SARS-CoV2 RNA Research Articles

The differential immune response in mild versus fatal SARS-CoV2 infection

This study compared the immune response in mild versus fatal SARS-CoV2 infection. Forty nasopharyngeal swabs with either productive mild infection (n = 20) or negative for SARS-CoV2 (n = 20) were tested along with ten lung sections from people who died of COVID-19 which contained abundant SARS-CoV2 and ten controls. There was a 25-fold increase in the CD3+T cell numbers in the viral positive nasopharyngeal swabs compared to the controls (p < 0.001) and no change in the CD3+T cell count in the fatal COVID-19 lungs versus the controls. CD11b + and CD206+ macrophage counts were significantly higher in the mild versus fatal disease (p = 0.002). In situ analysis for SARS-CoV2 RNA found ten COVID-19 lung sections that had no/rare detectable virus and also lacked the microangiopathy typical of the viral positive sections. These viral negative lung tissues when compared to the viral positive lung samples showed a highly significant increase in CD3+ and CD8 T cells (p < 0.001), equivalent numbers of CD163+ cells, and significantly less PDL1, CD11b and CD206+ cells (p = 0.002). It is concluded that mild SARS-CoV2 infection is marked by a much stronger CD3/CD8 T cell, CD11b, and CD206 macrophage response than the fatal lung disease where viral RNA is abundant.

POS-003 ACUTE KIDNEY INJURY IN CHILDREN WITH COVID-19 RELATED MULTISYSTEM INFLAMMATORY SYNDROME (MIS-C)

Although children with coronavirus disease 19 (COVID-19) generally experience a mild disease, a subset of them develop multisystem inflammatory syndrome (MIS-C) which can lead to multiorgan failure. There is relative rarity of literature regarding acute kidney injury (AKI) in MIS-C. We aim to characterize the clinical features, laboratory findings, and therapies for AKI in MIS-C in our setup. This was a 1 ½ year prospective study with patients from GMC Srinagar, its associated hospitals and Shifa Medical Centre, Srinagar. Children <21 years old who had AKI and met the criteria for MIS-C based on CDC guidelines were included in the study. A total of seven cases were included in the study ranging from 4 to 20 years (mean 11.4±5) with 4 females and 3 males. Persistent fever was present in all patients. Six children had vomiting/diarrhea along with rashes and/or swelling of hands. Myocardial involvement was seen in four, respiratory in two and musculoskeletal in one patient. Oropharyngeal swab for SARS-Cov2 RNA was negative in all the patients. Anticovid antibodies were positive in five patients and two had a history of contact with COVID-19 patients. AKI Stage 1 was present in 3, stage 2 and 3 in 2 patients each. Neutrophilia with lymphopenia was seen in all the patients and thrombocytopenia in 4 patients. Laboratory findings for inflammatory markers showed marked elevation of C-reactive protein (mean 87.6±72.1mg/l), ferritin (mean 810±224ng/ml), procalcitonin (mean 4.9±2.1ng/ml), ESR (mean 64.6±21.9mm/hr), fibrinogen (690±142.1mg/dl), LDH (mean 578.2±370.1U/L) and D-Dimer (mean 7.8±9.4 µgFFU/ml). The patients were treated with a combination of steroids, IVIg and inotropic support wherever needed. All of the patients recovered with a median duration hospital stay of 7 (IQR 5) days. Children with covid 19 infection should be carefully followed for MIS-C. Although children with MIS-C develop AKI, most of them have full clinical recovery. The long term prognosis of this syndrome is currently unknown until their follow up data becomes available in future.

Second round of an interlaboratory comparison of SARS-CoV2 molecular detection assays used by 45 veterinary diagnostic laboratories in the United States.

