BACKGROUND:Cigarette smoking (CS) is a major risk factor for cardiovascular disease through chronic inflammation. While its pulmonary effects are well established, the mechanisms linking lung inflammation to vascular injury remain unclear. Because neutrophils are early responders to CS-induced inflammation, we hypothesized that they drive systemic myelopoiesis and vascular inflammation via alarmin release. METHODS:Wild-type mice were exposed to inhaled CS or orally administered cigarette smoke extract. Immune cell composition in lung, bronchoalveolar lavage fluid, blood, spleen, and bone marrow was assessed by flow cytometry. Hematopoietic stem and progenitor cell proliferation, reactive oxygen species production, and S100A8/A9 (S100 calcium-binding protein A8/A9) release were quantified. Atherosclerosis progression was evaluated in Ldlr-/- (low-density lipoprotein receptor-deficient) mice fed a Western diet and treated with cigarette smoke extract. To define the role of neutrophil-derived S100A8/A9, bone marrow transplantation was performed using S100a9-/- or wild-type donors. RESULTS:CS exposure increased circulating monocytes and neutrophils through enhanced bone marrow myelopoiesis and elevated reactive oxygen species-dependent S100A8/A9 release. Oral cigarette smoke extract reproduced these effects, indicating direct activation of neutrophils independent of pulmonary inflammation or lipid changes. In Ldlr-/- mice, cigarette smoke extract accelerated atherosclerosis by promoting infiltration of inflammasome-primed neutrophils, increasing IL-1β (interleukin 1 beta) release, and impairing macrophage efferocytosis. Hematopoietic S100a9 deletion normalized myelopoiesis and reduced vascular inflammation and plaque burden. CONCLUSIONS:Ingested CS components directly activate neutrophils to release S100A8/A9, triggering myelopoiesis and vascular inflammation. These findings reveal that tobacco's cardiovascular toxicity extends beyond inhalation, implicating oral exposure as a driver of systemic inflammation and atherogenesis.

Background:Cigarette smoking (CS) is a major risk factor for cardiovascular disease (CVD) through chronic inflammation. While its pulmonary effects are well established, the mechanisms linking lung inflammation to vascular injury remain unclear. Because neutrophils are early responders to CS-induced inflammation, we hypothesized that they drive systemic myelopoiesis and vascular inflammation via alarmin release. Methods:Wild-type (WT) mice were exposed to inhaled CS or orally administered cigarette smoke extract (CSE). Immune cell composition in lung, bronchoalveolar lavage fluid (BALF), blood, spleen, and bone marrow (BM) was assessed by flow cytometry. Hematopoietic stem and progenitor cell (HSPC) proliferation, reactive oxygen species (ROS) production, and S100A8/A9 release were quantified. Atherosclerosis progression was evaluated in Ldlr ⁻ / ⁻ mice fed a Western diet and treated with CSE. To define the role of neutrophil-derived S100A8/A9, bone marrow transplantation was performed using S100a9 ⁻ / ⁻ or WT donors. Results:CS exposure increased circulating monocytes and neutrophils through enhanced BM myelopoiesis and elevated ROS-dependent S100A8/A9 release. Oral CSE reproduced these effects, indicating direct activation of neutrophils independent of pulmonary inflammation or lipid changes. In Ldlr ⁻ / ⁻ mice, CSE accelerated atherosclerosis by promoting infiltration of inflammasome-primed neutrophils, increased IL-1β release, and impaired macrophage efferocytosis. Hematopoietic S100a9 deletion normalized myelopoiesis and reduced vascular inflammation and plaque burden. Conclusions:Ingested CS components directly activate neutrophils to release S100A8/A9, triggering myelopoiesis and vascular inflammation. These findings reveal that tobacco's cardiovascular toxicity extends beyond inhalation, implicating oral exposure as a driver of systemic inflammation and atherogenesis. Novelty and Significance:What Is Known?: Cigarette smoking (CS) is a major risk factor for atherosclerosis, driving systemic inflammation and innate immune activation.Neutrophils and monocytes contribute to plaque progression, but the upstream mechanisms by which CS exacerbates their pathogenic roles remain incompletely understood.S100A8/A9 levels correlate with neutrophilia and cardiovascular risk in smokers, but their functional role in lesion biology is not fully defined.What New Information Does This Article Contribute?: Identifies S100A8/A9 as a key mediator linking CS exposure to enhanced medullary myelopoiesis, neutrophilia, and increased lesional infiltration of inflammasome-primed myeloid cells.Demonstrates that neutrophil-derived IL-1β impairs macrophage efferocytosis by downregulating phagocytosis receptors, thereby promoting plaque vulnerability.Reveals that CS drives atherosclerosis even in the absence of lipid perturbations or overt pulmonary injury, highlighting a novel oral exposure-vascular axis of disease propagation.

