Molecular docking aided machine learning for the identification of potential VEGFR inhibitors against renal cell carcinoma.

Renal cell carcinoma is a highly vascular tumor associated with vascular endothelial growth factor (VEGF) expression. The Vascular Endothelial Growth Factor -2 (VEGF-2) and its receptor was identified as a potential anti-cancer target, and it plays a crucial role in physiology as well as pathology. Inhibition of angiogenesis via blocking the signaling pathway is considered an attractive target. In the present study, 150 FDA-approved drugs have been screened using the concept of drug repurposing against VEGFR-2 by employing the molecular docking, molecular dynamics, grouping data with Machine Learning algorithms, and density functional theory (DFT) approaches. The identified compounds such as Pazopanib, Atogepant, Drosperinone, Revefenacin and Zanubrutinib shown the binding energy - 7.0 to - 9.5 kcal/mol against VEGF receptor in the molecular docking studies and have been observed as stable in the molecular dynamic simulations performed for the period of 500 ns. The MM/GBSA analysis shows that the value ranging from - 44.816 to - 82.582 kcal/mol. Harnessing the machine learning approaches revealed that clustering with K = 10 exhibits the relevance through high binding energy and satisfactory logP values, setting them apart from compounds in distinct clusters. Therefore, the identified compounds are found to be potential to inhibit the VEGFR-2 and the present study will be a benchmark to validate the compounds experimentally.

Navigating the Challenges of 3D Printing Personalized Medicine in Space Explorations: A Comprehensive Review.

Space exploration has undergone a paradigm shift in recent years, with a growing emphasis on long-duration missions and human habitation on other celestial bodies. Private aerospace businesses are at the forefront of advancing the next iteration of spacecraft, encompassing a wide range of applications such as deep space exploration (e.g., SpaceX) and cost-effective satellite deployments (e.g., Rocketlab). One of the critical challenges associated with prolonged space missions is the provision of personalized medical care. 3D printing technology has emerged as a potential solution, enabling the on-demand production of personalized medical devices and medications. However, the unique conditions of space pose substantial challenges to the successful implementation of 3D printing for personalized medicine. Tremendous scope for research exists in terms of resource utilization and waste management in space ecosystem, robotic and artificial intelligence (AI) enabled tool utilization, remote operability, interplanetary travel, space education and training tools, digital twins, space tourism and in many other aspects of 3D printing for personalized medicine in space explorations.

Spatiotemporal immune atlas of a clinical-grade gene-edited pig-to-human kidney xenotransplant.

Pig-to-human xenotransplantation is rapidly approaching the clinical arena; however, it is unclear which immunomodulatory regimens will effectively control human immune responses to pig xenografts. Here, we transplant a gene-edited pig kidney into a brain-dead human recipient on pharmacologic immunosuppression and study the human immune response to the xenograft using spatial transcriptomics and single-cell RNA sequencing. Human immune cells are uncommon in the porcine kidney cortex early after xenotransplantation and consist of primarily myeloid cells. Both the porcine resident macrophages and human infiltrating macrophages express genes consistent with an alternatively activated, anti-inflammatory phenotype. No significant infiltration of human B or T cells into the porcine kidney xenograft is detectable. Altogether, these findings provide proof of concept that conventional pharmacologic immunosuppression may be able to restrict infiltration of human immune cells into the xenograft early after compatible pig-to-human kidney xenotransplantation.

Performance evaluation of in-house developed Covid-19 IgG/IgM antibody rapid diagnostic kit.