The COVID-19 pandemic presents a continued public health challenge. Veterinary diagnostic laboratories in the United States use RT-rtPCR for animal testing, and many laboratories are certified for testing human samples; hence, ensuring that laboratories have sensitive and specific SARS-CoV2 testing methods is a critical component of the pandemic response. In 2020, the FDA Veterinary Laboratory Investigation and Response Network (Vet-LIRN) led an interlaboratory comparison (ILC1) to help laboratories evaluate their existing RT-rtPCR methods for detecting SARS-CoV2. All participating laboratories were able to detect the viral RNA spiked in buffer and PrimeStore molecular transport medium (MTM). With ILC2, Vet-LIRN extended ILC1 by evaluating analytical sensitivity and specificity of the methods used by participating laboratories to detect 3 SARS-CoV2 variants (B.1; B.1.1.7 [Alpha]; B.1.351 [Beta]) at various copy levels. We analyzed 57 sets of results from 45 laboratories qualitatively and quantitatively according to the principles of ISO 16140-2:2016. More than 95% of analysts detected the SARS-CoV2 RNA in MTM at ≥500 copies for all 3 variants. In addition, for nucleocapsid markers N1 and N2, 81% and 92% of the analysts detected ≤20 copies in the assays, respectively. The analytical specificity of the evaluated methods was >99%. Participating laboratories were able to assess their current method performance, identify possible limitations, and recognize method strengths as part of a continuous learning environment to support the critical need for the reliable diagnosis of COVID-19 in potentially infected animals and humans.

850 Potential role of skin in SARS-CoV-2 infection

It is well accepted that the main route of the SARS-CoV-2 entry to the body is via respiratory epithelial cells. However, skin cells also express ACE2 and TMPRSS2, the major factors that control SARS-CoV-2 host cell entrance. The dermatological lesions including chilblain-like acral lesions (COVID toes), vasculitis-like, urticaria-like lesions and maculopapular eruptions were reported in 1%–20% of COVID-19 patients, and SARS CoV-2 RNA was detected in skin biopsy and autopsy of some of COVID-19 patients.

SARS-CoV-2 Testing of Aircraft Wastewater Shows That Mandatory Tests and Vaccination Pass before Boarding Did Not Prevent Massive Importation of Omicron Variant into Europe.

Background: Most new SARS-CoV-2 epidemics in France occurred following the importation from abroad of emerging viral variants. Currently, the risk of new variants being imported is controlled based on a negative screening test (PCR or antigenic) and proof of up-to-date vaccine status, such as the International Air Transport Association travel pass. Methods: The wastewater from two planes arriving in Marseille (France) from Addis Ababa (Ethiopia) in December 2021 was tested by RT-PCR to detect SARS-CoV2 and screen for variants. These tests were carried out between landing and customs clearance and were then sequenced by MiSeq Illumina. Antigenic tests and sequencing by NovaSeq were carried out on respiratory samples collected from the 56 passengers on the second flight. Results: SARS-CoV-2 RNA suspected of being from the Omicron BA.1 variant was detected in the aircraft’s wastewater. SARS-CoV2 RNA was detected in 11 [20%) passengers and the Omicron BA.1 variant was identified. Conclusion: Our work shows the efficiency of aircraft wastewater testing to detect SARS-CoV-2 cases among travellers and to identify the viral genotype. It also highlights the low efficacy of the current control strategy for flights entering France from outside Europe, which combines a requirement to produce a vaccine pass and proof of a negative test before boarding.

What does the evidence for COVID-19 in dogs and cats point to? An investigation in the literature

In view of current COVID-19 pandemic scenario, uncertainties still remains around the role of domestic animals on COVID-19 epidemiology, this systematic review aims to present the scientific evidence available, so far, on dogs and cats epidemiological role in the COVID-19 pandemic. The systematic review was conducted in accordance with the recommendations of the Preferred Reporting Items for Systematic Reviews and Meta-Analyses – PRISMA. Selected studies reports negative results of dogs and cats both in serological tests, for the detection of neutralizing antibodies, and in tests that investigated the presence of the SARS-CoV2 RNA. We verified the lack of national studies, which serves as a motivation for more research to be carried out in Brazil. In Brazil, in the animals tested, 5.4 and 6.2% of cats and dogs were positive, respectively. Few studies, so far, have been carried out with stray animals or animals belonging to shelters. This absence of broad conducted studies on animal interface enphasizes the need for more research with animals in vulnerable conditions, since they can contribute to the spread of the disease if they are confirmed as positive for SARS-COV-2. The compilation of data, through this systematic review of the literature, to date suggests that dogs and cats are not factors of viral spread to humans. However, further studies are still recommended.

