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Coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has led to a global pandemic. New technologies have been utilized to develop several types of vaccines to prevent the spread of SARS-CoV-2 infection, including mRNA vaccines. Our group previously developed an effective DNA-based vaccine. However, emerging SARS-CoV-2 variants of concern (VOCs), such as the delta variant, have escaped mutations against vaccine-induced neutralizing antibodies. This suggests that modified vaccines accommodating VOCs need to be developed promptly. Here, we first modified the current DNA vaccine to enhance antigenicity. Compared with the parental DNA vaccine, the modified version (GP∆-DNA vaccine) induced rapid antibody production. Next, we updated the GP∆-DNA vaccine to spike glycoprotein of the delta variant (GP∆-delta DNA vaccine) and compared the efficacy of different injection routes, namely intramuscular injection using a needle and syringe and intradermal injection using a pyro-drive jet injector (PJI). We found that the levels of neutralizing antibodies induced by the intradermal PJI injection were higher than intramuscular injection. Furthermore, the PJI-injected GP∆-delta DNA vaccine effectively protected human angiotensin-converting enzyme 2 (hACE2) knock-in mice from delta-variant infection. These results indicate that the improved DNA vaccine was effective against emerging VOCs and was a potential DNA vaccine platform for future VOCs or global pandemics.

ObjectivesSpondyloarthritis (SpA) is known as series of immune-mediated inflammatory disease of the axial and peripheral joints. Human leucocyte antigen (HLA)-B27 is a genetic risk factor for SpA. Recent evidence suggests that the interleukin −17 (IL17) axis strongly contributes to SpA. This study aimed to assess the efficacy of an IL17A peptide-based vaccine on SpA manifestations in model rats.MethodsHLA-B27/human β2-microglobulin (hβ2M) transgenic rats were immunised with heat-inactivatedMycobacterium tuberculosis(MT) to develop spondylitis and arthritis as an experimental SpA model after immunisation with a keyhole limpet hemocyanin-conjugated IL17A peptide-based vaccine with an alum adjuvant three times. The IL17A antibody titre was assessed using ELISA, and arthritis score and joint thickness were monitored two times a week. Enzyme-linked immunospot (ELISpot) assays for IL4- and interferon-γ-secreting splenocytes were conducted to evaluate IL17A-specific T cell activation. We also evaluated the effect of IL17A vaccine in SpA therapeutic model.ResultsThe IL17A peptide-based vaccine with alum adjuvant successfully induced antibody production and suppressed the arthritis score and joint thickness. X-ray and histological analyses showed that enthesitis, bone destruction and new bone formation were inhibited by the IL17A vaccine. The ELISpot assay showed that the IL17A peptide-based vaccine did not elicit any IL17A-reactive T cell responses. IL17A vaccine tends to mitigate, but not significant, in SpA treatment model. These data showed that the peptide-based vaccine targeting IL17A alleviated the SpA phenotype in a heat-inactivated MT-induced SpA model in HLA-B27/hβ2M transgenic rats.ConclusionsIL17A peptide-based vaccine may be a therapeutic option for SpA treatment.

We conducted a nonrandomized, open-label phase I study to assess the safety and immunogenicity of an intradermal coronavirus disease 2019 (COVID-19) DNA vaccine (AG0302-COVID-19) administered using a pyro-drive jet injector at Osaka University Hospital between Yanagida November 2020 and December 2021. Twenty healthy volunteers, male or female, were enrolled in the low-dose (0.2 mg) or high-dose (0.4 mg) groups and administered AG0302-COVID19 twice at a 2-week interval. There were no adverse events that led to discontinuation of the study drug vaccination schedule. A serious adverse event (disc protrusion) was reported in one patient in the high-dose group, but the individual recovered, and the adverse event was not causally related to the study drug. In the analysis of the humoral immune response, the geometric mean titer (GMT) of serum anti-SARS-CoV-2 spike glycoprotein-specific antibody was low in both the low-dose and high-dose groups (246.2 (95% CI 176.2 to 344.1, 348.2 (95% CI 181.3 to 668.9)) at the 8 weeks after first vaccination. Regarding the analysis of the cellular immune, the number of IFN-γ-producing cells responsive to the SARS-CoV-2 spike glycoprotein increased with individual differences after the first dose and was sustained for several months. Overall, no notable safety issues were observed with the intradermal inoculations of AG0302-COVID19. Regarding immunogenicity, a cellular immune response was observed in some subjects after AG0302-COVID19 intradermal inoculation, but no significant antibody production was observed.

