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Introduction: Primary focal segmental glomerular sclerosis (FSGS) is a rare, likely immune-mediated disease. Rituximab (RTX) may play a role in management, although data in adults are scanty. Methods: We collected cases of RTX-treated primary FSGS within the Italian Society of Nephrology Immunopathology Working Group and explored response rate (24-hour proteinuria

BACKGROUND AND AIMS Rituximab is increasingly used to induce and maintain remission in ANCA-associated vasculitis (AAV). However, it remains unclear what the optimal dosing regimen is, especially in the setting of maintenance. Clinical trials have shown that vasculitis flares are unlikely as long as B-cell depletion in peripheral blood is sustained, with some evidence supporting a rituximab redosing strategy guided by B-cell-count monitoring. A better understanding of B-cell kinetics following rituximab treatment can facilitate personalized treatment approaches. This study aims to (1) describe B-cell repopulation in AAV patients treated with rituximab induction, and (2) identify clinical features predictive of B-cell repopulation. METHOD This is a single-centre, retrospective study, including AAV patients on no baseline immunosuppression who were treated with rituximab induction between December 2009 and March 2018 in the Nephrology Unit of Spedali Civili, Brescia. Only patients with available longitudinal B-cell monitoring for at least 6 months following rituximab were considered. B-cell counts were determined as CD19 + cells in peripheral blood using flow cytometry (Beckman Coulter Navios©). B-cell repopulation following rituximab was defined as CD19 + cell count >10 cells/μL, with a documented CD19 + count ≤ 10 cells/μL within the preceding year. For each patient, CD19 + cell counts were followed up to the earliest event among the following: (1) B-cell repopulation, (2) retreatment with rituximab or other induction agents, or (3) most recent observation available. Estimated glomerular filtration rate (eGFR) was calculated using the CKD-EPI equation. Flares were defined as active disease retreated with rituximab or cyclophosphamide and classified as major if one or more major items of the BVAS score v3 were present. Predictors of time to B-cell repopulation were assessed using uni- and multivariate analysis with log-rank test and Cox proportional hazards regression, respectively. RESULTS The study cohort included 64 patients. The main clinical characteristics are summarized in Table 1. During a median follow-up of 33.4 months (range 7.5–101.7 months), B-cell repopulation was observed in 36 patients (56.2%). Median time from rituximab induction to B-cell repopulation was 39.8 months [95% confidence interval (95% CI) 33.8–69.7]. Gender, ANCA subtype, rituximab dose, steroid pulses, plasma exchange and methotrexate/azathioprine maintenance were not significantly associated with time to B-cell repopulation. In contrast, B-cell repopulation was significantly slower in patients who were older (median time to repopulation 51.6 months in patients ≥ mean age of 66.8 years old versus 33.9 months in During sustained B-cell depletion, six vasculitis flares were observed, of which one was major (rise in creatinine > 30%) and the rest were minor (n = 4 worsening of urine abnormalities, n = 1 retro-orbital pain). CONCLUSION A substantial proportion of AAV patients treated with a single course of rituximab shows sustained B-cell depletion. Renal function emerged as the main independent predictor of B-cell repopulation, with delayed B-cell repopulation in people with kidney impairment. Only one major vasculitis flare was observed during sustained B-cell depletion. These data support the idea that longitudinal monitoring of B-cell counts can be a useful biomarker to guide rituximab dosing for maintenance in AAV, especially in patients with kidney impairment.

Objectives Rituximab has become the cornerstone of induction treatment in ANCA-associated vasculitis (AAV). B-cell depletion may increase the risk of hypogammaglobulinemia, potentially leading to severe infections. This study aims to assess factors associated with hypogammaglobulinemia in AAV patients treated with rituximab. Methods This retrospective cohort study included AAV patients treated with rituximab induction in 14 European centres. Severe adverse events (SAEs) were defined as episodes requiring hospitalization or intravenous antibiotics, malignancies, or death. Linear and logistic regression were used to identify predictors of IgG levels and of the risk of hypogammaglobulinemia, defined as IgG ≤7 g/l at 6 months. Results The study included 227 patients. IgG levels at 6 months were lower than baseline (P Conclusions In AAV patients treated with rituximab, hypogammaglobulinemia at 6 months after induction is common, and lower IgG levels are associated with serious infections. The risk of hypogammaglobulinemia in these patients increases with age and higher glucocorticoid doses.

