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Toxoplasma gondii is a single-celled parasite that causes a disease called toxoplasmosis. It can reach the central nervous system, but the mechanism of T. gondii disrupting the functioning of these brain regions occurs in bradyzoite stage of parasite, causing brain damage by forming tissue cysts in brain. In our study, the effects of T. gondii on locomotor activity, anxiety, learning and memory, and norepinephrine (NE), levodopa (L-DOPA), dopamine (DA) and 3,4-D-dihydroxyphenylacetic acid (DOPAC) catecholamines in amygdala, striatum, prefrontal cortex and hippocampus regions of the brain were investigated in bradyzoite stage. Twenty male Albino mice Mus musculus, 4–5 weeks old, weighing 20–25 g, were used. T. gondii inoculated to mice intraperitonealy with 48–50-hour passages of T. gondii RH Ankara strain. For intraperitoneal inoculation of mice 5x104 tachyzoites per mouse. No inoculation was made in control group (n: 20). Locomotor activity behavior in open field test (OFT), anxious behavior in elevated plus maze (EPM), and learning behavior in novel object recognition (NOR) tests were evaluated. NE, L-DOPA, DA and DOPAC were measured by HPLC in brain tissues of amygdala, striatum, prefrontal cortex and hippocampus. A decrease was observed in the locomotor activity, anxiety and learning values of the T. gondii group compared to the control group (p ;lt; 0.05). The heighten in NE and L-DOPA levels in amygdala tissue of T. gondii group compared to control group, an elevation in NE, L-DOPA, DA and DOPAC levels in striatum tissue, and an increase in levels of NE in prefrontal cortex tissue were detected in monoamine results. In hippocampus tissue, an increase was observed in DA levels, while a decrease was observed in NE, L-DOPA and DOPAC levels. In our study, it has been shown that T. gondii in bradyzoite stage reduces locomotor activity, causes learning and memory impairment, and has anxiogenic effects. © 2024 Elsevier B.V., School of Medicine, Duke University: 2021/09/03; Hitit Üniversitesi, HİTÜ: TIP19001.21.007

Introduction: Releasing of cerium oxide nanoparticles (nano-CeO2) to the nature has increased due to the widespread use in many fields ranging from cosmetics to the food industry. Therefore, nano-CeO2 has been included in the Organization for Economic Co-operation and Development's (OECD) priority list for engineering nanomaterials. In this study, the effects of nano-CeO2 on the freshwater mussels were investigated to reveal the impact on the freshwater systems on model organism. Methods: First, the chemical and structural properties of nano-CeO2 were characterized in details. Second, the freshwater mussels were exposed to environmentally relevant concentrations of nano-CeO2 as 10 mg, 25 mg and 50 mg/L during 48-h and 7-d. Third, after the exposure periods, hemolymph and tissue samples were taken to analyse the Total Hemocyte Counts (THCs) histology and oxidative stress parameters (total antioxidant status, glutathione, glutathione-S-transferase, and advanced oxidative protein products). Results: Significant decrease of the THCs was observed in the nano-CeO2 exposed mussels compared to the control group (P ;lt; 0.05). The histological results showed a positive association between nano-CeO2 exposure concentration in the water and level of tissue damage and histopathological alterations were detected in the gill and the digestive gland tissues. Oxidative stress parameters were slightly affected after exposure to nano-CeO2 (P ;gt; 0.05). In conclusion, this study showed that acute exposure of freshwater mussels to nano-CeO2 did not pose significant biological risk. However, it has been proven that mussels are able to accumulate nano-CeO2 significantly in their bodies. Conclusion: This suggests that nano-CeO2 may be a potential risk to other organisms in the ecosystem through trophic transfer in the food-web based on their habitat and niche in the ecosystem. © 2024 Elsevier GmbH

Introduction: Releasing of cerium oxide nanoparticles (nano-CeO2) to the nature has increased due to the widespread use in many fields ranging from cosmetics to the food industry. Therefore, nano-CeO2 has been included in the Organization for Economic Co-operation and Development's (OECD) priority list for engineering nanomaterials. In this study, the effects of nano-CeO2 on the freshwater mussels were investigated to reveal the impact on the freshwater systems on model organism. Methods: First, the chemical and structural properties of nano-CeO2 were characterized in details. Second, the freshwater mussels were exposed to environmentally relevant concentrations of nano-CeO2 as 10 mg, 25 mg and 50 mg/L during 48-h and 7-d. Third, after the exposure periods, hemolymph and tissue samples were taken to analyse the Total Hemocyte Counts (THCs) histology and oxidative stress parameters (total antioxidant status, glutathione, glutathione-S-transferase, and advanced oxidative protein products). Results: Significant decrease of the THCs was observed in the nano-CeO2 exposed mussels compared to the control group (P ;lt; 0.05). The histological results showed a positive association between nano-CeO2 exposure concentration in the water and level of tissue damage and histopathological alterations were detected in the gill and the digestive gland tissues. Oxidative stress parameters were slightly affected after exposure to nano-CeO2 (P ;gt; 0.05). In conclusion, this study showed that acute exposure of freshwater mussels to nano-CeO2 did not pose significant biological risk. However, it has been proven that mussels are able to accumulate nano-CeO2 significantly in their bodies. Conclusion: This suggests that nano-CeO2 may be a potential risk to other organisms in the ecosystem through trophic transfer in the food-web based on their habitat and niche in the ecosystem. © 2024 Elsevier GmbH

Chimeric Antigen Receptor (CAR) T cell therapy has produced revolutionary success in hematological cancers such as leukemia and lymphoma. Nonetheless, its translation to solid tumors faces challenges due to manufacturing complexities, short-lived in vivo persistence, and transient therapeutic impact. We introduce 'Drydux' - an innovative macroporous biomaterial scaffold designed for rapid, efficient in-situ generation of tumor-specific CAR T cells. Drydux expedites CAR T cell preparation with a mere three-day turnaround from patient blood collection, presenting a cost-effective, streamlined alternative to conventional methodologies. Notably, Drydux-enabled CAR T cells provide prolonged in vivo release, functionality, and enhanced persistence exceeding 150 days, with cells transitioning to memory phenotypes. Unlike conventional CAR T cell therapy, which offered only temporary tumor control, equivalent Drydux cell doses induced lasting tumor remission in various animal tumor models, including systemic lymphoma, peritoneal ovarian cancer, metastatic lung cancer, and orthotopic pancreatic cancer. Drydux's approach holds promise in revolutionizing solid tumor CAR T cell therapy by delivering durable, rapid, and cost-effective treatments and broadening patient accessibility to this groundbreaking therapy.

The transgenerational maternal effects of polycystic ovary syndrome (PCOS) in female progeny are being revealed. As there is evidence that a male equivalent of PCOS may exists, we ask whether sons born to mothers with PCOS (PCOS-sons) transmit reproductive and metabolic phenotypes to their male progeny. Here, in a register-based cohort and a clinical case-control study, we find that PCOS-sons are more often obese and dyslipidemic. Our prenatal androgenized PCOS-like mouse model with or without diet-induced obesity confirmed that reproductive and metabolic dysfunctions in first-generation (F1) male offspring are passed down to F3. Sequencing of F1–F3 sperm reveals distinct differentially expressed (DE) small non-coding RNAs (sncRNAs) across generations in each lineage. Notably, common targets between transgenerational DEsncRNAs in mouse sperm and in PCOS-sons serum indicate similar effects of maternal hyperandrogenism, strengthening the translational relevance and highlighting a previously underappreciated risk of transmission of reproductive and metabolic dysfunction via the male germline., CC BY 4.0© 2023 The Author(s)Correspondence: qiaolin.deng@ki.se (Q.D.), elisabet.stener-victorin@ki.se (E.S.-V.)We thank Zhiyi Zhao, Jacob Victorin, Sonja Edström, and Sara Pilström for technical assistance during animal work and molecular analysis; TSE Systems and the Metabolic Phenotyping Center at the Strategic Research program in Diabetes at the Karolinska Institutet; and the electron microscopy unit Emil at Huddinge University Hospital at the Karolinska Institutet. This work is supported by the Swedish Medical Research Council: project nos. 2018-02435 and 2022-00550 (E.S.-V.) and 2018-02557 and 2020-00253 (Q.D.); the Knut and Alice Wallenberg Foundation: 2019.0211 (Q.D.); Distinguished Investigator Grant – Endocrinology and Metabolism, Novo Nordisk Foundation: NNF22OC0072904 (E.S.-V.); the Diabetes Foundation:DIA2021-633 (E.S.-V.); the Novo Nordisk Foundation: NNF18OC0033992 and NNF19OC0056647 (E.S.-V.); the Strategic Research Program in Diabetes at the Karolinska Institutet (E.S.-V.); the Adlerbertska Research Foundation: GU 2019/86 (E.S.-V.); Karolinska Institutet KID funding: 2020-00990 (E.S.-V.); a Karolinska Instiutet faculty funded position (Q.D.); the Regional Agreement on Medical Training and Clinical Research between the Stockholm County Council and the Karolinska Institutet: 20190079 (E.S.-V.); O.E. och Edla Johanssons Stiftelse 2021 (S.R.); the Karolinska Institutet China scholarship council program (Q.L.); Magnus Bergvalls Stiftelse: 2020-03808 and 2021-04329 (S.R.); the Karolinska Institutet: 2020-02026 (S.R.); the National Fund for Scientific and Technological Development (FONDECYT): project no. 1151531 (T.S.P.); the FONDECYT: project no. 1201483 (B.E.); the National Commission for Scientific and Technological Research (CONICYT) (R.F.); HKH Kronprinsessan Lovisas förening för barnasjukvård (R.F.); and Stiftelsen Axel Tielmans minnesfond (R.F.)

