Simple Nano-Luciferase-Based Assay for the Rapid and High-Throughput Detection of SARS-CoV-2 3C-Like Protease.

In this study, we described an easy-to-perform nano-luciferase (nLuc) sensor for the rapid detection of 3-chymotrypsin-like protease (3CLpro) encoded by SARS-CoV-2. The technology is based on the cleavage reaction of recombinant-nLuc via 3CLpro. The nLuc-based assay is a general, one-step method and is naturally specific in detection. The stability, sensitivity, detection range, and response time are fully characterized. The application of 3CLpro detection in artificial and human saliva as well as antiviral drug screening demonstrates that the method can quantify 3CLpro with high sensitivity in one step. With its unique features, the nLuc-based assay may find broad applications in the auxiliary diagnosis of SARS-CoV-2, as well as other types of coronavirus infection.

Impact of the COVID-19 pandemic on out-of-hospital cardiac arrest outcomes in older adults in Japan.

Aim: The coronavirus disease (COVID-19) pandemic has negatively affected access to healthcare and treatment. This study aimed to explore the impact of the COVID-19 pandemic on older adults with out-of-hospital cardiac arrest (OHCA) in Japan, a country with a super-aging society. Methods: This secondary analysis of the All-Japan Utstein Registry included patients aged 65 years and older with bystander-witnessed OHCA between January 1, 2005, and December 31, 2020. Survival outcomes were compared by time period using multivariable logistic regression analyses. The primary outcome measured was the one-month survival rate with neurologically favorable outcomes. Results: Before the COVID-19 pandemic, survival outcomes were steadily improving, and 32,024 patients in 2019 and 31,894 in 2020 were eligible for analysis. The proportions of conventional cardiopulmonary resuscitation and shock by public-access automated external defibrillators were lower in 2020 than in 2019 (6.7% versus 5.7%, p < 0.001 and 2.5% versus 2.1%, p < 0.001, respectively). Compared to 2019, the one-month survival after OHCA and prehospital return of spontaneous circulation decreased significantly in 2020 than in 2019 (7.7% versus 6.6%, adjusted odds ratio [AOR]: 0.88, 95% confidence interval [CI]: 0.83-0.94, and 16.8% versus 14.9%, AOR: 0.87, 95% CI: 0.83-0.91, respectively). The proportion of neurologically favorable outcomes also decreased, but the decrease was not statistically significant (3.4% versus 2.8%, AOR: 0.92, 95% CI: 0.83-1.01). Conclusion: In this population-focused, bystander-witnessed study regarding OHCA, the analysis of nationwide registry data revealed that the COVID-19 pandemic was associated with reduced survival among older adults with OHCA in Japan.

Association of the COVID-19 Pandemic With Prehospital Characteristics and Outcomes of Pediatric Patients With Out-of-Hospital Cardiac Arrest in Japan, 2005-2020.

This cohort study investigates the association of the COVID-19 pandemic with prehospital characteristics and outcomes of pediatric patients with out-of-hospital-cardiac arrest in Japan.

Survival following an out-of-hospital cardiac arrest in Japan in 2020 versus 2019 according to the cause.

Aim: The coronavirus disease (COVID-19) pandemic has led to an increase in out-of-hospital cardiac arrests (OHCAs) and mortality. However, there has been no reports in Japan using nationwide registry data. We compared survival among patients with OHCAs and detailed information on the cause during the COVID-19 pandemic (2020), and during the pre-pandemic period (2019). Methods: Using a Japanese population-based retrospective cohort study design, we analyzed registry data on 39,324 and 39,170 patients with OHCAs in 2019 and 2020, respectively. We compared patient outcomes in 2019 and 2020 using univariable and multivariable logistic regression analyses. Results: The proportion of OHCAs of cardiac origin increased significantly from 61.6% in 2019 to 62.7% in 2020 (P = 0.001). The use of bystander CPR (6.9% versus 5.7%, P < 0.001) and publicaccess automated external defibrillator pads (3.7% versus 3.0%, P < 0.001) decreased significantly from 2019 to 2020. The 1-month survival for OHCA of cardiac origin (12.1% versus 10.7%; adjusted odds ratio [OR] 0.93, 95% confidence interval [CI] 0.87-1.00), asphyxia (10.9% versus 8.8%; adjusted OR 0.80, 95% CI 0.70-0.92), and external causes (adjusted OR 0.66; 95% CI 0.46-0.96), also decreased significantly from 2019 to 2020. Conclusions: In Japan, the 1-month survival after OHCA of cardiac origin, or due to asphyxia or external causes, decreased significantly during the COVID-19 pandemic period.

Clinical outcomes among out-of-hospital cardiac arrest patients treated by extracorporeal cardiopulmonary resuscitation: The CRITICAL study in Osaka.