Recent efforts have succeeded in surveying open chromatin at the single-cell level, but high-throughput, single-cell assessment of heterochromatin and its underlying genomic determinants remains challenging. We engineered a hybrid transposase including the chromodomain (CD) of the heterochromatin protein-1α (HP-1α), which is involved in heterochromatin assembly and maintenance through its binding to trimethylation of the lysine 9 on histone 3 (H3K9me3), and developed a single-cell method, single-cell genome and epigenome by transposases sequencing (scGET-seq), that, unlike single-cell assay for transposase-accessible chromatin with sequencing (scATAC-seq), comprehensively probes both open and closed chromatin and concomitantly records the underlying genomic sequences. We tested scGET-seq in cancer-derived organoids and human-derived xenograft (PDX) models and identified genetic events and plasticity-driven mechanisms contributing to cancer drug resistance. Next, building upon the differential enrichment of closed and open chromatin, we devised a method, Chromatin Velocity, that identifies the trajectories of epigenetic modifications at the single-cell level. Chromatin Velocity uncovered paths of epigenetic reorganization during stem cell reprogramming and identified key transcription factors driving these developmental processes. scGET-seq reveals the dynamics of genomic and epigenetic landscapes underlying any cellular processes.

Background Dialysis and kidney transplant patients with moderate-severe COVID-19 have a high mortality rate, around 30%, that is similar in the two populations, despite differences in their baseline characteristics. In these groups, the immunology of the disease has been poorly explored. Methods Thirty-two patients on dialysis or with kidney transplant and SARS-CoV-2 infection requiring hospitalization (COV group) were included in our study. Lymphocyte subsets, dendritic cell (DC) counts and monocyte activation were studied. SARS-CoV-2 anti-spike/anti-nucleocapsid were monitored, and baseline cytokines and chemokines were measured in 10 patients. Results The COV group, compared to healthy subjects and uninfected dialysis/kidney transplant controls, showed lower numbers of CD4 + and CD8 + T cells, Natural-Killer (NK), B cells, plasmacytoid and myeloid DCs, while the proportion of terminally differentiated B-cells was increased. IL6, IL10, IFN-α and chemokines involved in monocyte and neutrophil recruitment were higher in the COV group, compared to uninfected dialysis/kidney transplant controls. Patients with severe disease had lower CD4 + , CD8 + and B-cell counts and lower monocyte HLA-DR expression. Of note, when comparing dialysis and kidney transplant patients with COVID-19, the latter group presented lower NK and pDC counts and monocyte HLA-DR expression. Up to 60 days after symptom onset, kidney transplant recipients showed lower levels of anti-spike antibodies compared to dialysis patients. Conclusions During SARS-CoV-2 infection, dialysis and kidney transplant patients manifest immunophenotype abnormalities; these are similar in the two groups, however kidney transplant recipients show more profound alterations of the innate immune system and lower anti-spike antibody response. Graphical abstract Supplementary Information The online version contains supplementary material available at 10.1007/s40620-021-01214-8.

Tofu, one of the most important products made from soymilk, is obtained through a coagulation process performed with various coagulants (acids, salts and, enzymes). In this study, innovative tofu samples were produced using the grape pomace (GP) powders of different varieties (Barbera, Chardonnay, Moscato, and Pinot Noir) with different origins (fermented and distilled) at two concentration levels (2.5% and 5% w/v) as coagulants, and comparisons with traditional tofu were made. Physicochemical characteristics, phenolic contents, radical scavenging activity levels, textural properties, and consumer acceptability were evaluated. The moisture, protein content, and pH levels of GP tofu samples were slightly lower than those of traditional tofu. Regarding textural parameters, except for hardness, all other parameters were significantly lower in GP tofu samples, with differences due to GP concentration. The colours of GP tofu varied from amber-yellow to violet according to the GP origin. The blue-violet colours were observed predominantly in tofu samples obtained with Barbera and Pinot Noir GPs, while the other GP tofu samples showed amber-yellow colours. The concentrations of polyphenols were 2–10 times higher than in traditional tofu, while the radical scavenging activity levels were 9–80 times higher. The GP tofu samples were favoured by consumers, with small differences among the GP varieties.