In the interest of preventing the Coronavirus Disease 2019 (COVID-19) pandemic from spreading, it is crucial to promptly identify and confine afflicted patients. Serological antibody testing is a significant diagnostic technique that is increasingly employed in clinics, however its clinical use is still being investigated. The present study was carried out to scrutinize how well Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) antibody testing using in-house developed rapid antibody assay worked against the chemiluminescence (CLIA) assay. Either IgG positive (IgG + IgM-) or IgM positive (IgM + IgG-); both IgG and IgM positive (IgM + IgG+); and negatives (IgM- IgG-) have been evaluated. A total of 300 samples with diverse age and sexual identity data were included. The combined sensitivities for IgG + IgM+, IgM + IgG-, IgG + IgM- and IgG-IgM- were evaluated. More accurate diagnostic results may be obtained using molecular diagnostic tools. The Antibody Rapid Diagnostic kit's (in-house developed) performance was satisfactory for determining the presence of Covid-19 infection with IgG and IgM positivity. The IgG and IgM positivity helped evaluate the immune response in the individual for the COVID-19 infection. These results lend support to the additional utilisation of serological antibody tests in the COVID-19 diagnosis.

Circulating SARS-CoV-2+ megakaryocytes are associated with severe viral infection in COVID-19.

Several independent lines of evidence suggest that megakaryocytes are dysfunctional in severe COVID-19. Herein, we characterized peripheral circulating megakaryocytes in a large cohort of inpatients with COVID-19 and correlated the subpopulation frequencies with clinical outcomes. Using peripheral blood, we show that megakaryocytes are increased in the systemic circulation in COVID-19, and we identify and validate S100A8/A9 as a defining marker of megakaryocyte dysfunction. We further reveal a subpopulation of S100A8/A9+ megakaryocytes that contain severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) protein and RNA. Using flow cytometry of peripheral blood and in vitro studies on SARS-CoV-2-infected primary human megakaryocytes, we demonstrate that megakaryocytes can transfer viral antigens to emerging platelets. Mechanistically, we show that SARS-CoV-2-containing megakaryocytes are nuclear factor κB (NF-κB)-activated, via p65 and p52; express the NF-κB-mediated cytokines interleukin-6 (IL-6) and IL-1ß; and display high surface expression of Toll-like receptor 2 (TLR2) and TLR4, canonical drivers of NF-κB. In a cohort of 218 inpatients with COVID-19, we correlate frequencies of megakaryocyte subpopulations with clinical outcomes and show that SARS-CoV-2-containing megakaryocytes are a strong risk factor for mortality and multiorgan injury, including respiratory failure, mechanical ventilation, acute kidney injury, thrombotic events, and intensive care unit admission. Furthermore, we show that SARS-CoV-2+ megakaryocytes are present in lung and brain autopsy tissues from deceased donors who had COVID-19. To our knowledge, this study offers the first evidence implicating SARS-CoV-2+ peripheral megakaryocytes in severe disease and suggests that circulating megakaryocytes warrant investigation in inflammatory disorders beyond COVID-19.

Spatiotemporal immune atlas of the first clinical-grade, gene-edited pig-to-human kidney xenotransplant.

Pig-to-human xenotransplantation is rapidly approaching the clinical arena; however, it is unclear which immunomodulatory regimens will effectively control human immune responses to pig xenografts. We transplanted a gene-edited pig kidney into a brain-dead human recipient on pharmacologic immunosuppression and studied the human immune response to the xenograft using spatial transcriptomics and single-cell RNA sequencing. Human immune cells were uncommon in the porcine kidney cortex early after xenotransplantation and consisted of primarily myeloid cells. Both the porcine resident macrophages and human infiltrating macrophages expressed genes consistent with an alternatively activated, anti-inflammatory phenotype. No significant infiltration of human B or T cells into the porcine kidney xenograft was detected. Altogether, these findings provide proof of concept that conventional pharmacologic immunosuppression is sufficient to restrict infiltration of human immune cells into the xenograft early after compatible pig-to-human kidney xenotransplantation.

α2,6 Sialylation mediated by ST6GAL1 promotes glioblastoma growth.