COVID-19-associated pathological changes in placentae of third-trimester pregnant women

Background and aim Recently, many instances of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) [coronavirus disease 2019 (COVID-19)] placental invasion in late pregnancy have been reported, and this is still not completely clear. Controversy data are sequentially published, so more studies are needed to investigate placental pathology as it gives vital information on placental changes, mechanisms of maternal–fetal infection transmission, and viral effects on the placenta as inflammatory or vascular changes. We aimed to find out the placental pathological changes in term placentae and to detect SARS-CoV-2 RNA in specimens fixed in formalin and embedded in paraffin. Patients and methods A total of 30 placentae were obtained from COVID-19-infected pregnant women. They were examined for pathological changes by histopathology; immunohistochemistry for CD3, CD20, CD15, CD68, angiotensin-converting enzyme 2, cytokeratin, and PDL-1; and reverse transcription-PCR to recognize viral RNA. Results According to the COVID-19 status at the delivery time, four groups were identified. Group 1 (12 patients with COVID-19 at the delivery time) showed chronic villitis and placental maternal and fetal malperfusion. Group 2 (seven cases that turned negative by PCR of the nasopharyngeal swab prior to delivery by 5–8 days) showed maternal malperfusion but no villitis. Both groups were SARS-CoV-2 RNA positive by PCR of the nasopharyngeal swab. Group 3 (five cases that turned negative 1 day before delivery) showed maternal–fetal malperfusion and mild chronic subchorionitis and deciduitis. Only three had SARS-CoV RNA. Group 4 (six cases with a persistent negative swab for 20 days before delivery) showed focal villitis (much less than other groups) and maternal malperfusion. Only two had SARS-CoV-2 RNA. Immunophenotyping of cases in the four groups detected mixed inflammatory infiltrate with variable diffuse angiotensin-converting enzyme 2 immunostaining of villus and extravillous trophoblasts. Cytokeratin staining detected thinning and discontinuity of syncytiotrophoblast and strong PD-L1 expression was seen in villitis areas. Conclusion SARS-CoV-2-positive women exhibited a diverse range of results; thus, pregnant women should not be considered a homogeneous group as outcomes are dependent on a multitude of interdependent circumstances.

Histologic, viral, and molecular correlates of heart disease in fatal COVID-19

Cardiac manifestations are common in severe COVID-19. This study compared the histologic, viral, and molecular findings in cardiac tissue in fatal COVID-19 (n = 11) and controls (n = 11). In situ hybridization (SARS-CoV2 RNA) and immunohistochemistry for viral proteins and the host response were quantified for the samples and compared with qRTPCR and Western blot data. Control hearts showed a high resident population of macrophages that had variable ACE2 expression. Cardiac ACE2 expression was 10× greater in the heart tissues of cases and controls with obesity or type II diabetes. Multifocal endothelial cell swelling and degeneration, perivascular edema plus microvascular thrombi were unique to the cases. SARS-CoV2 RNA and nucleocapsid protein were rarely detected in situ in any COVID-19 heart. However, in each case abundant SARS-CoV-2 spike protein was evident. Co-expression experiments showed that the spike protein localized mostly to the ACE2+ interstitial macrophages/pericytes that were activated as evidenced by increased IL6 and TNFα expression. Western blots confirmed the presence of the viral spike protein, but not the nucleocapsid protein, in the cardiac homogenates. The intercalated disc proteins connexin 43, the primary cardiac gap junction protein, and NaV1.5, the predominant cardiac sodium channel, each showed marked lateral migration in the myocytes in the cases, which would increase the risk of reentrant arrhythmias. It is concluded that the viral spike protein, endocytosed by macrophages/pericytes, can induce a myocarditis with the possibility of conduction dysfunction due to abnormal localization of key intercalated disc proteins.

