Aging shapes infection profiles of influenza A virus and SARS-CoV-2 in human lung slices (preprint)

The recent coronavirus disease 2019 (COVID-19) outbreak revealed the susceptibility of elderly patients to respiratory virus infections, showing cell senescence or subclinical persistent inflammatory profiles and favouring the development of severe pneumonia. In our study, we evaluated the potential influence of lung aging on the efficiency of replication of influenza A virus (IAV) and severe acute respiratory syndrome coronavirus 2 (SARS-CoV 2), as well as determined the pro-inflammatory and antiviral responses of the distal lung tissue. Using precision-cut lung slices (PCLS) from donors of different ages, we found that pandemic H1N1 and avian H5N1 IAV replicated in the lung parenchyma with high efficacy. In contrast to these IAV strains, SARS-CoV-2 early isolate and Delta variant of concern (VOC) replicated less efficiently in PCLS. Interestingly, both viruses showed reduced replication in PCLS from older compared to younger donors, suggesting that aged lung tissue represents a sub optimal environment for viral replication. Regardless of the age-dependent viral loads, PCLS responded to infection with both viruses by an induction of IL-6 and IP-10/CXCL10 mRNAs, being highest for H5N1. Finally, while SARS-CoV-2 infection was not causing detectable cell death, IAV infection caused significant cytotoxicity and induced significant early interferon responses. In summary, our findings suggest that aged lung tissue might not favour viral dissemination, pointing to a determinant role of dysregulated immune mechanisms in the development of severe disease.

Effects of SARS-CoV-2 Infection on Pulmonary Function Tests and Exercise Tolerance

Introduction: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has showed to cause long-term pulmonary sequelae. Objects: The aim of this study was to evaluate the consequences on pulmonary function of the SARS-CoV-2 infection related to the severity of the disease and exercise tolerance. Method(s): a retrospective cohort study was performed at the "Policlinico Tor Vergata" Academic hospital (Rome, Italy) where 75 patients evaluated in the post-COVID outpatient clinics at the Respiratory units were included in the study. Complete pulmonary function tests, 6-minute walk tests and persistence of symptoms were evaluated. Result(s): among the 75 subjects, 23 were mild, 16 moderate, 26 severe and 10 very severe based on the WHO classification. Very severe patients had a lower FVC (100+/-10%pr) compared to the others groups (116+/-16%pr, 116+/-13%pr, 122+/-20%pr from mild to severe;p<0,05) and a lower TLC (94+/-13%pr) compared to the others (102+/-10%pr, 108+/-15%pr, 108+/-12%pr from mild to severe;p<0,05). DL'co and DL'co/VA were similar among groups. At the 6MWT, distance, basal and nadir SpO2 were similar among groups, but all groups presented a significant decrease of SpO2 from basal to the nadir (Basal SpO2: 97,0+/-1,0% vs Nadir SpO2: 93,6+/-2,7%, p.<0,01). A positive correlation was found between desaturation and delta SpO2 (basal-nadir) (R: 0.29, p<0,05) and the Distance Desaturation Product (R: 0.39, p<0,01). Conclusion(s): These results showed that, although the PFTs are within the normal range, there is a restrictive spirometric pattern in very severe subjects. Moreover, the only persistent pathological sequalae of SARS-CoV-2 infection were a mild desaturation at 6MWT, despite the severity.

Small Magnitude Events Highlight the Correlation Between Hydraulic Fracturing Injection Parameters, Geological Factors, and Earthquake Occurrence

Hydraulic fracturing (HF) operations are widely associated with induced seismicity in the Western Canadian Sedimentary Basin. This study correlates injection parameters of 12,903 HF stages in the Kiskatinaw area in northeast British Columbia with an enhanced catalog containing 40,046 earthquakes using a supervised machine learning approach. It identifies relevant combinations of geological and operational parameters related to individual HF stages in efforts to decipher fault activation mechanisms. Our results suggest that stages targeting specific geological units (here, the Lower Montney formation) are more likely to induce an earthquake. Additional parameters positively correlated with earthquake likelihood include target formation thickness, injection volume, and completion date. Furthermore, the COVID‐19 lockdown may have reduced the potential cumulative effect of HF operations. Our results demonstrate the value of machine learning approaches for implementation as guidance tools that help facilitate safe development of unconventional energy technologies.Alternate :Plain Language SummaryHydraulic fracturing (HF), a technique used in unconventional energy production, increases rock permeability to enhance fluid movement. Its use has led to an unprecedented increase of associated earthquakes in the Western Canadian Sedimentary Basin in the last decade, among other regions. Numerous studies have investigated the relationship between induced earthquakes and HF operations, but the connection between specific geological and operational parameters and earthquake occurrence is only partly understood. Here, we use a supervised machine learning approach with publicly available injection data from the British Columbia Oil and Gas Commission to identify influential HF parameters for increasing the likelihood of a specific operation inducing an earthquake. We find that geological parameters, such as the target formation and its thickness, are most influential. A small number of operational parameters are also important, such as the injected fluid volume and the operation date. Our findings demonstrate an approach with the potential to develop tools to help enable the continued development of alternative energy technology. They also emphasize the need for public access to operational data to estimate and reduce the hazard and associated risk of induced seismicity.

