Urinary extracellular vesicles (uEVs) and immune mediators have emerged as potential minimally invasive renal biomarkers. Even though active lupus nephritis (LN) is associated with immune complex deposition, tissue inflammation, and podocyte damage, it remains unclear how these parameters are simultaneously altered in systemic lupus erythematosus (SLE). Thus, we aimed to evaluate uEVs as biomarkers in LN, in association with urinary immune mediators. In this cross-sectional study, uEVs were isolated from SLE patients and healthy donors by differential centrifugation and characterized and/or quantified by electron microscopy, nanoscale flow cytometry, and nanoparticle tracking analysis (NTA). Urinary immune mediators were assessed by a multiplex assay. We included 82 patients (42.6 ± 11.3 years-old, 91.4% female), of whom 56.1% (n = 46) had LN, and 18 healthy donors (37.5 ± 8.2 years-old, 83.3% female). No differences were found for particle size/concentration by NTA, but higher counts of total (P = 0.03) and podocyte-derived (P = 0.01) uEVs were observed in SLE patients, especially in active LN (P = 0.02; P = 0.03). We also identified higher urinary levels of cytokines such as IL-6, IL-8, and CCL-2 according to SLE activity and LN (P < 0.05). Significant correlations were observed between uEVs, immune mediators, R-SLEDAI-2K, proteinuria, and albuminuria in active LN. Lastly, the combinatory analysis of podocyte uEVs, IL-6, IFN-γ, IL-8, uCCL-2 and CCL-3 showed a good predictive power to detect active LN (AUC = 0.88, P = 0.0009). Our results suggest that urinary podocyte-derived uEVs and cytokines are associated with LN activity, which may reflect podocyte injury mediated by inflammation. Thus, the combined application of these biomarkers could help to identify patients with podocyte damage and renal inflammation.

Kidney injury is an important outcome associated with COVID-19 severity. In this regard, alterations in urinary extracellular vesicles (uEVs) could be detected in the early phases of renal injury and may be reflective of the inflammatory process. This is an observational study performed with a case series of COVID-19 hospitalized patients presenting mild-to-critical disease. Total and podocyte-derived uEVs were identified by nanoscale flow cytometry, and urinary immune mediators were assessed by a multiplex assay. We studied 36 patients, where 24 (66.7%) were considered as mild/moderate and 12 (33.3%) as severe/critical. Increased levels of total uEVs were observed (p = 0.0001). Importantly, total uEVs were significantly higher in severe/critical patients who underwent hemodialysis (p = 0.03) and were able to predict this clinical outcome (AUC 0.93, p = 0.02). Severe/critical patients also presented elevated urinary levels (p < 0.05) of IL-1β, IL-4, IL-6, IL-7, IL-16, IL-17A, LIF, CCL-2, CCL-3, CCL-11, CXCL-10, FGFb, M-CSF, and CTAcK. Lastly, we observed that total uEVs were associated with urinary immune mediators. In conclusion, our results show that early alterations in urinary EVs could identify patients at higher risk of developing renal dysfunction in COVID-19. This could also be relevant in different scenarios of systemic and/or infectious disease.

In Brazil, the COVID-19 burden was substantial, and risk factors associated with higher in-hospital mortality rates have been extensively studied. However, information on short-term all-cause mortality and the factors associated with death in patients who survived the hospitalization period of acute SARS-CoV-2 infection is limited. We analyzed the six-month post-hospitalization mortality rate and possible risk factors of COVID-19 patients in a single center in Brazil. This is a retrospective cohort study focused on a six-month follow-up. The exclusion criteria were death during hospitalization, transference to another hospital, and age under 18. We collected data from the charts of all hospitalized patients from March 2020 to December 2020 with a positive RT-PCR test for SARS-CoV-2, resulting in a sample size of 106 patients. The main outcome was death after hospitalization, whereas comorbidities and demographics were evaluated as risk factors. The crude post-hospitalization death rate was 16%. The first 30 days of follow-up had the highest mortality rate. In a Cox regression model for post-hospitalization mortality, previous chronic kidney disease (HR, 4.06, 95%CI 1.46 – 11.30) and longer hospital stay (HR 1.01, 95%CI 1.00 – 1.02) were the only factors statistically associated with death. In conclusion, a high six-month all-cause mortality was observed. Within the six-month follow-up, a higher risk of death was observed for patients who had prior CKD and longer hospital stay. These findings highlight the importance of more intensive medical surveillance during this period.