The transgenerational maternal effects of polycystic ovary syndrome (PCOS) in female progeny are being revealed. As there is evidence that a male equivalent of PCOS may exists, we ask whether sons born to mothers with PCOS (PCOS-sons) transmit reproductive and metabolic phenotypes to their male progeny. Here, in a register-based cohort and a clinical case-control study, we find that PCOS-sons are more often obese and dyslipidemic. Our prenatal androgenized PCOS-like mouse model with or without diet-induced obesity confirmed that reproductive and metabolic dysfunctions in first-generation (F1) male offspring are passed down to F3. Sequencing of F1–F3 sperm reveals distinct differentially expressed (DE) small non-coding RNAs (sncRNAs) across generations in each lineage. Notably, common targets between transgenerational DEsncRNAs in mouse sperm and in PCOS-sons serum indicate similar effects of maternal hyperandrogenism, strengthening the translational relevance and highlighting a previously underappreciated risk of transmission of reproductive and metabolic dysfunction via the male germline., CC BY 4.0© 2023 The Author(s)Correspondence: qiaolin.deng@ki.se (Q.D.), elisabet.stener-victorin@ki.se (E.S.-V.)We thank Zhiyi Zhao, Jacob Victorin, Sonja Edström, and Sara Pilström for technical assistance during animal work and molecular analysis; TSE Systems and the Metabolic Phenotyping Center at the Strategic Research program in Diabetes at the Karolinska Institutet; and the electron microscopy unit Emil at Huddinge University Hospital at the Karolinska Institutet. This work is supported by the Swedish Medical Research Council: project nos. 2018-02435 and 2022-00550 (E.S.-V.) and 2018-02557 and 2020-00253 (Q.D.); the Knut and Alice Wallenberg Foundation: 2019.0211 (Q.D.); Distinguished Investigator Grant – Endocrinology and Metabolism, Novo Nordisk Foundation: NNF22OC0072904 (E.S.-V.); the Diabetes Foundation:DIA2021-633 (E.S.-V.); the Novo Nordisk Foundation: NNF18OC0033992 and NNF19OC0056647 (E.S.-V.); the Strategic Research Program in Diabetes at the Karolinska Institutet (E.S.-V.); the Adlerbertska Research Foundation: GU 2019/86 (E.S.-V.); Karolinska Institutet KID funding: 2020-00990 (E.S.-V.); a Karolinska Instiutet faculty funded position (Q.D.); the Regional Agreement on Medical Training and Clinical Research between the Stockholm County Council and the Karolinska Institutet: 20190079 (E.S.-V.); O.E. och Edla Johanssons Stiftelse 2021 (S.R.); the Karolinska Institutet China scholarship council program (Q.L.); Magnus Bergvalls Stiftelse: 2020-03808 and 2021-04329 (S.R.); the Karolinska Institutet: 2020-02026 (S.R.); the National Fund for Scientific and Technological Development (FONDECYT): project no. 1151531 (T.S.P.); the FONDECYT: project no. 1201483 (B.E.); the National Commission for Scientific and Technological Research (CONICYT) (R.F.); HKH Kronprinsessan Lovisas förening för barnasjukvård (R.F.); and Stiftelsen Axel Tielmans minnesfond (R.F.)

The transgenerational maternal effects of polycystic ovary syndrome (PCOS) in female progeny are being revealed. As there is evidence that a male equivalent of PCOS may exists, we ask whether sons born to mothers with PCOS (PCOS-sons) transmit reproductive and metabolic phenotypes to their male progeny. Here, in a register-based cohort and a clinical case-control study, we find that PCOS-sons are more often obese and dyslipidemic. Our prenatal androgenized PCOS-like mouse model with or without diet-induced obesity confirmed that reproductive and metabolic dysfunctions in first-generation (F1) male offspring are passed down to F3. Sequencing of F1–F3 sperm reveals distinct differentially expressed (DE) small non-coding RNAs (sncRNAs) across generations in each lineage. Notably, common targets between transgenerational DEsncRNAs in mouse sperm and in PCOS-sons serum indicate similar effects of maternal hyperandrogenism, strengthening the translational relevance and highlighting a previously underappreciated risk of transmission of reproductive and metabolic dysfunction via the male germline., CC BY 4.0© 2023 The Author(s)Correspondence: qiaolin.deng@ki.se (Q.D.), elisabet.stener-victorin@ki.se (E.S.-V.)We thank Zhiyi Zhao, Jacob Victorin, Sonja Edström, and Sara Pilström for technical assistance during animal work and molecular analysis; TSE Systems and the Metabolic Phenotyping Center at the Strategic Research program in Diabetes at the Karolinska Institutet; and the electron microscopy unit Emil at Huddinge University Hospital at the Karolinska Institutet. This work is supported by the Swedish Medical Research Council: project nos. 2018-02435 and 2022-00550 (E.S.-V.) and 2018-02557 and 2020-00253 (Q.D.); the Knut and Alice Wallenberg Foundation: 2019.0211 (Q.D.); Distinguished Investigator Grant – Endocrinology and Metabolism, Novo Nordisk Foundation: NNF22OC0072904 (E.S.-V.); the Diabetes Foundation:DIA2021-633 (E.S.-V.); the Novo Nordisk Foundation: NNF18OC0033992 and NNF19OC0056647 (E.S.-V.); the Strategic Research Program in Diabetes at the Karolinska Institutet (E.S.-V.); the Adlerbertska Research Foundation: GU 2019/86 (E.S.-V.); Karolinska Institutet KID funding: 2020-00990 (E.S.-V.); a Karolinska Instiutet faculty funded position (Q.D.); the Regional Agreement on Medical Training and Clinical Research between the Stockholm County Council and the Karolinska Institutet: 20190079 (E.S.-V.); O.E. och Edla Johanssons Stiftelse 2021 (S.R.); the Karolinska Institutet China scholarship council program (Q.L.); Magnus Bergvalls Stiftelse: 2020-03808 and 2021-04329 (S.R.); the Karolinska Institutet: 2020-02026 (S.R.); the National Fund for Scientific and Technological Development (FONDECYT): project no. 1151531 (T.S.P.); the FONDECYT: project no. 1201483 (B.E.); the National Commission for Scientific and Technological Research (CONICYT) (R.F.); HKH Kronprinsessan Lovisas förening för barnasjukvård (R.F.); and Stiftelsen Axel Tielmans minnesfond (R.F.)

The transgenerational maternal effects of polycystic ovary syndrome (PCOS) in female progeny are being revealed. As there is evidence that a male equivalent of PCOS may exists, we ask whether sons born to mothers with PCOS (PCOS-sons) transmit reproductive and metabolic phenotypes to their male progeny. Here, in a register-based cohort and a clinical case-control study, we find that PCOS-sons are more often obese and dyslipidemic. Our prenatal androgenized PCOS-like mouse model with or without diet-induced obesity confirmed that reproductive and metabolic dysfunctions in first-generation (F1) male offspring are passed down to F3. Sequencing of F1–F3 sperm reveals distinct differentially expressed (DE) small non-coding RNAs (sncRNAs) across generations in each lineage. Notably, common targets between transgenerational DEsncRNAs in mouse sperm and in PCOS-sons serum indicate similar effects of maternal hyperandrogenism, strengthening the translational relevance and highlighting a previously underappreciated risk of transmission of reproductive and metabolic dysfunction via the male germline., CC BY 4.0© 2023 The Author(s)Correspondence: qiaolin.deng@ki.se (Q.D.), elisabet.stener-victorin@ki.se (E.S.-V.)We thank Zhiyi Zhao, Jacob Victorin, Sonja Edström, and Sara Pilström for technical assistance during animal work and molecular analysis; TSE Systems and the Metabolic Phenotyping Center at the Strategic Research program in Diabetes at the Karolinska Institutet; and the electron microscopy unit Emil at Huddinge University Hospital at the Karolinska Institutet. This work is supported by the Swedish Medical Research Council: project nos. 2018-02435 and 2022-00550 (E.S.-V.) and 2018-02557 and 2020-00253 (Q.D.); the Knut and Alice Wallenberg Foundation: 2019.0211 (Q.D.); Distinguished Investigator Grant – Endocrinology and Metabolism, Novo Nordisk Foundation: NNF22OC0072904 (E.S.-V.); the Diabetes Foundation:DIA2021-633 (E.S.-V.); the Novo Nordisk Foundation: NNF18OC0033992 and NNF19OC0056647 (E.S.-V.); the Strategic Research Program in Diabetes at the Karolinska Institutet (E.S.-V.); the Adlerbertska Research Foundation: GU 2019/86 (E.S.-V.); Karolinska Institutet KID funding: 2020-00990 (E.S.-V.); a Karolinska Instiutet faculty funded position (Q.D.); the Regional Agreement on Medical Training and Clinical Research between the Stockholm County Council and the Karolinska Institutet: 20190079 (E.S.-V.); O.E. och Edla Johanssons Stiftelse 2021 (S.R.); the Karolinska Institutet China scholarship council program (Q.L.); Magnus Bergvalls Stiftelse: 2020-03808 and 2021-04329 (S.R.); the Karolinska Institutet: 2020-02026 (S.R.); the National Fund for Scientific and Technological Development (FONDECYT): project no. 1151531 (T.S.P.); the FONDECYT: project no. 1201483 (B.E.); the National Commission for Scientific and Technological Research (CONICYT) (R.F.); HKH Kronprinsessan Lovisas förening för barnasjukvård (R.F.); and Stiftelsen Axel Tielmans minnesfond (R.F.)

The transgenerational maternal effects of polycystic ovary syndrome (PCOS) in female progeny are being revealed. As there is evidence that a male equivalent of PCOS may exists, we ask whether sons born to mothers with PCOS (PCOS-sons) transmit reproductive and metabolic phenotypes to their male progeny. Here, in a register-based cohort and a clinical case-control study, we find that PCOS-sons are more often obese and dyslipidemic. Our prenatal androgenized PCOS-like mouse model with or without diet-induced obesity confirmed that reproductive and metabolic dysfunctions in first-generation (F1) male offspring are passed down to F3. Sequencing of F1–F3 sperm reveals distinct differentially expressed (DE) small non-coding RNAs (sncRNAs) across generations in each lineage. Notably, common targets between transgenerational DEsncRNAs in mouse sperm and in PCOS-sons serum indicate similar effects of maternal hyperandrogenism, strengthening the translational relevance and highlighting a previously underappreciated risk of transmission of reproductive and metabolic dysfunction via the male germline., CC BY 4.0© 2023 The Author(s)Correspondence: qiaolin.deng@ki.se (Q.D.), elisabet.stener-victorin@ki.se (E.S.-V.)We thank Zhiyi Zhao, Jacob Victorin, Sonja Edström, and Sara Pilström for technical assistance during animal work and molecular analysis; TSE Systems and the Metabolic Phenotyping Center at the Strategic Research program in Diabetes at the Karolinska Institutet; and the electron microscopy unit Emil at Huddinge University Hospital at the Karolinska Institutet. This work is supported by the Swedish Medical Research Council: project nos. 2018-02435 and 2022-00550 (E.S.-V.) and 2018-02557 and 2020-00253 (Q.D.); the Knut and Alice Wallenberg Foundation: 2019.0211 (Q.D.); Distinguished Investigator Grant – Endocrinology and Metabolism, Novo Nordisk Foundation: NNF22OC0072904 (E.S.-V.); the Diabetes Foundation:DIA2021-633 (E.S.-V.); the Novo Nordisk Foundation: NNF18OC0033992 and NNF19OC0056647 (E.S.-V.); the Strategic Research Program in Diabetes at the Karolinska Institutet (E.S.-V.); the Adlerbertska Research Foundation: GU 2019/86 (E.S.-V.); Karolinska Institutet KID funding: 2020-00990 (E.S.-V.); a Karolinska Instiutet faculty funded position (Q.D.); the Regional Agreement on Medical Training and Clinical Research between the Stockholm County Council and the Karolinska Institutet: 20190079 (E.S.-V.); O.E. och Edla Johanssons Stiftelse 2021 (S.R.); the Karolinska Institutet China scholarship council program (Q.L.); Magnus Bergvalls Stiftelse: 2020-03808 and 2021-04329 (S.R.); the Karolinska Institutet: 2020-02026 (S.R.); the National Fund for Scientific and Technological Development (FONDECYT): project no. 1151531 (T.S.P.); the FONDECYT: project no. 1201483 (B.E.); the National Commission for Scientific and Technological Research (CONICYT) (R.F.); HKH Kronprinsessan Lovisas förening för barnasjukvård (R.F.); and Stiftelsen Axel Tielmans minnesfond (R.F.)