Atrial Septal Defect Closure-Device Thrombosis after SARS-CoV2 Infection: Description of a Clinical Case

INTRODUCTION. We describe the case of a patient who underwent a percutaneous ostium secundum atrial septal defect (OS-ASD) closure and developed a SARS-CoV2 infection a few days later, with the early onset of intracardiac thrombosis on the closure-device. CLINICAL CASE An 11 year-old girl, who underwent an OS-ASD percutaneous closure with a 32 mm occluder, was diagnosed with a mild COVID19 a week after the procedure. After the recovery, at 1-month medical evaluation, transthoracic echocardiography (TTE) showed a large mobile mass attached to the right side of the well-positioned device, impinging on the tricuspid orifice, with no evidence of significant rigurgitation or functional stenosis. The patient was asymptomatic and on DAPT with aspirin and clopidogrel. She was hospitalized, clopidogrel was interrupted and warfarin was started. We excluded endocarditis. Transesophageal echocardiography (TEE) confirmed the diagnosis of intracardiac thrombosis, without evidence of residual interatrial shunt. Angio-pulmonary computed tomography excluded thromboembolism. Thrombophilic screening documented 3 heterozygote mutations (Factor V Leiden, MTHFR C677T/ A1298C) and no pathogenetic CYP2C19 polymorphisms emerged. Because of clinical stability, we opted for a conservative approach, continuing warfarin (INR target 2-3) for at least 3 months and close follow-up. CONCLUSION Thrombosis on ASD-closure devices is uncommon (<1%). This case suggests that the COVID19- triggered hypercoagulability state may lead to severe complications in patients who already have a prothrombotic predisposition. Thus, a vigilant workup is advisable.

The spike gene is a major determinant for the SARS-CoV-2 Omicron-BA.1 phenotype.

Variant of concern (VOC) Omicron-BA.1 has achieved global predominance in early 2022. Therefore, surveillance and comprehensive characterization of Omicron-BA.1 in advanced primary cell culture systems and animal models are urgently needed. Here, we characterize Omicron-BA.1 and recombinant Omicron-BA.1 spike gene mutants in comparison with VOC Delta in well-differentiated primary human nasal and bronchial epithelial cells in vitro, followed by in vivo fitness characterization in hamsters, ferrets and hACE2-expressing mice, and immunized hACE2-mice. We demonstrate a spike-mediated enhancement of early replication of Omicron-BA.1 in nasal epithelial cultures, but limited replication in bronchial epithelial cultures. In hamsters, Delta shows dominance over Omicron-BA.1, and in ferrets Omicron-BA.1 infection is abortive. In hACE2-knock-in mice, Delta and a Delta spike clone also show dominance over Omicron-BA.1 and an Omicron-BA.1 spike clone, respectively. Interestingly, in naïve K18-hACE2 mice, we observe Delta spike-mediated increased replication and pathogenicity and Omicron-BA.1 spike-mediated reduced replication and pathogenicity, suggesting that the spike gene is a major determinant of replication and pathogenicity. Finally, the Omicron-BA.1 spike clone is less well-controlled by mRNA-vaccination in K18-hACE2-mice and becomes more competitive compared to the progenitor and Delta spike clones, suggesting that spike gene-mediated immune evasion is another important factor that led to Omicron-BA.1 dominance.

Understanding the Role and Impact of Poly (Ethylene Glycol) (PEG) on Nanoparticle Formulation: Implications for COVID-19 Vaccines

Poly (ethylene glycol) (PEG) is a widely used polymer in a variety of consumer products and in medicine. PEGylation refers to the conjugation of PEG to drugs or nanoparticles to increase circulation time and reduce unwanted host responses. PEG is viewed as being well-tolerated, but previous studies have identified anti-PEG antibodies and so-called pseudoallergic reactions in certain individuals. The increased use of nanoparticles as contrast agents or in drug delivery, along with the introduction of mRNA vaccines encapsulated in PEGylated lipid nanoparticles has brought this issue to the fore. Thus, while these vaccines have proven to be remarkably effective, rare cases of anaphylaxis have been reported, and this has been tentatively ascribed to the PEGylated carriers, which may trigger complement activation in susceptible individuals. Here, we provide a general overview of the use of PEGylated nanoparticles for pharmaceutical applications, and we discuss the activation of the complement cascade that might be caused by PEGylated nanomedicines for a better understanding of these immunological adverse reactions.