Vibrio parahaemolyticus, an important food-borne pathogens found to be associated with seafoods and marine environs. It has been a topic of debate for many decades that most pathogens are known to enter a viable but nonculturable (VBNC) state under cold temperature and nutrient limited conditions. The present study examined the time required for the induction of VBNC state and the revival strategies of both the endemic O3:K6 and O1:K25 sporadic strains of V. parahaemolyticus. The results revealed that V. parahaemolyticus survived even after 55 days of incubation in nutrient starved media such as phosphate buffered saline (PBS) and Coastal Water (CW) and could be recovered by temperature upshift method, and compared the resuscitation using Dulbecco's Modified Eagle Medium (DMEM), sheep blood serum, chitin flakes with live Artemia salina, and the results suggests that chitin plays a significant role in regulating the VBNC state. It was also confirmed by Confocal Laser Scanning Microscopy (CLSM) and Scanning Electron Microscope (SEM) analysis that VBNC cells can alter their morphology to coccoid forms in order to survive in most extreme nutrient limited environment. Further data on the promoting factors and the exact mechanism that resuscitate VBNC V. parahaemolyticus in cold natural environments and frozen foods are needed to perform a robust risk assessment.

Still, it remains a debate after four decades of research on surviving cells, several bacterial species were naturally inducted and found to exist in a viable but non-culturable (VBNC) state, an adaptive strategy executed by most bacterial species under different stressful conditions. VBNC state are generally attributed when the cells lose its culturability on standard culture media, diminish in conventional detection methods, but retaining its viability, virulence and antibiotic resistance over a period of years and may poses a risk to marine animals as well as public health and food safety. In this present review, we mainly focus the VBNC state of Vibrios and other human bacterial pathogens. Exposure to several factors like nutrient depletion, temperature fluctuation, changes in salinity and oxidative stress, antibiotic and other chemical stress can induce the cells to VBNC state. The transcriptomic and proteomic changes during VBNC, modification in detection techniques and the most significant role of Rpf in conversion of VBNC into culturable cells. Altogether, detection of unculturable VBNC forms has significant importance, since it may not only regain its culturability, but also reactivate its putative virulence determinants causing serious outbreaks and illness to the individual.

Increase in neutrophil counts that are associated with cardiovascular risk factors including myocardial infarction (MI) and early development of heart attack is a major concern among all age groups in recent years. Neutrophil production in response to heart failure is mediated by various ways including recruitment of immune cells at the site of injury, release inflammatory molecules, and continuous supply of leucocytes. Recent evidences proved the importance of neutrophil-derived alarmin molecule S100A8/A9 in provoking inflammation after MI. Besides, clinical trials with increases in serum level of circulating calgranulins and major adverse cardiac events (MACE) in MI subjects were demonstrated which implicated that targeting neutrophils or their inflammatory alarmins could be effective in reducing heart attack. On the other hand, neutrophils also found to involved in the regression of inflammation and atherosclerotic plaques. In this review, we discuss the overview of S100 protein family in various metabolic diseases with a main focus on cardiac inflammation. We also discuss the recent evidences on neutrophil-derived S100A/A9 which triggers inflammation and improves cardiac function after MI. We also insist that neutrophils could be a better biomarker and therapeutic targets for various inflammatory diseases including cardiovascular disease.

