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1.
JAAD Case Rep ; 30: 35-37, 2022 Dec.
Article in English | MEDLINE | ID: covidwho-2069299
2.
Otolaryngology - Head and Neck Surgery ; 167(1 Supplement):P113, 2022.
Article in English | EMBASE | ID: covidwho-2064496

ABSTRACT

Introduction: The SARS-CoV-2 pandemic killed over 6 million people worldwide. Children were described to have predominantly mild or asymptomatic infections and to be less exposed to the virus, at least for the initial variants. In the present study, we describe how SARS-CoV-2 can silently infect tonsils and adenoids in children undergoing adenotonsillectomy. Method(s): In this cross-sectional study we assessed children who underwent adenotonsillectomy between October 2020 and September 2021 in a secondary hospital in Brazil. All the caregivers denied any symptom of acute viral upper airway infection in the month prior to surgery. Briefly, nasal cytobrush (NC), nasal wash (NW) and tonsillar tissue fragments posttonsillectomy were tested by RT-PCR, immunohistochemistry (IHC), in situ immunofluorescence (IF), and flow cytometry. Result(s): A total of 48 children (18 females, median age 5.5 years) were enrolled. None of them had been vaccinated against COVID-19 at the time of surgery. Only 2 had a history of previous COVID-19 diagnosis, 3 and 5 months, respectively, before surgery. SARS-CoV-2 RNA was detected in 25% (12) of patients-20% in palatine tonsils, 16.27% in the adenoids, 10.41% in NC, and 6.25% in NW. IHC labeling showed viral nucleoprotein presence in both adenoids and palatine tonsils, in epithelial surface and lymphoid cells from extrafollicular and follicular regions. In 5 out of 7 patients, in situ IF showed the expression of ACE2 and TMPRSS2 and viral spike protein in the tonsillar tissue. Flow cytometry revealed that SARS-CoV-2 is predominantly observed in CD123+ dendritic cells (10.57% of all tested sites), followed by CD14+ monocytes (6.32%). Conclusion(s): According to these results, the prevalence of SARS-CoV-2 infection seems to be higher than expected and underdiagnosed in children at this age group. Palatine tonsils and adenoids are important sites of infection and may be a reservoir for the virus. Nevertheless, it is still unclear the impact of these results on virus transmission.

3.
Chest ; 162(4):A2241, 2022.
Article in English | EMBASE | ID: covidwho-2060916

ABSTRACT

SESSION TITLE: Pulmonary Manifestations of Infections SESSION TYPE: Case Reports PRESENTED ON: 10/17/2022 03:15 pm - 04:15 pm INTRODUCTION: Diffuse alveolar hemorrhage (DAH) due to an undiagnosed autoimmune condition is rare and can be life-threatening. Veno-venous extracorporeal membrane oxygenation (VV-ECMO) has been described as a viable rescue therapy in severe cases, providing time to establish a diagnosis and begin remission induction therapy (1). We report a patient who presented during the Omicron surge with hypoxemic respiratory failure due to pulmonary hemorrhage ultimately diagnosed with antineutrophil cytoplasmic antibody (ANCA) associated vasculitis (AAV) who was supported with VV-ECMO without systemic anticoagulation. CASE PRESENTATION: A 46-year-old woman presented with subacute fatigue and two days of cough and brown sputum. She was found to have normocytic anemia (hemoglobin 3.5 g/dL), renal failure (serum creatinine 17.4 µmol/L), and bilateral pulmonary infiltrates on chest roentgenogram. Though vaccinated, nasal molecular testing detected SARS-CoV-2. She was intubated for progressive hypoxic respiratory failure and bronchoalveolar lavage fluid was consistent with DAH. She received empiric antibiotics, remdesivir, and pulse dose intravenous methylprednisolone as well as continuous renal replacement therapy and plasma exchange. Due to refractory hypoxemia she was cannulated for VV-ECMO. Systemic anticoagulation was deferred due to concerns that it may exacerbate her underlying pathology and due to a small subcortical bleed seen on computed tomography of the head. Perinuclear ANCA (titer >1:1280) was confirmed by immunofluorescence analysis with elevated myeloperoxidase serologies and cyclophosphamide was initiated. Glomeruli with cellular crescent formation consistent with AAV was later identified on renal biopsy. Her course was complicated by recurrent DAH while tapering steroids and an iliac vein thrombus, extracted during decannulation. Her respiratory failure resolved and she was discharged to rehab. DISCUSSION: Traditionally, VV-ECMO obligates systemic anticoagulation to prevent circuit thrombosis, however this may be viewed as a barrier to its use in patients with prohibitive bleeding risk and may contribute to the therapy's overall morbidity. Some institutions have begun to demonstrate the safety of ECMO with low- or prophylactic doses of anticoagulation (2), but this practice remains controversial. Detection of SARS-CoV-2 posed diagnostic and management challenges and its significance to this case remains uncertain. There are many past examples of infectious triggers for both DAH and AAV, and there is emerging evidence for an association between SARS-CoV-2 and ANCA (3). Concerns regarding the risk of B-cell depletion influenced the selection of remission induction therapy. CONCLUSIONS: In the case described, a patient with severe DAH was successfully supported with VV-ECMO. Withholding systemic anticoagulation did not prevent recurrent bleeding and may have contributed to a deep vein thrombosis. Reference #1: Arnold S, Deja M, Nitschke M, Bohnet S, Wallis S, Humrich JY, Riemekasten G, Steinhoff J, Lamprecht P. Extracorporeal membrane oxygenation in ANCA-associated vasculitis. Autoimmun Rev. 2021 Jan;20(1):102702. doi: 10.1016/j.autrev.2020.102702. Epub 2020 Nov 11. PMID: 33188916. Reference #2: Kurihara C, Walter JM, Karim A, et al. Feasibility of Venovenous Extracorporeal Membrane Oxygenation Without Systemic Anticoagulation. Ann Thorac Surg. 2020;110(4):1209-1215. doi:10.1016/j.athoracsur.2020.02.011 Reference #3: Kadkhoda, K., Laurita, K. Antineutrophil cytoplasmic antibodies and their association with clinical outcomes in hospitalized COVID-19 patients. Cell Death Discov. 7, 277 (2021). DISCLOSURES: No relevant relationships by Nathaniel Nelson No relevant relationships by Radu Postelnicu no disclosure on file for Antonio Velez;