As potential pandemic vaccines, DNA/RNA vaccines, viral vector vaccines and protein-based vaccines have been rapidly developed to prevent pandemic spread worldwide. In this study, we designed plasmid DNA vaccine targeting the SARS-CoV-2 Spike glycoprotein (S protein) as pandemic vaccine, and the humoral, cellular, and functional immune responses were characterized to support proceeding to initial human clinical trials. After intramuscular injection of DNA vaccine encoding S protein with alum adjuvant (three times at 2-week intervals), the humoral immunoreaction, as assessed by anti-S protein or anti-receptor-binding domain (RBD) antibody titers, and the cellular immunoreaction, as assessed by antigen-induced IFNγ expression, were up-regulated. In IgG subclass analysis, IgG2b was induced as the main subclass. Based on these analyses, DNA vaccine with alum adjuvant preferentially induced Th1-type T cell polarization. We confirmed the neutralizing action of DNA vaccine-induced antibodies by a binding assay of RBD recombinant protein with angiotensin-converting enzyme 2 (ACE2), a receptor of SARSCoV-2, and pseudo-virus assay, TCID assay with live SARS-CoV-2. Further B cell epitope mapping analysis using a peptide array showed that most vaccine-induced antibodies recognized the S2 and RBD subunits. Finally, DNA vaccine protected hamsters form SARSCoV-2 infection. In conclusion, DNA vaccine targeting the spike glycoprotein of SARS-CoV-2 might be an effective and safe approach to combat the COVID-19 pandemic. Funding: This study was supported by Project Promoting Support for Drug Discovery grants (JP20nk0101602 and JP21nf0101623h102) from the Japan Agency for Medical Research and Development and Panasonic Co. (Japan). To fight against the worldwide COVID-19 pandemic, the development of an effective and safe The Department of Health Development and Medicine is an endowed department supported by Anges, Daicel, and FunPep. The Department of Clinical Gene Therapy is financially supported by Novartis, AnGes, Shionogi, Boeringher, Fancl, Saisei Mirai Clinics, Rohto and Funpep. Declaration of Interest: R.M. is a stockholder of FunPep and Anges. T.O. T.K. and Y.S. are employees of Anges. R.I, A.T, H.K, S.K, E.T, S.M, and H.T are employees of FunPep. R.M, H.T, and A.T. are FunPep stockholders. All other authors declare no competing interests. Ethical Approval: All experiments were approved by the Ethical Committee for Animal Experiments of the Osaka University Graduate School of Medicine.

To fight against the worldwide COVID-19 pandemic, the development of an effective and safe vaccine against SARS-CoV-2 is required. As potential pandemic vaccines, DNA/RNA vaccines, viral vector vaccines and protein-based vaccines have been rapidly developed to prevent pandemic spread worldwide. In this study, we designed plasmid DNA vaccine targeting the SARS-CoV-2 Spike glycoprotein (S protein) as pandemic vaccine, and the humoral, cellular, and functional immune responses were characterized to support proceeding to initial human clinical trials. After intramuscular injection of DNA vaccine encoding S protein with alum adjuvant (three times at 2-week intervals), the humoral immunoreaction, as assessed by anti-S protein or anti-receptor-binding domain (RBD) antibody titers, and the cellular immunoreaction, as assessed by antigen-induced IFNγ expression, were up-regulated. In IgG subclass analysis, IgG2b was induced as the main subclass. Based on these analyses, DNA vaccine with alum adjuvant preferentially induced Th1-type T cell polarization. We confirmed the neutralizing action of DNA vaccine-induced antibodies via two different methods, a binding assay of RBD recombinant protein with angiotensin-converting enzyme 2 (ACE2), a receptor of SARS-CoV-2, and pseudovirus assay. Further B cell epitope mapping analysis using a peptide array showed that most vaccine-induced antibodies recognized the S2 and RBD subunits, but not the S1 subunit. In conclusion, DNA vaccine targeting the spike glycoprotein of SARS-CoV-2 might be an effective and safe approach to combat the COVID-19 pandemic.