AIM: Extracorporeal cardiopulmonary resuscitation (ECPR) is performed in refractory out-of-hospital cardiac arrest (OHCA) patients, and the eligibility has been conventionally determined based on three criteria (initial cardiac rhythm, time to hospital arrival within 45 minutes, and age <75 years) in Japan. Owing to limited information, this study descriptively determined neurological outcomes after applying the three criteria among OHCA patients who underwent ECPR. METHODS: This study conducted a post-hoc analysis of data from the Comprehensive Registry of Intensive Care for OHCA Survival (CRITICAL) study. This was a multi-institutional prospective observational study of OHCA patients in Osaka Prefecture, Japan. All adult (aged ≥18 years) OHCA patients with internal medical causes treated with ECPR between 1 July 2012 and 31 December 2019 were evaluated. We described one-month neurological favourable outcomes based on the three criteria (initial shockable, time to hospital arrival within 45 minutes, and age <75 years), and we compared them using the chi-square test. RESULTS: Among 18,379 patients screened from the CRITICAL study database, we included 517 OHCA patients treated by ECPR; 311 (60.2%) patients met all three criteria. Favourable neurological outcomes were as follows: patients meeting no or one criterion: 2.3% (1/43), those meeting two criteria: 8% (13/163), and those meeting all criteria: 16.1% (50/311) (P-value = 0.004). CONCLUSIONS: In this study, approximately 60% of patients treated by ECPR met the three criteria (initial shockable, time to hospital arrival within 45 minutes, and age <75 years), and the greater the number of criteria met, the better were the neurological outcomes achieved.

Clinical epidemiology and outcomes of COVID-19 patients with extracorporeal membrane oxygenation support in Japan: a retrospective study.

Objective: To elucidate the clinical epidemiology and outcomes of patients with COVID-19 who received extracorporeal membrane oxygenation (ECMO) in the Osaka Prefecture, Japan. Methods: We conducted a retrospective study of COVID-19 patients who received ECMO. Among patients diagnosed with COVID-19 between January 29 and November 9, 2020, we targeted patients who received ECMO. The outcome was all-cause mortality. The baseline characteristics of the COVID-19 patients who received ECMO were summarized according to the outcome. A univariable logistic regression model was applied to evaluate the association of each factor (sex, age group, city of residence, presence of comorbidities, presence of close contact, use of dialysis, and wave) with all-cause mortality. Odds ratios (ORs) with 95% confidence intervals were calculated. Results: Among the 14,864 patients diagnosed with COVID-19 during the study period, 39 patients received ECMO. Fourteen patients (35.9%) died. All patients aged 30-39 years survived, whereas all patients aged ≥80 years died. Higher mortality was observed among patients in the higher age group, and the P value for trend was significant (P value for trend: 0.04). Conclusions: Of the 14,864 COVID-19 patients in Osaka Prefecture until November 2020, 39 underwent ECMO. Of these, 14 died.

Characteristics and Outcomes of COVID-19 in Reproductive-Aged Pregnant and Nonpregnant Women in Osaka, Japan.

OBJECTIVE: To describe the clinical characteristics and outcomes of reproductive-aged female patients with coronavirus disease 2019 (COVID-19). METHODS: We conducted a retrospective study of female patients aged 10-49 years, diagnosed with COVID-19 in Osaka, Japan, between January and November 2020. We assessed their epidemiological and clinical characteristics according to their pregnancy status. RESULTS: A total of 4,156 patients were enrolled, of whom 29 (0.7%) were pregnant. Most patients exhibited mild symptoms, and 10.8% of the cases were asymptomatic. No moderate or severe cases were observed in pregnant women, whereas only 0.1% of the nonpregnant women had severe disease at diagnosis. No clusters were observed in the pregnant patients; however, most acquired the infection from a family member. Of the 29 pregnant women, 22 (75.9%) were hospitalized; whereas among the nonpregnant women, 579 (14.0%) were hospitalized (p < 0.001). No patients were admitted to the intensive care unit, and there were no deaths among women aged 10-49 years. CONCLUSIONS: Pregnant women accounted for 0.7% of the total cases of COVID-19 among women aged 10-49 years. Pregnant women were more likely to be hospitalized but generally had mild disease.

Aptamer/antibody sandwich method for digital detection of SARS-CoV2 nucleocapsid protein.

Nucleocapsid protein (N protein) is the most abundant protein in SARS-CoV2 and is highly conserved, and there are no homologous proteins in the human body, making it an ideal biomarker for the early diagnosis of SARS-CoV2. However, early detection of clinical specimens for SARS-CoV2 remains a challenge due to false-negative results with viral RNA and host antibodies based testing. In this manuscript, a microfluidic chip with femtoliter-sized wells was fabricated for the sensitive digital detection of N protein. Briefly, ß-galactosidase (ß-Gal)-linked antibody/N protein/aptamer immunocomplexes were formed on magnetic beads (MBs). Afterwards, the MBs and ß-Gal substrate fluorescein-di-ß-d-galactopyranoside (FDG) were injected into the chip together. Each well of the chip would only hold one MB as confined by the diameter of the wells. The MBs in the wells were sealed by fluorocarbon oil, which confines the fluorescent (FL) product generated from the reaction between ß-Gal and FDG in the individual femtoliter-sized well and creates a locally high concentration of the FL product. The FL images of the wells were acquired using a conventional inverted FL microscope. The number of FL wells with MBs (FL wells number) and the number of wells with MBs (MBs wells number) were counted, respectively. The percentage of FL wells was calculated by dividing (FL wells number) by (MBs wells number). The higher the percentage of FL wells, the higher the N protein concentration. The detection limit of this digital method for N protein was 33.28 pg/mL, which was 300 times lower than traditional double-antibody sandwich based enzyme-linked immunosorbent assay (ELISA).