4.
Kidney International Reports ; 7(9):S471, 2022.
Article in English | EMBASE | ID: covidwho-2041699

ABSTRACT

Introduction: Vaccination is a known trigger for the development of de-novo or flare of glomerular diseases. Here we present a case series of fourteen patients with COVID vaccine- associated glomerular diseases (CVAGD). Methods: Patients with new onset proteinuria, hematuria or renal failure after SARS- CoV2 vaccine were included in the study. Demographic and clinical details were collected and laboratory investigations including serum creatinine, albumin, urine microscopy and urine spot protein creatinine ratio were done. Renal biopsy specimens were subjected to light microscopy and immunofluorescence examination. Results: We cared for 14 patients with CVAGD. Of them, eight patients were males. The mean age was 25.7 years. Three patients had relapse of their previous disease while eleven patients had no previously detected renal diseases. Eleven patients had received COVISHIELD and three had received COVAXIN. All patients presented after the first vaccine dose. At presentation, seven patients had nephrotic syndrome, two patients had rapidly progressive renal failure and five patients had nephritic syndrome. The mean duration of symptom onset after vaccination was 18 days. Renal biopsy revealed IgA nephropathy in 3 patients, endocapillary proliferative glomerulonephritis in 2 patients, minimal change disease in 5 patients, pauci- immune glomerulonephritis (ANCA associated vasculitis) in one patient, lupus nephritis ISN/RPS class 3 in one and focal segmental glomerulosclerosis in two patients. There was no history of COVID infection in any of our patients. Three patients had renal failure at presentation but none required renal replacement therapy. The patients with MCD and FSGS were treated with steroids, patients with ANCA vasculitis and lupus nephritis were managed with the appropriate Cyclophosphamide and steroid regimens while the others were managed conservatively with anti-proteinuric medications. On follow up, five patients (One IgAN, three MCD, one endocapillary proliferative GN) achieved complete remission of proteinuria and resolution of renal failure, while the remaining eight patients achieved partial remission. One patient with MCD had a relapse of proteinuria 3 weeks after achieving partial remission, he responded well to steroid therapy. All 14 patients remain on close follow up. Conclusions: Although causality cannot be definitively established, there is a definite temporal association between the presentation of glomerular diseases and COVID vaccination, in the absence of other inciting factors. Hence, new-onset or relapse of glomerular diseases presenting post vaccination, although rare, should be observed as a possible adverse event. Intriguing questions such as how to proceed with the vaccination schedule in patients with CVAGD and would changing the vaccine type reduce the risk of relapse remain unanswered. No conflict of interest

5.
Journal of South China Agricultural University ; 41(5):27-35, 2020.
Article in Chinese | CAB Abstracts | ID: covidwho-2040361