OX40 boosts and sustains humoral and cellular immune responses to SARS-CoV2 spike protein and RNA vaccinations

Abstract To prevent SARS-CoV-2 infections and generate long-lasting immunity, vaccines need to generate both, a strong humoral immune response as well as viral-specific CD4 and CD8 T cells. Previous results from our lab have shown that immunization in the presence of an agonist antibody targeting OX40 led to higher antibody titers and increased numbers of antigen-specific CD4 and CD8 T cells. Using the same strategy, we explored the effect of OX40 co-stimulation in the prime and boost together with the SARS-CoV-2 spike protein + adjuvant in C57Bl/6 mice. Our results show that OX40 engagement led to a significant increase in long-lived antibody responses when compared to mice that did not receive additional co-stimulation. In addition, spike protein and peptide-specific proliferation were greatly increased for both CD4 and CD8 T cells, with augmented secretion of IFN-γ, TNF-α and IL-2. Booster immunizations (7 months post prime) did not lead to anergy, but instead further increased the antibody and T cell responses. In initial experiments, the self-amplifying RNA (saRNA) vaccination generated lower antibody titers, independent of OX40 co-stimulation. However, the saRNA vaccine did induce a robust expansion of antigen-specific CD8 T cells, that expressed high levels of GrzmB, which was further increased by OX40 administration. The strong immune responses and the differential effects of the protein and saRNA vaccines suggest that heterologous prime-boost approaches, as currently approved in adults, might be beneficial in boosting different arms of the anti-viral immune response against SARS-CoV2, with OX40 agonists enhancing both approaches. Further studies in animals will aid defining the effects and benefits of this approach.

Ophthalmic Manifestation of COVID-19 Disease

Coronavirus is the novel coronavirus known as severe acute respiratory syndrome coronavirus 2 (SARS-CoV2). It's a non-segmented enveloped positive sense RNA virus with 82 percent genetic similarity to the SARS coronavirus (SARSCoV), which sparked an outbreak in early 2003. Respiratory droplets are assumed to be the primary mechanism of transmission because the virus is found in respiratory secretions. Conjunctival transmission and aerosol transmission have also been proposed, however they are also problematic. Fever, tiredness, and dry mouth are the three most prevalent COVID-19 clinical symptoms. In several patients, however, conjunctivitis was the initial symptom. The disease's respiratory complications have been the focus of diagnostic and therapeutic efforts, However, a number of ocular concerns have surfaced. Infected patients' tears have been confirmed to contain SARSCov2 RNA, and studies indicate that the virus is spreading. For viral transmission, the ocular surface could serve as both an entry point and a reservoir. COVID-19 has been linked to mild conjunctivitis, which can be the disease's first and only symptom. Ocular symptoms can be treated with observation without therapy, antibacterial eye drops, antiviral eye drops, and antiallergic eye drops, according to clinical trials. As a result, the only recommended treatment for COVID-19 related ocular manifestations is close observation. The motivation behind this examination was to investigate the event of visual appearances in patients who had been determined to have COVID illness 2019 (coronavirus) because of serious intense respiratory condition.

COVID-19 pandemic and kidney disease in children: literature review and own regional observations

