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Recombinant protein polymers with repeat sequences of specific amino acids can be regarded as sustainable functional materials that can be designed using genetic engineering. However, synthesizing genes encoding these proteins is significantly time-consuming and labor-intensive owing to the difficulty of using common gene synthesis tools, such as restriction enzymes and PCR primers. To overcome these obstacles, a novel method is proposed herein: seamless cloning of rolling-circle amplicons (SCRCA). This method involves one-pot preparation of repetitive-sequence genes with overlapping ends for cloning, facilitating the easy construction of the desired recombinants. SCRCA is used to synthesize 10 genes encoding hydrophilic resilin-like and hydrophobic elastin-like repeat units that induce liquid-liquid phase separation. SCRCA shows higher transformation efficiency and better workability than conventional methods, and the time and budget required for SCRCA are comparable to those required for non-repetitive-sequence gene synthesis. Additionally, SCRCA facilitates the construction of a repeat unit library at a low cost. The library shows considerably higher diversity than that of the current state-of-the-art method. By combining this library construction with the directed evolution concept, an elastin-like protein polymer with the desired functions can be rapidly developed. SCRCA can greatly accelerate research on protein polymers., (© 2025 The Author(s). Advanced Science published by Wiley‐VCH GmbH.)

Background: The 6-min walk test (6MWT), used to monitor disease progression or exacerbation in interstitial lung disease, faces challenges such as requiring a 30-m walking path and difficulty assessing patients with gait disturbance. The 1-min sit-to-stand test (1STST) offers a convenient alternative, potentially addressing these issues. Despite its advantages, the effectiveness of the 1STST in patients with idiopathic pulmonary fibrosis (IPF) and progressive pulmonary fibrosis (PPF) still needs to be explored. We assessed 1STST's ability to detect exercise-induced desaturation in a randomized, crossover trial involving patients with IPF or PPF., Methods: Participants were divided into group A (6MWT to 1STST) and B (1STST to 6MWT), with a 30-min rest period between the tests. The primary endpoint was the difference in nadir oxygen saturation (SpO 2 ) between the groups throughout the study. Secondary endpoints included the percentage of participants with a nadir SpO₂ <88% during the tests, a decline of ≥4% in SpO 2, and the variation in Borg scores post-tests., Results: Twenty-three participants (91.3% male; mean age ± standard deviation: 77.2 ± 7.4 years) diagnosed with IPF and PPF were enrolled in this study. The difference in nadir SpO 2 between the 1STST and 6MWT was 1.14% (95% confidence interval: -0.18, 2.48), with the 95% confidence intervals falling within the predefined equivalence range. No significant differences were observed in the secondary endpoints., Conclusions: The results suggest that the 1STST is as effective as the 6MWT in detecting desaturation in patients with IPF and PPF., Trial Registration: This study was registered on the website of the Japan Registry of Clinical Trials (jRCT1032230037; URL: https://jrct.niph.go.jp/)., Competing Interests: Declaration of competing interest The authors have no conflicts of interest., (Copyright © 2025 The Japanese Respiratory Society. Published by Elsevier B.V. All rights reserved.)