ABSTRACT

Objective: To prepare monoclonal antibodies against porcine epidemic diarrhea virus (PEDV) N protein, and develop an indirect immuno-fluorescence assay method used for detecting PEDV. Method: The expressed recombinantly PEDV N protein was used as an immunogen and 8-week-old female BALB/c mice were immunized. Then their spleen cells with high antibody titer were isolated and fused with SP2/0 cells. The hybridoma cell lines secreting monoclonal antibodies against PEDV N protein were screened. In Vero cells infected with PEDV, monoclonal antibody of anti-PEDV N protein was used as the primary antibody and FITC-goat-anti-mouse IgG was used as the secondary antibody to develop indirect immuno-fluorescence assay method used for detecting PEDV. Result: The prepared hybridoma cell lines could stably secrete anti-PEDV N protein antibodies, ELISA antibody titer in cell supernatant was above 1:3 200, and in mouse ascites above 1:1 000 000. While monoclonal antibodies were applied in established indirect immuno-fluorescence assay, the optimal conditions were that cells were fixed with 80% () acetone at -20 degrees C for 30 min;The primary antibody was diluted 1 000 times by PBS buffer solution and incubated at 4 degrees C overnight;The secondary antibody was diluted 100 times by PBS buffer solution and incubated at 37 degrees C for 1 h. Transmissible gastroenteritis virus (TGEV), classical swine fever virus (CSFV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine reproductive virus (PRV), porcine enteric a corone virus (PEAV), porcine rotavirus (PoRV) and PEDV were detected by established indirect immuno-fluorescence assay method, only PEDV showed positive, all the else viruses showed negative.

6.
Biomedicines ; 10(9)2022 Sep 09.
Article in English | MEDLINE | ID: covidwho-2032840

ABSTRACT

High-throughput and rapid screening testing is highly desirable to effectively combat the rapidly evolving COVID-19 pandemic co-presents with influenza and seasonal common cold epidemics. Here, we present a general workflow for iterative development and validation of an antibody-based microarray assay for the detection of a respiratory viral panel: (a) antibody screening to quickly identify optimal reagents and assay conditions, (b) immunofluorescence assay design including signal amplification for low viral titers, (c) assay characterization with recombinant proteins, inactivated viral samples and clinical samples, and (d) multiplexing to detect a panel of common respiratory viruses. Using RT-PCR-confirmed SARS-CoV-2 positive and negative pharyngeal swab samples, we demonstrated that the antibody microarray assay exhibited a clinical sensitivity and specificity of 77.2% and 100%, respectively, which are comparable to existing FDA-authorized antigen tests. Moreover, the microarray assay is correlated with RT-PCR cycle threshold (Ct) values and is particularly effective in identifying high viral titers. The multiplexed assay can selectively detect SARS-CoV-2 and influenza virus, which can be used to discriminate these viral infections that share similar symptoms. Such protein microarray technology is amenable for scale-up and automation and can be broadly applied as a both diagnostic and research tool.

7.
Acta Veterinaria et Zootechnica Sinica ; 53(6):2024-2028, 2022.
Article in Chinese | CAB Abstracts | ID: covidwho-2025545

ABSTRACT

This study aimed to analyze the proliferation characteristics of porcine deltacoronavirus (PDCoV) in suspension cultured porcine kidney cells LLC-PK1, so as to provide Candidate cell for large-scale production of PDCoV inactivated vaccine. LLC-PK1 cells were suspended by gradually decreasing serum method. PDCoV adaptive monoclonal cell lines were screened by limited dilution method. Indirect immunofluorescence method was used to identify the infectivity of PDCoV. The initial cell density, MOI, time of receiving virus collection and TPCK pancreatin concentration were screened to determine the best suspension culture conditions. The suspension cell strain LLC-PK1Sa which can proliferate PDCoV efficiently was screened out;PDCoV can specifically infect LLC-PK1 cells;PDCoV inoculated LLC-PK1Sa cells with a density of 2 x 106 cells.mL-1 according to the MOI of 10-3, When the final concentration of TPCK pancreatin reached 7.5 g.mL-1, the titer of virus solution harvested 48 h after inoculation was the highest. In this study, the efficient proliferation of PDCoV in LLC-PK1Sa suspension cells was realized for the first time, and the suspension culture conditions were preliminarily optimized, which could provide theoretical reference for large-scale production of PDCoV inactivated vaccine.

8.
Wiener Tierarztliche Monatsschrift ; 109(Artikel 11), 2022.
Article in English | CAB Abstracts | ID: covidwho-2025202

ABSTRACT

We have evaluated the diagnostic performance of immunochromatographic point-of-care tests (POCT) for the detection of rotavirus, coronavirus, Escherichia (E.) coli F5, Cryptosporidium (C.) parvum, Clostridium (Cl.) perfringens and Giardia (G.) intestinalis in fresh and thawed faecal samples from calves aged up to six months with diarrhoea. We performed POCTs to detect rotavirus, coronavirus, E. coli F5, C. parvum, Cl. perfringens and G. intestinalis on fresh samples in a field study and re-evaluated the performance for C. parvum, Cl. perfringens and G. intestinalis using thawed samples. We calculated the performance based on the results of the reference methods, which were RT-qPCR for the detection of rota- and coronavirus and bacteriological culturing and PCR to detect E. coli F5 and Cl. perfringens a and ss2 toxins. C. parvum was detected by phase-contrast microscopy and G. intestinalis by immunofluorescence microscopy. We collected 177 faecal samples from diarrhoeic calves. We found good performance for the POCT targeting rotavirus (sensitivity (SE)=92.9%;specificity (SP)=95.6%) and C. parvum (SE=63.3%;SP=96.2%). For E. coli F5, the number of true positive samples (n=1) was too low to evaluate the performance. The POCT to detect coronavirus gave a poor performance (SE=3.3%;SP=96.6%) and the POCT to detect Cl. perfringens a moderate performance (SE=52.8%;SP=78.2%). G. intestinalis POCT showed a higher sensitivity to immunofluorescence microscopy in thawed than in fresh faecal samples (SE=43.9% versus SE=29.2%). There are substantial differences in diagnostic performance between the commercially available immunochromatographic POCTs. Still, POCT can make a valuable contribution to the diagnosis and prevention of calf diarrhoea.