Characteristics and outcomes of older patients with coronavirus disease 2019 in Japan.

AIM: The epidemiological characteristics, in-hospital treatments and outcomes of coronavirus disease 2019 among older patients have not been fully evaluated in Japan. METHODS: In this retrospective observational study carried out in Osaka Prefecture, Japan, we enrolled patients aged ≥60 years with laboratory-confirmed coronavirus disease 2019 from January to November 2020. The main outcome was mortality during the observation period, based on the Infectious Diseases Control Law. Cox regression analysis was used to evaluate the association between epidemiological factors and mortality among older patients with coronavirus disease 2019. RESULTS: Older patients accounted for 21.5% (3192/14 846) of the registered patients with coronavirus disease 2019. The number of patients according to age was as follows: 60-69 years, 1140 (35.7%); 70-79 years, 1058 (33.1%); 80-89 years, 749 (23.5%); and ≥90 years, 245 (7.7%). The proportion of deaths during the observation period was 8.5% (271/3192). The proportion of deaths increased with increasing age category (from 1.9% to 20.4%, P for trend <0.001). In multivariable Cox regression analysis, patients aged 70-79, 80-89 and ≥90 years had higher hazard ratios and 95% confidence intervals of death (2.62 [1.63-4.23], 5.99 [3.77-9.50] and 10.24 [6.03-17.40], respectively) than those aged 60-69 years. Factors such as male sex, presence of comorbidities, cluster cases in medical institutions and moderate/severe symptoms at diagnosis were also associated with mortality. CONCLUSIONS: This study shows the epidemiological characteristics of older patients with coronavirus disease 2019 in Osaka Prefecture, Japan. The proportion of deaths was 8.5% in total and increased with increasing age. Geriatr Gerontol Int 2021; 21: 629-635.

Characteristics and Survival of Intensive Care Unit Patients with Coronavirus Disease in Osaka, Japan: A Retrospective Observational Study.

The epidemiological and clinical characteristics, treatments, and outcomes of patients with coronavirus disease 2019 (COVID-19) who are admitted to the intensive care unit (ICU) have not been adequately evaluated in Japan. We analyzed the registry data of 205 patients with COVID-19 admitted to the ICU between February and November 2020, in Osaka Prefecture, Japan. A Cox proportional hazards model was used to assess the association between epidemiological factors and mortality among ICU patients. Of the 205 ICU patients, 161 (78.5%) were men and 149 (72.7%) were older than 60 years. A total of 117 patients (57.1%) had comorbidities. The most common symptoms at diagnosis were mild (n = 131, 63.9%). A total of 187 patients (91.2%) received mechanical ventilation, and 32 patients (15.6%) required extracorporeal membrane oxygenation. Patients were followed up for a median of 25 days after ICU admission. A total of 147 patients (71.7%) were alive at discharge, and 58 patients (28.3%) died. The hazard ratio for mortality among patients aged >80 years was 6.02 (95% confidence interval: 2.10-17.25) in the multivariable model, which was higher than that among those aged ≤59 years. These results are useful for recognizing the clinical course of this infection in ICU patients.

Characteristics and Outcomes of Pediatric COVID-19 Patients in Osaka, Japan.

The epidemiological information on characteristics, in-hospital treatments, and outcomes of the coronavirus disease 2019 (COVID-19) among pediatric patients has not been fully evaluated in Japan. This was a retrospective observational study conducted in the Osaka Prefecture, Japan, and we enrolled laboratory-confirmed COVID-19 patients aged ≤ 19 years old from January to November in 2020. Of 14,846 COVID-19 eligible patients, 1240 pediatric patients (8.4%) were registered during the study period; 329 were children aged 0-9 years (26.5%) and 911 were adolescents aged 10-19 years (73.5%). The majority of the patients exhibited mild symptoms at diagnosis (872, 70.3%), some were asymptomatic (296, 23.9%). Cluster infections occurred in child-care facilities (26, 7.9%) among children and in universities (27, 3.0%) and schools (18, 2.0%) among adolescents. The number of close-contact cases was 260 (69.0%) in children and 459 (50.4%) in adolescents. Sixty of the children (18.2%) and 90 of the adolescents (9.9%) were hospitalized. One patient received mechanical ventilation, and none underwent extracorporeal membrane oxygenation. One patient was admitted to the intensive care unit; there were no deaths. These results are useful for recognizing the clinical course from transmission route to outcomes of this infection in pediatric patients.