Background: Excessive infiltration of neutrophils to the ischemic heart after an acute myocardial infarction (MI) is mediated in-part by the S100A8/A9-NLRP3-IL-1β signaling axis in neutrophils. Despite upregulation of NLRP3 inflammasome signaling components in neutrophils, the serum levels of IL-1β, was not impacted by MI suggesting that IL-1β is not released systemically. We hypothesize that IL-1β is released locally within the bone marrow (BM) by inflammasome-primed and reverse-migrating neutrophils. Methods: Using parabiosis techniques, we first characterized the migration patterns of different blood cell types across the parabiotic barrier. We next induced MI in one of the parabiotic mice and examined the ability of injury-exposed neutrophils to permeate the parabiotic barrier and induce granulopoiesis in non-infarcted parabionts. Finally, we studied the molecular mechanisms that govern reverse migration and retention of primed neutrophils, IL-1β secretion and granulopoiesis. Cardiac function was assessed by echocardiography. Results: MI promotes granulopoiesis in the BM and introducing a time-dependent parabiotic barrier to the free movement of neutrophils inhibited granulopoiesis in the non-infarcted parabionts. MI also promotes greater accumulation/ retention of inflammasome-primed neutrophils in the BM in a neutrophil CXCR4 (C-X-C-motif chemokine receptor 4) and serum S100A8/A9 dependent manner. In the BM, the primed neutrophils secrete IL-1β through formation of gasdermin D pores and, promote granulopoiesis. Strategies aimed at inhibition of neutrophil homing or release of IL-1β in the BM markedly suppressed MI-induced granulopoiesis and, improved cardiac function. Conclusions: Our data reveals a new paradigm of how circulatory cells can deliver signaling molecules (e.g., IL-1β) directly to the site of action rather than through systemic release. The absolute requirement of signal-2 in the BM for IL-1β release is consistent with our model where neutrophils undergo programming in the infarcted heart and inertly return to the sites of production (i.e., BM) with cytokine cargo (i.e., IL-1β) to regulate granulopoiesis. We suggest that this pathway may exist to limit the off-target effects of systemic IL-1β release.

BACKGROUND:Acute myocardial infarction (MI) results in overzealous production and infiltration of neutrophils to the ischemic heart. This is mediated in part by granulopoiesis induced by the S100A8/A9-NLRP3-IL-1β signaling axis in injury-exposed neutrophils. Despite the transcriptional upregulation of the NLRP3 (Nod Like Receptor Family Pyrin Domain-Containing 3) inflammasome and associated signaling components in neutrophils, the serum levels of IL-1β (interleukin-1β), the effector molecule in granulopoiesis, were not affected by MI, suggesting that IL-1β is not released systemically. We hypothesize that IL-1β is released locally within the bone marrow (BM) by inflammasome-primed and reverse-migrating neutrophils.METHODS:Using a combination of time-dependent parabiosis and flow cytometry techniques, we first characterized the migration patterns of different blood cell types across the parabiotic barrier. We next induced MI in parabiotic mice by permanent ligation of the left anterior descending artery and examined the ability of injury-exposed neutrophils to permeate the parabiotic barrier and induce granulopoiesis in noninfarcted parabionts. Last, using multiple neutrophil adoptive and BM transplant studies, we studied the molecular mechanisms that govern reverse migration and retention of the primed neutrophils, IL-1β secretion, and granulopoiesis. Cardiac function was assessed by echocardiography.RESULTS:MI promoted greater accumulation of the inflammasome-primed neutrophils in the BM. Introducing a time-dependent parabiotic barrier to the free movement of neutrophils inhibited their ability to stimulate granulopoiesis in the noninfarcted parabionts. Previous priming of the NLRP3 inflammasome is not a prerequisite, but the presence of a functional CXCR4 (C-X-C-motif chemokine receptor 4) on the primed-neutrophils and elevated serum S100A8/A9 levels are necessary for homing and retention of the reverse-migrating neutrophils. In the BM, the primed-neutrophils secrete IL-1β through formation of gasdermin D pores and promote granulopoiesis. Pharmacological and genetic strategies aimed at the inhibition of neutrophil homing or release of IL-1β in the BM markedly suppressed MI-induced granulopoiesis and improved cardiac function.CONCLUSIONS:Our data reveal a new paradigm of how circulatory cells establish a direct communication between organs by delivering signaling molecules (eg, IL-1β) directly at the sites of action rather through systemic release. We suggest that this pathway may exist to limit the off-target effects of systemic IL-1β release.