In this paper we report about the demonstration of the nuclear track imaging capabilities of Ag-activated phosphate glass. A 375 nm laser and confocal laser scanning microscopy (CLSM) were respectively used for track excitation and detection. Specifically, the blue and orange radiophotoluminescent (RPL) tracks and dose distributions observed after irradiation with soft X-rays, gamma rays and heavy charged particles (HCPs) are examined. In addition, the origins of the reductions in RPL efficiency for high-dose X-ray irradiation and for irradiation with HCPs with high linear energy transfer (LET) values are investigated via a CLSM and a conventional fluorescent reader and discussed. © 2017 Elsevier B.V., Embargo Period 12 months

We investigate the physical and dosimetric characteristics of Ag-activated phosphate glass exposed to proton beams with various energies. We present time-resolved blue and orange radiophotoluminescent (RPL) spectra and examine the dependence of the peak wavelengths and RPL area intensities on gate time. Moreover, we consider various micro-scale 3D patterns written on Ag-activated glass by a focused proton beam and evaluate their orange and blue RPL axial-profiles using two-photon excitation microscopy. Finally, we discuss the remarkable broadening of the Bragg curve observed in Ag-activated glass and argue that it should be attributed to the saturation effect and/or non-linearity effect from the viewpoint of the relation between the dose delivered and the signal.

Silver-activated phosphate glass has long been used as passive-type solid state dosimeter for both individual and environmental dose monitoring. The principle is classified as radio-photoluminescence (RPL) whose mechanism is different from TSL and OSL. The RPL dosimeter has excellent characteristics such as stable sensitivity, negligible fading and reliable repeatability in radiation dose measurement. In Japan it is named “Glass Badge” and has come into wide use in individual dose monitoring services. In order to contribute to restoration of the contaminated area in Fukushima new-type glass dosimeters have been developed and the fruits of this work have led to various applications.

We investigate the characterisation and comparison of micro-scale patterns in silver-activated phosphate glass and lithium fluoride (LiF) single crystal written by a proton beam with an energy range of 1.7–4.5 MeV. A variety of micro-scale structures based on blue and orange luminescent Ag-related colour centres in Ag-activated glass and green and red luminescent F-aggregated colour centres in LiF are visualised by combining single- and two-photon confocal microscopes equipped with a blue continuous wave (cw) laser diode and a femtosecond (fs) laser in the near-infrared (NIR) region, respectively. Two- and three-dimensional (2D and 3D) reconstructed images are analytically evaluated using lateral/axial RPL intensity mapping. In addition, the origins of different RPL intensity profiles in a plateau region along the axial direction in both materials are examined and discussed.

We report the demonstration of a variable periodic time operated fibre-coupled dosimetry system using Ag-activated phosphate glass based on radiophotoluminescent (RPL) phenomenon. In order to overcome the inevitable “build-up” effect occurring in RPL glasses during and after X-ray irradiation, pertinent experiments are designed and carried out. The RPL intensities for four successive sequences, namely, the pre-dose, X-ray exposure, build-up and optical bleaching processes, are examined for the orange and blue RPL bands. Build-up curves and corresponding build-up rates are measured for different excitation wavelengths, analytically evaluated, and compared with those of a real-time fibre plastic scintillation dosimetry system. In addition, the orange RPL signal dependence on temperature and variable interval is examined for three successive sequences. The quick response of the dosimetry system developed here to the on/off switching operations is confirmed using seven successive sequences.

