Cellular Lesions Research Articles

Diffuse leptomeningeal glioneuronal tumour (DLGNT) in children: the emerging role of genomic analysis

Diffuse leptomeningeal glioneuronal tumours (DLGNT) represent rare enigmatic CNS tumours of childhood. Most patients with this disease share common radiological and histopathological features but the clinical course of this disease is variable. A radiological hallmark of this disease is widespread leptomeningeal enhancement that may involve the entire neuroaxis with predilection for the posterior fossa and spine. The classic pathologic features include low- to moderate-density cellular lesions with OLIG2 expression and evidence of ‘oligodendroglioma-like’ appearance. The MAPK/ERK signaling pathway has recently been reported as a potential driver of tumourigenesis in up to 80% of DLGNT with KIAA1549:BRAF fusions being the most common event seen. Until now, limited analysis of the biological drivers of tumourigenesis has been undertaken via targeted profiling, chromosomal analysis and immunohistochemistry. Our study represents the first examples of comprehensive genomic sequencing in DLGNT and shows that it is not only feasible but crucial to our understanding of this rare disease. Moreover, we demonstrate that DLGNT may be more genomically complex than single-event MAPK/ERK signaling pathway tumours.

Potential Use of Biomarker γ-H2AX on Peripheral Blood Patient Who Underwent Radioiodine Ablation Treatment in Indonesia

Thyroid cancer is one of the most common endocrine malignancies. Although the 10-year survival rate of differentiated thyroid cancer (DTC) is about 90% after conventional treatments, a small proportion of patients still suffer from tumor recurrence or drug resistance. DNA doublestrand breaks (DSBs) are important cellular lesions that can result from ionizing radiation exposure. The biomarker for DSB formation is the phosphorylated form of the histone H2 variant H2AX (γ-H2AX). We propose the use of γ-H2AX as a DNA DSB biomarker in thyroid cancer patients receiving radioiodine treatment as a possibility to detect the potential of instability genome after receiving the treatment. Evaluating DNA DSB damage with γ-H2AX biomarker might be important in managing thyroid cancer.

Understanding Beta-Lactam-Induced Lysis at the Single-Cell Level.

Mechanical rupture, or lysis, of the cytoplasmic membrane is a common cell death pathway in bacteria occurring in response to β-lactam antibiotics. A better understanding of the cellular design principles governing the susceptibility and response of individual cells to lysis could indicate methods of potentiating β-lactam antibiotics and clarify relevant aspects of cellular physiology. Here, we take a single-cell approach to bacterial cell lysis to examine three cellular features—turgor pressure, mechanosensitive channels, and cell shape changes—that are expected to modulate lysis. We develop a mechanical model of bacterial cell lysis and experimentally analyze the dynamics of lysis in hundreds of single Escherichia coli cells. We find that turgor pressure is the only factor, of these three cellular features, which robustly modulates lysis. We show that mechanosensitive channels do not modulate lysis due to insufficiently fast solute outflow, and that cell shape changes result in more severe cellular lesions but do not influence the dynamics of lysis. These results inform a single-cell view of bacterial cell lysis and underscore approaches of combatting antibiotic tolerance to β-lactams aimed at targeting cellular turgor.

Pharmacokinetics and Target Attainment of SQ109 in Plasma and Human-Like Tuberculosis Lesions in Rabbits.

ABSTRACTSQ109 is a novel well-tolerated drug candidate in clinical development for the treatment of drug-resistant tuberculosis (TB). It is the only inhibitor of the MmpL3 mycolic acid transporter in clinical development. No SQ109-resistant mutant has been directly isolated thus far in vitro, in mice, or in patients, which is tentatively attributed to its multiple targets. It is considered a potential replacement for poorly tolerated components of multidrug-resistant TB regimens. To prioritize SQ109-containing combinations with the best potential for cure and treatment shortening, one must understand its contribution against different bacterial populations in pulmonary lesions. Here, we have characterized the pharmacokinetics of SQ109 in the rabbit model of active TB and its penetration at the sites of disease—lung tissue, cellular and necrotic lesions, and caseum. A two-compartment model with first-order absorption and elimination described the plasma pharmacokinetics. At the human-equivalent dose, parameter estimates fell within the ranges published for preclinical species. Tissue concentrations were modeled using an “effect” compartment, showing high accumulation in lung and cellular lesion areas with penetration coefficients in excess of 1,000 and lower passive diffusion in caseum after 7 daily doses. These results, together with the hydrophobic nature and high nonspecific caseum binding of SQ109, suggest that multiweek dosing would be required to reach steady state in caseum and poorly vascularized compartments, similar to bedaquiline. Linking lesion pharmacokinetics to SQ109 potency in assays against replicating, nonreplicating, and intracellular M. tuberculosis showed SQ109 concentrations markedly above pharmacokinetic-pharmacodynamic targets in lung and cellular lesions throughout the dosing interval.