10.
Annals of the Rheumatic Diseases ; 81:332-333, 2022.
Article in English | EMBASE | ID: covidwho-2009011

ABSTRACT

Background: During the COVID-19 pandemic, it remains a major concern whether patients with rheumatic musculoskeletal disease treated with conventional (cs) or biologic (b) disease modifying drugs (DMARDs) exhibit an adequate immune response to the currently available SARS-CoV2 vaccines. There remains an urgent need for more data on SARS-CoV-2 vaccine efficacy to inform healthcare providers on the efficiency of the applied vaccination, potential need of and period for booster and/or re-vaccination. Objectives: To assess and compare the efficacy of the SARS-CoV2 vaccines BNT162b2 vaccine (Pfzer/BioNTech) and mRNA-1273 vaccine (Moderna). (The vaccines were administered as part of the Danish vaccine roll out and offered each with two doses and approximately four weeks apart). Patients' SARS-CoV2 IgG serum level was used as proxy to determine vaccination response. Methods: We established the 'Detection of SARS-CoV2 antibodies in Danish Infammatory Rheumatic Outpatients' study (DECODIR) as a longitudinal prospective cohort study. Patients with rheumatoid arthritis (RA), spondyloarthrop-athies (SpA) or psoriatic arthritis (PsA) receiving their outpatient treatment and monitored in the Danish DANBIO registry at the Danish Hospital for Rheumatic Diseases (DG), Sonderborg were included (April-June 2021). Bloods, patient reported outcome measurements (PROMS), clinical data and treatment information (cs/bDMARD) were collected at baseline (prior to vaccination) and after six weeks and six months. SARS-CoV2 IgG levels in serum were assessed by ELISA (ThermoFischer), and manufacturer's cut-off (>=10 EliA U/mL) selected as defnition of sufficient IgG response. Associations between antibody response, age, gender, disease (RA/PsA/SpA), treatment with no or cs/bDMARDs and disease activity were tested using proportional odds regression and bootstrapped tests of medians. Results were reported using mean, median (IqR) and bootstrapped 95% confdence interval (CI) of the median. Results: A total of 243 patients were included at baseline and after six weeks;at six months' follow-up data were available for 233 patients. After six weeks, vaccination was followed by a signifcant increase in IgG levels (median of <0.7 EliA U/mL at baseline versus 36.5 EliA U/mL). Patients treated with a combination of both cDMARD and bDMARD had signifcantly lower IgG levels compared to patients without any DMARD treatment (8,2 EliA U/mL vs 19.5 EliA U/mL (p<0.001)). Patients treated with oral prednisolone (any dose) also showed signifcantly lower median IgG levels compared to patients without DMARD treatment (3,8 EliA U/mL vs 19.5 EliA U/mL (p<0.01)). The actual measurements six months after baseline demonstrated a signifcant decrease of IgG levels for the whole study population (median of 16 EliA U/mL at six month vs 36.5 EliA U/mL at six weeks, p < 0.001) (Figure 1). Similar to week 6, lowest response rates were found in patients treated with prednisolone or combination of csDMARD and bDMARD. After 6 months, the proportional odds model revealed signifcantly lower median IgG antibody level in patients who received Pfzer compared to Moderna (median 15 EliA U/mL (95%CI: 13-18) vs 44.5 EliA U/mL (95%CI: 36-83) (p<0.001). Conclusion: IgG levels decreased markedly six months after the initial double dose regimen. Patients treated with a combination of cs/bDMARD or oral pred-nisolone are at higher risk of inadequate vaccine response as measured by IgG level. Our results support the decision for the need of a third booster vaccine in patients with infammatory rheumatic diseases, especially in the case of cs/bDMARD combination treatment and prednisolone. The data may indicate a need for further revaccination in these patients.

