Senri Kinran University

Letermovir and ganciclovir are used to prevent cytomegalovirus (CMV) infection, but the generation of resistant viruses and the interactions between wild and resistant CMV have not been studied. We evaluated the effect of letermovir/ganciclovir on the release of the wild-type and letermovir/ganciclovir-resistant CMV in their coinfected or superinfected cells. We analysed the extracellular CMV infectivity released from CMV-infected cells treated with letermovir and ganciclovir. Subsequently, we characterized growth of letermovir/ganciclovir-resistant viruses in wild-type CMV-infected cells in the presence of letermovir/ganciclovir, respectively. The number of infectious cells resuming viral replication after letermovir/ganciclovir removal decreased over time, and the CMV-infected cells treated with letermovir or ganciclovir remained infectious for 1 month. During the treatment course, letermovir-/ganciclovir-resistant viruses arise from CMV infection foci, and we investigated the proliferation of resistant viruses in the presence of letermovir/ganciclovir. Letermovir inhibited superinfected CMV release by inhibiting the superinfected CMV from using the DNA terminase-packaging complex pathway occupied by a prior infected CMV. Co-infection and superinfection of ganciclovir-resistant CMV in wild-type CMV-infected cells inhibited viral release by inhibiting DNA synthesis by ganciclovir phosphorylated by wild-type UL97. These results suggested that letermovir-resistant CMV emerging in wild-type CMV-infected cells would not easily spread to the surrounding wild-type CMV-infected cells in CMV-infected patients under letermovir. Despite different mechanisms of action, both drugs inhibited the spread of newly emerging resistant viruses around wild-type CMV-infected lesions during letermovir/ganciclovir treatment, suggesting the practical use of both drugs and strategies for treating drug-resistant CMV.

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease with an unknown pathogenesis. It has been reported that mutations in the gene for Cu/Zn superoxide dismutase (SOD1) cause familial ALS. Mutant SOD1 undergoes aggregation and forms amyloid more easily, and SOD1-immunopositive inclusions have been observed in the spinal cords of ALS patients. Because of this, SOD1 aggregation is thought to be related to the pathogenesis of ALS. Some core regions of amyloid have been identified, but the issue of whether these regions form aggregates in living cells remains unclear, and the mechanism responsible for intracellular SOD1 aggregation also remains unclear. The findings reported in this study indicate that the aggregation of the ALS-linked mutant SOD1-EGFP was significantly enhanced when the BioID2 gene was fused to the N-terminus of the mutant SOD1-EGFP plasmid for cellular expression. Expression of a series of BioID2-(C-terminal deletion peptides of SOD1)-EGFP permitted us to identify 1-35 as a minimal N-terminal sequence and Ile35 as an essential amino acid residue that contributes to the intracellular aggregation of SOD1. The findings also showed that an additional substitution of Ile35 with Ser into the ALS mutant SOD1 resulted in the significant suppression of aggregate formation. The fact that no Ile35 mutations have been reported to date in ALS patients indicates that all ALS mutant SOD1s contain Ile35. Taken together, we propose that Ile35 plays a pivotal role in the aggregation of the ALS-linked SOD1 and that this study will contribute to our understanding of the mechanism responsible for SOD1 aggregation.

Although neonates are commonly exposed to vaginal herpes simplex virus (HSV)-2, neonatal herpes is rare. Therefore, we analyzed paired infant and maternal HSV-2 isolates from two cases of mother-to-infant transmission to identify viral factors contributing to vertical transmission. Sixteen infant isolates with neonatal herpes and 27 genital isolates in their third trimester were included. The infant isolates were significantly more temperature-independent than the maternal isolates. Sequence comparison revealed viral UL13 protein kinase (UL13-PK) mutation in the infant isolates in both cases. In the expanded cohort, infant isolates (5/18) had significantly more UL13-PK mutations than genital isolates (1/29). Isolates within 8 days post-birth (3/4) had a significantly higher frequency of UL13-PK mutation than those after 9 days (2/14), suggesting a close association between UL13-PK mutations and vertical transmission. Elongation factor 1-delta was identified as a target of UL13-PK by proteomic analysis of UL13-PK-positive and -negative HepG2 cells. The mixed infant isolates with the intact and mutated UL13-PK conferred altered cell tropism, temperature independence adapting to fetal temperature, and better growth properties in Vero and hepatoblastoma HepG2 cells than in HSV-2 with intact and mutated UL13-PK alone, indicating that viral UL13-PK mutation is essential for vertical HSV-2 transmission.