Genomic, Microbial and Immunological Microenvironment of Colorectal Polyps.

Simple SummaryColorectal cancers (CRC) initiate from small cell clusters known as polyps. Colonoscopic surveillance and removal of polyps is an important strategy to prevent CRC progression. Recent advances in sequencing technologies have highlighted genetic mutations in polyps that potentially contribute to CRC development. However, CRC might be considered more than a genetic disease, as emerging evidence describes early changes to immune surveillance and gut microbiota in people with polyps. Here, we review the molecular landscape of colorectal polyps, considering their genomic, microbial and immunological features, and discuss the potential clinical utility of these data.Colorectal cancer (CRC) develops from pre-cancerous cellular lesions in the gut epithelium, known as polyps. Polyps themselves arise through the accumulation of mutations that disrupt the function of key tumour suppressor genes, activate proto-oncogenes and allow proliferation in an environment where immune control has been compromised. Consequently, colonoscopic surveillance and polypectomy are central pillars of cancer control strategies. Recent advances in genomic sequencing technologies have enhanced our knowledge of key driver mutations in polyp lesions that likely contribute to CRC. In accordance with the prognostic significance of Immunoscores for CRC survival, there is also a likely role for early immunological changes in polyps, including an increase in regulatory T cells and a decrease in mature dendritic cell numbers. Gut microbiotas are under increasing research interest for their potential contribution to CRC evolution, and changes in the gut microbiome have been reported from analyses of adenomas. Given that early changes to molecular components of bowel polyps may have a direct impact on cancer development and/or act as indicators of early disease, we review the molecular landscape of colorectal polyps, with an emphasis on immunological and microbial alterations occurring in the gut and propose the potential clinical utility of these data.

Abstract 1910: Augmented secretion of IL-1α from mouse oral squamous cell carcinoma (OSCC) cells caused by the serum deprivation and hypoxia promotes immune-suppressive activity of mesenchymal stromal cells

Abstract Objectives; We have already reported that mouse oral squamous carcinoma (OSCC), Sq-1979-1, cells produce IL-1α which specifically enhances the immunosuppressive activity of co-cultured mesenchymal stromal, 10T1/2, cells (Morimoto-Ito, Open Dent J. 2019). In this report, we evaluated conditions promoting the expression of IL-1α of Sq-1979-1 cells to further enhance immunosuppressive function of the 10T1/2 cells, and also examined the expression in OSCC tissues. Methods; Expression of IL-1α in Sq-1979-1 cells was evaluated by RT-PCR, western blots and enzyme-linked immune sorbent assay (ELISA). Conditioned mediums (CMs) from Sq-1979-1 cells were co-cultured with 10T1/2 cells and anti-CD3 antibody-stimulated mouse spleen cells. The production of interferon (IFN)- γ and IL-1α was evaluated using ELISA. The function of IL-1α in CM was examined by anti- IL-1α antibody. Immunohistochemical analysis of OSCC tissues was performed. Results; The production of IL-1α from Sq-1979-1 cells was specifically enhanced in lower serum (0.5% or 1.0%FBS), or under hypoxia (1.0% oxygen) compared to that in the standard growth condition supplemented with 10% FBS under normoxia. Compared to standard CM, the CMs prepared in 0.5% and 1.0% FBS, or under hypoxia significantly reduced IFN-γ production from stimulated spleen cells co-cultured with 10T1/2 cells. These functions of the CMs completely abolished by anti- IL-1α antibody. Expression of IL-1α in OSCC tissues was prominent in the midst of carcinomatous cellular lesion, or nearby necrotic lesion, where the supply deficiency could be occurred. Conclusions; The promotion of immunosuppressive activity of mesenchymal cells by Sq-1979-1 cells was specifically augmented in lower serum and hypoxic conditions, mediated by increased IL-1α production. Citation Format: Nobuo Kondoh, Akihiro Matsunami, Masako Mizuno-Kamiya, Naoki Umemura, Eiji Takayama, Harumi Kawaki, Kyouhei Ueno, Hiroe Morimoto-Ito, Yasunori Muramatsu, Shinichiro Sumitomo. Augmented secretion of IL-1α from mouse oral squamous cell carcinoma (OSCC) cells caused by the serum deprivation and hypoxia promotes immune-suppressive activity of mesenchymal stromal cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1910.