11.
Annals of the Rheumatic Diseases ; 81:374, 2022.
Article in English | EMBASE | ID: covidwho-2008943

ABSTRACT

Background: The relevance of studying immune response after SARS-CoV-2 vaccination in patients with infammatory immune-mediated diseases (IMIDs) represents a deep concern regarding the risk estimation and management of patients with these diseases on immunomodulatory drugs. It is well known that certain treatments as anti CD20 therapies results in a diminished immunogenicity against common vaccines but it is a scarce data regarding the cellular protection obtained upon vaccination between patients with different IMID and between different treatments. Objectives: To compare a potential detriment on cellular and antibody-mediated protection upon SARS-CoV-2 vaccination in patients with IMIDs treated with immunosuppressive drugs. Methods: We recruited 73 patients with rheumatoid arthritis-RA-(n=49), spondy-larthritis-SpA-(n=19), infammatory bowel disease-IBD-(n=5), idiopathic juvenile arthritis-IJA-(n=2) and heterogenous group composed of sclerodermia, lupus, uveitis(n=6). They were treated mainly with rituximab (n=27), TNFi (n=37) or JAKi (n=3). We collected data of age,sex, csDMARDs, previous SARS-CoV-2 infection, last RTX infusion and prednisone use. After one month of vaccination, we assessed the humoral response performing the Thermo Scientific EliA SARS-CoV-2-Sp1 IgG Test (positivity cut-off >0.70 IU/ml) which was also compared with the data with of 35 healthy controls. In addition, in 40 patients who had serum antibody levels under 100UI/ml, we analysed the cellular response by the use of the QuantiFERON SARS-CoV-2 Starter Pack (Quiagen). A cut-off value of 0.15 IU/ml discriminate between positive or negative cell-mediated immune responses. We compared differences among the different IMIDs and between the different immu-nosuppressive treatments through non-parametric test (p<0.05) Results: Regarding demographic characteristics of patients, older patients (>56 years) and female sex were factors which were associated with low titles of serum antibodies. Anti-spike IgG antibodies were present in an 86% of the IMIDs patients and in 100% healthy controls with signifcant different IgG titre (median [IQR]): 51[11-184] vs 700[440-940];p<0.0001. The differences between (median [IQR]) serum antibody levels were statistically different between IMID type: 33[1-138] in RA vs 94[34-191] in SpA vs 204[187-204] in IBD vs 133[61-204] in IJA vs 13[1.5-31.8] in the rest;p=0.04. Remarkably, patients with IBD who had the highest antibodies titles were the youngest compared with the other patients. Target of the therapy played also an important role in serum antibody levels being these: 3.6 [0.7-51] in RTX patients vs 156 [45-204] in TNFi vs 40 [18-58] in JAKi patients;p<0.0001. In those patients who the last infusion of rituximab was, at least, one year before vaccination presented CD19+ B cells detected by fow cytometry and anti-spike IgG antibodies as well. Cell-mediated responses to SARS-CoV-2 were positive in 33% of IMIDs patients, indeterminated in 3% and negative in 65% of the patients. Strikingly, out of the 33% positive patients, 85% were treated with RTX. A 61% of the RTX patients had inducible cell-mediated responses vs 14% of the patients treated with TNFi;p<0.01. On the other hand, there were not differences in cell-mediated responses between positive and negative antibody patients. Conclusion: Titres of serum antibodies against spike protein of SARS-CoV-2 were lower in IMIDs patients than in controls. Patients with RTX had lower rates of positivity humoral response as well as lower serum titles than patients treated with other therapies regardless the patients 'age. Neverthless, in those patients in whom RTX infusion was delayed because of vaccination they conserved a humoral response. On the other hand, more patients treated with RTX had inducible cell-mediated responses compared with patients with TNFi.

12.
Human Reproduction ; 37:i29, 2022.
Article in English | EMBASE | ID: covidwho-2008565

ABSTRACT

Presence of SARS-CoV-2, the virus responsible for COVID-19, has been reported in numerous organs and tissues of infected patients, including the reproductive system. The effects of COVID-19 on human reproduction remain poorly understood. While cases of intrauterine transmission between expectant mother and fetus have been documented, the impact of SARS-CoV-2 infection on early embryogenesis and establishment of a pregnancy are not known. This prompted us to ask if SARS-CoV-2 can infect embryos, since such an event could impact embryo viability and affect a subsequent pregnancy. We used a three-pronged approach to investigate this possibility: 1) Using RNA-seq and immunofluorescence, we learned that ACE2 and TMPRSS2, two factors required on target cells for SARS-CoV-2 entry, are coexpressed in cells of the trophectoderm in blastocyst-stage preimplantation embryos;2) Using fluorescent reporter virions pseudotyped with Spike (S) glycoprotein from SARS-CoV-2, we observed robust infection of trophectoderm cells, and this permissiveness could be attenuated with blocking antibodies targeting S or ACE2;and 3) Exposing human blastocysts to live, fully infectious SARS-CoV-2, we detected cases of infection that compromised embryo health. Therefore, we identify a new human target tissue for SARS-CoV-2 with potential medical implications for reproductive health during the COVID-19 pandemic and its aftermath.