Introduction: Members of the Herpesviridae family have been described in patients with systemic lupus erythematous (SLE), but the clinical impact on renal function is not well known. Methods: HSV1, HSV2, VZV, EBV, CMV, HHV-6, HHV-7, and HHV-8 were evaluated by molecular biology on admission in blood samples from 40 consecutive SLE patients hospitalized for lupus activity. Results: Patients were 90.0% female, 77.5% non-white, with average age of 32.7 ± 13.6 years. We found positivity for EBV (65.0%), CMV (30.0%), HSV-1 (30.0%), HHV-6 (12.5%), and HHV-7 (7.5%). For all viruses, age, SLEDAI, hematological tests, ferritin, LDH, C-reactive protein, and erythrocyte sedimentation rate (ESR) were not significant. However, EBV positivity was a significant factor for higher serum creatinine (3.0 ± 2.8 vs. 0.9 ± 0.8; P = 0.001) and urea (86 ± 51 vs. 50 ± 46; P = 0.03). Moreover, positive cases for EBV only or with combined co-infections (66.7%-CMV; 58.3%-HSV-1) or negative for EBV only were evaluated by Kruskal-Wallis test again showed statistical significance for serum creatinine and urea (both P ≤ 0.01), with posttest also showing statistical differences for renal dysfunction and EBV presence (alone or in combined co-infections). The presence of EBV viral load was also significant for nephrotic-range proteinuria, renal flare, and the need for hemodialysis. Conclusion: Members of the Herpeviridae family (mainly EBV, HSV-1 and CMV) are common on hospital admission of SLE patients, reaching 65% for EBV, which seems to be associated with renal dysfunction and could reflect a previous association or overlapping disease, which is not well understood.

This study aimed to evaluate the hematological and coagulation parameters according to the clinical outcomes of coronavirus disease (COVID-19). We analyzed the hematological and coagulation parameters of hospitalized patients with COVID-19 at admission, and two and three weeks during hospitalization. To assess the performance of these parameters in predicting poor outcomes, receiver operating characteristic (ROC) curves were created. We studied 128 patients with COVID-19 (59.2±17.7 years, 56% male). Non-survivors (n=54, 42%) presented significant alterations in hematological and coagulation parameters at admission, such as increased in white blood cells (WBC), neutrophil, and band cell counts, as well as elevated prothrombin time (PT), activated partial thromboplastin time, and D-dimer levels. During follow-up, the same group presented a gradual increase in D-dimer and PT levels, accompanied by a reduction in PT activity, hemoglobin, and red blood cell count (RBC). ROC curves showed that WBC, neutrophil, and band cell counts presented the best area under the curve (AUC) values with sensitivity and specificity of >70%; however, a logistic regression model combining all the parameters, except for RBC, presented an AUC of 0.89, sensitivity of 84.84%, and specificity of 77.41%. Our study shows that significant alterations in hematological and coagulation tests at admission could be useful predictors of disease severity and mortality in COVID-19.

Introduction: In systemic lupus erythematosus (SLE), an inflammatory state leads to oxidative stress and excessive formation of reactive oxygen species, increasing cellular damage, mutations and compromised DNA repair. Therefore, our study aimed to evaluate the performance of the alkaline comet assay to investigate DNA damage in women with SLE.Methods: This is a cross-sectional study that included Brazilian women with SLE attended at an university hospital (Rio de Janeiro, Brazil) from 2015 to 2018. Disease activity was assessed using the SLE Disease Activity Index 2000 (SLEDAI-2K). Diagnosis of CMVand EBV infections were performed by qPCR. DNA damage was determined by alkaline comet assay, which is a electrophoresis techinique used to evaluate DNA fragmentation in leukocytes, which is expressed as arbitrary units (AU). Results: We studied 34 SLE patients with a median age of 34.5 years-old. Hospitalized patients (n=11, 32.3%) presented higher SLEDAI-2K, where 9 (81.8%) had SLEDAI-2K ≥ 5 (p=0.038). We did not identify significant differences between healthy and SLE; however, hospitalized patients showed higher median AU, but with no statistical significance (p=0.095). Also, no differences were in DNA damage according to SLEDAI-2, lupus nephritis, use of immunossupressants, or CMV/EBV infections. DNA damage was not associated with levels of anti-dsDNA. Lastlty, ROC curves demonstrated a poor predictive power of DNA damage to differentiate groups (AUC < 0.7, p > 0,05). Conclusion: Our results indicate that DNA damage measured by alkaline comet assay is not associated with SLE severity and the presence of viral infections