One of the least-investigated areas of brain pathology research is glycosylation, which is a critical regulator of cell surface protein structure and function. ß-Galactoside α2,6-sialyltransferase (ST6GAL1) is the primary enzyme that α2,6 sialylates N-glycosylated proteins destined for the plasma membrane or secretion, thereby modulating cell signaling and behavior. We demonstrate a potentially novel, protumorigenic role for α2,6 sialylation and ST6GAL1 in the deadly brain tumor glioblastoma (GBM). GBM cells with high α2,6 sialylation exhibited increased in vitro growth and self-renewal capacity and decreased mouse survival when orthotopically injected. α2,6 Sialylation was regulated by ST6GAL1 in GBM, and ST6GAL1 was elevated in brain tumor-initiating cells (BTICs). Knockdown of ST6GAL1 in BTICs decreased in vitro growth, self-renewal capacity, and tumorigenic potential. ST6GAL1 regulates levels of the known BTIC regulators PDGF Receptor ß (PDGFRB), Activated Leukocyte Cell Adhesion Molecule, and Neuropilin, which were confirmed to bind to a lectin-recognizing α2,6 sialic acid. Loss of ST6GAL1 was confirmed to decrease PDGFRB α2,6 sialylation, total protein levels, and the induction of phosphorylation by PDGF-BB. Thus, ST6GAL1-mediated α2,6 sialylation of a select subset of cell surface receptors, including PDGFRB, increases GBM growth.

Megakaryocytes are a Novel SARS-CoV-2 Infection Target and Risk Factor for Mortality and Multi-Organ Failure

Discovery of a biomarker for patients at high risk of progression to severe Coronavirus Disease 2019 (COVID-19) is critical for clinical management, particularly in areas of the world where widespread vaccine distribution and herd immunity have yet to be achieved. Herein, we characterize peripheral blood from 218 COVID-19 patients with flow cytometry and provide evidence that megakaryocytes are a target for infection by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). We demonstrate a positive correlation between infected megakaryocytes expressing the protein calprotectin (also called S100A8/A9), a known marker of COVID-19 severity. Additionally, we show that infected, calprotectin expressing megakaryocytes are correlated with COVID-19 severity and are a prognostic indicator of 30-day clinical outcomes including respiratory failure, thrombotic events, acute kidney injury (AKI), ICU admission, and mechanical ventilation. These findings represent a novel SARS-CoV-2 infection target with significant clinical implications as a biomarker for clinical outcomes associated with severe COVID-19.

Comparative Clinical Evaluation of Semilunar Coronally Positioned Flap Alone and Semilunar Coronally Positioned Flap in Conjunction with Free Gingival Graft for Root Coverage.

Gingival recession accounts for apical migration of the gingival margin, resulting in exposure of the cementoenamel junction and root surface, with exposure of the root surface linked to deteriorated esthetic appearance and increased dentinal hypersensitivity. Various surgical techniques have been used to correct labial gingival recession defects. The present study evaluated and compared the results of semilunar coronally positioned flap (SCPF) alone and in conjunction with free gingival graft (FGG) for the treatment of Miller Class I and II gingival recession defects in maxillary anterior teeth. A total of 20 bilateral Miller Class I and II gingival recession sites were included and randomly allocated (n = 10 sites/group) to either the semilunar coronally positioned flap technique alone (SCPF group; control) or with FGG (SCPF+FGG group; test). Longitudinal alterations in probing depth (PD), recession width (RW), recession height (RH), width of keratinized tissue (WKT), and clinical attachment level (CAL) were measured and analyzed for both groups at 1-, 3-, 6-, and 12-month follow-ups. Both groups saw a significant decrease in RH, RW, and CAL and a significant increase in WKT. No statistically significant difference was observed in the final root coverage outcome between both groups in terms of RH, RW, and CAL, but a significant increase in WKT was seen with SCPF+FGG. Both techniques demonstrated optimal results without significant differences in the final root coverage outcomes except for WKT, which had a statistically significant increase in the SCPF+FGG group.

Strategy of Isolating 'Primed' Tumor Initiating Cells Based on Mitochondrial Transmembrane Potential.