Background and objectiveSystemic small vessel vasculitides carry an increased risk of acute arterial and venous thromboembolic events (AVTE); however, this risk has not been systematically explored in Eosinophilic Granulomatosis with Polyangiitis (EGPA). This study assessed the occurrence and main risk factors of AVTE among EGPA patients as compared to the general community from the population-based Bruneck cohort.MethodsWe conducted a retrospective multicenter cohort study on 573 EGPA patients. Clinical and serological data were collected at diagnosis. Occurrence of AVTE and time to the first AVTE after EGPA diagnosis were recorded. Age-standardized event rate (SER) of AVTE as compared to the reference cohort was assessed. Cox regression was applied to identify AVTE predictors.Results129 EGPA patients (22.5%) had AVTE, considered as potentially life-threatening in 55.8%. Seventy patients experienced an AVTE prior to diagnosis (of whom 58.6% in the two years before diagnosis) and 75 following EGPA diagnosis, of whom 56% in the two subsequent years. The SER of AVTE as compared to the reference cohort was 2.10 (95% CI 1.67-2.63). This risk was particularly increased in patients with history of AVTE and with a Birmingham Vasculitis Activity Score ≥20 at diagnosis. Patients receiving immunosuppression within 2 months of diagnosis were at lower risk, while antiplatelet or anticoagulant treatment did not confer measurable benefit.ConclusionEGPA is associated with AVTE in approximately one quarter of patients, particularly around diagnosis. Immunosuppressants seemed to exert a protective effect, while anticoagulant and antiplatelet agents did not.

Background and Aims Idiopathic Focal Segmental Glomerulosclerosis (FSGS) is a rare glomerulonephritis often complicated by a chronic relapsing course frequently characterized by dependency or resistance to immunosuppressive treatment. Moreover, about half of the patients with active disease would develop end-stage renal disease within 10 years from the diagnosis, highlighting the need of novel therapeutic approaches. Rituximab (RTX), a chimeric monoclonal antibody against CD20, showed promising results in pediatric steroid-dependent/frequently relapsing FSGS and in post-transplantation recurrence. However, evidence about its role in the FSGS of the adult is still lacking with small case series suggesting conflicting results. In this study we assess the efficacy of RTX in the largest cohort of adults with FSGS currently available in literature. Methods Adults with biopsy proven idiopathic FSGS treated with RTX were retrospectively identified among several Italian nephrology units. Response to RTX was evaluated at 3, 6, 12 months and, when available, during the long-term follow-up. A positive response (POR) was defined as: (1) proteinuria 50% compared to baseline, (2) stable renal function (3) decreased or stable dose of glucocorticoids and other immunosuppressants. Severe Adverse Events (SAEs) have been recorded. Results 31 patients have been identified: 18 steroid-dependent, 11 steroid-resistant, and 2 patients with major contraindication to steroid therapy. RTX has been administered at a median of 87 months (IQR 54–96) from the diagnoses using heterogeneous schedules of administration. Overall, the POR rate at 6 months was 52% (steroid-dependent=69%; steroid-resistant=22%). At univariate analyses, POR to RTX at 6 months was associated to the steroid-dependent status (p=0.0347) and a proteinuria at RTX Conclusion RTX may be an option in the FSGS of the adult, especially in the steroid-dependent patients and the ones with less severe nephrotic syndrome. In the responders, a RTX based maintenance therapy may be required.

ANCA-associated vasculitis (AAV) are the prototype of a disease characterised by the presence of a biomarker; ANCA positivity in fact is recorded in 90% of cases of GPA and MPA. The role of ANCA in the management of AAV ranges from diagnostic to prognostic purposes with also therapeutic implications. Changes in clinical practice with the increased use of rituximab have drawn attention to B-cells as a biomarker able to contribute to patient management. Cytokines and other circulating factors, although still at a research stage, may represent future biomarkers of interest or even therapeutic options. From the point of view of renal involvement in AAV, proteinuria and microhematuria are still the biomarkers employed in everyday clinical practice with a proposed role for emerging ones (MCP1, CD163 and CD25). The aim of this review is to discuss the role of well-known biomarkers in everyday clinical management of AAV patients as well as future perspectives for those that are still at a research stage with attention to therapeutic implications.

The case of a 35-year-old man presenting microscopic hematuria allows the following topics to be examined in depth: 1. How is hematuria defined and classified? 2. What are the causes of hematuria? 3. Hematuria is an important sign for the diagnosis of cancer of the urinary system. What are the risk factors for bladder carcinoma? 4. What is the role of the urinary dipstick test in the diagnosis of hematuria? 5. What is the role of the examination of urine sediment in the diagnosis of hematuria? 6. Once microscopic hematuria is confirmed, how can we proceed to reach a final diagnosis?