Copy number alteration profiling facilitates differential diagnosis between ossifying fibroma and fibrous dysplasia of the jaws

Ossifying fibroma (OF) and fibrous dysplasia (FD) are two fibro-osseous lesions with overlapping clinicopathological features, making diagnosis challenging. In this study, we applied a whole-genome shallow sequencing approach to facilitate differential diagnosis via precise profiling of copy number alterations (CNAs) using minute amounts of DNA extracted from morphologically correlated microdissected tissue samples. Freshly frozen tissue specimens from OF (n = 29) and FD (n = 28) patients were obtained for analysis. Lesion fibrous tissues and surrounding normal tissues were obtained by laser capture microdissection (LCM), with ~30–50 cells (5 000–10 000 µm2) per sample. We found that the rate of recurrent CNAs in OF cases was much higher (44.8%, 13 of 29) than that in FD cases (3.6%, 1 of 28). Sixty-nine percent (9 of 13) of the CNA-containing OF cases involved segmental amplifications and deletions on Chrs 7 and 12. We also identified eight CNA-associated genes (HILPDA, CALD1, C1GALT1, MICALL2, PHF14, AIMP2, MDM2, and CDK4) with amplified expression, which was consistent with the copy number changes. We further confirmed a jaw lesion with a previous uncertain diagnosis due to its ambiguous morphological features and the absence of GNAS mutation as OF based on the typical Chr 12 amplification pattern in its CNA profile. Moreover, analysis of a set of longitudinal samples collected from an individual with a cellular lesion in suspicion of OF at the first surgery, recurrence and the latest malignant transformation revealed identical CNA patterns at the three time points, suggesting that copy number profiling can be used as an important tool to identify borderline lesions or lesions with malignant potential. Overall, CNA profiling of fibro-osseous lesions can greatly improve differential diagnosis between OF and FD and help predict disease progression.

CD25 Enables Oncogenic BCR-Signaling and Represents a Therapeutic Target in B Cell Malignancies

Background and hypothesis: B cells critically depend on continuous survival and proliferation signals from a functional B cell receptor (BCR). In >50% of B cell malignancies, the tumor clone is driven by an oncogenic BCR-mimic. Oncogenic mimics of BCR-signaling include BCR-ABL1 (Ph+ ALL) and other ABL1-fusion genes (Ph -like ALL), viral oncoproteins (e.g. EBV; KSHV), BRAF- (hairy cell leukemia) and NF-κB-pathway lesions (Hodgkin's lymphoma, primary mediastinal B cell lymphoma). B cell tumors driven by oncogenic BCR-signaling represent a highly heterogeneous group of various cellular origins, genetic lesions and divergent clinical outcomes. Despite this heterogeneity, our correlative analyses showed that these B cell malignancies are uniformly characterized by high expression levels of CD25.Features of CD25 function in B cell malignancies: We found out that adults with Ph+ ALL (ECOG; MDACC) and children with Ph -like ALL (P9906) with high CD25 expression at the time of diagnosis have a particularly poor outcome (n=416; P =0.005). While CD25 mediates IL2 signaling on T cells and is highly expressed on regulatory T cells (Tregs), CD25 on B-lineage cells did not pair with IL2Rβ and was not responsive to IL2. Our genetic studies showed that BCR signaling rapidly induces CD25 cell surface translocation by PKCδ-dependent phosphorylation of the CD25 cytoplasmic tail (S268), which is further supported by transcriptional upregulation of CD25 by activation of NF-κB. Antibody mediated stimulation of surface exposed CD25 in response to BCR signaling shuttles inhibitory phosphatases (e.g. SHIP1) from the cytoplasm to the cell membrane. CD25-mediated membrane recruitment of inhibitory phosphatases limits kinase signaling (e.g. SYK, SRC and ERK) downstream of the BCR or its oncogenic mimics. Consistent with a role as negative feedback regulator, our genetic studies showed that CD25 deficient pre-B cells (Il2ra-/- ) have autonomous pre-BCR signals, resulting in uncoordinated Ca2+ oscillations of variable duration. Genetic ablation of CD25 subverted the ability of B cell leukemia cells to balance oncogenic signal strength, resulting in p53 checkpoint activation, cell cycle arrest, therefore failed initiation of B cell leukemia in transplant recipients. Importantly, reconstitution of myristoylated CD25 tail but not a mutant construct lacking the serine/threonine motif (S268A/T271A) rescued proliferation and survival defects of Il2ra-/- ALL cells.CD25 as novel therapeutic target in BCR-mimic B cell malignancies: Despite heterogeneity of B cell malignancies, we found that CD25 is an integrated biomarker of tumor clones driven by oncogenic BCR-mimics. BCR-dependent B cell tumors further exposed CD25 on the surface in response to the PKCδ activator, PEP005 (also known as Ingenol Mebutate) with rapid phosphorylation of PKCδT505, followed by CD25S268. As a strategy to eradicate CD25-expressing B cell malignancies, combined treatment of PEP005 with ADCT-301, an antibody drug conjugate (ADC) targeting CD25 with a pyrrolobenzodiazepine (PBD) dimer as the cytotoxic warhead, synergistically delayed leukemia development in recipient mice xenografted with patient-derived Ph+ ALL cells. Notably, synergistic effects of ADCT-301 were further enhanced by treatment with AZD5363, an AKT-inhibitor, which further upregulates CD25 on patient-derived Ph+ ALL cells by imitating signaling pressure leading to differentiation of large circulating pre-B to small pre-B cell fraction.Conclusion: Based on these finding, we propose CD25 as an integrated biomarker of BCR-dependent B cell tumors that are highly sensitive to established BCR signaling antagonists (e.g. Ibrutinib, Entospletinib) and as a therapeutic target in refractory B cell tumors. DisclosuresZammarchi:ADC Therapeutics: Employment. Van Berkel:ADC Therapeutics: Employment. Muschen:AbbVie: Research Funding; Pfizer: Research Funding.