BackgroundPost-implant right heart failure (RHF) has been recognized as a crucial prognostic factor in patients receiving left ventricular assist devices (LVADs), and its management has long attracted attention from cardiologists and surgeons.Case presentationThis report described an 18-year-old female with acutely deteriorating heart failure due to dilated cardiomyopathy who underwent paracorporeal pulsatile-flow LVAD and developed early post-implant RHF. At postoperative day (POD) six, she was almost asymptomatic at rest on 2.5 mg/kg/min of dobutamine; however, the echocardiogram, performed as part of the daily postoperative care, revealed a severely enlarged right ventricle with a decompressed left ventricle, implying the development of post-implant RHF. Bolus infusion of saline and reduction of pump flow (6.0 L/min to 3.0 L/min) led to normalization of both ventricular shapes in 30 s, suggesting that RHF could be managed without surgical interventions. Milrinone was started on POD six, followed by sildenafil administration on POD seven. Fluid balance was strictly adjusted under the close observation of daily echocardiograms. Milrinone and dobutamine were discontinued on PODs 18 and 21, respectively. The patient was listed for a heart transplant on POD 40. Despite reduced right ventricular function (right ventricular stroke work index of 182.34 mmHg*ml/m− 2, body surface area 1.5 m2), she was successfully converted to implantable LVAD on POD 44 with no recurrence of post-implant RHF thereafter for four years.ConclusionsIn post-implant RHF management, early detection, together with proper and prompt medical management, is crucial to avoiding any surgical intervention. Close observation of daily echocardiograms might be helpful in detecting subclinical RHF and is useful for post-implant medical management.

Introduction: Emergency life-saving technicians (ELSTs), who are certified specialists trained for prehospital emergency care in Japan, are the principal decision maker to provide medical care to severely neurologically damaged patients. The aim of this study is to clarify of awareness and attitude of ELSTs toward organ transplantation and donation. Method: In the present study, a web based questionnaire was originally designed for ESLTs in a certain prefecture in Japan containing questions about awareness of organ transplantation and donation itself and donor cards, and administered to all 345 ESLTs who worked in the fire departments, which are currently main emergent service in Japan, in a certain prefecture. Submission of this questionnaire indicated the subject’s consent to participate in the study. Valid responses were obtained from 144 (42%) participants. Results: 58 (40.3%) had an interest in organ transplantation and donation, but 21 (14.6%) had no interest and the others (45.1%) didn’t make decision. Their interest rate in organ transplantation and donation was lower than that of normal public (65.5%) reported by the national survey conducted by the Cabinet Office in 2021. 96 (66.7%) recognized that the symbol of organ transplantation was green ribbon and 138 (95.5%) recognized the donor card delivered by Japan Organ Transplantation Network. Most participants recognized the method to present their living willingness to donate their organs, such as a driver license (87.5％) and a donor card (77.1%). Possession rate of their own donor card in the participants was 21.5%, which was greater than that of the normal public (13.3%). The rate of participants who found a patient’s donor card during daily prehospital emergency care, was 16.5%, which was greater than that the possession rate of normal public reported by national survey (13.3%). Regarding education experience, only 18 (12.6％) received professional education at the training program for in-hospital transplant procurement coordinators organized by the prefectural procurement coordinator in 4 and training schools or colleges in 10. This rate was greater than that of national pubic (5%) but lower than other medical staffs, such as physicians and nurses. 56 (39.2%) wanted to have an opportunity to learn organ transplantation and donation, but 15 (10.5%) didn’t and the others (50.3%) didn’t make decision. Conclusion: Although the possession rate of their own donor card was greater than normal publics, the interest rate in organ transplantation and donation in participants were lower than normal publics, probably because they had little opportunities to learn organ transplantation and donation before and after becoming ELSTs. To improve their awareness and attitude towards organ transplantation and donation, we need to establish education program of organ transplantation and donation for ELSTs and increase opportunity for them to learn organ transplantation and donation.