Development of nucleic acid delivery systems requires the use of adequate tissue analysis methods, especially when aiming tissue-targeted drug delivery. In this chapter, a protocol is presented for analyzing a reporter signal from the lung tissue. Because lung is an important target tissue from the clinical point of view, yet represents a challenge from the histological point of view, this protocol can be used in any lung-targeting drug delivery project.

Traumatic brain injury (TBI) is a major public health problem. Here, we developed a novel model of non-invasive TBI induced by laser irradiation in the telencephalon of adult zebrafish (Danio rerio) and assessed their behavior and neuromorphology to validate the model and evaluate potential targets for neuroreparative treatment. Overall, TBI induced hypolocomotion and anxiety-like behavior in the novel tank test, strikingly recapitulating responses in mammalian TBI models, hence supporting the face validity of our model. NeuN-positive cell staining was markedly reduced one day, but not seven days, after TBI, suggesting increased neuronal damage immediately after the injury, and its fast recovery. The brain-derived neurotrophic factor (Bdnf) level in the brain dropped immediately after the trauma, but fully recovered seven days later. A marker of microglial activation, Iba1, was elevated in the TBI brain, albeit decreasing from Day 3. The levels of hypoxia-inducible factor 1-alpha (Hif1a) increased 30 min after the injury, and recovered by Day 7, further supporting the construct validity of the model. Collectively, these findings suggest that our model of laser-induced brain injury in zebrafish reproduces mild TBI and can be a useful tool for TBI research and preclinical neuroprotective drug screening. © 2022 by the authors.

Acute lung injury remains a challenging clinical condition, necessitating the development of novel, safe and efficient treatments. The prevention of macrophage M1-polarization is a viable venue to tackle excessive inflammation. We performed a phenotypic screening campaign to identify azolopyrimidine compounds that effectively inhibit LPS-induced NO synthesis and interleukin 6 (IL-6) secretion. We identified lead compound 9g that inhibits IL-6 secretion with IC50 of 3.72 µM without apparent cytotoxicity and with minimal suppression of macrophage phagocytosis in contrast to dexamethasone. In a mouse model of LPS-induced acute lung injury, 30 mg/kg i.p. 9g ameliorated anxiety-like behavior, inhibited IL-6 release, and limited neutrophil infiltration and pulmonary edema. A histological study confirmed the protective activity of 9g. Treatment with compound 9g prevented the migration of CD68+ macrophages and the incidence of hemorrhage. Hence, we have identified a promising pharmacological approach for the treatment of acute lung injury that may hold promise for the development of novel drugs against cytokine-mediated complications of bacterial and viral infections. © 2022 by the authors. Licensee MDPI, Basel, Switzerland.

The agonistic potentials of therapeutic anti-CD40 antibodies have been profiled in relation to antibody isotype and epitope specificity. Still, clinical impact relies on a well-balanced clinical efficacy versus target-mediated toxicity. As CD40-mediated immune activation must rely on a combination of stimulation of antigen-presenting cells (APCs) alongside antigen presentation, for efficient T cell priming, alternative approaches to improve the therapeutic outcome of CD40-targeting strategies should focus on providing optimal antigen presentation together with CD40 stimulation. Herein, a bispecific antibody targeting CD40 as a means to deliver cargo (i.e., synthetic peptides) into APCs through a non-covalent, high-affinity interaction between the antibody and the cargo peptide, further referred to as the Adaptable Drug Affinity Conjugate (ADAC) technology, has been developed. The ADAC platform demonstrated a target-specific CD4+ and CD8+ T cell expansion in vitro and significantly improved peptide-specific CD8+ T cell proliferation in vivo. In addition, the strategy dramatically improved the in vitro and in vivo half-life of the synthetic peptides. Future applications of ADAC involve pandemic preparedness to viral genetic drift as well as neoepitope vaccination strategies where the bispecific antibody is an off-the-shelf product, and the peptide antigen is synthesized based on next-generation sequencing data mining., QC 20230308

Studying tissue composition and function in non-human primates (NHPs) is crucial to understand the nature of our own species. Here we present a large-scale cell transcriptomic atlas that encompasses over 1 million cells from 45 tissues of the adult NHP Macaca fascicularis. This dataset provides a vast annotated resource to study a species phylogenetically close to humans. To demonstrate the utility of the atlas, we have reconstructed the cell–cell interaction networks that drive Wnt signalling across the body, mapped the distribution of receptors and co-receptors for viruses causing human infectious diseases, and intersected our data with human genetic disease orthologues to establish potential clinical associations. Our M. fascicularis cell atlas constitutes an essential reference for future studies in humans and NHPs., QC 20230116

Osteoclast (OC) abnormalities represent osteoporosis's critical mechanism (OP). OCs undergo multiple processes that range from monocytic to functional. Different drugs target OCs at different developmental stages; however, almost no Suitable drug-targeted delivery systems exist. Therefore, we designed two dual-targeting nanoparticles to target OCs at different functional stages. Using the calcitonin gene-related peptide receptor (CGRPR), which OC precursors highly express, and specific TRAPpeptides screened in the bone resorption lacuna, where mature OCs function, respectively, two types of dual-targeted nanoparticles were constructed. Afterwards, nanoparticles were grafted with hyaluronic acid (HA), which specifically binds to CD44 on the surface of the OCs. In vivo and in vitro experiments show that both nanoparticles have noticeable targeting effects on OCs. This suggests that dual-targeting nanoparticles designed for different functional periods of OC can be well targeted to the corresponding OC, and further promote the more precise delivery of drugs used to treat OP., QC 20221214

The mammalian brain contains many specialized cells that develop from a thin sheet of neuroepithelial progenitor cells. Single-cell transcriptomics revealed hundreds of molecularly diverse cell types in the nervous system, but the lineage relationships between mature cell types and progenitor cells are not well understood. Here we show in vivo barcoding of early progenitors to simultaneously profile cell phenotypes and clonal relations in the mouse brain using single-cell and spatial transcriptomics. By reconstructing thousands of clones, we discovered fate-restricted progenitor cells in the mouse hippocampal neuroepithelium and show that microglia are derived from few primitive myeloid precursors that massively expand to generate widely dispersed progeny. We combined spatial transcriptomics with clonal barcoding and disentangled migration patterns of clonally related cells in densely labeled tissue sections. Our approach enables high-throughput dense reconstruction of cell phenotypes and clonal relations at the single-cell and tissue level in individual animals and provides an integrated approach for understanding tissue architecture., QC 20221109

The importance of lubrication between oral surfaces provided by the salivary film is most acutely apparent when it is disrupted, a prevalent consequence of salivary gland hypofunction experienced with aging, a symptom of certain diseases, or a side effect of some medical interventions. Sufferers report difficulty with speech and oral food processing and collectively is detrimental to quality of life. Polyethylene glycol (PEG) is widely employed as a successful biocompatible boundary lubricant in engineering and biomedical applications. It is hypothesized that the immobilization of PEG to biological materials such as oral epithelial cells and tissue can mimic the salivary film and provide durable relief from the symptoms of mucosal dryness. To do so, PEG is functionalized with a sugar binding lectin (wheat germ agglutinin) to enhance epithelial adhesion through lectin-sugar interactions. Retention and lubricity are characterized on an ex vivo oral tissue tribology rig. WGA-PEG coats and retains on mucin films, oral epithelial cells, and porcine tongue tissue, and offers sustained reduction in coefficient of friction (COF). WGA-PEG could be developed into a useful topical treatment for reducing oral friction and the perception of dry mouth., QC 20220615

Neurodevelopmental disorders of genetic origin delay the acquisition of normal abilities and cause disabling phenotypes. Nevertheless, spontaneous attenuation and even complete amelioration of symptoms in early childhood and adolescence can occur in many disorders, suggesting that brain circuits possess an intrinsic capacity to overcome the deficits arising from some germline mutations. We examined the molecular composition of almost a trillion excitatory synapses on a brain-wide scale between birth and adulthood in mice carrying a mutation in the homeobox transcription factor Pax6, a neurodevelopmental disorder model. Pax6 haploinsufficiency had no impact on total synapse number at any age. By contrast, the molecular composition of excitatory synapses, the postnatal expansion of synapse diversity and the acquisition of normal synaptome architecture were delayed in all brain regions, interfering with networks and electrophysiological simulations of cognitive functions. Specific excitatory synapse types and subtypes were affected in two key developmental age-windows. These phenotypes were reversed within 2-3 weeks of onset, restoring synapse diversity and synaptome architecture to the normal developmental trajectory. Synapse subtypes with rapid protein turnover mediated the synaptome remodeling. This brain-wide capacity for remodeling of synapse molecular composition to recover and maintain the developmental trajectory of synaptome architecture may help confer resilience to neurodevelopmental genetic disorders., QC 20230614