Background: Spinal muscular atrophy (SMA) is an autosomal recessive neuromuscular disease. Most patients with SMA have a mutation in the survival motor neuron 1 (SMN1) gene on chromosome 5q. With current genetic testing, SMN1 copy number is determined; a diagnosis is reached when the copy number is zero. When the SMN1 copy number is 1, exons and intron/exon boundaries of the allele are examined for single-nucleotide variants (SNVs). Genetically undiagnosed cases of SMA exist when 2 copies of SMN1 exist or when a SNV is in the deep intron. Furthermore, SMN1 is highly homologous to SMN2; therefore, it is expected that many SNVs have not been elucidated., Methods: This retrospective observational study conducted in Japan used pre-collected DNA samples from patients with clinically diagnosed SMA. Enrollment period was January 28, 2020 to September 30, 2021. SNV analysis of SMN1 (exon 1-8 and intron 1-7) was conducted by long-range polymerase chain reaction and next-generation sequencing., Results: From 336 DNA samples collected from patients, 62 patient samples were included in the SNV analysis. Two patients have been genetically diagnosed (a heterozygous variant in intron 6 with 1 copy of SMN1; a homozygous missense mutation in exon 3 with 2 copies of SMN1). Three SNVs in intron 6, c.834+1506A>G (n = 9), c.834+1751G>A (n = 2), and c.835-367C>A (n = 5) were identified; all were numerically, and c.834+1506A>G and c.835-367C>A were significantly, more frequent in patients with 0 copies versus those with ≥ 1 copy of exon 7 in SMN1. We confirmed 3 hybrid SMN gene types in 5 patients that contained SMN2 gene sequence (aaTgg) flanked by upstream "t" and downstream "G" SMN1 sequence., Conclusions: In this study of patients with clinically diagnosed SMA, 2 cases with genetic SMN types were identified that would not have been identified through current genetic testing, which examines SMN1 deletions only. Furthermore, for 1 patient with a homozygous SMN1 missense mutation, SMA was not suspected by the current copy number screening method. This study demonstrated the importance of performing full-length sequencing for clinically diagnosed SMA to complement current screening methods., Trial Registration: University Hospital Medical Information Network Clinical Trials Registry (Number: UMIN000040095)., Competing Interests: Declarations. Ethics approval and consent to participate: The study protocol and the informed consent form were approved by the Ethics Committee of Tokyo Women’s Medical University on September 17, 2019 (approval no. 393B). Written informed consent form was obtained by patients prior to data collection. This study was conducted in accordance with the protocol, the Declaration of Helsinki, and all relevant regulations and guidelines. Consent for publication: Not applicable. Competing interests: TK, MY, YO, KM, and YK have no conflicts of interest to declare. TH is an employee of Biogen Japan Ltd and owns shares in Biogen USA. KS has received research funds from Biogen Japan Ltd., and received lecture fees from Biogen Japan Ltd, Novartis AG, and Chugai Pharmaceutical Co. Ltd., (© 2025. The Author(s).)

Background: Mechanical cardiopulmonary resuscitation (CPR) devices address the limitations of manual CPR, but their impact on intrathoracic injuries during extracorporeal CPR (ECPR) remains unclear. This study investigated the relationship between mechanical CPR and severe intrathoracic hemorrhage during ECPR compared to manual CPR., Methods: We conducted a single-center retrospective study of consecutive patients who underwent ECPR from April 2017 to March 2024 according to a standard institutional protocol. Patients were divided into a mechanical CPR group (piston-driven compressions before veno-arterial extracorporeal membrane oxygenation [VA-ECMO]) and a manual CPR group. The primary outcome was intrathoracic hemorrhage requiring transcatheter arterial embolization (TAE). Secondary outcomes included other intrathoracic injuries and 180-day survival., Results: A total of 91 patients were enrolled (mechanical n  = 48, manual n  = 43). Intrathoracic hemorrhage requiring TAE occurred more frequently in the mechanical CPR group (18.8% vs. 2.3%, p  = 0.030). On multivariate analysis, mechanical CPR was independently associated with this outcome (adjusted odds ratio 6.29; 95% confidence interval 1.20-65.10). In the mechanical group, older age and larger thoracic transverse diameter were significantly related to intrathoracic hemorrhage requiring TAE. Mediastinal hematoma (18.8% vs. 2.3%, p  = 0.030) and hemothorax (20.8% vs. 4.7%, p  = 0.049) were also more frequent in the mechanical group. The 180-day survival rates did not differ significantly between groups (27.7% vs. 25.0%, log-rank p  = 0.540)., Conclusions: Mechanical CPR during ECPR is associated with an increased risk of severe intrathoracic hemorrhage. While mechanical CPR devices may provide benefits in certain scenarios, clinicians should carefully consider individual patient characteristics and closely monitor for complications., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2025 The Author(s).)