Introduction. Currently, the coronavirus infection pandemic caused by the SARS-CoV-2 virus continues around the world. Research data from domestic and foreign authors indicate that the kidneys are a target organ for a new infection, lesions vary from proteinuria and hematuria to acute kidney injury.Aim of the study – to determine the frequency and nature of kidney damage in children with confirmed coronavirus infection.Materials and methods. A retrospective and prospective analysis of cases of confirmed COVID-19 infection in children (n = 441) admitted to the Samara Regional Children’s Infectious Diseases Hospital from March 2020 to July 2021 was carried out. SARSCoV-2 RNA was detected in all patients by a one-step reverse transcription reaction combined with a polymerase chain reaction. The changes in the kidneys that occurred in 57 children were studied. The research results were processed using the Statistica 7.0 software (StatSoft, USA).Results. The involvement of the kidneys in the infectious process was detected in every 8 children with COVID-19 (12.9%), more often in the form of isolated urinary syndrome, the detection rate of which correlated with the severity of the course of coronavirus infection: in severe cases, proteinuria was detected in 31.6% of patients, hematuria – in 21%, acute kidney injury – in 10.5%, diabetic nephropathy – in 5.3%. Kidney damage was combined with damage to the respiratory and gastrointestinal tract, characterized by rapid recovery of urine output and azotemia parameters without special renal therapy. A clinical case of the onset of nephrotic syndrome that developed 2 weeks after suffering a coronavirus infection is described.Conclusions. Children with COVID-19 require kidney function monitoring for early detection and correction in case of impairment. Patients with isolated urinary syndrome in the acute period require long-term observation in order to detect latent renal pathology.

Case series of acute kidney injury in children with SARS-CoV2 associated multisystem inflammatory syndrome

PurposeIn the first part of 2021 in Khimki regional hospital was eight-times increase of hemolytic-uremic syndrome (HUS) in children vs 2020. There are some data on association of HUS and COVID-19 in the literature. Our aim to describe case series of HUS in children with SARS-Cov2 associated multisystem inflammatory syndrome (MIS-C).Methods & MaterialsWe developed 8 children, age 1-11 yrs. (Me 4,5), 3M/5F. All had anemia (Hb 58 - 105, Me 83,8 g/l), thrombocytopenia (45 -136, Me 84*109/l) and kidney injury (azotemia, diuresis decrease, reduced GFR).ResultsIn anamnesis nobody had COVID-19, SARS-Cov2 RNA (oropharyngeal swab) was negative. 6 had anti-SARS-Cov2 IgG. 3 patients had antibody inversion during hospitalization. That means, they developed acute kidney injury (AKI) in early convalescence period of COVID-19. 6 children had fever, vomiting and diarrhea in anamnesis. 1 had only myalgia, 1 had transient hemorrhagic rush.All children had SIRS: ESR acceleration (15-43 mm/h), CRP increase in 6 cases (not more 5 norms), elevated ferritin (307-648 mkg/l) and D-dimer (3,24-8 mkg/ml). Mixed urinary syndrome was noted: proteinuria (0,1 - 6 g/l), hematuria (RBC 6-50). 100% had azotemia: urea (12,7-65,3, Me 35 mmol/l), creatinine (74-1450,2, Me 499 mkmol/l). GFR by Schwartz formula was reduced in all cases (1,7-58,1 ml/min/1,73 m2). 3 had anuria, 2 oliguria. Renal replacement therapy (RRT) performed in 5 cases: 3 hemodiafiltration (HDF) only; 2 - HDF + plasma exchange (1 - 7 procedures). All children were complex treated and discharged with normal clinical urine test and creatinine level, diuresis 1,6-2,4 ml/kg/h.ConclusionIn MIS-C pathogenesis thrombotic microangiopathia is one of the key points. We saw elevation of HUS in children in the COVID-19-pandemia period. 6 from 8 described cases had connection of AKI with previous COVID-19. In addition, they all had SIRS markers, corresponding diagnostic criteria of SARS-Cov2 associated multisystem inflammatory syndrome.

SARS-CoV2-associated multisystem inflammatory syndrome in children: data from Khimki hospital