Dendritic spines are submicron, subcellular compartments whose shape is defined by actin filaments and associated proteins. Accurately mapping the cytoskeleton is a challenge, given the small size of its components. It remains unclear whether the actin-associated structures analyzed in dendritic spines of neurons in vitro apply to dendritic spines of intact, mature neurons in situ. Here, we combined advanced preparative methods with multitilt serial section electron microscopy (EM) tomography and computational analysis to reveal the full three-dimensional (3D) internal architecture of spines in the intact brains of male mice at nanometer resolution. We compared hippocampal (CA1) pyramidal cells and cerebellar Purkinje cells in terms of the length distribution and connectivity of filaments, their branching-angles and absolute orientations, and the elementary loops formed by the network. Despite differences in shape and size across spines and between spine heads and necks, the internal organization was remarkably similar in both neuron types and largely homogeneous throughout the spine volume. In the tortuous mesh of highly branched and interconnected filaments, branches exhibited no preferred orientation except in the immediate vicinity of the cell membrane. We found that new filaments preferentially split off from the convex side of a bending filament, consistent with the behavior of Arp2/3-mediated branching of actin under mechanical deformation. Based on the quantitative analysis, the spine cytoskeleton is likely subject to considerable mechanical force in situ., QC 20230613

In our overall goal to develop anti-Parkinson drugs, we designed novel diketopiperazines (DKP1-6) aiming to both reach the blood-brain barrier and counteract the oxidative stress related to Parkinson's Disease (PD). The anti-Parkinson properties of DKP 1–6 were evaluated using neurotoxin-treated PC12 cells, as in vitro model of PD, while their cytotoxicity and genotoxicity potentials were investigated in newborn rat cerebral cortex (RCC) and primary human whole blood (PHWB) cell cultures. The response against free radicals was evaluated by the total antioxidant capacity (TAC) assay. Comet assay was used to detect DNA damage while the content of 8-hydroxyl-2′-deoxyguanosine (8-OH-dG) was determined as a marker of oxidative DNA damage. PAMPA-BBB and Caco-2 assays were employed to evaluate the capability of DKP1-6 to cross the membranes. Stability studies were conducted in simulated gastric and intestinal fluids and human plasma. Results showed that DKP5-6 attenuate the MPP + -induced cell death on a nanomolar scale, but a remarkable effect was observed for DKP6 on Nrf2 activation that leads to the expression of genes involved in oxidative stress response thus increasing glutathione biosynthesis and ROS buffering. DKP5-6 resulted in no toxicity for RCC neurons and PHWB cells exposed to 10–500 nM concentrations during 24 h as determined by MTT and LDH assays and TAC levels were not altered in both cultured cell types. No significant difference in the induction of DNA damage was observed for DKP5-6. Both DKPs resulted stable in simulated gastric fluids (t1/2 > 22h). In simulated intestinal fluids, DKP5 underwent immediate hydrolysis while DKP6 showed a half-life higher than 3 h. In human plasma, DKP6 resulted quite stable. DKP6 displayed both high BBB and Caco-2 permeability confirming that the DKP scaffold represents a useful tool to improve the crossing of drugs through the biological membranes., QC 20230613

New variants in the SARS-CoV-2 pandemic are more contagious (Alpha/Delta), evade neutralizing antibodies (Beta), or both (Omicron). This poses a challenge in vaccine development according to WHO. We designed a more universal SARS-CoV-2 DNA vaccine containing receptor-binding domain loops from the huCoV-19/WH01, the Alpha, and the Beta variants, combined with the membrane and nucleoproteins. The vaccine induced spike antibodies crossreactive between huCoV-19/WH01, Beta, and Delta spike proteins that neutralized huCoV-19/WH01, Beta, Delta, and Omicron virus in vitro. The vaccine primed nucleoprotein-specific T cells, unlike spike-specific T cells, recognized Bat-CoV sequences. The vaccine protected mice carrying the human ACE2 receptor against lethal infection with the SARS-CoV-2 Beta variant. Interestingly, priming of cross-reactive nucleoprotein-specific T cells alone was 60% protective, verifying observations from humans that T cells protect against lethal disease. This SARS-CoV vaccine induces a uniquely broad and functional immunity that adds to currently used vaccines., QC 20230523

Rhipicephalus sanguineus sensu lato and Rhipicephalus turanicus are very prevalent in Israel and are known to be vectors of human and animal diseases. The aim of this study was to identify the pathogens found in questing ticks and such parasitizing domestic and wild animals. Ticks were collected from 16 localities in Israel with the flagging technique and by examining dogs, hedgehogs, a badger and a tortoise. Bacterial and protozoal pathogens were analyzed by PCR and sequencing. Overall, 374 R. sanguineus s.l. specimens were collected, out of which 142 by flagging and 132 from six dogs. Rickettsia africae, Rickettsia massiliae, Rickettsia conorii subsp. israelensis, and Anaplasma sp. were identified in ticks collected by flagging, Rickettsia aeschlimannii was found only in specimens collected from dogs, while Ehrlichia sp., Coxiella burnetii, Hepatozoon canis and Leishmania infantum were recorded in ticks collected by flagging and from dogs. Out of 226 specimens of R. turanicus, 124 were collected by flagging, while additional 33 from eight dogs, 64 from seven southern white-breasted hedgehogs (Erinaceus concolor), two from a European badger (Meles meles) and one from a Greek tortoise (Testudo graeca). Out of 65 R. sanguineus s.l. pools 17 (26.2%) had pathogens, while seven of them were positive for one pathogen, and 10 for two pathogens. In 43 R. turanicus pools, R. aeschlimannii R. africae, Rickettsia barbariae, R. massiliae, Anaplasma sp., Ehrlichia sp. and C. burnetii, as well as Babesia microti, B. vogeli, Hepatozoon felis, and L. infantum was detected, while Listeria monocytogenes, Bartonella sp. and Toxoplasma gondii were negative in all R. sanguineus s.l. and R. turanicus pools examined. In conclusion, Babesia microti is reported for the first time in Israel, R. africae, R. aeschlimannii, C. burnetii and L. infantum are reported for the first time in R. sanguineus s.l. and R. turanicus, while H. felis is reported for the first time from R. turanicus in the c

Rhipicephalus sanguineus sensu lato and Rhipicephalus turanicus are very prevalent in Israel and are known to be vectors of human and animal diseases. The aim of this study was to identify the pathogens found in questing ticks and such parasitizing domestic and wild animals. Ticks were collected from 16 localities in Israel with the flagging technique and by examining dogs, hedgehogs, a badger and a tortoise. Bacterial and protozoal pathogens were analyzed by PCR and sequencing. Overall, 374 R. sanguineus s.l. specimens were collected, out of which 142 by flagging and 132 from six dogs. Rickettsia africae, Rickettsia massiliae, Rickettsia conorii subsp. israelensis, and Anaplasma sp. were identified in ticks collected by flagging, Rickettsia aeschlimannii was found only in specimens collected from dogs, while Ehrlichia sp., Coxiella burnetii, Hepatozoon canis and Leishmania infantum were recorded in ticks collected by flagging and from dogs. Out of 226 specimens of R. turanicus, 124 were collected by flagging, while additional 33 from eight dogs, 64 from seven southern white-breasted hedgehogs (Erinaceus concolor), two from a European badger (Meles meles) and one from a Greek tortoise (Testudo graeca). Out of 65 R. sanguineus s.l. pools 17 (26.2%) had pathogens, while seven of them were positive for one pathogen, and 10 for two pathogens. In 43 R. turanicus pools, R. aeschlimannii R. africae, Rickettsia barbariae, R. massiliae, Anaplasma sp., Ehrlichia sp. and C. burnetii, as well as Babesia microti, B. vogeli, Hepatozoon felis, and L. infantum was detected, while Listeria monocytogenes, Bartonella sp. and Toxoplasma gondii were negative in all R. sanguineus s.l. and R. turanicus pools examined. In conclusion, Babesia microti is reported for the first time in Israel, R. africae, R. aeschlimannii, C. burnetii and L. infantum are reported for the first time in R. sanguineus s.l. and R. turanicus, while H. felis is reported for the first time from R. turanicus in the c

BACKGROUND: Myeloperoxidase ANCA-associated vasculitis is a major cause of ESKD. Efficacy of anti-CD20 mAb treatment was tested in a mouse model of the disease. METHOD(S): MPO immunization induced anti-MPO autoimmunity, and a subnephritogenic dose of sheep anti-mouse GBM globulin triggered GN. RESULT(S): Anti-CD20 mAb treatment increased the numbers and immunomodulatory capacity of MPO-specific T regulatory cells (Tregs) and attenuated T cell-mediated and humoral anti-MPO autoimmunity and GN. Disabling of Tregs negated the therapeutic benefit of anti-CD20 treatment. The mechanism of enhancement of Treg activity could be attributed to anti-CD20 mAb effects on inducing B cell apoptosis. Administering anti-CD20 mAb-induced apoptotic splenocytes to mice developing anti-MPO GN was as effective as anti-CD20 mAb treatment in inducing Tregs and attenuating both anti-MPO autoimmunity and GN. A nonredundant role for splenic macrophages in mediating the anti-CD20 mAb-induced immunomodulation was demonstrated by showing that administration of anti-CD20 mAb ex vivo-induced apoptotic splenocytes to unmanipulated mice attenuated autoimmunity and GN, whereas deletion of splenic marginal zone macrophages prevented anti-CD20 mAb-induced immunomodulation and treatment efficacy. Six days after administering anti-CD20 mAb to mice with murine anti-MPO GN, cell-mediated anti-MPO responses and GN were attenuated, and Tregs were enhanced, but ANCA levels were unchanged, suggesting humoral autoimmunity was redundant at this time point. CONCLUSION(S): Collectively, these data suggest that, as well as reducing humoral autoimmunity, anti-CD20 mAb more rapidly induces protective anti-MPO Treg-mediated immunomodulation by splenic processing of anti-CD20-induced apoptotic B cells.Copyright © 2021 by the American Society of Nephrology.

