An infectivity-enhancing site on the SARS-CoV-2 spike protein targeted by antibodies.

Antibodies against the receptor-binding domain (RBD) of the SARS-CoV-2 spike protein prevent SARS-CoV-2 infection. However, the effects of antibodies against other spike protein domains are largely unknown. Here, we screened a series of anti-spike monoclonal antibodies from coronavirus disease 2019 (COVID-19) patients and found that some of antibodies against the N-terminal domain (NTD) induced the open conformation of RBD and thus enhanced the binding capacity of the spike protein to ACE2 and infectivity of SARS-CoV-2. Mutational analysis revealed that all of the infectivity-enhancing antibodies recognized a specific site on the NTD. Structural analysis demonstrated that all infectivity-enhancing antibodies bound to NTD in a similar manner. The antibodies against this infectivity-enhancing site were detected at high levels in severe patients. Moreover, we identified antibodies against the infectivity-enhancing site in uninfected donors, albeit at a lower frequency. These findings demonstrate that not only neutralizing antibodies but also enhancing antibodies are produced during SARS-CoV-2 infection.

Deep heat therapy system with resonant cavity applicator for articular cartilage in knee osteoarthritis.

[Purpose] Heat can prevent cartilage degeneration when applied to articular cartilage, but the size of the human knee joint makes it difficult to target cartilage during heat treatment. In this study, we aimed to establish a heat therapy method capable of safely applying heat to deep intra-articular tissues utilizing a resonant cavity applicator and to confirm the extent of cartilage heating in the human knee when using this system. [Participants and Methods] Heating experiments were carried out on the knees of healthy three volunteers using a resonant cavity applicator and a microwave diathermy system. After heat application, temperature distributions inside the knee were measured noninvasively using our measurement method based on ultrasound imaging techniques. [Results] We observed an increase in the temperature around the cartilage tissue in the knees of the volunteers using an ultrasonic thermometer; there was no increase in temperature in the overlying layers. During heating with up to 20 W of power, none of the volunteers experienced adverse reactions. [Conclusion] This study indicates the potential safety and effectiveness of the resonant cavity heat therapy system for knee osteoarthritis in a clinical setting.

A pilot study comparing the efficacy of radiofrequency and microwave diathermy in combination with intra-articular injection of hyaluronic acid in knee osteoarthritis.

[Purpose] This study aimed to compare the efficacy of radiofrequency diathermy with that of microwave diathermy in combination with intra-articular injection of hyaluronic acid into the knee of patients with osteoarthritis (OA). [Subjects] A total of 17 patients with knee OA were enrolled. The participants were randomly divided into two groups: a radiofrequency diathermy group (RF group, 9 subjects), and a microwave diathermy group (MW group, 8 subjects). [Methods] Subjects received radiofrequency or microwave thermal therapy 3 times at 1-week intervals. Intra-articular injection of hyaluronic acid was administered 10 min before every thermal therapy session. The outcome was evaluated using the Japan Orthopaedic Association (JOA) and the Lequesne Index (LI) at baseline, at weeks 1 (1 week after the first thermal therapy) and 3 (1 week after the last thermal therapy). [Results] The JOA scale increased significantly after three sessions of thermal therapy in the RF group, while no significant increase was observed in the MW group. LI decreased significantly after 3 weeks in the RF group. In the MW group, there was no significant difference in LI between the two time points. [Conclusion] This study revealed that symptom relief in patients with knee OA was greater with radiofrequency diathermy than with microwave diathermy with concurrent use of hyaluronic acid injection, presumably due to the different heating characteristics of the two methods.

Efficacy of a Deep Thermal Therapy System for Osteoarthritis of the Knee.