Augmented secretion of IL-1α from mouse oral squamous cell carcinoma (OSCC) vcells caused by serum deprivation and hypoxia promotes immune suppressive activity of mesenchymal stromal cells

ObjectivesWe have previously reported that mouse oral squamous carcinoma (OSCC), Sq-1979-1, produces interleukin (IL)-1α, which specifically enhances the immunosuppressive activity of co-cultured mesenchymal stromal 10T1/2 cells. This study assessed the conditions promoting the production of IL-1α in Sq-1979-1 cells, which could further enhance the immunosuppressive function of 10T1/2 cells, and evaluated its expression in OSCC tissues. MethodsThe expression of IL-1α was examined by RT-PCR, western blotting, and enzyme-linked immune sorbent assay (ELISA). The interferon (IFN)- γ-producing capability of anti-CD3 antibody-stimulated mouse spleen cells co-cultured with 10T1/2 cells and conditioned medium (CM) from Sq-1979-1 cells was examined by ELISA. The function of IL-1α was examined using an anti-IL1α antibody. Immunohistochemical analysis of the OSCC tissues was performed. ResultsThe production of IL-1α from Sq-1979-1 cells was synergistically enhanced in lower serum (0.5% or 1.0% FBS) at the transcriptional level, and under hypoxia (1.0% oxygen) at the release level compared to that in the control medium supplemented with 10% FBS under normoxia. The IFN-γ-producing capability of stimulated spleen cells co-cultured with 10T1/2 cells was significantly reduced in the CMs prepared with the lower serum or under hypoxia. These functions of CMs were completely abolished by the anti-IL-1α antibody. The expression of IL-1α in OSCC tissues was prominent in the midst of a carcinomatous cellular lesion or a nearby necrotic lesion, where a supply deficiency could occur. Conclusions: IL-1α production by Sq-1979-1 cells was synergistically augmented under low serum and hypoxic conditions, which could promote the immunosuppressive activity of mesenchymal cells.

ROLE OF ADVANCED MAGNETIC RESONANCE IMAGING TECHNIQUES IN DISTINCTION BETWEEN CEREBRAL ABSCESSES AND HIGH-GRADE NEOPLASMS.