The spike gene is a major determinant for the SARS-CoV-2 Omicron-BA.1 phenotype (preprint)

Variant of concern (VOC) Omicron-BA1 has achieved global predominance in early 2022. Therefore, surveillance and comprehensive characterization of Omicron-BA.1 in advanced primary cell culture systems and multiple animal models is urgently needed. Here, we characterized Omicron-BA.1 and recombinant Omicron-BA.1 spike gene mutants in comparison with VOC Delta in well-differentiated primary human nasal and bronchial epithelial cells in vitro, followed by in vivo fitness characterization in naive hamsters, ferrets and hACE2-expressing mice, and in immunized hACE2-mice. We demonstrate a spike mediated enhancement of early replication of Omicron-BA.1 in nasal epithelial cultures, but limited replication in bronchial epithelial cultures. In Syrian hamsters, Delta showed dominance over Omicron-BA.1 and in ferrets, Omicron-BA.1 infection was abortive. In mice expressing the authentic hACE2-receptor, Delta and a Delta spike clone also showed dominance over Omicron-BA.1 and an Omicron-BA.1 spike clone, respectively. Interestingly, in naive K18-hACE2 mice, we observed Delta spike-mediated increased replication and pathogenicity and Omicron-BA.1 spike-mediated reduced replication and pathogenicity, suggesting that the spike gene is a major determinant of both Delta and Omicron-BA.1 replication and pathogenicity. Finally, the Omicron-BA.1 spike clone was less well controlled by mRNA-vaccination in K18-hACE2-mice and became more competitive compared to the progenitor and Delta spike clones, suggesting that spike gene-mediated immune evasion is another important factor that led to Omicron-BA.1 dominance.

Generation of precision-cut slice cultures of human placenta.

We present a protocol to generate an advanced ex vivo model of human placenta. We use a vibrating tissue slicer to obtain precision-cut slices representative of the entire thickness of human placenta. This approach delivers standardized cultures with a preserved microstructure and cellular composition comparable to the native tissue. We applied this system to study SARS-CoV-2 infection at the maternal-fetal interface. Moreover, this system can be used to investigate the basic functions of the human placenta in health and disease. For complete details on the use and execution of this protocol, please refer to Fahmi et al. (2021).

Immune-mediated Hepatitis associated with SARS-CoV-2 mRNA vaccination

In a fraction of SARS-CoV-2 vaccinees, hepatitis compatible with features of autoimmune hepatitis (AIH) have been observed. However, it remains unclear whether the association is coincidental, reflects drug-induced liver injury, or involves vaccine-induced antigen-specific immune activation. Here, we report a case of a 52-year-old male developing a transient hepatitis after the first mRNA vaccination and severe AIH-compatible hepatitis after the second. The intrahepatic immune cell infiltrate was analysed by highly multiplexed imaging mass cytometry broadly covering key immune cell populations. Liver and longitudinal blood samples were analysed for the presence and phenotype of SARS-CoV-2 Spike-specific CD8 T cells using MHC class I tetramer technology. Additionally, Serum titers against SARS-Cov2-Spike antibodies were assessed. We identified a panlobular CD8 T cell dominant immune cell infiltrate in the liver without significant plasma cell components. Spike-specific CD8 T cells were highly enriched within the intrahepatic CD8 T cell population expressing activation markers and a tissue-resident phenotype. The activation phenotype correlated with the circulating Spike-specific CD8 T cell profile and longitudinal analysis revealed a rapid decline of T cell activation after the initiation of budesonid therapy. However, the patient experienced a mild relapse under therapy that was paralleled by the peripheral activation of Spike-specific CD8 T cells and was controlled under systemic steroid therapy. Collectively, our results indicate that an immune-mediated hepatitis after COVID19 vaccination can present with typical clinical features of an AIH but can be pathophysiologically separated from a classical AIH. Whether a long-term immunosuppressive regimen will be required remains to be determined.

Diversity regained: Precautionary approaches to COVID-19 as a phenomenon of the total environment.