BACKGROUND: This study sought to assess the efficacy of a deep-tissue thermal therapy system with a resonant cavity applicator (DTT-RCA), which safely heats deep joint tissue for treating osteoarthritis (OA) of the knee. METHODS: Two groups of participants were recruited. The DTT-RCA group comprised 20 knees. Kellgren-Lawrence (K-L) grade was I and II in 8 knees (DTT-RCA I/II group) [mean age 73.3 years (standard deviation 11.4) ], III and VI in 12 knees (DTT-RCA III/IV group) [75.4 (8.6) years]. The control group comprised 13 knees [68.2 (10.8) years]. K-L grade was I in 7 knees and II in 6 knees. This group received exercise therapy. The DTT-RCA I/II group and the control group were imaged by MRI T2 mapping at baseline and 6 months to determine the area of cartilage degeneration. RESULTS: Visual Analogue Scale improved only in the DTT-RCA I/II post-intervention (p < 0.01). Japanese Orthopedic Association knee rating scores (DTT-RCA I/II: p < 0.01, control group: p < 0.01), the Japanese Knee Osteoarthritis Measure (DTT-RCA I/II: p < 0.05, control: p < 0.01), and the Knee injury and Osteoarthritis Outcome Score (DTT-RCA I/II: p < 0.01, DTT-RCA III/IV: p < 0.05, control: p < 0.01) post-intervention. The magnitude of change did not differ significantly between the three groups. The area of cartilage degeneration did not change significantly post-intervention in the DTT-RCA I/II group, not even relative to the control group. CONCLUSIONS: This was the first study to test a DTT-RCA system in patients with knee OA. The system reduced the clinical symptoms of knee OA and could potentially be effective for conservative therapy.

Validation of Noninvasive Ultrasound Temperature Measurement System Through Experiments Using Resonant Cavity Applicator.

We developed a resonant cavity applicator for noninvasive deep hyperthermia treatment of osteoarthritis. In previous studies, we confirmed its viability by heating agar phantoms and conducting computer simulations. To evaluate the heating performance of this system on human subjects, it is necessary to develop a new noninvasive temperature measurement system. In this study, we developed a new temperature measurement system utilizing ultrasound imaging techniques. After heating an agar phantom with our resonant cavity applicator, temperature measurements from both our custom-made noninvasive ultrasonic thermometer and an optical fiber thermometer were collected and compared. Both temperature distributions showed a reliable trend, with heat energy concentrated at the center of the agar phantom. Average error rates were less than 13% for temperatures. The results of this study describe the viability of the temperature measurement system and the resonant cavity applicator for clinical applications.

ME3738 protects against lithocholic acid-induced hepatotoxicity, which is associated with enhancement of biliary bile acid and cholesterol output.

ME3738 (22beta-methoxyolean-12-ene-3beta, 24(4beta)-diol), a derivative of soyasapogenol, attenuates liver disease in several models of chronic liver inflammation. In the present study, we have investigated a protective effect of ME3738 in a typical bile acid-induced cholestatic liver model, lithocholate (LCA) feeding mouse. Co-administration of ME3738 resulted in decreases in plasma alanine aminotransferase (ALT) and alkaline phosphatase (ALP) activities and hepatic bile acid level, and increases in biliary outputs of bile acid and cholesterol, as compared with the results in mice treated with LCA alone. LCA sulfation by hydroxysteroid sulfotransferase 2a and hydroxylation have been reported to be involved in protection against LCA-induced hepatotoxicity. ME3738-treatment, however, had no clear influence on the hydroxysteroid sulfotransferase 2a protein level and LCA 6alpha-, 6beta- and 7alpha-hydroxylase activities, but increased biliary cholesterol output. Cholate (CA)-treatment has been shown to induce hepatotoxicity in farnesoid X receptor-null mice, which is scarcely dependent on bile acid sulfation and hydroxylation but associated with decreased biliary bile acid output. Co-administration of ME3738 decreased the ALT and ALP activities and hepatic bile acid level, and increased biliary outputs of bile acid and cholesterol in farnesoid X receptor-null mice, as compared with the results in the mice treated with CA. Moreover, a clear correlation between biliary outputs of cholesterol and bile acid was observed in these two bile acid-induced hepatotoxicity mouse models. These results suggest that ME3738 protects against bile acid-induced hepatotoxicity through increased biliary bile acid output that is not related to bile acid metabolism but associated with cholesterol output.

Novel transgenic rat for in vivo genotoxicity assays using 6-thioguanine and Spi- selection.

