The Prevalence and Evolutionary Characteristics of Bovine Coronavirus in China (preprint)

Four viral intestinal pathogens such as bovine coronavirus (BCoV), bovine rotavirus (BRV), bovine viral diarrhea virus (BVDV) and bovine astrovirus (BoAstV) are the most common causes of adult cattle diarrhea. In this study, 797 samples were collected from January 2020 to February 2023. Among them, BCoV is major pathogen we detected and the positive rate was 48.18% (471/794). Farther more, among 307 diarrhea samples and 487 asymptomatic samples, 159 and 312 were BCoV positive, respectively. The results reveal that BCoV is associated with bovine diarrhea (p=0.001). In addition, we also amplified and analyzed Hemagglutinin-esterase (HE), Spike (S) and whole genome of partially positive sample. Getting 8 HE complete sequence, 8 S complete sequence and 5 whole genomes. The molecular characterization provides reveal that 5 strains were branched Chinese strains, Japan and part of American strains in GⅡb，and Korea strains branched in GⅡa. BCoV-Neimeng S in this study was mutated 143H/A to 143R, which is uncommon in S gene. As a residue on the side chain of the core binding site of S1-NTD, this mutation changed the tertiary structure and may be possible to affect the glycoside binding spectrum. The noteworthy that BCoV-SJZ2 and BCoV-YZ1 have four amino acid insertions on HE, which is the same as SWUN/A10/2018. We also analysis 4.8kDa and 4.9kDa non-structural protein large-scale truncation and membrane protein mutation. This article will discuss epidemiology and genome analysis.

Cross-lagged analysis of the relationship between risk perception, physical activity, and adolescent mental health (preprint)

Background Anxiety symptoms were prevalent in teenagers during the new coronavirus pandemic at 31% and depression symptoms were at 34%, both significantly higher than they were before the pandemic, according to research on global adolescent mood disorders during the epidemic. It is unclear, nevertheless, if physical activity still promotes mental health given the perceived danger of sickness, even though it has been demonstrated to be useful in reducing teenage mental health issues. Therefore, this study looked into the relationship between teenage mental health, risk perception, and physical activity. Methods In December 2022 and January 2023, two surveys were given to the same pupils in five high schools. During the New Crown pandemic, the risk perception scores, physical activity levels, and mental health of adolescents were examined in the study N=344. Results For adolescents' risk perceptions, there were significant gender differences (P＜0.01), with gender difference effects of 0.255 (d = 0.416) and 0.195 (d = 0.402) for the two measurements, respectively. For mental health, there were gender differences, with gender difference effects of 0.159 (d = 0.262) and 0.179 (d = 0.278) for the two measurements. The levels of risk perception, physical activity, and teenage mental health met persistent connections across months with contemporaneous correlations, however gender differences in physical activity levels were not significant (p > 0.05); Contrarily, in the cross-lagged study, males had higher levels of physical activity and mental health than females, and teenagers' risk perception was higher. Physical activity and mental health were both predicted by T1 ( values of 0.28, 0.19, and P＜0.01, respectively). Risk perception T1 physical exercise T2 mental health T2 (mediating value impact of 0.012, the Z value of 0.112), as well as the indirect effect of Bootstrap, were all able to predict mental health T2 (β= 0.33, P ＜0.01). Indicating a substantial mediating role for physical activity between risk perception and mental health is the 95% CI, not the 0 value from this pathway. Conclusion Raising awareness of physical activity among adolescents according to gender is important for improving mental health and there is a longitudinal causal relationship between perceived risk of disease, physical activity, and mental health among adolescents, suggesting that physical activity still has a role in mental health in the presence of risk perception.

Morphological, cellular, and molecular basis of brain infection in COVID-19 patients.

Although increasing evidence confirms neuropsychiatric manifestations associated mainly with severe COVID-19 infection, long-term neuropsychiatric dysfunction (recently characterized as part of "long COVID-19" syndrome) has been frequently observed after mild infection. We show the spectrum of cerebral impact of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, ranging from long-term alterations in mildly infected individuals (orbitofrontal cortical atrophy, neurocognitive impairment, excessive fatigue and anxiety symptoms) to severe acute damage confirmed in brain tissue samples extracted from the orbitofrontal region (via endonasal transethmoidal access) from individuals who died of COVID-19. In an independent cohort of 26 individuals who died of COVID-19, we used histopathological signs of brain damage as a guide for possible SARS-CoV-2 brain infection and found that among the 5 individuals who exhibited those signs, all of them had genetic material of the virus in the brain. Brain tissue samples from these five patients also exhibited foci of SARS-CoV-2 infection and replication, particularly in astrocytes. Supporting the hypothesis of astrocyte infection, neural stem cell-derived human astrocytes in vitro are susceptible to SARS-CoV-2 infection through a noncanonical mechanism that involves spike-NRP1 interaction. SARS-CoV-2-infected astrocytes manifested changes in energy metabolism and in key proteins and metabolites used to fuel neurons, as well as in the biogenesis of neurotransmitters. Moreover, human astrocyte infection elicits a secretory phenotype that reduces neuronal viability. Our data support the model in which SARS-CoV-2 reaches the brain, infects astrocytes, and consequently, leads to neuronal death or dysfunction. These deregulated processes could contribute to the structural and functional alterations seen in the brains of COVID-19 patients.