As COVID-19 emerged as a phenomenon of the total environment, and despite the intertwined and complex relationships that make humanity an organic part of the Bio- and Geospheres, the majority of our responses to it have been corrective in character, with few or no consideration for unintended consequences which bring about further vulnerability to unanticipated global events. Tackling COVID-19 entails a systemic and precautionary approach to human-nature relations, which we frame as regaining diversity in the Geo-, Bio-, and Anthropospheres. Its implementation requires nothing short of an overhaul in the way we interact with and build knowledge from natural and social environments. Hence, we discuss the urgency of shifting from current to precautionary approaches to COVID-19 and look, through the lens of diversity, at the anticipated benefits in four systems crucially affecting and affected by the pandemic: health, land, knowledge and innovation. Our reflections offer a glimpse of the sort of changes needed, from pursuing planetary health and creating more harmonious forms of land use to providing a multi-level platform for other ways of knowing/understanding and turning innovation into a source of global public goods. These exemplary initiatives introduce and solidify systemic thinking in policymaking and move priorities from reaction-based strategies to precautionary frameworks.

Rethinking hospital psychiatry in Italy in light of COVID-19 experience.

BACKGROUND: Italy retains a distinctive organization of mental health services according to a community-based model of care with a multidisciplinary team serving a well-defined catchment area under the coordination of the local department of mental health. The coronavirus disease 2019 (COVID-19) pandemic is forcing Italian mental health services to develop new organizational strategies at all levels of care in order to face the associated challenges. AIM: To explore factors associated with changes in psychiatric admissions to an inpatient psychiatric unit located in Lombardia Region, Italy. METHODS: All hospital admissions (n = 44) were recorded to an inpatient psychiatric unit during a three month national lockdown in Italy in 2020 and compared with those occurring over the same time period in 2019 (n = 71). For each admission, a 20-item checklist was completed to identify factors leading to admission. Statistical analyses were performed using Statistical Package for Social Sciences for Windows, release 11.0. Chi-square test (or Fisher's exact test) and Mann-Whitney U-test were applied, where appropriate. RESULTS: Hospital admissions dropped by 38% during the COVID-19 pandemic. No significant differences were found in demographics, clinical variables associated with hospital admissions and length of stay between 2019 and 2020. Compared with 2019, a significantly greater proportion of hospital admissions in 2020 were related to difficulties in organizing care programs outside the hospital (chi-square = 4.91, df 1, one-way P = 0.035) and in patients' family contexts (chi-square = 3.71, df 1, one-way P = 0.049). On the other hand, logistic and communication difficulties pertaining to residential facilities and programs were significantly more common in 2019 than in 2020 (chi-square = 4.38, df 1, one-way P = 0.032). CONCLUSION: Admissions to the inpatient psychiatric unit dropped significantly during the COVID-19 pandemic in 2020, with difficulties in organizing care programs outside the hospital and in patients' family contexts occurring more frequently compared with 2019.

Von willebrand factor and ADAMTS13 activity as endothelial injury markers and their prognostic value in COVID-19

Background : The coronavirus disease 2019 (COVID-19) increases thrombotic risk. The mechanisms that lead to this prothrombotic state are unclear. Aims : The main aim was to evaluate the von Willebrand factor (VWF) antigen and plasma ADAMTS13 activity as endothelial injury markers in COVID-19 and their prognostic value in COVID-19 evolution. Methods : We present a prospective study in COVID-19 patients recruited in our institution. The patients were divided into 2 groups depending on whether hospitalization was needed. Inpatients were subclassified into ward patients and those requiring intensive care. Thirty non-COVID-19 inpatients and 30 non-COVID-19 healthy individuals were recruited. VWF antigen, ADAMTS13 activity, D-dimer, and fibrinogen were measured during the first week once COVID-19 was diagnosed. Quantitative data were expressed as median (p25-p75) and qualitative data as percentage. Results : Fifty COVID-19 inpatients (44% in the intensive care unit [ICU]) and 102 COVID-19 outpatients were enrolled. Inpatients were older and had a higher incidence of hypertension and diabetes. The COVID-19 inpatients had higher D-dimer, fibrinogen, and VWF antigen levels and a lower ADAMTS13 activity compared with the COVID-19 outpatients ( P < 0.05). ICU patients had higher D-dimer and VWF antigen levels compared with the ward patients and the lowest ADAMTS13 activity ( P < 0.05). An imbalance in VWF antigen/ ADAMTS13 activity ratio was observed in COVID-19, reaching the highest in ICU patients. In contrast to other acute inflammatory diseases, a significative reduction in ADAMTS13 activity was observed in all COVID-19 patients. Conclusions : There is an increase in VWF antigen and an ADAMTS13 activity reduction in COVID-19 related to disease severity and could predict poor clinical outcomes. The ADAMTS13 activity reduction could be a marker associated with COVID-19 in contrast to other inflammatory conditions.