• Distinguishing between cerebral abscess and rim-enhancing tumor is a problem that is encountered frequently by radiologists. Conventional contrast-enhanced MRI reveals ring enhancement of a brain abscess that is similar to the ring enhancement of a necrotic high-grade glioma or metastasis.• Advanced MRI techniques (DWI, MRS and DSC MR perfusion) may be used to differentiate between cerebral abscesses and high grade brain neoplasms.• DWI is an advanced MRI technique that provides the physiological nature of the disease based on motion of water molecules across cell membrane.• DWI allows assessment of the cellularity of the lesion non-invasively, as cellular and subcellular elements significantly impede the mobility of water molecules, thus densely cellular lesions show restricted diffusion and hence low ADC values. • Magnetic Resonance spectroscopy (MRS) is a non-invasive diagnostic test for measuring biochemical changes in the brain, it MRS is capable of differentiating normal from pathologic brain.• Dynamic susceptibility contrast (DSC) perfusion MRI is a technique that allows derivation of relative cerebral blood volume (rCBV) maps on the basis of the susceptibility effect related to passage of MRI contrast material through tissue.• It provides noninvasive physiologic measurements of tumor vascularity and relative cerebral blood volume (rCBV) maps, which can be used to identify and quantify areas of neovascularization.• We try to evaluate the role of advanced MRI techniques (DWI, MRS, DSC PERFUSION) to reach a final diagnosis in a case of brain abscess versus a cystic or necrotic tumor.

Upregulated SOCC and IP3R calcium channels and subsequent elevated cytoplasmic calcium signaling promote nonalcoholic fatty liver disease by inhibiting autophagy.

Nonalcoholic fatty liver disease (NAFLD) is related to elevated cytoplasmic calcium signaling in hepatocytes, which may be mediated by store-operated calcium channel (SOCC) and inositol triphosphate receptor (IP3R). However, the regulatory effect of calcium signaling on lipid accumulation and degeneration in hepatocytes and the underlying molecular mechanism remain unknown. Autophagy inhibition promotes lipid accumulation and steatosis in hepatocytes. However, the association between elevated calcium signaling and autophagy inhibition in hepatocytes and its effect on hepatocyte fatty lesions remain unclear. Here, we established a mouse hepatocyte fatty gradient model using oleic acid. SOCC and IP3R channel opening and cytoplasmic calcium levels gradually increased with the hepatocyte pimelosis degree, whereas autophagy gradually decreased. We also established an optimal oleic acid (OOA) hepatocyte model, observing significantly increased SOCC and IP3R channel opening and calcium influx alongside significantly decreased autophagy and aggravated cellular fatty lesion. Calcium channel blockers (CCBs) and calcium channel gene silencing reagents (CCGSRs), respectively, reversed these effects, indicating that elevated cytoplasmic calcium signaling promotes NAFLD occurrence and the development by inhibiting hepatocyte autophagy. In the OOA model, upregulated extracellular regulated protein kinases 1/2 (ERK1/2), which can be regulated by SOCC and IP3R proteins transient receptor potential canonical 1 (TRPC1)/IP3R with elevated cytoplasmic calcium signaling, over-inhibited forkhead/winged helix O (FOXO) signaling and over-activated mammalian target of rapamycin complex 1 (mTORC1) signaling. Over-inhibited FOXO signaling significantly downregulated autophagy-related gene 12, which inhibits autophagosome maturation, while over-activated mTORC1 signaling over-inactivated Unc-51 like autophagy activating kinase 1, which inhibits preautophagosome formation. CCBs and CCGSRs recovered autophagy by significantly downregulating ERK1/2 to block abnormal changes in FOXO and mTORC1 signaling. Our findings indicate that upregulated SOCC and IP3R channels and subsequent elevated cytoplasmic calcium signaling in hepatocyte fatty lesions inhibits hepatocyte autophagy through (TRPC1/IP3R)/ERK/(FOXO/mTORC1) signaling pathways, causes lipid accumulation and degeneration in hepatocytes, and promotes NAFLD occurrence and development.

WWP2 and PPP1R3A are abnormally regulated in arrhythmia-induced cardiac damage.