Background: Myocardial infarction (MI) triggers myelopoiesis, resulting in heightened production of neutrophils. However, the mechanisms that sustain their production and recruitment to the injured heart are unclear. Methods: Using a mouse model of the permanent ligation of the left anterior descending artery and flow cytometry, we first characterized the temporal and spatial effects of MI on different myeloid cell types. We next performed global transcriptome analysis of different cardiac cell types within the infarct to identify the drivers of the acute inflammatory response and the underlying signaling pathways. Using a combination of genetic and pharmacological strategies, we identified the sequelae of events that led to MI-induced myelopoiesis. Cardiac function was assessed by echocardiography. The association of early indexes of neutrophilia with major adverse cardiovascular events was studied in a cohort of patients with acute MI. Results: Induction of MI results in rapid recruitment of neutrophils to the infarct, where they release specific alarmins, S100A8 and S100A9. These alarmins bind to the Toll-like receptor 4 and prime the nod-like receptor family pyrin domain-containing 3 inflammasome in naïve neutrophils and promote interleukin-1β secretion. The released interleukin-1β interacts with its receptor (interleukin 1 receptor type 1) on hematopoietic stem and progenitor cells in the bone marrow and stimulates granulopoiesis in a cell-autonomous manner. Genetic or pharmacological strategies aimed at disruption of S100A8/A9 and their downstream signaling cascade suppress MI-induced granulopoiesis and improve cardiac function. Furthermore, in patients with acute coronary syndrome, higher neutrophil count on admission and after revascularization correlates positively with major adverse cardiovascular disease outcomes. Conclusions: Our study provides novel evidence for the primary role of neutrophil-derived alarmins (S100A8/A9) in dictating the nature of the ensuing inflammatory response after myocardial injury. Therapeutic strategies aimed at disruption of S100A8/A9 signaling or their downstream mediators (eg, nod-like receptor family pyrin domain-containing 3 inflammasome, interleukin-1β) in neutrophils suppress granulopoiesis and may improve cardiac function in patients with acute coronary syndrome.

Myocardial infarction (MI) triggers myelopoiesis resulting in heightened number of neutrophils in the circulation. However, the mechanism that sustain their number and recruitment to the infarcted heart are unclear. Here, we show that in a mouse model of MI (permanent ligation of LAD), neutrophils are rapidly recruited to the infarct, where they release specific alarmins, S100A8 and S100A9. These alarmins acting either in an autocrine or paracrine manner, primed the Nod Like Receptor (NLR) family Pyrin Domain Containing 3 (Nlrp3) inflammasome in naïve neutrophils via their interaction with the Toll Like Receptor (TLR) 4. The interaction did not result in the release of IL1β systemically. However, the primed neutrophils, loaded with pro-interleukin-1β (IL-1β) returned to the bone marrow (BM) in a CXCR4 (C-X-C-motif chemokine receptor 4)- dependent manner. While at the BM, the primed-neutrophils released IL-1β through Gasdermin D pores and, stimulated granulopoiesis in a cell-autonomous manner. Strategies aimed at preventing the Nlrp3 inflammasome-priming or re-entry of the primed neutrophils to the BM dampened MI-induced granulopoiesis and markedly improved cardiac function. In subjects with acute ST-elevation myocardial infarction (STEMI), the number of neutrophils in the circulation increased both at the time of admission and following revascularization. Most importantly, patients with higher peak neutrophil counts demonstrated significantly higher incidence of major adverse cardiovascular events (MACE) during the one year follow up period. Similar to mouse data, the plasma levels of IL-1β did not change in STEMI patients at any time. However, the circulating neutrophils carried greater amounts of pro-IL-1β confirming our mouse data that granulopoiesis is likely not induced by systemic but locally delivered IL-1β by reverse migrating neutrophils. These data reveal a new paradigm of how circulatory cells establish direct communication between organs by delivering signaling molecules directly at the sites of action rather through systemically.

We sought to delineate the retinal features associated with the high-fat diet (HFD) mouse, a widely used model of obesity. C57BL/6 mice were fed either a high-fat (60% fat; HFD) or low-fat (10% fat; LFD) diet for up to 12 months. The effect of HFD on body weight and insulin resistance were measured. The retina was assessed by electroretinogram (ERG), fundus photography, permeability studies, and trypsin digests for enumeration of acellular capillaries. The HFD cohort experienced hypercholesterolemia when compared to the LFD cohort, but not hyperglycemia. HFD mice developed a higher body weight (60.33 g vs. 30.17g, p < 0.0001) as well as a reduced insulin sensitivity index (9.418 vs. 62.01, p = 0.0002) compared to LFD controls. At 6 months, retinal functional testing demonstrated a reduction in a-wave and b-wave amplitudes. At 12 months, mice on HFD showed evidence of increased retinal nerve infarcts and vascular leakage, reduced vascular density, but no increase in number of acellular capillaries compared to LFD mice. In conclusion, the HFD mouse is a useful model for examining the effect of prediabetes and hypercholesterolemia on the retina. The HFD-induced changes appear to occur slower than those observed in type 2 diabetes (T2D) models but are consistent with other retinopathy models, showing neural damage prior to vascular changes.

