SARS-CoV-2 RNA Wastewater Settled Solids Surveillance Frequency and Impact on Predicted COVID-19 Incidence Using a Distributed Lag Model

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA concentrations in wastewater settled solids correlate well with coronavirus disease 2019 (COVID-19) incidence rates (IRs). Here, we develop distributed lag models to estimate IRs using concentrations of SARS-CoV-2 RNA from wastewater solids and investigate the impact of sampling frequency on model performance. SARS-CoV-2 N gene and pepper mild mottle virus (PMMoV) RNA concentrations were measured daily at four wastewater treatment plants in California. Artificially reduced data sets were produced for each plant with sampling frequencies of once every 2, 3, 4, and 7 days. Sewershed-specific models that related daily N/PMMoV to IR were fit for each sampling frequency with data from mid-November 2020 through mid-July 2021, which included the period of time during which Delta emerged. Models were used to predict IRs during a subsequent out-of-sample time period. When sampling occurred at least once every 4 days, the in- and out-of-sample root-mean-square error changed by <7 cases/100 000 compared to daily sampling across sewersheds. This work illustrates that real-time, daily predictions of IR are possible with small errors, despite changes in circulating variants, when sampling frequency is once every 4 days or more. However, reduced sampling frequency may not serve other important wastewater surveillance use cases. Distributed lag models predicted COVID-19 incidence rates with relatively small errors using wastewater solids surveillance of SARS-CoV-2 N genes.

Influence of traditional Chinese medicines on the in vivo metabolism of lopinavir/ritonavir based on UHPLC-MS/MS analysis.

A fast, reliable, and cost-effective liquid chromatography-tandem mass spectrometry method was established to determine the effects of the traditional Chinese medicine employed to treat coronavirus disease 2019, namely, Lianhua Qingwen granules, Huoxiang Zhengqi capsules, Jinhua Qinggan granules, Shufeng Jiedu capsules, and Angong Niuhuang pills, on the pharmacokinetics of lopinavir/ritonavir in rats. Blood samples were prepared using the protein precipitation method and atazanavir was selected as the internal standard (IS). Separation was performed on an Agilent ZORBAX eclipse plus C18 (2.1 mm × 50 mm, 1.8 µm) column using acetonitrile and water containing 0.1% formic acid as the mobile phase for gradient elution. The flow rate was 0.4 mL/min and the injection volume was 2 µL. Agilent Jet Stream electrospray ionization was used for mass spectrometry detection under positive ion multiple reaction monitoring mode at a transition of m/z 629.3→447.3 for lopinavir, m/z 721.3→296.1 for ritonavir, and m/z 705.4→168.1 for the IS. The method showed good linearity in the concentration range of 25-2500 ng/mL (r=0.9981) for lopinavir and 5-500 ng/mL (r=0.9984) for ritonavir. The intra-day and inter-day precision and accuracy were both within ±15%. Items, such as dilution reliability and residual effect, were also within the acceptable limits. The method was used to determine the effects of five types of traditional Chinese medicines on the pharmacokinetics of lopinavir/ritonavir in rats. The pharmacokinetic results showed that the half-life of ritonavir in the groups administered Lianhua Qingwen granules and Huoxiang Zhengqi capsules combined with lopinavir/ritonavir was prolonged by approximately 1.5- to 2-fold relative to that in the control group. Similarly, the pharmacokinetic parameters of lopinavir were altered. Overall, the results of this study offer important theoretical parameters for the effective clinical use of five types of traditional Chinese medicines combined with lopinavir/ritonavir to reduce the occurrence of clinical adverse reactions.

T-Cell Repertoire Characteristics of Asymptomatic and Re-Detectable Positive COVID-19 Patients.

The prevention of the COVID-19 pandemic is highly complicated by the prevalence of asymptomatic and recurrent infection. Many previous immunological studies have focused on symptomatic and convalescent patients, while the immune responses in asymptomatic patients and re-detectable positive cases remain unclear. Here we comprehensively analyzed the peripheral T-cell receptor (TCR) repertoire of 54 COVID-19 patients in different courses, including asymptomatic, symptomatic, convalescent, and re-detectable positive cases. We identified a set of V-J gene combinations characterizing the upward immune responses through asymptomatic and symptomatic courses. Furthermore, some of these V-J combinations could be awakened in the re-detectable positive cases, which may help predict the risk of recurrent infection. Therefore, TCR repertoire examination has the potential to strengthen the clinical surveillance and the immunotherapy development for COVID-19.

High-Frequency, High-Throughput Quantification of SARS-CoV-2 RNA in Wastewater Settled Solids at Eight Publicly Owned Treatment Works in Northern California Shows Strong Association with COVID-19 Incidence.