13.
Kidney International Reports ; 7(6):S442, 2022.
Article in English | EMBASE | ID: covidwho-2004049

ABSTRACT

Introduction: Evidence regarding thrombotic microangiopathy related to covid-19 is reported in the literature, particularly in severe cases. We describe a case recovered from previous asymptomatic covid-19, presenting with acute renal failure, hemolytic anemia, and low platelets. Thrombotic microangiopathy (TMA) was confirmed by renal biopsy, without immunofluorescence staining for C3c and C1q, suggesting this case is not complement-mediated. Anticoagulant therapy led to kidney function improvement. Methods: Case report. Results: A 72-year-old women with a past medical history of primary hypertension was referred to the hospital for the diagnosis of acute renal failure. Three days prior to admission, she suffered abdominal pain, decreased urine output, her blood test revealed elevated serum creatinine of 393 umol/L, and low platelets of 43.6 K/uL. She denied history of hematologic or renal disorders, yet mentioned that she found asymptomatic covid-19 one month before admission. On admission, the vital signs was significant for a blood pressure of 140/80 mmHg. Physical examination was noted with facial oedema, upper abdominal pain, otherwise unremarkable. Laboratory test confirmed acute renal failure with the ongoing increase of serum urea 30.4 mmol/L and creatinine 575 umol/L. Her total blood count discovered thrombocytopenia and anemia, with the platelet count of 50 k/uL, and the hemoglobin of 94 g/L. Lactate dehydrogenase was in upper limit of 434 U/L, and the bilirubin level was in normal range. The peripheral blood smear showed “fragmented” RBCs. Coombs’ test was negative for both direct and indirect method. Stool examination failed to detect either red or white blood cell. Haptoglobin level was 1.14 g/L, which was in normal range (0.41-2.58 g/L). Ddimer was elevated 1376 ng/mL, and the fibrinogen 6.37 g/L. Immunology investigation was conducted with the result of normal level for both complement C3 and C4, negative reaction for anti-cardiolipin IgM and IgG, anti MPO, anti PR3, RF, anti-streptolysin O. Bone marrow aspiration did not show abnormalities. There were Forrest III gastric ulcers found by gastric endoscopy (two ulcers with diameter of 9mm and 10mm, with pseudo-membrane covered). Initially she was treated symptomatically with amlodipin, intravenous PPI, and IV furosemide. As the kidney function was getting worse, hemodialysis was initiated at day 1, day 3, day 6, and day 10 of admission. Renal biopsy was performed and showed active thrombotic microangiopathy. Given the normal complement profile, and negative C3c staining on immunofluorescence of renal biopsy investigation, complement mediated TMA may not be the pathogenesis of this case. The patient was started for anticoagulant therapy, initially subcutaneous low molecular weight heparin and then oral anti-vitamin K. She obtained dramatic recovery with dialysis off, increased urine output, normalized platelets and red cell count, and serum urea and creatinine back to nearly normal range at discharge. Conclusions: Complement related thrombotic microangiopathy is a rare and severe condition. This case of TMA after covid-19 reveals a non-complement mediated pathogenesis, with different treatment. Anticoagulation is an effective therapy in hypercoagulation induced TMA. No conflict of interest

14.
Yaoxue Xuebao ; 57(6):1808-1815, 2022.
Article in Chinese | EMBASE | ID: covidwho-1998089

ABSTRACT

To investigate the effect of Fufang yinhua jiedu (FFYH) granules against coronavirus and its potential mechanism, we used Huh7, Huh7.5, H460, and C3A cell lines as in vitro models to evaluate the cytotoxicity and antiviral activity of FFYH by observation of cell pathogenic effect (CPE);and then the inhibitory effect of FFYH on the transcription expression of coronavirus RNA and inflammatory factor mRNA were evaluated by quantitive reverse transcription PCR (qRT-PCR);finally, the inhibitory effect of FFYH on the expression of coronavirus protein and its underlying mechanism against coronavirus were investigated by Western blot and immunofluorescence. Our results indicated that 50% toxic concentration (TC50) FFYH on Huh7, Huh7.5, H460, and C3A cells were 2 035.21, 5 245.69, 2 935.28 and 520 µg·mL-1, respectively;50% inhibitory concentration (IC50) of FFYH on HCoV-229E in Huh7 and Huh7.5 cells were 438.16 and 238.54 µg·mL-1 with safety index (SI) of 4.64 and 21.99, respectively;IC50 of FFYH on HCoV-OC43 in H460 cells was 165.13 µg·mL-1 with SI of 17.78. Moreover, FFYH not only could inhibit the replication of coronaviruses (HCoV-OC43 and HCoV-229E) through inhibiting the transcription of viral RNA and the expression of viral protein, but also effectively suppress the expression of inflammatory factors interleukin-6 (IL-6), tumor necrosis factor α (TNF-α) and interleukin-8 (IL-8) at mRNA level caused by coronaviruses, which might be associated with the inhibitory effect of FFYH on mitogen-activated protein kinase (MAPK) pathway and the nuclear translocation of nuclear transcription factor-κB (NF-κB). In summary, our results demonstrated that FFYH exhibited a good in vitro anti-coronavirus effect, which provides a theoretical basis for its clinical use in the treatment of anti-coronavirus pneumonia.