PurposeMultisystem inflammatory syndrome in children (MIS-C) – the most severe status, associated with SARS-COV-2 infection. Clinical and laboratory characteristics of MIS-C, and treatment optimization are topical problems on current stage. Our aim to analyze main features of the MIS-C.Methods & MaterialsWe evaluated 41 children (21M/18F) 1 - 16 yrs. (8,45±0,677 yrs.) hospitalized in Khimki Regional Hospital with MIS-C from June 2020 to April 2021. 7 children had background disease (allergy, nephroblastoma, ulcerative colitis, diabetes mellitus, urologic pathology). 5 children were mongoloid race (12%). Excessive weight had 27%, according to BMI; 17% had high growth to age. 58% had A(II) blood group, Rh(+)-positive were 74%.ResultsAll children at the admission had fever more than 3 days. Anti-SARS-COV-2 IgM and IgG had 1 patient, IgG only - 25 (61%); 2 patients were positive SARS-Cov2 RNA in the oropharyngeal swab; 27% had family contact to СOVID-19. By severity 66% children were hospitalized to the ICU. Clinically skin and mucosal signs had 31 (76 %), gastro-intestinal symptoms – 22 (54%), pneumonia – 20 (49%), kidney injury –15 (40%), cardiovascular damage - 10 (24%), CNS –6 (15%). Simultaneous damage of 2 systems had 37% children, 3 – 46%, 4 - 12%, 5 – 1 patient, 6 – 1 patient. 5 cases developed acute kidney injury like hemolytic-uremic syndrome (HUS), as the main MIS-C performance.Laboratory features: increased ESR (Мe 40,0 (IQR 30-50) мм/ч), elevated CRP (Мe 118,9 (IQR 71,5-129,4)), ferritin (Мe 471,0 (IQR 214,08-990,28)), D-dimer (Мe 2,81 (IQR 1,76 – 4,55)), LDH (Мe 594,0 (IQR 511,0-663,0)), CK (Мe 112,0 (IQR 61,35-288,7)), CK-MB (Мe 44,95 (IQR 33,0-80,1)), decreased albumin (Мe 27,4 (IQR 23,0-33,0)). In 76% patients were treated with methylprednisolone pulse, IVIG in 20%, standard doses of corticosteroids – 10%. All children discharged home with recovery.ConclusionMIS-C is the most severe form of COVID-19 in children. In Khimki Regional Hospital 41 patient were successfully treated. The main features are severe status (66%-ICU), 63% children had 3+ affected systems; the main symptoms are skin and mucosal damage, gastrointestinal signs and headache, high BMI. There were 5 patients with HUS, during SARS-Cov2 associated MIS-C.

COVID-19 IgG/IgM patterns, early IL-6 elevation and long-term radiological sequelae in 75 patients hospitalized due to interstitial pneumonia followed up from 3 to 12 months.

BackgroundCOVID-19 pandemic resulted in about 165 million infections and 3.4 million deaths all over the world across 15 months. The most severe clinical presentation of COVID-19 diseases is interstitial pneumonia.MethodsIn this paper we describe clinical outcomes based on radiological features as well as the pattern of haematochemical parameters and IgG/IgM antibodies in 75 patients hospitalized due to COVID-related interstitial pneumonia not requiring intensive care assistance. Each patient underwent routine laboratory tests, including inflammatory markers and coagulation profile at baseline. Computed Tomography (CT) was performed at baseline and after 3 months to assess the persistence of radiological sequelae. A Generalized Linear Model (GLM) was used to test for each patient the association between individual haematochemical parameters at the time of hospital admission and the subsequent radiological features after three months. The presence of IgG antibodies was quantitatively determined in 70 patients at the time of hospital admission and after 3 months. A subgroup of 49 and 21 patients underwent additional dosage of IgG after 6 and 12 months, respectively. IgM serological antibodies were available for 17 patients at baseline and 61 at T3, with additional follow-up for 51 and 20 subjects after 6 and 12 months, respectively.ResultsOnly 28 out of 75 patients discharged from the hospital were totally healed after 3 months, while 47 patients (62.7%) still presented radiological sequelae. According to the GLM model, specific haematochemical baseline parameters—such as IL-6, GPT, platelets and eosinophil count—showed a statistically significant association with the presence of radiological sequelae at month 3 highlighting an OR = 0.5, thus meaning that subjects completely healed after 3 months presented half levels of IL-6 at baseline compared to patients with sequelae. In general, IgG serum levels were always higher than IgM at the time of hospitalization (75% at T0; n = 12 out of 16 patients with data available in both visits), after 3 months (72.1%; n = 44 out of 61 pts.), after 6 months (56.8%; 25 out of 44 pts.), and one year after hospitalization (60%; 12 out of 20 pts.). Overall, IgG and IgM serum levels presented a statistically significant decreasing trend from the baseline to month 3, 6 and 12. One patient presented an increase in IgM between baseline and month 3 but negative PCR test for SARS-COV2 on throat swab.ConclusionsAs supported by our findings on 75 patients, COVID-related interstitial pneumonia triggers early IgG levels (higher than IgM) that gradually decrease over 12 months. Mid-term sequelae are still detectable at lung Computed Tomography after 3 months from the hospital admission. Occasionally, it is possible to observe increase of IgM levels in presence of low concentrations of IgG and negative PCR ELISA tests for SARS-COV2 RNA. Baseline levels of IL-6 could be proposed as predictor of radiological mid/long-term sequelae after COVID-related interstitial pneumonia.