A number of recent retrospective studies have demonstrated that severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA concentrations in wastewater are associated with coronavirus disease 2019 (COVID-19) cases in the corresponding sewersheds. Implementing high-resolution, prospective efforts across multiple plants depends on sensitive measurements that are representative of COVID-19 cases, scalable for high-throughput analysis, and comparable across laboratories. We conducted a prospective study across eight publicly owned treatment works (POTWs). A focus on SARS-CoV-2 RNA in solids enabled us to scale up our measurements with a commercial lab partner. Samples were collected daily, and results were posted to a website within 24 h. SARS-CoV-2 RNA in daily samples correlated with the incidence of COVID-19 cases in the sewersheds; a 1 log10 increase in SARS-CoV-2 RNA in settled solids corresponds to a 0.58 log10 (4×) increase in sewershed incidence rate. SARS-CoV-2 RNA signals measured with the commercial laboratory partner were comparable across plants and comparable to measurements conducted in a university laboratory when normalized by pepper mild mottle virus (PMMoV) RNA. Results suggest that SARS-CoV-2 RNA should be detectable in settled solids for COVID-19 incidence rates of >1/100,000 (range, 0.8 to 2.3 cases per 100,000). These sensitive, representative, scalable, and comparable methods will be valuable for future efforts to scale up wastewater-based epidemiology. IMPORTANCE Access to reliable, rapid monitoring data is critical to guide response to an infectious disease outbreak. For pathogens that are shed in feces or urine, monitoring wastewater can provide a cost-effective snapshot of transmission in an entire community via a single sample. In order for a method to be useful for ongoing COVID-19 monitoring, it should be sensitive for detection of low concentrations of SARS-CoV-2, representative of incidence rates in the community, scalable to generate data quickly, and comparable across laboratories. This paper presents a method utilizing wastewater solids to meet these goals, producing measurements of SARS-CoV-2 RNA strongly associated with COVID-19 cases in the sewershed of a publicly owned treatment work. Results, provided within 24 h, can be used to detect incidence rates as low as approximately 1/100,000 cases and can be normalized for comparison across locations generating data using different methods.

Effects of a Web-Based Parent-Child Physical Activity Program on Mental Health in Parents of Children with ASD.

Parents of children with ASD experience a higher incidence of mental health difficulties, including stress, depression, and anxiety, than parents of children without ASD. According to studies related to ASD, parent-child physical activity programs are an effective approach to encourage both parents and their children with ASD to exercise together, thus improving the mental health of parents due to this interactive family activity. The purpose of the present study was to explore the effects of this web-based parent-child physical activity program on the mental health of parents of children with ASD. A total of 94 parent-child pairs consented to participate in this study, and 75 parent-child pairs completed the study. Three instruments-DASS-21, PSI-4-SF, and WHOQOL-26-were used to measure mental health, parental stress, and quality of life, respectively. A randomized controlled trial design was implemented to examine the effectiveness of the 10-week web-based parent-child physical activity program on improving the mental health of parents of children with ASD. The results showed that after the 10-week parent-child physical activity program, there were significant differences in overall DASS-21 and PSI-4-SF for the experimental group, compared with control group (p < 0.05), which indicated that the parent-child physical activity program has a positive influence on mental health in parents of children with ASD. One sub-area of WHOQOL-26 between the experimental and control groups across pre-/post-testing intervals also showed greater reductions in the item of psychological health (p < 0.05). In conclusion, the findings demonstrated the efficacy of the web-based parent-child physical activity program for improving mental health in parents of children with ASD.

Management of Follow-Up With Preterm Infants During the Outbreak in China.

Introduction: Coronavirus disease 2019 (COVID-19) swept Wuhan in January 2020. Other cities in China also suffered during the pandemic. Routine medical services were conducted in the Neonatal Intensive Unit (NICU) as usual, but the follow-up after discharge was seriously affected. Objective: To investigate the feasibility and effectiveness of a combination of online and face-to-face follow-up for preterm infants during the COVID-19 epidemic and to explore a follow-up pattern that can provide follow-up services while maximizing the protection of preterm infants and soothing the fear of their parents. Methods: Preterm infants (n = 35) whose first follow-up appointment was scheduled from February 1 to April 30, 2020, and preterm infants (n = 43) in the NICU follow-up group who were discharged from January 1, 2018, to January 31, 2020, who had a second or later routine follow-up appointment scheduled from February 1 to April 30, 2020, were enrolled. We provided a combination of online and face-to-face follow-up for preterm infants surveyed with the Wenjuanxing platform before and after the online follow-up and compared the first-time follow-up rate between the outbreak and the same period of the previous year. Results: Feeding and oral medicine and supplements were the most concerning problems of the parents of preterm infants. The anxiety level of the family was significantly decreased after online follow-up (P < 0.05). A total of 96.8% of parents were satisfied or very satisfied with online follow-up, and 95.2% of parents thought that online follow-up had answered all their questions. Only 35.5% of parents thought online follow-up could replace face-to-face follow-up. Conclusion: The combination of online and face-to-face follow-up alleviated the anxiety of the parents during the outbreak and achieved a similar first-time follow-up rate as the same period in 2019.

Pre-optimized phage therapy on secondary Acinetobacter baumannii infection in four critical COVID-19 patients.

Phage therapy is recognized as a promising alternative to antibiotics in treating pulmonary bacterial infections, however, its use has not been reported for treating secondary bacterial infections during virus pandemics such as coronavirus disease 2019 (COVID-19). We enrolled 4 patients hospitalized with critical COVID-19 and pulmonary carbapenem-resistant Acinetobacter baumannii (CRAB) infections to compassionate phage therapy (at 2 successive doses of 109 plaque-forming unit phages). All patients in our COVID-19-specific intensive care unit (ICU) with CRAB positive in bronchoalveolar lavage fluid or sputum samples were eligible for study inclusion if antibiotic treatment failed to eradicate their CRAB infections. While phage susceptibility testing revealed an identical profile of CRAB strains from these patients, treatment with a pre-optimized 2-phage cocktail was associated with reduced CRAB burdens. Our results suggest the potential of phages on rapid responses to secondary CRAB outbreak in COVID-19 patients.