Background and Aim: Diagnosis and management of inflammatory bowel disease (IBD) requires a clinical approach, combining information from histology and other modalities for clinical decision making. However, histology is costly due to specimen processing, and intra- and interobservational variation and the significance of subtle findings remain unclear when translating into clinical practice. Biospectroscopy involves combining microscopy with spectroscopy and provides chemical and imaging identification for complex biological specimens, which, as an advantage, do not require any chemical processing techniques or staining. Briefly, infrared (IR) or laser energies vibrate biochemical bonds in biospecimens (fluids or tissue), which are then recorded as absorbance intensities in large hyperspectral datasets. Regression and classification algorithms used in multivariate analysis for machine learning are then applied for modeling and predictive capacity. To date, there has been little published research using this technology for assessing IBD. We aimed to undertake an explorative analysis of intestinal inflammation using Fourier-transform infrared microspectroscopy, develop a model for assessing severity of colitis, and identify potential spectral biomarkers for colitis. This study was a proof-of-concept study using a gold-standard murine colitis model to inform further research in human work. Method(s): We used formalin-fixed paraffin-embedded intestinal sections from C57/BL6 mice (30 mice, three sections per mouse) with chronic colitis induced by 3% oral dextran sodium sulfate (n = 27), of which a subset (n = 14) were treated with intravenous human amnion epithelial cells and showed a reduction in histological severity of inflammation. Tissue sections were placed on a reflective slide, and an adjacent 5 mum section was stained with hematoxylin and eosin for comparison. Background scans and regions of interest from the intestinal mucosa were imaged using the Agilent Cary

Cardiac injury and dysfunction occur in COVID-19 patients and increase the risk of mortality. Causes are ill defined but could be through direct cardiac infection and/or inflammation-induced dysfunction. To identify mechanisms and cardio-protective drugs, we use a state-of-the-art pipeline combining human cardiac organoids with phosphoproteomics and single nuclei RNA sequencing. We identify an inflammatory "cytokine-storm", a cocktail of interferon gamma, interleukin 1beta, and poly(I:C), induced diastolic dysfunction. Bromodomain-containing protein 4 is activated along with a viral response that is consistent in both human cardiac organoids (hCOs) and hearts of SARS-CoV-2-infected K18-hACE2 mice. Bromodomain and extraterminal family inhibitors (BETi) recover dysfunction in hCOs and completely prevent cardiac dysfunction and death in a mouse cytokine-storm model. Additionally, BETi decreases transcription of genes in the viral response, decreases ACE2 expression, and reduces SARS-CoV-2 infection of cardiomyocytes. Together, BETi, including the Food and Drug Administration (FDA) breakthrough designated drug, apabetalone, are promising candidates to prevent COVID-19 mediated cardiac damage.Copyright © 2021 Elsevier Inc.

Objective: To evaluate the efficacy of acellular collagen scaffold (ACS) in combination with basic fibroblast growth factor (bFGF) for the repair of traumatic tympanic membrane (TM) perforation in a rat model. Study Design: A prospective controlled animal study in a rat model of traumatic TM perforation. Setting(s): Tertiary medical center. Subjects and Methods: Sprague-Dawley rats (N = 84) with unilateral traumatic perforation of the right TMs were randomized to receive ACS, bFGF, ACS in combination with bFGF (ACS/bFGF), or nothing (spontaneous healing without any interventions as a control group). The healing outcomes were evaluated by otoscopy, optical coherence tomography, histology, and transmission electron microscopy at 1, 2, and 4 weeks postoperatively. The hearing outcomes were assessed with auditory brainstem response testing. Result(s): ACS/bFGF resulted in higher perforation closure rates at an earlier stage than spontaneous healing, ACS, and bFGF. Based on histology, optical coherence tomography, and transmission electron microscopy, a trilaminar structure and uniform thickness with mature, densely packed collagen fibers were seen in the ACS/bFGF group. Auditory brainstem response evaluation also showed that ACS/bFGF treatment promoted faster functional hearing recovery as compared with the control group. Conclusion(s): ACS is an effective TM scaffold and a carrier for bFGF. ACS/bFGF improves the TM closure rate, results in better-reconstructed TMs, and improves hearing. ACS/bFGF serves as a potential substitute for TM perforations in clinical settings.Copyright © American Academy of Otolaryngology-Head and Neck Surgery Foundation 2020.

Oxaliplatin is a platinum-based alkylating chemotherapeutic agent used for cancer treatment. Neurotoxicity is one of its major adverse effects that often demands dose limitation. However, the effects of chronic oxaliplatin on the toxicity of the autonomic nervous system regulating cardiorespiratory function and adaptive reflexes are unknown. Male Sprague Dawley rats were treated with intraperitoneal oxaliplatin (3 mg kg-1 per dose) 3 times a week for 14 days. The effects of chronic oxaliplatin treatment on baseline mean arterial pressure (MAP); heart rate (HR); splanchnic sympathetic nerve activity (sSNA); phrenic nerve activity (PNA) and its amplitude (PNamp) and frequency (PNf); and sympathetic reflexes were investigated in anaesthetised, vagotomised and artificially ventilated rats. The same parameters were evaluated after acute oxaliplatin injection, and in the chronic treatment group following a single dose of oxaliplatin. The amount of platinum in the brain was determined with atomic absorption spectrophotometry. Chronic oxaliplatin treatment significantly increased MAP, sSNA and PNf and decreased HR and PNamp, while acute oxaliplatin had no effects. Platinum was accumulated in the brain after chronic oxaliplatin treatment. In the chronic oxaliplatin treatment group, further administration of a single dose of oxaliplatin increased MAP and sSNA. The baroreceptor sensitivity and somatosympathetic reflex were attenuated at rest while the sympathoexcitatory response to hypercapnia was increased in the chronic treatment group. This is the first study to reveal oxaliplatin-induced alterations in the central regulation of cardiovascular and respiratory functions as well as reflexes that may lead to hypertension and breathing disorders which may be mediated via accumulated platinum in the brain.Copyright © 2020, Springer-Verlag GmbH Germany, part of Springer Nature.

Aim: To investigate the effect of latent cytomegalovirus (CMV) infection on experimental anti-myeloperoxidase (MPO) autoimmunity and glomerulonephritis (GN). Background(s): Autoimmunity to MPO results in anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) and GN. CMV is present, lifelong, in a latent state after primary infection. Latent CMV infection causes increased peripheral blood CD28- T cells that contribute to mortality in AAV, but this is mechanistically unclear. Method(s): Mice were infected with mouse CMV (K181delm157). Seven weeks later, when viral latency was established (by organ CMV plaque assay, including kidneys), mice were immunised with MPO and autoimmunity assessed 10- and 28-days after immunization. Anti-MPO GN was induced by immunising mice with MPO, GN was triggered by low dose anti-mouse basement membrane globulin, and disease assessed 4 days post trigger (day 21). Result(s): Latent CMV infection induced heightened anti-MPO autoimmunity with increased lymph node MPO-stimulated IL-17A and IFNgamma production (ELISPOT day 10, sustained at day 21 in anti-MPO GN). Latent CMV infection did not significantly increase MPO-ANCA titres, but autoreactive B cell numbers were increased. In mice with GN, latent infection resulted in increased intrarenal macrophages, CD4 + and CD8+ T cells. Intrarenal CMVspecific CD8+ T cells were detected in CMV infected mice with GN by MHCIpeptide tetramer staining. CD28-CD4 + and CD8 + populations were expanded, while intrarenal CD4+ and CD8+ T cells in CMV infected mice with GN exhibited heightened TNF and IFNgamma production as well as increased cytotoxic potential, measured by surface CD107a expression. CMV infected mice with GN developed increased proteinuria, though short-term histological glomerular injury was similar. Conclusion(s): Experimentally, latent CMV infection results in heightened anti-MPO autoimmunity and renal inflammation; and may directly influence the course of MPO-AAV in humans.

Purpose: Pancreatic ductal adenocarcinoma (PDAC) has the lowest five-year survival rate of all cancers in the United States. Programmed death 1 receptor (PD-1)-programmed death ligand 1 (PD-L1) immune checkpoint inhibition has been unsuccessful in clinical trials. Myeloid-derived suppressor cells (MDSCs) are known to block anti-tumor CD8+ T cell immune responses in various cancers including pancreas. This has led us to our objective that was to develop a clinically relevant in vitro organoid model to specifically target mechanisms that deplete MDSCs as a therapeutic strategy for PDAC. Method(s): Murine and human pancreatic ductal adenocarcinoma (PDAC) autologous organoid/immune cell co-cultures were used to test whether PDAC can be effectively treated with combinatorial therapy involving PD-1 inhibition and MDSC depletion. Result(s): Murine in vivo orthotopic and in vitro organoid/immune cell co-culture models demonstrated that polymorphonuclear (PMN)-MDSCs promoted tumor growth and suppressed cytotoxic T lymphocyte (CTL) proliferation, leading to diminished efficacy of checkpoint inhibition. Mouse-and human-derived organoid/immune cell co-cultures revealed that PD-L1-expressing organoids were unresponsive to nivolumab in vitro in the presence of PMN-MDSCs. Depletion of arginase 1-expressing PMN-MDSCs within these co-cultures rendered the organoids susceptible to anti-PD-1/PD-L1-induced cancer cell death. Conclusion(s): Here we use mouse-and human-derived autologous pancreatic cancer organoid/immune cell co-cultures to demonstrate that elevated infiltration of polymorphonuclear (PMN)-MDSCs within the PDAC tumor microenvironment inhibit T cell effector function, regardless of PD-1/PD-L1 inhibition. We present a pre-clinical model that may predict the efficacy of targeted therapies to improve the outcome of patients with this aggressive and otherwise unpredictable malignancy.Copyright © 2020 by the authors. Licensee MDPI, Basel, Switzerland.

Aim: To assess whether ASK1 blockade inhibits the progression of experimental diabetic kidney disease by reducing stress-induced premature senescence (SIPS) and its consequences. Background(s): The development of SIPS in kidney tubular epithelial cells (TEC) results in the release of stress-associated secretory proteins (SASPs), including proinflammatory and profibrotic factors which promote progression of kidney diseases. Activation of p38 MAPK is a key mechanism promoting SIPS. Therefore, we hypothesized that blockade of ASK1-p38 MAPK signalling would inhibit kidney diseases by reducing SIPS and SASPs. Method(s): Development of SIPS and production of SASPs were assessed in archived tissue from a model of progressive fibrotic kidney disease- streptozotocin (STZ)-induced diabetic kidney disease (DKD) in hypertensive endothelial nitric oxide synthase (Nos3) deficient mice. Mice with DKD were treated with an ASK1 inhibitor (GS-444217) as either an early intervention (2-8 weeks after STZ) or a late intervention (8-15 weeks after STZ). Result(s): STZ-induced diabetes resulted in the development of senescent TEC in Nos3-/- mice. Treatment with GS-444217 blocked activation of the p38 MAPK pathway without affecting hyperglycaemia. This was associated with reduced levels of cellular senescence (Cdkn2A) and SASPs (CCL2, TNF-alpha, TGF-beta1), which correlated with diminished levels of kidney fibrosis and renal function impairment. Therapeutic blockade of ASK1 was effective in reducing SIPS as both an early intervention (Cdkn2A #51%; P = 0.0005) and as a late intervention (Cdkn2A #45%; P = 0.02) in the DKD model. Conclusion(s): Our data show that ASK1 inhibition reduces SIPS and SASPs independent of the underlying diabetic state. This may contribute to the overall benefit seen with ASK1 blockade in DKD. Further studies are warranted to explore this mechanism.

