0 µg/cm 2 for FD (Lid/AG = 1/1), suggesting that membrane permeability was inhibited. Conclusion These findings suggest that complex formation occurred in FD Lid/ AG; this enhanced the solubility, membrane permeability was inhibited of this dispersion. Graphical Abstract

Background: There are few data verifying the utility of the CHADS-P2A2RC score in comparison with the CHADS2 score for estimating net adverse clinical events (NACE) in real-world settings. Methods: We performed analysis for a total of 3,985 chronic coronary syndrome (CCS) patients without atrial fibrillation (AF) who underwent percutaneous coronary intervention (PCI) between April 2013 and March 2019 for whom information was obtained from the CLIDAS (Clinical Deep Data Accumulation System)-PCI database. The primary endpoint was NACE defined as the composite of 3-point major adverse cardiovascular events (3P-MACE) (cardiovascular death, non-fatal myocardial infarction, and non-fatal stroke) and GUSTO moderate/severe bleeding events. Results: Kaplan-Meier analysis showed that both the CHADS-P2A2RC and CHADS2 scores stratified the risks. The incidences of NACE were stratified well by the very-high-risk category, which was uniquely defined as a CHADS-P2A2RC score of ≥6 (hazard ratio: 2.38, 95% CI=1.91-2.97, p-value <0.001). The area under the curve (AUC) in estimating NACE within 3 years was higher when the CHADS-P2A2RC score was used than when the CHADS2 score was used (0.67 vs. 0.62, p=0.003). This was mainly due to the accuracy in estimating bleeding events (0.66 vs. 0.60, p=0.006). Conclusions: The accuracy in estimating NACE after PCI for CCS patients without AF was higher when the CHADS-P2A2RC score was used than when the CHADS2 score was used, mainly due to the accuracy in predicting bleeding risk. Higher incidences of endpoints were well-stratified by a very-high-risk category defined as a CHADS-P2A2RC score of ≥6.

Programmed death 1 ligand (PD-L1), an important immune checkpoint molecule, is mainly expressed on cancer cells and has been shown to exert an immunosuppressive effect on T-cell function by binding to programmed cell death 1 (PD-1) expressed on T-cells. Recently, immune checkpoint inhibitors using antibody drugs such as nivolumab and atezolizumab have attracted attention. However, clinical challenges, including limitations to the scope of their application, are yet to be addressed. In this study, we developed a novel immune checkpoint inhibitor that targets PD-L1 using lipid-siRNA conjugates (lipid-siPDL1s). The inhibitory effect of lipid-siPDL1s on PD-L1 expression was evaluated and found to strongly suppress mRNA expression. Notably, lipid-siPDL1s exerted a significantly stronger effect than unmodified siPDL1. Interestingly, lipid-siPDL1s strongly inhibited PD-L1 expression despite cancer cell stimulation by interferon-gamma, which induced the overexpression of PD-L1 genes. These results strongly suggest that lipid-siPDL1s could be used as novel immune checkpoint inhibitors.

ABSTRACT Objectives Neuropathic pain is characterized by long-lasting, intractable pain. Sciatic nerve ligation is often used as an animal model of neuropathic pain, and the spared nerve injury (SNI) model, in which the common peroneal nerve (CPN) and tibial nerve (TN) are ligated, is widely used. In the present study, we evaluated the analgesic effect of a cholinergic agonist, carbachol, on a neuropathic pain model prepared by sural nerve (SN) ligation in mice. Methods The SN was tightly ligated as a branch of the sciatic nerve. Mechanical and thermal allodynia, and hyperalgesia were assessed using von Frey filaments and heat from a hot plate. The analgesic effects of intracerebroventricularly-administered morphine and carbachol were compared. Results SN ligation resulted in a significant decrease in pain threshold for mechanical stimulation 1 day after ligation. In response to thermal stimulation, allodynia was observed at 50°C and hyperalgesia at 53 and 56°C 3 days after ligation. Content of thiobarbituric acid reactive substances (TBARS) in the spinal cord increased significantly at 6 and 12 h after ligation. Acetylcholine content of the spinal cord also increased at 5 and 7 days after ligation. Intracerebroventricular administration of carbachol at 7 days after ligation produced a marked analgesic effect against mechanical and thermal stimuli, which was stronger and longer-lasting than morphine at all experimental time points. Conclusion These findings suggest that cholinergic nerves are involved in allodynia and hyperalgesia of the SN ligation neuropathic pain model.

Objectives: Perennial allergic rhinitis (PAR) is common in Japan. Second-generation antihistamines (SGAs) are commonly used for its treatment; however, it remains unclear which SGA is the most cost-effective. Additionally, the pharmacoeconomics of Japanese Kampo shoseiryuto (which was traditionally prescribed to treat PAR in Japan) remains poorly understood. In this study, we aimed to investigate the effectiveness of various SGAs and shoseiryuto for the treatment of PAR in Japanese outpatients, from the healthcare payer's perspective. Methods: The most cost- and clinically effective SGAs were determined from a list of 6 SGAs (bepotastine, 10 mg; cetirizine, 10 mg; ebastine, 10 mg; epinastine, 20 mg; loratadine, 10 mg; and olopatadine, 5 mg) together with shoseiryuto, using the overall improvement rate through a model-based analysis. The time horizon was 28 days. Costs were determined based on the Medical Fee Index in 2020. Deterministic and probabilistic sensitivity analyses were conducted to address the uncertainty of the base-case results. Results: Overall, bepotastine (10 mg) and ebastine (10 mg) were cost-effective. Shoseiryuto was less cost-effective than ebastine (10 mg) (dominated). Ebastine (10 mg) was the most cost-effective option based on deterministic and probabilistic sensitivity analyses. Conclusions: Ebastine (10 mg) was the most cost-effective treatment strategy for PAR among the agents evaluated in this study. This insight could aid in establishing an appropriate formulary for treating PAR in hospitals and communities.