Various stem cells have been found to be dependent on mitochondrial energetics. The role of mitochondria in regulating the self-renewal of normal stem cells and stem-like tumor initiating cells (TICs) is increasingly being appreciated. We proposed that TIC populations have a sub population of cells that are "primed" by mitochondria for self-renewal. Using ovarian cancer model, we have developed a protocol to identify and isolate these "primed" cells using Fluorescence-Assisted Cell Sorting (FACS). We combined live cell stains for a functional marker of TICs and for mitochondrial transmembrane potential to enrich TICs with higher mitochondrial potential that form in vitro spheroids 10-fold more than the other TICs with lower mitochondrial potential. This protocol can be directly used or modified to be used in various cell types. Thus, this protocol is anticipated to be invaluable for the basic understanding of mitochondrial and energetic heterogeneity within stem cell population, and may also prove valuable in translational studies in regenerative medicine and cancer biology.

Role of Arthroscopy in Various Ankle Disorders.

INTRODUCTION: Ankle arthroscopy has come a long way since it was thought, it is not feasible because of tight joint and anatomical characteristics of ankle joint. The same anatomical features like capsular attachment and safe accessory portals are used to access the whole joint even with a rigid arthroscope. Ankle distraction method was routinely used to access the anterior ankle. However, nowadays, anterior arthroscopy is done in dorsiflexion as this increases the anterior ankle joint volume, and thereby easy access to various anatomical structures. On the other hand, intermittent traction is used to access the posterior ankle. Initially used as a diagnostic tool, ankle arthroscopy is now used extensively as a therapeutic and reconstruction tool. New evidence is published for all inside ligament reconstructions, effective management of impingement syndromes, and osteochondral lesions. The indications are being extended to fracture management and arthrodesis. METHODOLOGY: This narrative review was performed following a literature search in the Pubmed database and Medline using the following keywords: ankle arthroscopy, portals, ankle OCD, functional outcome. Related articles were then reviewed. CONCLUSION: Complications rate is reduced with a better understanding of the relative anatomy of surrounding neurovascular structures and tendons with regard to the position of ankle joint. This review on ankle arthroscopy focuses on anatomy, indications, and complications. Ankle arthroscopy is a safe and elegant tool as any other joint arthroscopy.

Awareness of General Practitioners toward Hypertension Management.

INTRODUCTION: The silent killer, hypertension (HTN), is a significant risk factor for cardiovascular disease. In India, HTN has a major public health effect on cardiovascular health and health-care systems. AIM: The present study was aimed to evaluate the awareness of general practitioners (GPs) toward HTN management and also their approach toward management. MATERIALS AND METHODS: The present study was a questionnaire-based assessment study. A total of 100 GPs were included in the study as our study sample. All the participants were well informed about the study and after that those who were willing to participate were enrolled after obtaining a written informed consent. RESULTS: The common presenting symptoms of hypertensive patients were predominantly morning headache (71%), dizziness (41%), palpitation (39%), and fatigability (29%). Majority (84%) practiced cuff placement method covering about 2/3rd of the arm at heart level. The preferred position while blood pressure (BP) examination of patient by majority of practitioners was while sitting (53%). The number of readings usually taken for measuring BP for each patient was as follows: one (3%), two (42%), and three (55)%. CONCLUSION: Although GPs in our study are well informed and up to date on certain aspects of HTN diagnosis and treatment, they may still lack an appropriate approach to HTN history taking, diagnosis, and treatment.

Association of State Stay-at-Home Orders and State-Level African American Population With COVID-19 Case Rates.