Author Block: *Michael Dattilo, Brian Samuels, C Ethier. “Tree Shrew Spontaneous Retinal Venous Pulsation Changes Due to Changes in the Translaminar Pressure Difference.” Visual Impairment and Multi-Morbidity West 221/222 4:00pm – 5:45pm Moderators: Dawn K DeCarlo (University of Alabama at Birmingham), Ava K Bittner (Nova Southeastern University, College of Optometry), Sharon Ann Bentley (Queensland University of Technology), Walter Wittich (University of Montreal). Biochemistry and Molecular Mechanisms of Diabetic Retinopathy West Exhibition Hall 4:00pm – 5:45pm Posterboard Number: B0203 Abstract Author Block: *Samantha Prabakaran, Eleni Beli, Yaqian Duan, Sergio Li Calzi, Maria Grant. “Altered Circadian Metabolites in Type 2 Diabetic Mice with Diabetic Retinopathy.” Visual Fields, Psychophysics, and Electrophysiology West Exhibition Hall 4:00pm – 5:45pm Posterboard Number: A0627 Abstract Author Block: *Cyril Nyankerh, Sampson Abu, Lyne Racette. “No Racial Differences Exist in Visual Field Test-Retest Variability in Healthy Eyes.” Diabetic Retinopathy: Pathogenic Mechanisms West Exhibition Hall 4:00pm – 5:45pm Posterboard Number: B0219 Abstract Author Block: *Bright Asare-Bediako, Sunil Nooti, Sergio Li Calzi, Baskaran Athmanathan, Cristiano Pedrozo Vieira, Ana Leda Longhini, Mariana Dupont, Dibyendu Chakraborty, Prabhakara Nagareddy, Maria Grant. “In Vivo Retinal Structural Lesions, Functional Deficits and Peripheral Blood Cell Phenotype in Mice with High-Fat Diet-Induced Retinopathy.” Diabetic Retinopathy: Pathogenic Mechanisms West Exhibition Hall 4:00pm – 5:45pm Posterboard Number: B0235 Abstract Author Block: *Delaney McFarland, Sandra Hammer, Maria Grant, Julia Busik “Control of Retinal Cholesterol Levels by Fasting-Induced Activation of SIRT1-LXR Pathway in Diabetic Retinopathy.” New Theraputic Avenues in Diabetic Retinopathy West Exhibition Hall 4:00pm – 5:45pm Posterboard Number: B0250 Abstract Author Block: *Dibyendu Chakraborty, Cristiano Pedrozo Vieira, Sergio Calzi, Ana L. Longhini, Michael Boulton, Maria Grant. “Human Induced Pluripotent Stem Cell (hiPSC)Derived Mesoderm Promotes Retinal Function in a Type 2 Diabetes (db/db) Model.”Author Block: *Dibyendu Chakraborty, Cristiano Pedrozo Vieira, Sergio Calzi, Ana L. Longhini, Michael Boulton, Maria Grant. “Human Induced Pluripotent Stem Cell (hiPSC)Derived Mesoderm Promotes Retinal Function in a Type 2 Diabetes (db/db) Model.” New Theraputic Avenues in Diabetic Retinopathy West Exhibition Hall 4:00pm – 5:45pm Posterboard Number: B0259 Abstract Author Block: *Sandra Hammer, Julia Busik, Maria Grant. “The Role of the Sirt1/Lxr Signaling Axis in Retinal Endothelial Cell Inflammation and Metabolism.” Tuesday, April 30, 2019 Hyperreflective Intraretinal Spots in Macular Diseases: From Bench to Bedside West 223/224 7:00am – 8:30am Special Interest Group Abstract Author Block: *Elisabetta Pilotto, *David Sarraf, *Christine Curcio, *Ursula SchmidtErfurth, *Edoardo Midena. AMD Translational Studies West Exhibition Hall 8:45am – 10:30am Posterboard Number: A0269 Abstract Author Block: *Ankita Kotnala, David Anderson, Nathan Patterson, Jeffery Messinger, Christine Curcio, Kevin Schey. “Sample Preparation Effects on Retina Lipid Analysis by MALDI Imaging and LC-MS Technologies.” Diabetic Retinopathy East 2/3 10:00am – 10:15am Presentation Number: 2788 Abstract Author Block: *Cristiano Pedrozo Vieira, Ana Leda Longhini, Sergio Calzi, Bright AsareBediako, Mariana Dupont, Julia Busik, Maria Grant. “Targeting LXR Using N, N-dimethyl-3ßhydroxycholenamide (DMHCA) Provides Protection from Development of Diabetic Retinopathy