Morphologically, immunohistochemically and PCR proven lymphocytic viral peri-, endo-, myocarditis in patients with fatal COVID-19

BackgroundDespite a reported cardiac injury in patients with new coronavirus infection, the possibility and specifics of genuine viral myocarditis in COVID-19 remains not fully clear.PurposeTo study the presence of SARS-CoV-2 in the myocardium and the morphological properties of myocarditis in patients with severe coronavirus infection (COVID-19).MethodsAutopsy data of eight elderly patients (75.6 ± 7.4 years), four male and four female, with severe new coronavirus infection were studied. The lifetime diagnosis of COVID-19 is based on a positive result of the PCR study. The inclusion criterion was the presence of morphological signs of myocarditis according to the Dallas criteria. A standard histological examination included staining by hematoxylin and eosin, toluidin blue and Van Gieson. An immunohistochemical study was performed using antibodies to CD3, CD 68, CD20, perforin, toll-like receptor (TLR) types 4 and 9. PCR in real-time was performed to determine the viral RNA in the myocardium.ResultsAll patients had severe bilateral viral pneumonia. In all cases, myocarditis was not clinically diagnosed. Morphological examination of the heart found signs of active lymphocytic myocarditis. PCR identified the SARS-Cov2 RNA in all cases. There were also signs of destructive coronaritis in all cases, thrombovasculitis, lymphocytic pericarditis (in 3 cases) and endocarditis (in 2 cases). The absence of neutrophils confirms the aseptic nature of inflammation. An immunohistochemical study showed the CD3-positive T lymphocytes in the infiltrates. Increased expression of TLR type 4 and less 9 was also detected.ConclusionMorphological and immunohistochemical evidence of myocarditis in COVID-19 was presented. Lymphocytic infiltrations and positive PCR confirm the viral nature of inflammation. Myocarditis in COVID-19 is also characterized by coronaritis with microvascular thrombosis and associated with lymphocytic endo- and pericarditis.

T-Cup: A Cheap, Rapid, and Simple Home Device for Isothermal Nucleic Acid Amplification.

During the SARS‐CoV2 pandemic, it has become clear that centralized testing suffers from multiple bottlenecks. Logistics, number of machines, and people available to run the diagnostic tests are limited. A solution to those bottlenecks would be a fully decentralized system, where people can test themselves at home and only report back the outcome of the test in a centralized database. Here a noninstrumental device capable of achieving isothermal conditions useful for detecting the SARS‐CoV2 RNA using loop mediated amplification (LAMP) tests is presented. This device, compared to others reported in literature or present on the market, is cheap, easy to produce and use, and has little impact on the environment. Using a simple aluminum coffee capsule, a phase change material, and a 3D printed holder, this device, when placed in boiling water, is able to maintain a temperature of 65 °C for 25 min, required for running the LAMP reaction. In principle, this device can be applied to any LAMP reaction, and hence employed for many different applications, and can be deployed in large quantities in short amount of time.