Importance: To cope with the continuing coronavirus disease 2019 (COVID-19) pandemic, state and local officials need information on the effectiveness of policies aimed at curbing disease spread, as well as state-specific characteristics, like the racial mix, associated with increased risks related to the disease. Objective: To investigate whether state-imposed stay-at-home orders (SAHOs) and the proportion of African American population in a state were associated with the state-level COVID-19 cases. Design, Setting, and Participants: This cross-sectional study used daily, state-level data on COVID-19 cases, tests, and fatalities from the COVID Tracking Project. Data from March 1 to May 4, 2020, for all states (except Washington state) as well as the District of Columbia were used. Exposures: The key exposure variables were state-level SAHO (1 if in place, 0 otherwise), and proportion of state population who are African American. Main Outcomes and Measures: The primary outcome was daily cumulative COVID-19 case rates. A secondary outcome was subsequent COVID-19 fatality rates, derived using mean cumulative fatality rates 21 to 28 days after each date. Multivariate regression models were estimated. Results: The final sample included 3023 pooled state- and day-level observations. The mean (SD) cumulative positive case rate was 103.186 (200.067) cases per 100 000 state population, the mean (SD) cumulative test rate was 744.23 (894.944) tests per 100 000 state population, and the mean (SD) subsequent cumulative fatality rate was 12.923 (21.737) deaths per 100 000 state population. There was a negative association of SAHOs with cumulative case rates (ß = -1.166; 95% CI, -1.484 to -0.847; P < .001) and subsequent fatality rates (ß = -0.204; 95% CI, -0.294 to -0.113; P < .001). Estimation analyses indicated that expected cumulative case rates would have been more than 200% higher and fatality rates approximately 22% higher if there were no SAHOs, as compared with SAHOs fully in place. A higher proportion of African American population was associated with higher case rates (ß = 0.045; 95% CI, 0.014 to 0.077; P = .001) and fatality rates (ß = 0.068; 95% CI, 0.044 to 0.091; P < .001). Conclusions and Relevance: In this cross-sectional study, SAHOs were associated with reductions in COVID-19 case rates. These findings could help inform policy makers to address the continued COVID-19 pandemic in the US. The proportion of African American population was positively associated with COVID-19 case rates, and this state-level finding adds to evidence from existing ecological studies using county-level data on racial disparities in COVID-19 infection rates and underlines the urgency of better understanding and addressing these disparities.

Stay-at-Home Orders, African American Population, Poverty and State-level Covid-19 Infections: Are there associations?

ImportanceTo cope with the continuing COVID-19 pandemic, state and local health officials need information on the effectiveness of policies aimed at curbing contagion, as well as area-specific socio-demographic characteristics that can portend vulnerability to the disease. ObjectiveTo investigate whether state-imposed stay-at-home orders, African American population in the state, state poverty and other state socio-demographic characteristics, were associated with the state-level incidence of COVID-19 infection. Design, Setting, ParticipantsState-level, aggregated, publicly available data on positive COVID-19 cases and tests were used. The period considered was March 1st-May 4th. All U.S. states except Washington were included. Outcomes of interest were daily cumulative and daily incremental COVID-19 infection rates. Outcomes were log-transformed. Log-linear regression models with a quadratic time-trend and random intercepts for states were estimated. Covariates included log-transformed test-rates, a binary indicator for stay-at-home, percentage of African American, poverty, percentage elderly, state population and prevalence of selected comorbidities. Binary fixed effects for date each state first started reporting test data were included. ResultsStay-at-home orders were associated with decreases in cumulative ({beta}:-1.23; T-stat: - 6.84) and daily ({beta}:-0.46; T-stat: -2.56) infection-rates. Predictive analyses indicated that expected cumulative infection rates would be 3 times higher and expected daily incremental rates about 60% higher in absence of stay-at-home orders. Higher African American population was associated with higher cumulative ({beta}: 0.08; T-stat: 4.01) and daily ({beta}: 0.06; T-stat: 3.50) rates. State poverty rates had mixed results and were not robust to model specifications. There was strong evidence of a quadratic daily trend for cumulative and daily rates. Results were largely robust to alternate specifications. ConclusionsWe find evidence that stay-at-home orders, which were widely supported by public-health experts, helped to substantially curb COVID-19 infection-rates. As we move to a phased re-opening, continued precautions advised by public-health experts should be adhered to. Also, a larger African American population is strongly associated with incidence of COVID-19 infection. Policies and resources to help mitigate African American vulnerability to COVID-19 is an urgent public health and social justice issue, especially since the ongoing mass protests against police brutality may further exacerbate COVID-19 contagion in this community. Key PointsO_ST_ABSQuestionC_ST_ABSDid the stay-at-home orders, African American population and other socio demographic factors across states have any associations with COVID-19 infection rates across states? FindingsMultivariate log-linear regression models using daily state level data from March-May found evidence that when stay-at-home orders were implemented, they helped reduce state COVID-19 cumulative and daily infection rates substantially. Further, we found that states with larger African-American population had higher COVID-19 infection rates. MeaningResults suggest that state-level stay-at-home orders helped reduce COVID-19 infection rates substantially, and also that African American populations may be especially vulnerable to COVID-19 infection.