15.
Front Immunol ; 13: 951254, 2022.
Article in English | MEDLINE | ID: covidwho-1987497

ABSTRACT

Neutrophil extracellular traps (NETs) are web-like structures extruded by neutrophils after activation or in response to microorganisms. These extracellular structures are decondensed chromatin fibers loaded with antimicrobial granular proteins, peptides, and enzymes. NETs clear microorganisms, thus keeping a check on infections at an early stage, but if dysregulated, may be self-destructive to the body. Indeed, NETs have been associated with autoimmune diseases such as systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), antiphospholipid syndrome (APS), psoriasis, and gout. More recently, increased NETs associate with COVID-19 disease severity. While there are rigorous and reliable methods to quantify NETs from neutrophils via flow cytometry and immunofluorescence, the accurate quantification of NETs in patient plasma or serum remains a challenge. Here, we developed new methodologies for the quantification of NETs in patient plasma using multiplex ELISA and immunofluorescence methodology. Plasma from patients with SLE, non-genotyped healthy controls, and genotyped healthy controls that carry either the homozygous risk or non-risk IRF5-SLE haplotype were used in this study. The multiplex ELISA using antibodies detecting myeloperoxidase (MPO), citrullinated histone H3 (CitH3) and DNA provided reliable detection of NETs in plasma samples from SLE patients and healthy donors that carry IRF5 genetic risk. An immunofluorescence smear assay that utilizes only 1 µl of patient plasma provided similar results and data correlate to multiplex ELISA findings. The immunofluorescence smear assay is a relatively simple, inexpensive, and quantifiable method of NET detection for small volumes of patient plasma.


Subject(s)
COVID-19 , Extracellular Traps , Lupus Erythematosus, Systemic , Humans , Interferon Regulatory Factors/metabolism , Neutrophils
16.
Biochemical and Cellular Archives ; 22(1):603-608, 2022.
Article in English | EMBASE | ID: covidwho-1980678

ABSTRACT

The novel coronavirus 2019 (COVID-19) was first discovered in Wuhan city (China) in patients with severe respiratory disorders and the disease which it cause named scientifically as Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). Since, its discovery, more than 1,338,604 of COVID-19 case was confirmed between Iraqi peoples a while 30 June 2021 with nearly 17,156 death cases. In order to detect the association of COVID-19 with elevated IL-6 and others blood parametres as D-dimer, C-reactive protein, Ferritin and Lactate dehydrogenase and some other risk factors as age, gender, smoking and place of residence, the study conducted 70 people were devided into three groups. Under 30 year, between 30 to 40 year and above 40 year and also 20 healthy subject were taken as a control group. Levels of IL-6 and the other hematological parameters were estimeated using Automated fluorescent immunoassay system (AFIAS-6) technique. The result showed a significant increase in the level of IL-6 and the other hematological parameters studied in all age groups of COVID-19 patients in comparison with the healthy subjects group. The results also showed that patients with blood group type (o+) were the most frequent in the disease in the study by percent (35.71%) followed by patients with AB+, A+ and B+ and A- blood group type and amounted (25.71%, 18.57%, 14.9% and 5.72%), respectively. Results also showed that the age groups above 40 years were most frequent with the disease in comparison with group of between 30 to 40 and group of under 30 year. The disease was more common in males compared to females in this study and also was more common among non-smokers and between urban residents more than among city residents.

17.
J Cutan Pathol ; 2022 Aug 03.
Article in English | MEDLINE | ID: covidwho-1968081

ABSTRACT

We present a case of eosinophil-rich linear IgA bullous disease (LABD) following the administration of a messenger RNA COVID-19 booster vaccine. A 66-year-old man presented to the emergency department with a 3-week history of a pruritic blistering rash characterized by fluid-filled bullae and multiple annular and polycyclic plaques. He was initially diagnosed with bullous pemphigoid based on a biopsy showing a subepidermal blister with numerous eosinophils. However, direct immunofluorescence studies showed linear IgA and IgM deposition along the basement membrane zone with no immunoreactivity for C3 or IgG. Additionally, indirect immunofluorescence was positive for IgA basement membrane zone antibody. The patient was subsequently diagnosed with LABD and initiated on dapsone therapy with resolution of his lesions at 3-month follow-up. This case illustrates the growing number of autoimmune blistering adverse cutaneous reactions from vaccination. Dermatopathologists should be aware that features of autoimmune blistering diseases can overlap and may not be distinguishable based on these histopathological findings alone. Confirmation with direct immunofluorescence and/or serological studies may be necessary for accurate diagnosis.