The histologic and molecular correlates of liver disease in fatal COVID-19 including with alcohol use disorder

Hepatic disease is common in severe COVID-19. This study compared the histologic/molecular findings in the liver in fatal COVID-19 (n = 9) and age-matched normal controls (n = 9); three of the fatal COVID-19 livers had pre-existing alcohol use disorder (AUD). Controls showed a high resident population of sinusoidal macrophages that had variable ACE2 expression. Histologic findings in the cases included periportal/lobular inflammation. SARS-CoV2 RNA and nucleocapsid protein were detected in situ in 2/9 COVID-19 livers in low amounts. In 9/9 cases, there was ample in situ SARS-CoV-2 spike protein that co-localized with viral matrix and envelope proteins. The number of cells positive for spike/100× field was significantly greater in the AUD/COVID-19 cases (mean 5.9) versus the non-AUD/COVID-19 cases (mean 0.4, p < 0.001) which was corroborated by Western blots. ACE2+ cells were 10× greater in AUD/COVID-19 livers versus the other COVID-19/control liver samples (p < 0.001). Co-expression experiments showed that the spike protein localized to the ACE2 positive macrophages and, in the AUD cases, hepatic stellate cells that were activated as evidenced by IL6 and TNFα expression. Injection of the S1, but not S2, subunit of spike in mice induced hepatic lobular inflammation in activated macrophages. It is concluded that endocytosed viral spike protein can induce hepatitis in fatal COVID-19. This spike induced hepatitis is more robust in the livers with pre-existing AUD which may relate to why patients with alcohol abuse are at higher risk of severe liver disease with SARS-CoV2 infection.

Respiratory virus monitoring in patients with community-acquired pneumonia during COVID-19 pandemic in Khabarovsk in 2020

Introduction. Emergence and spread of new coronavirus SARS-CoV-2 among population of the Khabarovsk krai influenced the growth of reported cases of community-acquired pneumonia in year 2020. Aim. To determine proportion of other respiratory viruses in development of viral pneumonia epidemic process in the Khabarovsk city in year 2020 during COVID-19 pandemic. Materials and methods. Sputum of 346 patients with community-acquired pneumonia that were hospitalized with suspected diagnosis of COVID-19 was analyzed during year 2020 in Khabarovsk city. Identification of viral agents was performed via real-time reverse-transcriptase polymerase chain reaction. Results. SARSCOV-2 RNA was identified in 183 (52.9%) out of 346 patients. Among other respiratory viruses parainfluenza virus type 3 and rhinoviruses were dominant mostly in SARS-CoV-2 negative examined people. It should be noted that etiology of pneumonia was identified only in 12.9% of all cases in this group (163 people). Co-infection with SARS-CoV-2 and other respiratory viruses such as parainfluenza virus type 3 virus, other coronaviruses and adenovirus was detected only in 2.2% of the cases. Conclusion. Low level of respiratory viruses detection in sputum can be caused by poor technique of sample collection in the hospital, disruption of storage and transportation conditions as well as development of secondary bacterial infection in certain patients. In order to evaluate influence of other respiratory viruses on the course of COVID-19 with underlying coinfection further investigation including analysis of patients’ clinical data is needed.

RNA regulation in immunity.

RNA regulation in immunity.

SARS-CoV2 Infection and the Importance of Potassium Balance.

SARS-CoV2 infection results in a range of symptoms from mild pneumonia to cardiac arrhythmias, hyperactivation of the immune response, systemic organ failure and death. However, the mechanism of action has been hard to establish. Analysis of symptoms associated with COVID-19, the activity of repurposed drugs associated with lower death rates or antiviral activity in vitro and a small number of studies describing interventions, point to the importance of electrolyte, and particularly potassium, homeostasis at both the cellular, and systemic level. Elevated urinary loss of potassium is associated with disease severity, and the response to electrolyte replenishment correlates with progression toward recovery. These findings suggest possible diagnostic opportunities and therapeutic interventions. They provide insights into comorbidities and mechanisms associated with infection by SARS-CoV2 and other RNA viruses that target the ACE2 receptor, and/or activate cytokine-mediated immune responses in a potassium-dependent manner.