Photoprotective Properties of Vitamin D and Lumisterol Hydroxyderivatives.

We have previously described new pathways of vitamin D3 activation by CYP11A1 to produce a variety of metabolites including 20(OH)D3 and 20,23(OH)2D3. These can be further hydroxylated by CYP27B1 to produce their C1α-hydroxyderivatives. CYP11A1 similarly initiates the metabolism of lumisterol (L3) through sequential hydroxylation of the side chain to produce 20(OH)L3, 22(OH)L3, 20,22(OH)2L3 and 24(OH)L3. CYP11A1 also acts on 7-dehydrocholesterol (7DHC) producing 22(OH)7DHC, 20,22(OH)27DHC and 7-dehydropregnenolone (7DHP) which can be converted to the D3 and L3 configurations following exposure to UVB. These CYP11A1-derived compounds are produced in vivo and are biologically active displaying anti-proliferative, anti-inflammatory, anti-cancer and pro-differentiation properties. Since the protective role of the classical form of vitamin D3 (1,25(OH)2D3) against UVB-induced damage is recognized, we recently tested whether novel CYP11A1-derived D3- and L3-hydroxyderivatives protect against UVB-induced damage in epidermal human keratinocytes and melanocytes. We found that along with 1,25(OH)2D3, CYP11A1-derived D3-hydroxyderivatives and L3 and its hydroxyderivatives exert photoprotective effects. These included induction of intracellular free radical scavenging and attenuation and repair of DNA damage. The protection of human keratinocytes against DNA damage included the activation of the NRF2-regulated antioxidant response, p53-phosphorylation and its translocation to the nucleus, and DNA repair induction. These data indicate that novel derivatives of vitamin D3 and lumisterol are promising photoprotective agents. However, detailed mechanisms of action, and the involvement of specific nuclear receptors, other vitamin D binding proteins or mitochondria, remain to be established.

Starter consortia for on-farm cocoa fermentation and their quality attributes.

A starter consortium of Saccharomyces cerevisiae (Y), Lactobacillus plantarum (LAB), and Acetobacter aceti (AAB) was defined to ferment the Cocoa beans (Theobroma cacao). Emphasis was laid to optimize the microbial concentration with a functional ratio of selected cultures. A central composite rotatable design (CCRD) was employed to study the effect of inoculum size (0-20% w/v) with alcohol, titrable acidity, polyphenols, anthocyanin, cut test, and sensory as response variables. The significant (p < 0.05) response surface models with high coefficients of determination values (R2) ranging from 0.82 to 0.93 were considered for the experimental data, which represented the polynomial response models for describing the constraints. Based on the design, the concentration of consortia ranged 9.03X103 CFU/g of Y, 5.9X104 CFU/g of LAB, and 7.0X104 CFU/g of AAB. The graphical optimization of superimposed contour plots fulfilled the desired metabolites; alcohol (Y1) ≤ 11 mg/g, titrable acidity (Y2) ≥ 0.25%, polyphenols (Y3) ≤ 4.0 mg/g, anthocyanin (Y4) ≤ 14 mg/g, sensory (Y5) ≥ 6.0, and cut test (Y6)≥95%. Thus, validation through a field trial was confirmed to adopt the techno-economic feasibility on-farm process with precise inoculums. The effect of starter consortia on Cocoa fermentation and quality was found to be significant.

Increased flux through the mevalonate pathway mediates fibrotic repair without injury.