SARS-CoV-2 can infect and propagate in human placenta explants.

The ongoing SARS-CoV-2 pandemic continues to lead to high morbidity and mortality. During pregnancy, severe maternal and neonatal outcomes and placental pathological changes have been described. We evaluate SARS-CoV-2 infection at the maternal-fetal interface using precision-cut slices (PCSs) of human placenta. Remarkably, exposure of placenta PCSs to SARS-CoV-2 leads to a full replication cycle with infectious virus release. Moreover, the susceptibility of placental tissue to SARS-CoV-2 replication relates to the expression levels of ACE2. Viral proteins and/or viral RNA are detected in syncytiotrophoblasts, cytotrophoblasts, villous stroma, and possibly Hofbauer cells. While SARS-CoV-2 infection of placenta PCSs does not cause a detectable cytotoxicity or a pro-inflammatory cytokine response, an upregulation of one order of magnitude of interferon type III transcripts is measured. In conclusion, our data demonstrate the capacity of SARS-CoV-2 to infect and propagate in human placenta and constitute a basis for further investigation of SARS-CoV-2 biology at the maternal-fetal interface.

Von Willebrand factor and ADAMTS13 activity as clinical severity markers in patients with COVID-19.

The coronavirus disease 2019 (COVID-19) increases thrombotic risk. The mechanisms that lead to this prothrombotic state are not well established. The main aim was to evaluate the von Willebrand factor (VWF) antigen and plasma ADAMTS13 activity as endothelial injury markers in COVID-19. We present a prospective study in COVID-19 patients recruited in our institution. VWF antigen, ADAMTS13 activity, D-dimer, and fibrinogen were measured during the first week once COVID-19 was diagnosed. Fifty COVID-19 inpatients [44% in the intensive care unit (ICU)] and 102 COVID-19 outpatients were enrolled. Thirty age and gender matched non-COVID-19 ward inpatients and 30 non-COVID-19 healthy individuals were recruited. The COVID-19 inpatients had higher D-dimer, fibrinogen, and VWF antigen levels and a lower ADAMTS13 activity compared with the COVID-19 outpatients (p < 0.05). ICU patients had higher D-dimer and VWF antigen levels compared with the ward patients and the lowest ADAMTS13 activity (p < 0.05). An imbalance in VWF antigen/ADAMTS13 ratio was observed in COVID-19, reaching the highest in ICU patients. In contrast to other ward non-COVID-19 inpatients, a significative reduction in ADAMTS13 activity was observed in all COVID-19 patients. There is an increase in VWF antigen and an ADAMTS13 activity reduction in COVID-19 related to disease severity and could predict poor clinical outcomes. The ADAMTS13 activity reduction could be a marker associated with COVID-19 compared to other non-critical medical conditions.

mRNA vaccines manufacturing: Challenges and bottlenecks.

Vaccines are one of the most important tools in public health and play an important role in infectious diseases control. Owing to its precision, safe profile and flexible manufacturing, mRNA vaccines are reaching the stoplight as a new alternative to conventional vaccines. In fact, mRNA vaccines were the technology of choice for many companies to combat the Covid-19 pandemic, and it was the first technology to be approved in both United States and in Europe Union as a prophylactic treatment. Additionally, mRNA vaccines are being studied in the clinic to treat a number of diseases including cancer, HIV, influenza and even genetic disorders. The increased demand for mRNA vaccines requires a technology platform and cost-effective manufacturing process with a well-defined product characterisation. Large scale production of mRNA vaccines consists in a 1 or 2-step in vitro reaction followed by a purification platform with multiple steps that can include Dnase digestion, precipitation, chromatography or tangential flow filtration. In this review we describe the current state-of-art of mRNA vaccines, focusing on the challenges and bottlenecks of manufacturing that need to be addressed to turn this new vaccination technology into an effective, fast and cost-effective response to emerging health crises.

A New Chemical Probe Challenges the Broad Cancer Essentiality of CK2.

Casein kinase 2 (CK2) is highly expressed in cancer and has been considered a potential therapeutic target. Wells and colleagues developed and characterized the new CK2 inhibitor SGC-CK2-1. Unexpectedly, this potent and highly selective chemical probe does not show broad antiproliferative activity in cancer cells.