The present work aimed to identify the roles of WWP2 (an E3 ubiquitin-protein ligase) and protein phosphatase 1 regulatory subunit 3A (PPP1R3A) in different pathological stages of cardiac arrhythmia development. Leptin-deficient mice (C57BLKS-Leprdb/Leprdb) were used for the development of initial and severe stages of cardiac arrhythmia. Histology, ECG, immunohistochemistry and Western blotting were used to analyse cardiac arrhythmia, WWP2 and PPP1R3A expression. Histopathological studies of 4-month-old mice showed cardiac degeneration, cellular lesions, and swollen tissue structure with loss of tissue elasticity, indicative of the initial condition of cardiac arrhythmia. The leptin-deficient 7-month-old mice showed cardiac tissue hardening with increased secretion of extracellular matrix. The development of initial- and severe-cardiac arrhythmia was further evident with electrocardiogram studies, which showed more PP interval variations as the disease progressed. At the molecular level, WWP2 showed marginal upregulation in the initial stages of arrhythmia and was predominantly expressed within nuclei. WWP2 was overexpressed 6.6-fold in the severe stage of cardiac arrhythmia and was spread throughout the tissue layer. Interestingly, PPP1R3A was significantly overexpressed in initial cardiac arrhythmia conditions, but was downregulated and restricted to more nuclear expression in advanced cardiac arrhythmia. Silencing of PPP1R3A, enhances the expression of WWP2 to 5.3-fold in initial stages, but remarkable variation not observed in advanced cardiac arrhythmia conditions. Our results suggest that PPP1R3A had a control over WWP2 in the initial stages of cardiac arrhythmia. In particular, PPP1R3A overexpression implies its potential protective effect in initial cardiac arrhythmia stages.

Treatment of Anti-Body Mediated Rejection Complicated by Cardiogenic Shock with a Temporary Left Ventricular Assist Device as a Bridge to Recovery

<h3>Introduction</h3> Anti-body mediated rejection (AMR) after cardiac transplantation is a difficult entity to diagnose and treat. Mortality rates in AMR remain high and treatment strategies vary but consist of therapies that aim to remove circulating antibodies, decrease antibody production, suppress T and B-cell responses, and inhibit complement. We report a case of AMR associated cardiogenic shock supported on Impella 5.0. <h3>Case Report</h3> : A 51-year-old man who received a cardiac transplant in 2018 was admitted for chest pain and dyspnea. Six weeks prior, he had similar symptoms with a right heart catherization (RHC) showing normal hemodynamics. Endomyocardial biopsy did not show cellular or AMR. On admission, he was found to be in cardiogenic shock SCAI stage D with severe bi-ventricular failure. RHC confirmed elevated right and left-sided filling pressures and low cardiac output (cardiac power output 0.7). Biopsy showed grade 1R cellular rejection and infiltrative quilty lesions with focal endocardial inflammation and C3d positive capillaries. He was intubated for hypoxemic respiratory failure and an Impella 5.0 was placed for left ventricular unloading and tissue perfusion in addition to milrinone and epinephrine drips. Labs were significant for new class II DSA (DQ7, DR53 immunodominant antibodies). He was treated with intravenous solumedrol, plasmapheresis, and antithymocyte globulin-rabbit (rATG, Thymoglobulin). He received multiple sessions of bortezomib infusion with plasmapheresis while supported with Impella 5.0. He improved over the following three weeks and was extubated with explant of the Impella 5.0 on day 16. His DSA's continued to decrease and graft function improved with ejection fraction up to 50% (Figure 1). He was discharged with plans for further bortezomib and plasmapheresis sessions. <h3>Summary</h3> This case demonstrates the successful treatment of cardiogenic shock precipitated by AMR with an Impella 5.0 as a bridge to recovery.

Consciousness, Anesthesia, and Acetylcholine.

Consciousness, Anesthesia, and Acetylcholine.

Nephrotic syndrome with focal segmental glomerular lesions unclassified by Columbia classification; Pathology and clinical implication.