Systemic lupus erythematosus (lupus) is an autoimmune disease characterized by autoantibodies that form immune complexes with self-antigens, which deposit in various tissues, leading to inflammation and disease. The etiology of disease is complex and still not completely elucidated. Dysregulated inflammation is an important disease feature, and the mainstay of lupus treatment still utilizes nonspecific anti-inflammatory drugs. Granulocyte colony-stimulating factor (G-CSF) is a growth, survival, and activation factor for neutrophils and a mobilizer of hematopoietic stem cells, both of which underlie inflammatory responses in lupus. To determine whether G-CSF has a causal role in lupus, we genetically deleted G-CSF from Lyn-deficient mice, an experimental model of lupus nephritis. Lyn-/-G-CSF-/- mice displayed many of the inflammatory features of Lyn-deficient mice; however, they had reduced bone marrow and tissue neutrophils, consistent with G-CSF's role in neutrophil development. Unexpectedly, in comparison to aged Lyn-deficient mice, matched Lyn-/-G-CSF-/- mice maintained neutrophil hyperactivation and exhibited exacerbated numbers of effector memory T cells, augmented autoantibody titers, and worsened lupus nephritis. In humans, serum G-CSF levels were not elevated in patients with lupus or with active renal disease. Thus, these studies suggest that G-CSF is not pathogenic in lupus, and therefore G-CSF blockade is an unsuitable therapeutic avenue.Copyright © 2021 The Authors. Immunity, Inflammation and Disease published by John Wiley & Sons Ltd

Aim: To investigate IL-7Ralpha signalling in experimental anti-myeloperoxidase (MPO) glomerulonephritis (GN) by inhibiting IL-7Ralpha using a neutralising, non-depleting monoclonal antibody. Background(s): Microarray profiling of T cells from patients with anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) has demonstrated that signalling through the memory T cell receptor IL-7Ralpha predicts relapse and overall severity, but whether IL-7Ralpha has any direct functional role is unknown. Method(s): Anti-MPO GN was induced by immunising mice with MPO, then GN triggered with low dose anti-mouse basement membrane globulin 4 days before the end of experiments. Autoimmunity was either acute (immunizations days 0, 7; ending day 21) or chronic (immunizations days 0, 7, 42; ending day 56). Anti-IL-7Ralpha (or control) antibodies were administered (ip) after the onset of autoimmunity, one day before GN was triggered. In a further experiment in the chronic model, anti-IL7Ralpha treatment commenced on day 35. Treatment was administered every second day. Result(s): Control ovalbumin-immunized mice given anti-basement membrane globulin developed minimal injury. Compared with mice with anti- MPO GN given control antibodies, anti-IL-7Ralpha antibodies administered after the induction of anti-MPO autoimmunity (day 17) attenuated GN at day 21 with reduced albuminuria, glomerular segmental necrosis, tubulointerstitial injury, glomerular and interstitial leukocytes and intrarenal chemokines, with no corresponding increase in intrarenal PD-1 expression. Anti-IL-7Ralpha therapy in the 56-day model dampened systemic proinflammatory immune responses and renal leukocyte infiltration, without affecting albuminuria or glomerular histology. Prophylactic therapy prior to the induction of disease in the chronic model resulted in a similar outcome. Conclusion(s): These findings mechanistically validate a role for the IL-7Ralpha in AAV and suggest that neutralizing anti-IL-7Ra

The impact of treatment on the risk of lymphoma in patients with rheumatoid arthritis (RA) is unclear. Here, we aimed to assess if the risk of lymphoma differs according to the type of Tumor Necrosis factor inhibitor (TNFi), comparing monoclonal anti-TNF antibodies (Ab) to the soluble TNF receptor. We used BAFF-transgenic (Tg) mice as a model of autoimmunity-associated lymphoma. Six-month aged BAFF-Tg mice were treated with TNFi for 12 months. Histological examination of the spleen, assessment of the cellular composition of the spleen by flow cytometry and assessment of B cell clonality were performed at sacrifice. Crude mortality and incidence of lymphoma were significantly higher in mice treated with monoclonal anti-TNF Ab compared to both controls and mice treated with the soluble TNF receptor, even at high dose. Flow cytometry analysis revealed decreased splenic macrophage infiltration in mice treated with monoclonal anti-TNF Ab. Overall, this study demonstrates, for the first time, that a very prolonged treatment with monoclonal anti-TNF Ab increase the risk of lymphoma in B cell-driven autoimmunity. This data suggests a closer monitoring for lymphoma development in patients suffering from B cell-driven autoimmune disease with long-term exposure to monoclonal anti-TNF Ab.Copyright This article is protected by copyright. All rights reserved.

Aim: To investigate whether recombinant follistatin can suppress renal interstitial fibrosis in a mouse model of folic acid nephropathy (FAN). Background(s): Renal interstitial fibrosis is a common pathologic feature in progressive kidney disease, but we lack effective anti-fibrotic therapies. Activin A, a member of the transforming growth factor beta superfamily of cytokines, is upregulated in animal models of renal fibrosis including unilateral ureter obstruction and diabetic kidney disease. Follistatin is a secreted protein that binds to and inhibits the function of Activin A. We evaluated the therapeutic potential of recombinant follistatin (FST288) in the FAN model. Method(s): Male C57BL/6 mice (n = 8/9 group) were administered 250 mg/ kg folic acid. Urine was collected on day 3. Mice were treated once daily with 2 mg/kg FST288 (Paranta Biosciences) or vehicle by intraperitoneal injection from day 6 until being killed on day 14. Normal mice were controls. Renal function, inflammation and fibrosis were assayed. Result(s): Vehicle-treated animals developed renal function impairment (12.1 +/- 2.4 vs 7.2 +/- 1.7 mumol/L plasma creatinine in normals; P < 0.01), with focal areas of the cortex exhibiting tubular damage, macrophage and myofibroblast infiltration and collagen IV deposition. RT-PCR analysis of kidney tissue showed increased levels of Collagen I, CD68, alpha-SMA and Activin A mRNA. Treatment with FST288 did not affect renal dysfunction (10.5 +/- 1.8 mumol/L plasma creatinine; P=NS). Similarly, FST288 treatment did not affect histologic damage or reduce mRNA levels of collagen I, CD68, alpha-SMA or ActA (all P=NS). The day 3 urine KIM-1 to creatinine ratio was not different between FST288 and vehicle treated groups, confirming an equivalent induction of acute kidney injury. Conclusion(s): FST288 treatment did not provide protection against renal interstitial inflammation or fibrosis in the FAN model.

Cyclophilins have important homeostatic roles, but following tissue injury, cyclophilin A (CypA) can promote leukocyte recruitment and inflammation, while CypD can facilitate mitochon-drial-dependent cell death. This study investigated the therapeutic potential of a selective cyclo-philin inhibitor (GS-642362), which does not block calcineurin function, in mouse models of tubular cell necrosis and renal fibrosis. Mice underwent bilateral renal ischemia/reperfusion injury (IRI) and were killed 24 h later: treatment with 10 or 30 mg/kg/BID GS-642362 (or vehicle) began 1 h before surgery. In the second model, mice underwent unilateral ureteric obstruction (UUO) surgery and were killed 7 days later; treatment with 10 or 30 mg/kg/BID GS-642362 (or vehicle) began 1 h before surgery. GS-642362 treatment gave a profound and dose-dependent protection from acute renal failure in the IRI model. This protection was associated with reduced tubular cell death, including a dramatic reduction in neutrophil infiltration. In the UUO model, GS-642362 treatment signifi-cantly reduced tubular cell death, macrophage infiltration, and renal fibrosis. This protective effect was independent of the upregulation of IL-2 and activation of the stress-activated protein kinases (p38 and JNK). In conclusion, GS-642362 was effective in suppressing both acute kidney injury and renal fibrosis. These findings support further investigation of cyclophilin blockade in other types of acute and chronic kidney disease.Copyright © 2020 by the authors.

STUDY QUESTION: How is endometrial epithelial receptivity, particularly adhesiveness, regulated at the luminal epithelial surface for embryo implantation in the human? SUMMARY ANSWER: Podocalyxin (PCX), a transmembrane protein, was identified as a key negative regulator of endometrial epithelial receptivity; specific downregulation of PCX in the luminal epithelium in the mid-secretory phase, likely mediated by progesterone, may act as a critical step in converting endometrial surface from a non-receptive to an implantation-permitting state. WHAT IS KNOWN ALREADY: The human endometrium must undergo major molecular and cellular changes to transform from a non-receptive to a receptive state to accommodate embryo implantation. However, the fundamental mechanisms governing receptivity, particularly at the luminal surface where the embryo first interacts with, are not well understood. A widely held view is that upregulation of adhesion-promoting molecules is important, but the details are not well characterized. STUDY DESIGN, SIZE, DURATION: This study first aimed to identify novel adhesion-related membrane proteins with potential roles in receptivity in primary human endometrial epithelial cells (HEECs). Further experiments were then conducted to determine candidates' in vivo expression pattern in the human endometrium across the menstrual cycle, regulation by progesterone using cell culture, and functional importance in receptivity using in vitro human embryo attachment and invasion models. PARTICIPANTS/MATERIALS, SETTING, METHODS: Primary HEECs (n=9) were isolated from the proliferative phase endometrial tissue, combined into three pools, subjected to plasma membrane protein enrichment by ultracentrifugation followed by proteomics analysis, which led to the discovery of PCX as a novel candidate of interest. Immunohistochemical analysis determined the in vivo expression pattern and cellular localization of PCX in the human endometrium across the menstrual cycle (n=23).