SARS-CoV-2 productively infects primary human immune system cells in vitro and in COVID-19 patients.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection is associated with a hyperinflammatory state and lymphocytopenia, a hallmark that appears as both signature and prognosis of disease severity outcome. Although cytokine storm and a sustained inflammatory state are commonly associated with immune cell depletion, it is still unclear whether direct SARS-CoV-2 infection of immune cells could also play a role in this scenario by harboring viral replication. We found that monocytes, as well as both B and T lymphocytes, were susceptible to SARS-CoV-2 infection in vitro, accumulating double-stranded RNA consistent with viral RNA replication and ultimately leading to expressive T cell apoptosis. In addition, flow cytometry and immunofluorescence analysis revealed that SARS-CoV-2 was frequently detected in monocytes and B lymphocytes from coronavirus disease 2019 (COVID-19) patients. The rates of SARS-CoV-2-infected monocytes in peripheral blood mononuclear cells from COVID-19 patients increased over time from symptom onset, with SARS-CoV-2-positive monocytes, B cells, and CD4+ T lymphocytes also detected in postmortem lung tissue. These results indicated that SARS-CoV-2 infection of blood-circulating leukocytes in COVID-19 patients might have important implications for disease pathogenesis and progression, immune dysfunction, and virus spread within the host.

SARS-Cov-2 pneumonia phenotyping on imaging exams of patients submitted to minimally invasive autopsy.

Background: Correlation between pathology and imaging of the new SARS-Cov-2 disease (COVID-19) is scarce. This study aimed to characterize SARS-Cov-2 pneumonia on imaging of patients submitted to minimally invasive autopsy (MIA). Methods: This unicentric retrospective observational study included 46 consecutive patients with confirmed COVID-19 who underwent MIA. All clinical chest images were reviewed and classified for the presence and grade of viral pneumonia, as well as disease evolution. On CT, phenotypes were described as consistent with mild, moderate, or severe viral pneumonia, with or without radiological signs of organizing pneumonia (OP). In severe pneumonia, CT could also be classified as diffuse progressive OP or radiological diffuse alveolar damage (DAD). Specific features on CT were noted, including fibroproliferative signs that could indicate potential or initial fibrosis. Results: MIA showed a heterogeneous panel of alterations, with a high prevalence of OP and acute fibrinous and organizing pneumonia (AFOP). Also, signs of interstitial fibrosis corresponded to the most prevalent pathological feature. Initial chest radiography (CXR) findings were mainly consistent with moderate or severe viral pneumonia. Most patients showed stability or improvement (reduction of opacities) on imaging. CTs were performed on 15 patients. Consolidations were found in most patients, frequently showing features consistent with an OP phenotype. Fibroproliferative changes were also prevalent on CT. Conclusions: In this study, SARS-Cov-2 pneumonia showed heterogeneous radiological and pathological patterns. Signs of organization and potential or initial fibrosis were prevalent on both imaging and pathology. Imaging phenotyping may help to predict post-infection fibrosing interstitial pneumonitis in COVID-19.

Inflammasomes are activated in response to SARS-CoV-2 infection and are associated with COVID-19 severity in patients.

Severe cases of COVID-19 are characterized by a strong inflammatory process that may ultimately lead to organ failure and patient death. The NLRP3 inflammasome is a molecular platform that promotes inflammation via cleavage and activation of key inflammatory molecules including active caspase-1 (Casp1p20), IL-1ß, and IL-18. Although participation of the inflammasome in COVID-19 has been highly speculated, the inflammasome activation and participation in the outcome of the disease are unknown. Here we demonstrate that the NLRP3 inflammasome is activated in response to SARS-CoV-2 infection and is active in COVID-19 patients. Studying moderate and severe COVID-19 patients, we found active NLRP3 inflammasome in PBMCs and tissues of postmortem patients upon autopsy. Inflammasome-derived products such as Casp1p20 and IL-18 in the sera correlated with the markers of COVID-19 severity, including IL-6 and LDH. Moreover, higher levels of IL-18 and Casp1p20 are associated with disease severity and poor clinical outcome. Our results suggest that inflammasomes participate in the pathophysiology of the disease, indicating that these platforms might be a marker of disease severity and a potential therapeutic target for COVID-19.

SARS-CoV-2-triggered neutrophil extracellular traps mediate COVID-19 pathology.

Severe COVID-19 patients develop acute respiratory distress syndrome that may progress to cytokine storm syndrome, organ dysfunction, and death. Considering that neutrophil extracellular traps (NETs) have been described as important mediators of tissue damage in inflammatory diseases, we investigated whether NETs would be involved in COVID-19 pathophysiology. A cohort of 32 hospitalized patients with a confirmed diagnosis of COVID-19 and healthy controls were enrolled. The concentration of NETs was augmented in plasma, tracheal aspirate, and lung autopsies tissues from COVID-19 patients, and their neutrophils released higher levels of NETs. Notably, we found that viable SARS-CoV-2 can directly induce the release of NETs by healthy neutrophils. Mechanistically, NETs triggered by SARS-CoV-2 depend on angiotensin-converting enzyme 2, serine protease, virus replication, and PAD-4. Finally, NETs released by SARS-CoV-2-activated neutrophils promote lung epithelial cell death in vitro. These results unravel a possible detrimental role of NETs in the pathophysiology of COVID-19. Therefore, the inhibition of NETs represents a potential therapeutic target for COVID-19.