BackgroundThe Columbia classification is widely used for diagnosis of focal segmental glomerulosclerosis (FSGS). In practice, we occasionally encounter segmental glomerular lesions unclassified as Columbia classification. We analyzed the clinical implication of unclassified segmental lesions comparing with Columbia-classified FSGS.MethodsA retrospective cohort study from 13 local hospitals in Japan. From 172 biopsy cases diagnosed with FSGS or minimal change disease (MCD)/FSGS spectrum with unclassified segmental lesions, adult patients with nephrotic syndrome who received immunosuppressive therapies were included. The cases are classified by pathology, i.e., typical FSGS lesions sufficiently classified into subgroups of Columbia classification: collapsing (COL), tip (TIP), cellular (CEL), perihilar (PH), and not otherwise specified (NOS), and unclassified by the Columbia classification into three subgroups: “endothelial damage,”; “simple attachment,”; and “minor cellular lesion,”. The response to immunosuppressive treatment and 30% decline of eGFR were compared.ResultsAmong 48 eligible cases, all were Japanese, 34 were typical FSGS; 13 TIP, 15 CEL, 6 NOS, and no COL or PH cases. Fourteen were unclassified cases: endothelial damage (n = 6), simple attachment (n = 5), and minor cellular lesion (n = 3). The median age of overall patients was 60 years old and the median of eGFR and urinary protein creatinine ratio was 51.5 mL/min/1.73m2 and 7.35, respectively. They received similar therapeutic regimen. Kaplan-Meier analysis revealed no significant difference in treatment response between typical FSGS and unclassified cases. Evaluating among the subgroups, endothelial damage, simple attachment and minor cellular lesion showed similar treatment response to TIP or CEL. No significant difference was also observed in the 30% decline of eGFR.ConclusionsJapanese adult patients with nephrotic syndrome showing unclassified segmental lesions as Columbia classification may be equivalent clinical impact as Columbia classification of FSGS.

Femoral marrow MRI is a non-invasive, non-irradiated and useful tool for detecting bone marrow involvement in non-Hodgkin lymphoma.

Femoral marrow magnetic resonance imaging (MRI) is a non-invasive, non-irradiated and useful modality for evaluating bone marrow (BM) conditions. Human adult femoral BM is almost uniformly fatty marrow and has the largest volume of a single bone. MRI has an extremely high resolution for fat and water, which allows high-contrast imaging of cellular infiltration into fat tissue. In hematological diseases, femoral BM MRI can clearly detect cell infiltration, which is symmetrically imaged from the proximal to the distal direction of abnormal signal areas. Thus, we investigated the significance of femoral MRI for non-Hodgkin lymphoma (NHL). We analyzed the data of 69 NHL patients who received femoral MRI at diagnosis in this single-center retrospective cohort study. The median patient age was 73 years. MRI patterns were mainly classified as uniform patterns or nonuniform patterns. We also classified the range of cellular marrow as high-grade or low-grade based on whether it had spread to over half of the femur. Both overall survival (OS) and progression-free survival (PFS) were significantly influenced by abnormal femoral marrow MRI. In particular, the patients with cellular femoral marrow lesions had a worse OS and PFS based on log-rank tests. Multivariable analyses with the Cox proportional hazards model revealed that OS and PFS were significantly influenced by cellular marrow diagnosed by femoral MRI. We concluded that femoral marrow MRI is a useful tool for detecting BM involvement and an independent prognostic factor in NHL patients.

Increased Reactive Oxygen Species and Distinct Oxidative Damage in Resveratrol-suppressed Glioblastoma Cells.

Background and Aim: Glioblastoma multiforme (GBM) is a highly aggressive brain malignancy that lacks reliable treatments. Resveratrol possesses anti-cancer effects, but its activity against glioblastoma cells is variable for unknown reasons. One mechanism through which anti-cancer drugs exert their effects is oxidative damage caused by increased reactive oxygen species (ROS) production. Thus, the present study examined the relationship between oxidative stress and sensitivity to resveratrol in glioblastoma cells.Methods: Two GBM cell lines (U251 and LN428) were exposed to 100 μM resveratrol for 48 h, and proliferation and apoptosis were assessed. ROS generation was evaluated using 2′-7′-dichlorodihydrofluorescein diacetate-based flow cytometry and fluorescent microscopy. Immunocytochemical staining and western blotting were conducted at regular intervals to profile the expression patterns of superoxide dismutase-2 (SOD2), catalase, caspase-9, caspase-3, and sulfotransferases (SULTs) in untreated and resveratrol-treated GBM cells.Results: Resveratrol-treated U251 cells, but not resveratrol-treated LN428 cells, exhibited remarkable growth arrest and extensive apoptosis accompanied by elevated intracellular ROS levels and attenuated SOD2 and catalase expression. Mitochondrial impairment and more distinct increases in the expression of activated caspase-9 and caspase-3 were detected in U251 cells following resveratrol treatment. The levels of resveratrol metabolic enzymes (SULT1A1 and SULT1C2) were lower in U251 cells than in LN428 cells.Conclusions: Resveratrol increased ROS generation and induced oxidation-related cellular lesions in U251 cells by activating an ROS-related mitochondrial signal pathway. The levels of SULTs and ROS may indicate the therapeutic outcomes of resveratrol treatment in GBM.