Purpose: Oocyte quality and reproductive outcome are negatively affected by advanced maternal age, ovarian stimulation and method of oocyte maturation during assisted reproduction; however, the mechanisms responsible for these associations are not fully understood. The aim of this study was to compare the effects of ageing, ovarian stimulation and in-vitro maturation on the relative levels of transcript abundance of genes associated with DNA repair during the transition of germinal vesicle (GV) to metaphase II (MII) stages of oocyte development. Method(s): The relative levels of transcript abundance of 90 DNA repair-associated genes was compared in GV-stage and MII-stage oocytes from unstimulated and hormone-stimulated ovaries from young (5-8-week-old) and old (42-45-week-old) C57BL6 mice. Ovarian stimulation was conducted using pregnant mare serum gonadotropin (PMSG) or anti-inhibin serum (AIS). DNA damage response was quantified by immunolabeling of the phosphorylated histone variant H2AX (gammaH2AX). Result(s): The relative transcript abundance in DNA repair genes was significantly lower in MII oocytes compared to GV oocytes in young unstimulated and PMSG stimulated but was higher in AIS-stimulated mice. Interestingly, an increase in the relative level of transcript abundance of DNA repair genes was observed in MII oocytes from older mice in unstimulated, PMSG-stimulated and AIS-stimulated mice. Decreased gammaH2AX levels were found in both GV oocytes (82.9%) and MII oocytes (37.5%) during ageing in both ovarian stimulation types used (PMSG/AIS; p < 0.05). Conclusion(s): In conclusion, DNA repair relative levels of transcript abundance are altered by maternal age and the method of ovarian stimulation during the GV-MII transition in oocytes.Copyright © 2020, Springer Science+Business Media, LLC, part of Springer Nature.

Aim: To develop a simplified model of renal ischaemia/reperfusion (I/R) injury. Background(s): The transition of acute kidney injury (AKI) to chronic kidney disease is recognized as a major clinical problem. However, a limitation of current animal models of renal I/R injury is the requirement for near fatal injury for the subsequent development of renal fibrosis. Therefore, we investigated a modified gradient technique of I/R injury to facilitate the development of renal fibrosis while maintaining overall normal kidney function. Method(s): Groups of 6-8 male C57BL/6 mice were anaesthetized with sodium pentobarbital, the left kidney removed, and then a silk thread run under the lower renal branch and the right kidney raised under tension of a 20 g weight using a pulley system for 30, 40 or 60 min. Then the kidney was lowered, the wound sutured and intraperitoneal saline given. Groups of mice were killed after 1, 3, 7 or 28 days of reperfusion. Result(s): In the raised kidney, the lower pole showed complete cessation of blood flow while the upper pole showed an incomplete cessation. An ischaemic period of 60 min, but not 30 or 40 min, induced significant renal impairment (187.0 +/- 65.3 vs 17.9 +/- 4.8 mumol/L serum creatinine in normal; P < 0.001). Tubular necrosis was more severe in the lower pole than the upper pole. Renal function returned to normal on day 3 and remained normal to day 28. On days 3 and 7, the lower pole showed more severe tubular damage compared to the upper pole, and on day 28 the lower pole showed substantial fibrosis whereas the upper pole was relatively normal. Conclusion(s): We have developed a model of clinically 'silent' renal fibrosis.

Introduction/Aim. A major cause of mortality and morbidity in COPD are acute exacerbations (AECOPD), most frequently triggered by infections, such as influenza A virus (IAV). However, the identity of key factors of the immune system which contribute to AECOPD remain ill-defined. Innate immune responses to pathogens depend on the activation of inflammasomes, which facilitate the maturation and release of proinflammatory cytokines such as IL-1beta. Recently, clinical studies and human in vitro data have demonstrated excessive activation of inflammasomes in AECOPD. The aim of this study was to investigate a novel functional link between inflammasome activation and IAV infection in the pathogenesis of AECOPD. Methods. We employed a murine COPD model with acute IAV infection. Specifically, 6-week-old mice were exposed to cigarette smoke (CS) for 8 weeks. After 8 weeks, mice were infected with IAV and culled 5 days post infection. We also used COPD patient serum during stable disease, with their own corresponding IAV-induced acute exacerbation. Results. We discovered excessive productions of genes and proteins encoding AIM2 which forms a cytoplasmic DNA-sensing inflammasome, and IL-1beta in a murine COPD model with acute IAV infection. Nevertheless, the first-line therapy for AECOPD is glucocorticosteroids, which is limited in its efficacy. In that respect, Pirfenidone (PFD) is clinically used anti-inflammatory therapeutic. Importantly, our compelling data supported that PFD can alleviate the mRNA expressions of Aim2, and Il1beta in COPD model following acute IAV infection. We also discovered an elevated IL-1beta protein levels in serum of AECOPD patients compared to their stable status. Conclusion. Collectively, we define for the first time that hyperactivation of AIM2/IL-1beta axis plays a role in AECOPD. Finally, our data inform the future clinical utility (repurposing) of PFD to target inflammasomes in AECOPD and this would represent an enormous change in the therapeu

In recent years, many light-microscopy protocols have been published for visualization of nanoscale structures in the kidney. These protocols present researchers with new tools to evaluate both foot process anatomy and effacement, as well as protein distributions in foot processes, the slit diaphragm and in the glomerular basement membrane. However, these protocols either involve the application of different complicated super resolution microscopes or lengthy sample preparation protocols. Here, we present a fast and simple, five-hour long procedure for three-dimensional visualization of kidney morphology on all length scales. The protocol combines optical clearing and tissue expansion concepts to produce a mild swelling, sufficient for resolving nanoscale structures using a conventional confocal microscope. We show that the protocol can be applied to visualize a wide variety of pathologic features in both mouse and human kidneys. Thus, our fast and simple protocol can be beneficial for conventional microscopic evaluation of kidney tissue integrity both in research and possibly in future clinical routines., QC 20211215

The application of histological methods in meat composition analysis and identification of prohibited tissues and organs added to meat and meat products is still in the research phase, although there have been some promising results. The aim of this work was to assess the possibility of using histological and histochemical methods for analysis of kulen composition. In this research, six samples of kulen were examined, one of which was produced in domestic conditions, while the rest were commercial products sampled from local markets. The samples were carried through classical histological preparation. The obtained slides were stained with haematoxylin/eosin, Masson-Goldner, toluidine blue and periodic acid-Schiff/alcian blue. The content of muscle, fat and connective tissue was evaluated using histomorphometric analysis. Histological analysis of kulen composition determined the presence of the following structures: muscle, adipose and connective tissues, blood vessels, glandular epithelium, peripheral nerve, cartilage and plant tissue. The histomorphometric analysis showed that the kulen products contained on average 56±2.52% muscle tissue, 7.27±1.38% connective tissue and 19.82±3.24% adipose tissue. The results show that by applying histological methods it is possible to identify different permitted and prohibited animal tissues in kulen, and hence, it is possible to analyse the composition of kulen. It is also possible to confirm the presence of various plant tissues, but for their precise identification, additional histological methods are needed., Primena histoloških metoda u analizi sastava mesa i utvrđivanju prisustva nedozvoljenih tkiva u mesu i proizvodima od mesa je još uvek u fazi istraživanja, iako su postojeći rezultati u ovoj oblasti obeć avajuć i. Cilj našeg istraživanja bio je ispitivanje moguć nosti korišć enja histoloških i histohemijskih metoda u analizi sastava fermentisane kobasice u tipu kulena. U ovom istraživanju ispitano je šest uzoraka kulena, od kojih je jedan proizveden u domać im uslovima, dok su ostali komercijalni proizvodi poticali iz lokalnih marketa. Uzorci su sprovedeni klasičnom histološkom procedurom. Dobijeni isečci bojeni su metodama: hematoksilin/ eosin,Masson-Goldner, toluidin plavo i perjodna kiselina/Schiff/alcijan plavo. Sadržaj mišić nog, masnog i vezivnog tkiva procenjen je primenom histomorfometrijskih analiza. Histološkom analizom sastava kulena utvrđeno je prisustvo sledeć ih struktura: mišić no, masno i vezivno tkivo, krvni sudovi, epitel žlezdanog tipa, periferni nerv, hrskavica i biljno tkivo. Histomorfometrijska analiza je pokazala da proizvodi sadrže u proseku 56±2,52% mišić nog tkiva, 7,27±1,38% vezivnog tkiva i 19,82±3,24% masnog tkiva. Rezultati pokazuju da je primenom histoloških metoda moguć e identifikovati različita dozvoljena i nedozvoljena životinjska tkiva u kulenu, pa je samim tim moguće analizirati sastav kulena. Takođe je moguć e potvrditi prisustvo različitih biljnih tkiva, ali je za njihovu preciznu identifikaciju neophodno primeniti dodatne metode.

Aim: To investigate the effect of latent cytomegalovirus (CMV) infection on experimental anti-myeloperoxidase (MPO) autoimmunity and glomerulonephritis (GN). Background(s): Autoimmunity to MPO results in anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) and GN. CMV is present, lifelong, in a latent state after primary infection. Latent CMV infection causes increased peripheral blood CD28- T cells that contribute to mortality in AAV, but this is mechanistically unclear. Method(s): Mice were infected with mouse CMV (K181delm157). Seven weeks later, when viral latency was established (by organ CMV plaque assay, including kidneys), mice were immunised with MPO and autoimmunity assessed 10- and 28-days after immunization. Anti-MPO GN was induced by immunising mice with MPO, GN was triggered by low dose anti-mouse basement membrane globulin, and disease assessed 4 days post trigger (day 21). Result(s): Latent CMV infection induced heightened anti-MPO autoimmunity with increased lymph node MPO-stimulated IL-17A and IFNgamma production (ELISPOT day 10, sustained at day 21 in anti-MPO GN). Latent CMV infection did not significantly increase MPO-ANCA titres, but autoreactive B cell numbers were increased. In mice with GN, latent infection resulted in increased intrarenal macrophages, CD4 + and CD8+ T cells. Intrarenal CMVspecific CD8+ T cells were detected in CMV infected mice with GN by MHCIpeptide tetramer staining. CD28-CD4 + and CD8 + populations were expanded, while intrarenal CD4+ and CD8+ T cells in CMV infected mice with GN exhibited heightened TNF and IFNgamma production as well as increased cytotoxic potential, measured by surface CD107a expression. CMV infected mice with GN developed increased proteinuria, though short-term histological glomerular injury was similar. Conclusion(s): Experimentally, latent CMV infection results in heightened anti-MPO autoimmunity and renal inflammation; and may directly influence the course of MPO-AAV in humans.