Macrophages are important in mounting an innate immune response to injury as well as in repair of injury. Gene expression of Rho proteins is known to be increased in fibrotic models; however, the role of these proteins in idiopathic pulmonary fibrosis (IPF) is not known. Here, we show that BAL cells from patients with IPF have a profibrotic phenotype secondary to increased activation of the small GTPase Rac1. Rac1 activation requires a posttranslational modification, geranylgeranylation, of the C-terminal cysteine residue. We found that by supplying more substrate for geranylgeranylation, Rac1 activation was substantially increased, resulting in profibrotic polarization by increasing flux through the mevalonate pathway. The increased flux was secondary to greater levels of acetyl-CoA from metabolic reprogramming to ß oxidation. The polarization mediated fibrotic repair in the absence of injury by enhancing macrophage/fibroblast signaling. These observations suggest that targeting the mevalonate pathway may abrogate the role of macrophages in dysregulated fibrotic repair.

NOX4 modulates macrophage phenotype and mitochondrial biogenesis in asbestosis.

Macrophage activation is implicated in the development of pulmonary fibrosis by generation of profibrotic molecules. Although NADPH oxidase 4 (NOX4) is known to contribute to pulmonary fibrosis, its effects on macrophage activation and mitochondrial redox signaling are unclear. Here, we show that NOX4 is crucial for lung macrophage profibrotic polarization and fibrotic repair after asbestos exposure. NOX4 was elevated in lung macrophages from subjects with asbestosis, and mice harboring a deletion of NOX4 in lung macrophages were protected from asbestos-induced fibrosis. NOX4 promoted lung macrophage profibrotic polarization and increased production of profibrotic molecules that induce collagen deposition. Mechanistically, NOX4 further augmented mitochondrial ROS production and induced mitochondrial biogenesis. Targeting redox signaling and mitochondrial biogenesis prevented the profibrotic polarization of lung macrophages by reducing the production of profibrotic molecules. These observations provide evidence that macrophage NOX4 is a potentially novel therapeutic target to halt the development of asbestos-induced pulmonary fibrosis.

Susceptibility of microtubule-associated protein 1 light chain 3ß (MAP1LC3B/LC3B) knockout mice to lung injury and fibrosis.

Insufficient autophagy has been reported in idiopathic pulmonary fibrosis (IPF) lungs. Specific roles of autophagy-related proteins in lung fibrosis development remain largely unknown. Here, we investigated the role of autophagy marker protein microtubule-associated protein 1 light chain 3ß (LC3B) in the development of lung fibrosis. LC3B-/- mice upon aging show smaller lamellar body profiles, increased cellularity, alveolar epithelial cell type II (AECII) apoptosis, surfactant alterations, and lysosomal and endoplasmic reticulum stress. Autophagosomal soluble N-ethylmaleimide-sensitive factor attachment protein receptor syntaxin 17 is increased in the AECII of aged LC3B-/- mice and patients with IPF. Proteasomal activity, however, remained unaltered in LC3B-/- mice. In vitro knockdown of LC3B sensitized mouse lung epithelial cells to bleomycin-induced apoptosis, but its overexpression was protective. In vivo, LC3B-/- mice displayed increased susceptibility to bleomycin-induced lung injury and fibrosis. We identified cathepsin A as a novel LC3B binding partner and its overexpression in vitro drives MLE12 cells to apoptosis. Additionally, cathepsin A is increased in the AECII of aged LC3B-/- mice and in the lungs of patients with IPF. Our study reveals that LC3B mediated autophagy plays essential roles in AECII by modulating the functions of proteins like cathepsin A and protects alveolar epithelial cells from apoptosis and subsequent lung injury and fibrosis.-Kesireddy, V. S., Chillappagari, S., Ahuja, S., Knudsen, L., Henneke, I., Graumann, J., Meiners, S., Ochs, M., Ruppert, C., Korfei, M., Seeger, W., Mahavadi, P. Susceptibility of microtubule-associated protein 1 light chain 3ß (MAP1LC3B/LC3B) knockout mice to lung injury and fibrosis.