Les lésions histologiques associées à l’infection par le SARS-CoV-2

L’infection liée au SARS-CoV-2 entraîne des lésions qui prédominent largement, mais pas exclusivement, au niveau de l’arbre respiratoire. Leur analyse a largement bénéficié des études autopsiques. Ainsi, des lésions ont pu être observées dans les différents organes, tissus ou cellules de l’organisme. Ces observations ont permis d’améliorer rapidement la connaissance des différents mécanismes physiopathologiques associés à cette maladie émergente. Le virus et ses différentes composantes peuvent être détectés à partir de tissus fixés et inclus en paraffine, par immunohistochimie, hybridation in situ, biologie moléculaire et/ou microscopie électronique. Plusieurs inconnues persistent sur le rôle direct joué par le SARS-CoV-2 dans les différentes lésions observées, notamment au niveau cardiaque, cérébral, hépatique, digestif, cutané et rénal. Il est donc primordial de multiplier les analyses tissulaires et cellulaires chez ces patients afin de mieux comprendre les conséquences de la maladie liée à la COVID-19, en particulier selon les facteurs épidémiologiques et les co-morbidités. La connaissance de cette maladie doit permettre, plus particulièrement, de participer au développement des stratégies thérapeutiques. Cette revue décrit les principales lésions histologiques et cellulaires associées à l’infection par le SARS-CoV-2.

Effects of Oxygen on Renal Function and Oxidative Stress During Hypothermic Machine Perfusion in an Experimental Porcine Model of Kidney Donation after Cardiac Death

The efficacy and safety of using high concentrations of oxygen during hypothermic machine perfusion (HMP) have not been fully elucidated to date. This study investigated the impact of administering high concentrations of oxygen on renal function during HMP in a porcine donation after circulatory death (DCD), as well as the metabolic and biochemical effects of this method. A randomized nonblinded cohort study was established in a porcine transplant (KT) model mimicking Maastricht type III DCD under oxygen-supplemented HMP (Ox-HMP) compared to non-supplemented (nOx-HMP) (LifePort® kidney transporter) conditions. The primary endpoint was evolution of renal function post-KT, whereas secondary endpoints included changes in perfusion dynamics, miRNA expression and cellular lesion measured by LDH and lactate levels in perfusate, lipid peroxidation in kidney biopsies, ATP generation, epithelial mesenchymal transition (EMT) and oxidative gene expression in cell cultures and histology evaluation. ATP generation and oxidative stress, as measured by lipid peroxidation, increased simultaneously after warm ischemia in the Ox-HMP group. Ox-HMP did not exhibit a significant effect on kidney function or animal survival. A significant increase in lipid peroxidation was observed in the Ox-HMP group. This resulted in a greater expression of the genes responsible for producing superoxide dismutase 1 (SOD-1) and catalase oxygenation enzymes, although only SOD-1 showed statistical significance. Respiratory chain dysfunction was maintained in the Ox-HMP group with a non-significant decrease in ATP production, increased proton leakage, and a decrease in respiratory reserve. Regarding epithelial-mesenchymal transition, an upward trend in the expression of vimentin, fibronectin, and collagen genes was observed only in the Ox-HMP group. Finally, the expression levels of miR-101 and miR-126, related to characteristic functions of the tubular epithelium, were significantly modified (miRNA levels expressed as DCT. A brief bubble Ox-HMP treatment did not show a clear positive effect on renal function and oxidative stress markers. The role and safety of adding oxygen during HMP still need to be elucidated. Currently, this Ox-HMP method cannot be considered standard practice.

Aging, senescence and mitochondria: the PGC-1/ERR axis.

Aging is a degenerative process that results from the accumulation of cellular and tissue lesions, leading progressively to organ dysfunction and death. Although the biological basis of human aging remains unclear, a large amount of data points to deregulated mitochondrial function as a central regulator of this process. Mounting years of research on aging support the notion that the engendered age-related decline of mitochondria is associated with alterations in key pathways that regulate mitochondrial biology. Particularly, several studies in the last decade have emphasized the importance of the estrogen-related receptor (ERR) family of nuclear receptors, master regulators of mitochondrial function, and their transcriptional coactivators PGC-1s in this context. In this review, we summarize key discoveries implicating the PGC-1/ERR axis in age-associated mitochondrial deregulation and tissue dysfunction. Also, we highlight the pharmacological potential of targeting the PGC-1/ERR axis to alleviate the onset of aging and its adverse effects.