18.
JAAD Case Rep ; 26: 27-29, 2022 Aug.
Article in English | MEDLINE | ID: covidwho-1966833
19.
Gastroenterology ; 162(7):S-277, 2022.
Article in English | EMBASE | ID: covidwho-1967262

ABSTRACT

Background: Although respiratory failure is the hallmark of severe disease, it is increasingly clear that Coronavirus Disease 2019 (COVID-19) is a multi-system disorder. The presence of gastrointestinal (GI) involvement by Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has been suggested by epidemiological, clinical, non-human primate, in-vitro (enteroid) and ex-vivo (human biopsy) studies. Having recently documented persistence of SARS-CoV-2 within the intestinal epithelium 7 months after infection, here we aimed to study mucosal immune cell abnormalities in individuals with prior history of COVID-19. Methods: Individuals with previous COVID-19 diagnosis (by either RT–PCR or seroconversion) and controls (without RT-PCR or serological evidence of prior COVID-19 infection) undergoing endoscopic evaluation were recruited into the study (Table 1). Colonic and small intestinal (duodenal and ileal) biopsies were analyzed by multiparameter flow cytometry for mucosal immune cell populations including myeloid cells (classical and non-classical monocytes, dendritic cell subsets), T cells (subsets and activation state), B cells (including plasma cells) and NK cells. Persistence of viral antigens was determined by immunofluorescence microscopy (n=30) using a previously published anti-nucleocapsid (NP) antibody. Results: Thirty subjects with a previous history of COVID-19 (post-COVID), median of 4 months from diagnosis (range 1-10 months), were recruited and compared with 40 normal volunteer (NV) controls. Relative to controls, post-COVID subjects displayed higher frequencies of classical (CD14+) monocytes in both, the colon and the small bowel, while significantly higher frequencies of conventional dendritic cells (cDC)1 (lin-HLA-DRhiCD14- CD11c+CD141+) and cDC2 (lin-HLA-DRhiCD14-CD11c+CD1c+) were noted in the colon. Among NK subsets, CD56bright CD16- NK cells were significantly higher in the colon of post-COVID subjects. Among T cell subsets, CD8+ tissue resident memory T cells (CD8+CD69+CD103+) were significantly increased in colon of post-COVID subjects compared to NV. Among B cell subsets, plasma cells (CD3-CD27+CD38hi) trended higher (p= 0.06), while mucosal B cells (CD3-CD19+) were significantly lower in the terminal ileum of post-COVID subjects compared to NV. Finally, with IF, we detected SARS-CoV-2 NP in 10 out of 30 (33%) of post-COVID subjects (Figure 1). Conclusion: Innate and adaptive immune cell abnormalities persist in the intestinal mucosa of post-COVID subjects for up to 10 months and may reflect viral persistence or immune cell dysregulation in the intestines. These findings have major implications for understanding the pathogenesis of long-term sequelae of COVID-19, including long-haul COVID.(Table Presented)(Figure Presented)

20.
British Journal of Dermatology ; 186(6):e254-e255, 2022.
Article in English | EMBASE | ID: covidwho-1956709

ABSTRACT

We present the case of a 68-year-old woman who presented with a blistering skin eruption 5 days after the administration of the first dose of Pfizer-BioNTech mRNA COVID-19 vaccine. Examination revealed tense bullae in a localized distribution confined to the dorsal aspect of her hands, forearms and ears only. This was preceded by severe pruritus. She had no mucosal involvement and was otherwise systemically well. She had a background of chronic obstructive pulmonary disease and hypercholesterolaemia with no previous history of COVID-19. Skin biopsy revealed a subepidermal bulla containing numerous eosinophils in keeping with bullous pemphigoid (BP). The diagnosis was confirmed with a positive direct immunofluorescence (IF) which showed linear IgG and C3 deposition at the basement membrane zone. Indirect IF was positive for anti-BP180 and anti-BP230. The patient was treated with oral prednisolone and doxycycline to good effect She proceeded to have the second dose of the Pfizer-BioNTech vaccine while on treatment and did not experience a flare of BP. However, a week later, she developed erythematous annular plaques with milia over the dorsi of her hands. Skin biopsy revealed multiple milia within the papillary dermis in keeping with milia en plaque. To to our knowledge, this is the first case of a patient developing BP with subsequent milia en plaque following the Pfizer-BioNTech mRNA COVID-19 vaccine (Damiani G, Pacifico A, Pelloni F, Iorizzo M. The first dose of COVID-19 vaccine may trigger pemphigus and bullous pemphigoid flares: is the second dose therefore contraindicated? J Eur Acad Dermatol Venereol 2021;35: e645-7). She has since been weaned off systemic treatment for BP;however, she continues to require ongoing input for the management of milia en plaque.

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