Silver-activated phosphate glass (Ag+-glass), which is commonly employed as a radiophotoluminscence (RPL) dosimeter, was applied as a fluorescent nuclear track detector (FNTD). The heavy ions from C to Fe ions were detected as fluorescent nuclear tracks under a confocal microscope. The detection linear energy transfer (LET) threshold was estimated to be 5 keV/μm in water. The fading effect was observed in which the fluorescent intensity decreases with repeated laser scans. The Ag+-glass has good potential applicability as the FNTD for the high LET range (5–200 keV/μm) in radioimmunotherapy and space radiation dosimetry uses.

Silver (Ag+) doped phosphate glass after exposure to ionizing radiation has an intense luminescence by stimulating with ultra-violet light. This phenomenon is called radiophotoluminescence (RPL). The RPL intensity was increased linearly with increasing X-ray irradiation dose up to about 1000 Gy. Small amount of bead-type RPL glass as large as about 0.05mm was well prepared with a jet flame system. It was confirmed that the RPL glass beads could be used as passive-type radiation dosimeters. Visualization and monitoring system of radiation dose distribution utilizing RPL phenomenon in the Ag+-doped phosphate glass are developed in this study. In the presentation, the basic luminescence properties are reported as well as the application of Ag+-doped phosphate glass beads to radiation dose distribution monitoring.

Silver ion-activated phosphate glass (Ag

Ag-activated phosphate glass based on the radiophotoluminescence (RPL) phenomenon has been used as the most commonly known RPL material and as an accumulated-type passive detector. In this work, the transient-state evaluation of the dose distributions achieved by X- and gamma-ray irradiations within the Ag-activated phosphate glass was performed using a time-resolved technique for the first time. Specifically, the blue RPL intensity ascribed to the electron-trapped Ag0 centres as a function of the depth at the vicinity of the surface was investigated for different types of radiation and a wide range of energies. In addition, the dose distributions at each layer within the glass confirmed by the time-resolved measurement were compared with those reconstructed by a disk-type transparent glass detector based on the blue RPL with a diameter of 100 mm. © 2015 Elsevier B.V. All rights reserved., 24 months Embargo

Silver ion-activated phosphate glass (Ag+-glass) has a good potential for application to radiation dosimetry in various radiation fields due to its multifunctional properties as a detector. The Ag+-glass provides three independent signals of radiophotoluminescence, optical absorption, and nuclear track. The combination of these signals allows the dynamic range of the measured dose (10 μGy–10 kGy) and linear energy transfer ( 1 MeV/μm) to be widened.Silver ion-activated phosphate glass (Ag+-glass) has a good potential for application to radiation dosimetry in various radiation fields due to its multifunctional properties as a detector. The Ag+-glass provides three independent signals of radiophotoluminescence, optical absorption, and nuclear track. The combination of these signals allows the dynamic range of the measured dose (10 μGy–10 kGy) and linear energy transfer ( 1 MeV/μm) to be widened.

The conjugate addition of allyl cyanide (II) to α,β-unsaturated thioamides (I) is catalyzed by soft Lewis acid/hard Broensted base/hard Lewis base cooperation to give the adducts (III) or (VI), which undergo cyclization into isothiazoles such as (IV).

The conjugate addition of allyl cyanide (II) to α,β-unsaturated thioamides (I) is catalyzed by soft Lewis acid/hard Broensted base/hard Lewis base cooperation to give the adducts (III) or (VI), which undergo cyclization into isothiazoles such as (IV).

Ag-activated phosphate glass, which is the most commonly known radiophotoluminescent (RPL) material, has the capability to operate not only dosimeters but also two- and three-dimensional (2D and 3D) dose imaging detectors in the same host. This passive detector is based on radiation-induced, optically active nano-scale defects. In this work, the transient-state optical properties of the blue and orange RPL were investigated using a time-resolved spectrum technique for 137Cs and 60Co gamma-ray-irradiated Ag-activated phosphate glass. Specifically, the blue RPL intensity with a decay time of 5 ns as a function of the depth at the vicinity of the surface was systematically examined to clarify an accurate dose distribution within the glass. Moreover, a feasibility study into the use of an RPL Ag-activated phosphate glass detector for fluorescent nuclear track imaging was demonstrated using a confocal fluorescence image microscope for the first time.