Background: The forced oscillation technique (FOT) is a minimally invasive test to evaluate asthma during resting ventilation. However, its role in longitudinal assessments, such as clinical remission, remains unclear., Objective: To longitudinally assess asthma clinical remission and identify parameters that predict clinical remission at 12 months from baseline FOT., Methods: Adult patients with asthma at our university hospital between April 2022 and May 2023 were enrolled in this prospective observational study. They were evaluated for 12 months after enrollment to determine whether they met the following clinical remission criteria: asthma control test score of more than or equal to 20 at enrollment and 12 months, no asthma exacerbations for 12 months, and no regular oral corticosteroid use during the 12 months. FOT parameters at enrollment were analyzed for associations with clinical remission., Results: A total of 94 patients with asthma completed the study and were categorized into clinical and nonclinical remission groups. Comparison of pulmonary function tests, including the FOT, between the 2 groups revealed significant differences in resistance at 5 Hz and resistance at 20 Hz (R20) but not in forced expiratory volume in 1 second. Multivariate logistic regression analysis revealed that R20 was associated with clinical remission, with adjusted odds ratios of 0.32 (95% CI: 0.12-0.91, P = .033) for R20., Conclusion: R20 can be a useful predictor of future exacerbations in patients with asthma. These findings may assist in evaluating adult patients with asthma and normal forced expiratory volume in 1 second., Competing Interests: Disclosures The authors have no conflicts of interest to report., (Copyright © 2024 American College of Allergy, Asthma & Immunology. Published by Elsevier Inc. All rights reserved.)

Objective: Tixagevimab/cilgavimab is a cocktail of two long-acting monoclonal antibodies approved for pre-exposure prophylaxis (PrEP) of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection (cause of coronavirus disease 2019 [COVID-19]) in immunocompromised (IC) or high-risk patients. We investigated the patient characteristics and clinical outcomes of IC patients administered tixagevimab/cilgavimab for PrEP in real-world use in Japan., Methods: This observational study used anonymous secondary data from Real-World Data Co., Ltd. for IC patients aged ≥12 years administered tixagevimab/cilgavimab between September 2022 and September 2023. We analyzed the baseline characteristics and event-rates of COVID-19-related clinical outcomes within 6 months of administration., Results: Data were analyzed for 397 IC patients. About half (53.4 %) were male and the median age was 71.0 (interquartile range 61.0, 77.0) years. Malignancy (97.2 %), cardiovascular disease (71.3 %), and diabetes (66.5 %) were frequent comorbidities. Systemic corticosteroids and immunosuppressants were prescribed to 87.4 % and 24.9 %, respectively. The two most common target clinical conditions were active therapy for hematologic malignancies (88.2 %) and treatment with B cell-depleting therapies (57.4 %). The event-rates per 100 person-months (95 % confidence interval; number) for medically attended COVID-19, COVID-19 hospitalization, in-hospital mortality due to COVID-19, and all-cause death were 4.14 (3.06-5.48; n = 49), 1.74 (1.09-2.64; n = 22), 0.07 (0.00-0.42; n = 1), and 0.60 (0.26-1.17; n = 8), respectively., Conclusion: This is the first report using a multicenter database to describe the clinical characteristics and COVID-19-related outcomes of IC patients administered with tixagevimab/cilgavimab in real-world settings in Japan. This cohort of IC patients who received tixagevimab/cilgavimab included many elderly patients with comorbidities., Competing Interests: Declaration of competing interest Toshibumi Taniguchi has received lecture fees from AstraZeneca K.K. Tomoyuki Homma, Yoichi Tamai, Yoshifumi Arita, Masakazu Fujiwara, Naho Kuroishi, Keiji Sugiyama, Shinichi Kanazu and Atsushi Maruyama are employees of AstraZeneca K.K., (Copyright © 2024 Japanese Society of Chemotherapy, Japanese Association for Infectious Diseases, and Japanese Society for Infection Prevention and Control. Published by Elsevier Ltd. All rights reserved.)

Using the free latissimus dorsi muscle flap for dynamic reconstruction for established facial paralysis helps reconstruct various facial dysfunctions, depending on nerve and muscle arrangements. The authors divided the latissimus dorsi muscle into 3 flaps, 2 placed in the cheek area to reconstruct a "voluntary and spontaneous smile" and 1 fixed to the fascia implanted in the eyelid to treat lagophthalmos in a 1-stage procedure in a 61-year-old woman who underwent tumor resection for a left facial nerve schwannoma and developed complete facial paralysis. Fascia lata and free latissimus dorsi muscle grafting were performed. Spontaneous smile was reconstructed using the contralateral facial nerve as a motor source. Voluntary smile and eyelid closure were reconstructed using the ipsilateral masseter nerve, dividing the latissimus dorsi muscle flap into 3 muscle bodies. This procedure restored her smile and eyelid closure ability, with a nearly normal overall facial impression, making it a valuable technique for 1-term reconstruction of smiles and eyelid closure., Competing Interests: The authors report no conflicts of interest., (Copyright © 2024 by Mutaz B. Habal, MD.)