Recently, there has been an increase in the demand for gluten-free bread due to health reasons. One of the flours used to produce gluten-free bread is rice flour; flour characteristics are very important for breadmaking. Although a study has shown that the addition of high-temperature water can improve the quality of rice flour bread, studies are yet to consider different rice properties. Therefore, the aim of this study was to investigate the effect of adding high-temperature water and rice flour characteristics on the quality of rice flour bread using six commercially available rice flours. The rice flours used in the sample had amylose content from 12.1% to 24.5%, damaged starch content from 2.4% to 5.5%, mode diameter from 16.3 to 63.3µm, protein content from 5.4% to 6.1%, and moisture content in the range of 12.0%-15.0%. The results showed that regardless of the rice characteristics, breads prepared at the optimum watering temperature were puffier and softer than those prepared using cold water (5°C). For rice flours with similar particle size, the optimal water temperature and degree of gelatinization for breadmaking increased with rice flours with lower amylose content. Furthermore, the rheological properties of dough prepared at the optimum water addition temperature were stable, with loss modulus (G″) being dominant over the entire frequency range in the frequency sweep test. Since the water temperature added to the dough affects breadmaking properties more than the characteristics of the rice flour, adjusting the water temperature may enable the production of high-quality bread even with rice flour unsuitable for making. PRACTICAL APPLICATION: Presently, the addition of high-temperature water to rice flour has been shown to improve the bread quality. In this study, we investigated the effects of high-temperature water addition on the quality of rice flour bread using rice flour varieties with different flour characteristics. Even in rice flour with small particle size and low amylose content, which is not suitable for breadmaking, bread quality can be improved by adding hot water at around 70°C. This is a simple and practical method to improve the quality of gluten-free rice flour bread without adding thickeners.

Penelitian ini bertujuan untuk meningkatkan hasil belajar IPA siswa kelas V UPT SD Negeri 76 Pinrang, dengan jenis penelitian tindakan kelas. Subjek dalam penelitian ini adalah siswa kelas V UPT SD Negeri 76 Pinrang tahun ajaran 2020/2021 dengan jumlah siswa 12 orang terdiri dari 6 siswa laki-laki dan 6 siswa perempuan. Data hasil penelitian diperoleh melalui tes, observasi, dan dokumentasi. Teknik analisis data menggunakan analisis deskriptif kualitatif. Berdasarkan hasil analisis data menunjukkan peningkatan proses dan hasil belajar dari siklus I ke siklus II dengan masing-masing memperoleh nilai dari aspek guru pada siklus I 67% dan siklus II 87% sedangkan aspek siswa pada siklus I 62% dan siklus II 80%, serta hasil ketuntasan belajar siklus I 67% dengan nilai rata-rata 70 dan siklus II ketuntasan belajar mencapai 80% dengan nilai rata-rata 81. Dengan demikian, dapat disimpulkan bahwa penerapan pendekatan STEAM (Science, Technology, Engineering, Arts and Mathematics) dapat meningkatkan hasil belajar IPA siswa kelas V UPT SD Negeri 76 Pinrang. Kata Kunci: STEAM; Hasil Belajar, IPA

Increasing number of individuals worldwide are consuming gluten-free products, for example, bread, for health and other reasons. However, gluten-free products are currently expensive and/or their preparation involves the use of specialist machinery or food additives. In this study, we focused on the thickening effect of starch gelatinization and attempted to develop a novel method for gluten-free rice flour bread production without the use of additives. We aimed to determine the effect of adding high-temperature water to gluten-free rice flour on the properties of the resulting batter, primarily gelatinization. The water was tested at temperatures between 50 and 80°C, in 2°C increments. For comparison, control bread from gluten-free rice flour was made using cold (5°C) water. The addition of water at a temperature between 66 and 70°C significantly improved the specific volume and firmness of bread (p<0.05, Dunnett's test; compared with control). Additionally, maintaining the gelatinization temperature of the bread batter for approximately 1-10 s and the degree of gelatinization of batter, approximately 5%-10%, were crucial for obtaining good-quality bread. Further, the addition of water at a temperature above 78°C adversely affected the bread-making properties. This simple method developed for making high-quality bread from gluten-free rice flour will make gluten-free bread products more widely available to and acceptable by the consumers. PRACTICAL APPLICATION: Currently, making high-quality bread from gluten-free rice flour involves the use of food additives or special rice flour. Here, we present a simple method for producing high-quality bread by manipulating the temperature of water added during the preparation of rice flour batter. We optimized the method by analyzing the gelatinization properties of the batter and determined the optimal water temperature suitable for bread making. This method yields high-quality gluten-free bread and is cost-effective and simple to implement.