Transgenic rodents are valuable models for investigating the genotoxicity of chemicals in vivo. Here, we report the establishment of a novel transgenic rat for genotoxicity analysis. In this model, about 10 copies of lambdaEG10 DNA carrying the gpt gene of E. coli and the red/gam genes of lambda phage are integrated per haploid genome of Sprague-Dawley rats at position 4q24-q31. After recovery of lambdaEG10 phage, point mutations in the gpt gene and deletions in the red/gam genes are identified by 6-thioguanine and Spi(-) selection, respectively. To examine the suitability of these rats for performing in vivo mutagenicity assays, rats were treated with single intraperitoneal injections of ethylnitrosourea (ENU; 100 mg/kg) or benzo[a]pyrene (B[a]P; 62.5 and 125 mg/kg), and the mutant frequencies (MFs) in the liver were determined 7 days after the treatment. ENU enhanced the gpt MF about 7-fold over the control while it did not significantly increase the Spi(-) MF. B[a]P increased both the gpt and Spi(-) MFs several-fold in a dose-dependent manner. To examine the kinetics of MF, ENU was administered (50 mg/kg/day for 5 successive days) and gpt MFs in the liver were determined 7, 21, 35, and 70 days after the last injection. The MF increased to 8-fold and 13-fold over the control at 7 and 35 days, respectively, after the last injection and then slightly declined at 70 days. These kinetics are similar to those reported for ENU-treated lacZ transgenic mice. This novel transgenic rat could be useful for investigating species differences between rats and mice in their response to genotoxic agents.

Comparative assessment of prurifloxacin, sparfloxacin, gatifloxacin and levofloxacin in the rabbit model of proarrhythmia.

The administration of certain quinolone antibiotics has been associated with a prolongation of the QT interval on electrocardiogram, and in rare cases ventricular arrhythmias such as torsades de pointes. In this in vivo study using a rabbit arrhythmia model, we assessed the proarrhythmic effects and changes in the QT interval elicited by the administration of NM394 (UFX), an active metabolite of the new quinolone antibiotic prulifloxacin, and three representative quinolones, sparfloxacin (SPFX), gatifloxacin (GFLX) and levofloxacin (LVFX). Chloralose-anesthetized rabbits were co-administered a continuous infusion of methoxamine (15 microg/kg/min) together with NaOH (vehicle, 0.2 mol/L), SPFX (2, 3, 4 mg/kg/min), GFLX (4 mg/kg/min), LVFX (4 mg/kg/min) or UFX (4 mg/kg/min) via the ear vein, and then the effects on electrocardiogram were examined. SPFX and GFLX both prolonged the QT and QTc intervals. GFLX also induced premature ventricular contractions in all 6 rabbits that received it, and subsequently it induced torsades de pointes (TdP) in 3 of the 6 rabbits. SPFX infused at the dose of 4 mg/ kg/min induced conduction blocks without inducing TdP, whereas that infused at the lower dose of 3 mg/ kg/min induced both conduction blocks and TdP. The infusions with LVFX and UFX did not elicit remarkable prolongations in the QT interval, and none of the animals infused with the agents developed arrhythmia. These findings suggested that LVFX and UFX were less potent than SPFX and GFLX in prolonging the QT interval and inducing life-threatening arrhythmias.

SAR analysis of the improved resonant cavity applicator with electrical shield and water bolus for deep tumors by a 3-D FEM.

This paper discusses the improvements of the re-entrant resonant cavity applicator, such as an electromagnetic shield and a water bolus for concentrating heating energy on deep tumors in an abdominal region of the human body. From our previous study, it was found that the proposed heating system using the resonant cavity applicator, was effective for heating brain tumors and also for heating other small objects. However, when heating the abdomen with the developed applicator, undesirable areas such as the neck, arm, hip and breast were heated. Therefore, we have improved the resonant cavity applicator to overcome these problems. First, a cylindrical shield made of an aluminum alloy was installed inside the cavity. It was designed to protect non-tumorous areas from concentrated electromagnetic fields. Second, in order to concentrate heating energy on deep tumors inside the human body, a water bolus was installed around the body. Third, the length of the lower inner electrode was changed to control the heating area. In this study, to evaluate the effectiveness of the proposed methods, specific absorption rate (SAR) distributions were calculated by FEM with the 3-D anatomical human body model reconstructed from MRI images. From these results, it was confirmed that the improved heating system was effective to non-invasively heat abdominal deep tumors.