Although exercise derived activation of Nrf2 signaling augments myocardial antioxidant signaling, the molecular mechanisms underlying the benefits of moderate exercise training (MET) in the heart remain elusive. Here we hypothesized that exercise training stabilizes Nrf2-dependent antioxidant signaling, which then protects the myocardium from isoproterenol-induced damage. The present study assessed the effects of 6 weeks of MET on the Nrf2/antioxidant function, glutathione redox state, and injury in the myocardium of C57/BL6J mice that received isoproterenol (ISO; 50 mg/kg/day for 7 days). ISO administration significantly reduced the Nrf2 promoter activity (p < 0.05) and downregulated the expression of cardiac antioxidant genes (Gclc, Nqo1, Cat, Gsr, and Gst-μ) in the untrained (UNT) mice. Furthermore, increased oxidative stress with severe myocardial injury was evident in UNT+ISO when compared to UNT mice receiving PBS under basal condition. Of note, MET stabilized the Nrf2-promoter activity and upheld the expression of Nrf2-dependent antioxidant genes in animals receiving ISO, and attenuated the oxidative stress-induced myocardial damage. Echocardiography analysis revealed impaired diastolic ventricular function in UNT+ISO mice, but this was partially normalized in the MET animals. Interestingly, while there was a marginal reduction in ubiquitinated proteins in MET mice that received ISO, the pathological signs were attenuated along with near normal cardiac function in response to exercise training. Thus, moderate intensity exercise training conferred protection against ISO-induced myocardial injury by augmentation of Nrf2-antioxidant signaling and attenuation of isoproterenol-induced oxidative stress.

S100A8/A9 represents a novel biomarker and therapeutic target in sterile inflammatory diseases. Among the various S100 proteins, S100A8 and S100A9 have been shown to be the most important of all the damage-associated molecular pattern (DAMP) proteins in sterile inflammatory conditions such as diabetes, cardiovascular disease, autoimmune disorders, etc. We present here methods to quantify S100A8/A9 expression in various tissues in mouse models of myocardial infarction (MI) using flow cytometry (FC), immunofluorescence, quantitative real-time polymerase chain reaction (q-RT-PCR), and enzyme-linked immunosorbent assays (ELISA).

Rationale: There is incomplete knowledge of the impact of bone marrow cells on the gut microbiome and gut barrier function. Objective: We postulated that diabetes mellitus and systemic ACE2 (angiotensin-converting enzyme 2) deficiency would synergize to adversely impact both the microbiome and gut barrier function. Methods and Results: Bacterial 16S rRNA sequencing and metatranscriptomic analysis were performed on fecal samples from wild-type, ACE2(-/y), Akita (type 1 diabetes mellitus), and ACE2(-/y)-Akita mice. Gut barrier integrity was assessed by immunofluorescence, and bone marrow cell extravasation into the small intestine was evaluated by flow cytometry. In the ACE2(-/y)-Akita or Akita mice, the disrupted barrier was associated with reduced levels of myeloid angiogenic cells, but no increase in inflammatory monocytes was observed within the gut parenchyma. Genomic and metatranscriptomic analysis of the microbiome of ACE2(-/y)-Akita mice demonstrated a marked increase in peptidoglycan-producing bacteria. When compared with control cohorts treated with saline, intraperitoneal administration of myeloid angiogenic cells significantly decreased the microbiome gene expression associated with peptidoglycan biosynthesis and restored epithelial and endothelial gut barrier integrity. Also indicative of diabetic gut barrier dysfunction, increased levels of peptidoglycan and FABP-2 (intestinal fatty acid-binding protein 2) were observed in plasma of human subjects with type 1 diabetes mellitus (n=21) and type 2 diabetes mellitus (n=23) compared with nondiabetic controls (n=23). Using human retinal endothelial cells, we determined that peptidoglycan activates a noncanonical TLR-2 (Toll-like receptor 2) associated MyD88 (myeloid differentiation primary response protein 88)-ARNO (ADP-ribosylation factor nucleotide-binding site opener)-ARF6 (ADP-ribosylation factor 6) signaling cascade, resulting in destabilization of p120-catenin and internalization of VE-cadherin as a mechanism of deleterious impact of peptidoglycan on the endothelium. Conclusions: We demonstrate for the first time that the defect in gut barrier function and dysbiosis in ACE2(-/y)-Akita mice can be favorably impacted by exogenous administration of myeloid angiogenic cells.