Aim: To investigate whether recombinant follistatin can suppress renal interstitial fibrosis in a mouse model of folic acid nephropathy (FAN). Background(s): Renal interstitial fibrosis is a common pathologic feature in progressive kidney disease, but we lack effective anti-fibrotic therapies. Activin A, a member of the transforming growth factor beta superfamily of cytokines, is upregulated in animal models of renal fibrosis including unilateral ureter obstruction and diabetic kidney disease. Follistatin is a secreted protein that binds to and inhibits the function of Activin A. We evaluated the therapeutic potential of recombinant follistatin (FST288) in the FAN model. Method(s): Male C57BL/6 mice (n = 8/9 group) were administered 250 mg/ kg folic acid. Urine was collected on day 3. Mice were treated once daily with 2 mg/kg FST288 (Paranta Biosciences) or vehicle by intraperitoneal injection from day 6 until being killed on day 14. Normal mice were controls. Renal function, inflammation and fibrosis were assayed. Result(s): Vehicle-treated animals developed renal function impairment (12.1 +/- 2.4 vs 7.2 +/- 1.7 mumol/L plasma creatinine in normals; P < 0.01), with focal areas of the cortex exhibiting tubular damage, macrophage and myofibroblast infiltration and collagen IV deposition. RT-PCR analysis of kidney tissue showed increased levels of Collagen I, CD68, alpha-SMA and Activin A mRNA. Treatment with FST288 did not affect renal dysfunction (10.5 +/- 1.8 mumol/L plasma creatinine; P=NS). Similarly, FST288 treatment did not affect histologic damage or reduce mRNA levels of collagen I, CD68, alpha-SMA or ActA (all P=NS). The day 3 urine KIM-1 to creatinine ratio was not different between FST288 and vehicle treated groups, confirming an equivalent induction of acute kidney injury. Conclusion(s): FST288 treatment did not provide protection against renal interstitial inflammation or fibrosis in the FAN model.

BACKGROUND: Myeloperoxidase ANCA-associated vasculitis is a major cause of ESKD. Efficacy of anti-CD20 mAb treatment was tested in a mouse model of the disease. METHOD(S): MPO immunization induced anti-MPO autoimmunity, and a subnephritogenic dose of sheep anti-mouse GBM globulin triggered GN. RESULT(S): Anti-CD20 mAb treatment increased the numbers and immunomodulatory capacity of MPO-specific T regulatory cells (Tregs) and attenuated T cell-mediated and humoral anti-MPO autoimmunity and GN. Disabling of Tregs negated the therapeutic benefit of anti-CD20 treatment. The mechanism of enhancement of Treg activity could be attributed to anti-CD20 mAb effects on inducing B cell apoptosis. Administering anti-CD20 mAb-induced apoptotic splenocytes to mice developing anti-MPO GN was as effective as anti-CD20 mAb treatment in inducing Tregs and attenuating both anti-MPO autoimmunity and GN. A nonredundant role for splenic macrophages in mediating the anti-CD20 mAb-induced immunomodulation was demonstrated by showing that administration of anti-CD20 mAb ex vivo-induced apoptotic splenocytes to unmanipulated mice attenuated autoimmunity and GN, whereas deletion of splenic marginal zone macrophages prevented anti-CD20 mAb-induced immunomodulation and treatment efficacy. Six days after administering anti-CD20 mAb to mice with murine anti-MPO GN, cell-mediated anti-MPO responses and GN were attenuated, and Tregs were enhanced, but ANCA levels were unchanged, suggesting humoral autoimmunity was redundant at this time point. CONCLUSION(S): Collectively, these data suggest that, as well as reducing humoral autoimmunity, anti-CD20 mAb more rapidly induces protective anti-MPO Treg-mediated immunomodulation by splenic processing of anti-CD20-induced apoptotic B cells.Copyright © 2021 by the American Society of Nephrology.

Human amnion epithelial cells (hAECs) exert potent antifibrotic and anti-inflammatory effects when transplanted into preclinical models of tissue fibrosis. These effects are mediated in part via the secretion of soluble factors by hAECs which modulate signaling pathways and affect cell types involved in inflammation and fibrosis. Based on these reports, we hypothesized that these soluble factors may also support liver regeneration during chronic liver injury. To test this, we characterized the effect of both hAECs and hAEC-conditioned medium (CM) on liver repair in a mouse model of carbon tetrachloride (CCl4)-induced fibrosis. Liver repair was assessed by liver fibrosis, hepatocyte proliferation, and the liver progenitor cell (LPC) response. We found that the administration of hAECs or hAEC-CM reduced liver injury and fibrosis, sustained hepatocyte proliferation, and reduced LPC numbers during chronic liver injury. Additionally, we undertook in vitro studies to document both the cell-cell and paracrine-mediated effects of hAECs on LPCs by investigating the effects of co-culturing the LPCs and hAECs and hAEC-CM on LPCs. We found little change in LPCs co-cultured with hAECs. In contrast, hAEC-CM enhances LPC proliferation and differentiation. These findings suggest that paracrine factors secreted by hAECs enhance liver repair by reducing fibrosis while promoting regeneration during chronic liver injury.Copyright © The Author(s) 2020.

Amyotrophic lateral sclerosis (ALS) results in progressive impairment of upper and lower motor neurons. Increasing evidence from both in vivo and ex vivo studies suggest that iron accumulation in the motor cortex is a neuropathological hallmark in ALS. An in vivo neuroimaging marker of iron dysregulation in ALS would be useful in disease diagnosis and prognosis. Magnetic resonance imaging (MRI), with its unique capability to generate a variety of soft tissue contrasts, provides opportunities to image iron distribution in the human brain with millimeter to sub-millimeter anatomical resolution. Conventionally, MRI T1-weighted, T2-weighted, and T2*-weighted images have been used to investigate iron dysregulation in the brain in vivo. Susceptibility weighted imaging has enhanced contrast for para-magnetic materials that provides superior sensitivity to iron in vivo. Recently, the development of quantitative susceptibility mapping (QSM) has realized the possibility of using quantitative assessments of magnetic susceptibility measures in brain tissues as a surrogate measurement of in vivo brain iron. In this review, we provide an overview of MRI techniques that have been used to investigate iron dysregulation in ALS in vivo. The potential uses, strengths, and limitations of these techniques in clinical trials, disease diagnosis, and prognosis are presented and discussed. We recommend further longitudinal studies with appropriate cohort characterization to validate the efficacy of these techniques. We conclude that quantitative iron assessment using recent advances in MRI including QSM holds great potential to be a sensitive diagnostic and prognostic marker in ALS. The use of multimodal neuroimaging markers in combination with iron imaging may also offer improved sensitivity in ALS diagnosis and prognosis that could make a major contribution to clinical care and treatment trials. Level of Evidence: 2. Technical Efficacy: Stage 3.Copyright © 2021 International Society for M

Cardiac injury and dysfunction occur in COVID-19 patients and increase the risk of mortality. Causes are ill defined but could be through direct cardiac infection and/or inflammation-induced dysfunction. To identify mechanisms and cardio-protective drugs, we use a state-of-the-art pipeline combining human cardiac organoids with phosphoproteomics and single nuclei RNA sequencing. We identify an inflammatory "cytokine-storm", a cocktail of interferon gamma, interleukin 1beta, and poly(I:C), induced diastolic dysfunction. Bromodomain-containing protein 4 is activated along with a viral response that is consistent in both human cardiac organoids (hCOs) and hearts of SARS-CoV-2-infected K18-hACE2 mice. Bromodomain and extraterminal family inhibitors (BETi) recover dysfunction in hCOs and completely prevent cardiac dysfunction and death in a mouse cytokine-storm model. Additionally, BETi decreases transcription of genes in the viral response, decreases ACE2 expression, and reduces SARS-CoV-2 infection of cardiomyocytes. Together, BETi, including the Food and Drug Administration (FDA) breakthrough designated drug, apabetalone, are promising candidates to prevent COVID-19 mediated cardiac damage.Copyright © 2021 Elsevier Inc.

Objective: To evaluate the efficacy of acellular collagen scaffold (ACS) in combination with basic fibroblast growth factor (bFGF) for the repair of traumatic tympanic membrane (TM) perforation in a rat model. Study Design: A prospective controlled animal study in a rat model of traumatic TM perforation. Setting(s): Tertiary medical center. Subjects and Methods: Sprague-Dawley rats (N = 84) with unilateral traumatic perforation of the right TMs were randomized to receive ACS, bFGF, ACS in combination with bFGF (ACS/bFGF), or nothing (spontaneous healing without any interventions as a control group). The healing outcomes were evaluated by otoscopy, optical coherence tomography, histology, and transmission electron microscopy at 1, 2, and 4 weeks postoperatively. The hearing outcomes were assessed with auditory brainstem response testing. Result(s): ACS/bFGF resulted in higher perforation closure rates at an earlier stage than spontaneous healing, ACS, and bFGF. Based on histology, optical coherence tomography, and transmission electron microscopy, a trilaminar structure and uniform thickness with mature, densely packed collagen fibers were seen in the ACS/bFGF group. Auditory brainstem response evaluation also showed that ACS/bFGF treatment promoted faster functional hearing recovery as compared with the control group. Conclusion(s): ACS is an effective TM scaffold and a carrier for bFGF. ACS/bFGF improves the TM closure rate, results in better-reconstructed TMs, and improves hearing. ACS/bFGF serves as a potential substitute for TM perforations in clinical settings.Copyright © American Academy of Otolaryngology-Head and Neck Surgery Foundation 2020.

Oxaliplatin is a platinum-based alkylating chemotherapeutic agent used for cancer treatment. Neurotoxicity is one of its major adverse effects that often demands dose limitation. However, the effects of chronic oxaliplatin on the toxicity of the autonomic nervous system regulating cardiorespiratory function and adaptive reflexes are unknown. Male Sprague Dawley rats were treated with intraperitoneal oxaliplatin (3 mg kg-1 per dose) 3 times a week for 14 days. The effects of chronic oxaliplatin treatment on baseline mean arterial pressure (MAP); heart rate (HR); splanchnic sympathetic nerve activity (sSNA); phrenic nerve activity (PNA) and its amplitude (PNamp) and frequency (PNf); and sympathetic reflexes were investigated in anaesthetised, vagotomised and artificially ventilated rats. The same parameters were evaluated after acute oxaliplatin injection, and in the chronic treatment group following a single dose of oxaliplatin. The amount of platinum in the brain was determined with atomic absorption spectrophotometry. Chronic oxaliplatin treatment significantly increased MAP, sSNA and PNf and decreased HR and PNamp, while acute oxaliplatin had no effects. Platinum was accumulated in the brain after chronic oxaliplatin treatment. In the chronic oxaliplatin treatment group, further administration of a single dose of oxaliplatin increased MAP and sSNA. The baroreceptor sensitivity and somatosympathetic reflex were attenuated at rest while the sympathoexcitatory response to hypercapnia was increased in the chronic treatment group. This is the first study to reveal oxaliplatin-induced alterations in the central regulation of cardiovascular and respiratory functions as well as reflexes that may lead to hypertension and breathing disorders which may be mediated via accumulated platinum in the brain.Copyright © 2020, Springer-Verlag GmbH Germany, part of Springer Nature.