Syndecans are transmembrane proteoglycans that, like integrins, bind to components of the extracellular matrix. Previously, we showed significant associations of genetic variants in the Syndecan-4 (SDC4) gene with intra-abdominal fat, fasting plasma glucose levels, and insulin sensitivity index in children, and with fasting serum triglyceride levels in healthy elderly subjects. An independent study also reported a correlation between SDC4 and the risk of coronary artery disease in middle-aged patients. Here, we investigated whether deletion of Sdc4 promotes metabolic derangements associated with diet-induced obesity by feeding homozygous male and female Sdc4-deficient (Sdc4-/-) mice and their age-matched wild-type (WT) mice a high-fat diet (HFD). We found that WT and Sdc4-/- mice gained similar weight. However, while no differences were observed in males, HFD-fed female Sdc4-/- mice exhibited a higher percentage of body fat mass than controls and displayed increased levels of plasma total cholesterol, triglyceride, and glucose, as well as reduced whole-body insulin sensitivity. Additionally, they had an increased adipocyte size and macrophage infiltration in the visceral adipose tissue, and higher triglyceride and fatty acid synthase levels in the liver. Together with our previous human genetic findings, these results provide evidence of an evolutionarily conserved role of SDC4 in adiposity and its complications.

Ischemic myocardial damage triggers granulopoiesis resulting in heightened production and recruitment of neutrophils to the ischemic heart. However, little is known about the myocardial molecular drivers that orchestrate with the BM and spleen to stimulate granulopoiesis. To investigate the mechanisms, we first mapped the abundance of different myeloid cell types in mice following myocardial ischemia (MI) and found a dominance of neutrophils in the blood and heart on day 1. This was not due to demargination or apoptosis but rather enhanced proliferation of granulocyte precursor cells in the BM and spleen. Attracted by cellular debris and release of DAMPs (e.g. S100A8/A9) from necrotic cells including the first wave of neutrophils, they rapidly undergo Nlrp3 inflammasome priming orchestrated by S100A8/A9 binding to TLR4 on incoming neutrophils in an autocrine/ paracrine manner. The primed neutrophils fully loaded with pro-IL-1b return to the BM in CXCR4-dependent manner, release IL-1b locally via gasdermin D pore formation in close proximity to IL-1b sensing stem and progenitor cells and stimulate granulopoiesis in a non-cell autonomous manner. Genetic and/ or pharmacological strategies aimed at disruption of S100A8/A9 or its downstream signaling cascade suppressed MI-induced granulopoiesis and improved cardiac remodelling and function. In patients with acute coronary syndrome, higher neutrophil count correlated positively with major adverse cardiovascular disease outcomes. These studies uncovered a rather complex network of signaling pathways harbored by neutrophils that self-govern their production following ischemic myocardial injury.

Ischemic myocardial damage triggers granulopoiesis resulting in heightened production and recruitment of neutrophils to the ischemic heart. However, little is known about the myocardial molecular d...

An impaired wound healing is one of the major health related problem in diabetic and non-diabetic patients around the globe. The pathogenic bacteria play a predominant role in delayed wound healing, owing to interaction in the wound area. In our previous work we developed green chemistry mediated copper oxide nanoparticles using Ficus religiosa leaf extract. In the present study we make an attempt to evaluate the anti-bacterial, and wound healing activity of green synthesized copper oxide nanoparticles in male Wistar Albino rats. The agar well diffusion assay revealed copper oxide nanoparticles have substantial inhibition activity against human pathogenic strains such as Klebsiella pneumoniae, Shigella dysenteriae, Staphylococcus aureus, Salmonella typhimurium and Escherichia coli, which were responsible for delayed wound healing process. Furthermore, the analyses results of wound closure, histopathology and protein profiling confirmed that the F. religiosa leaf extract tailored copper oxide nanoparticles have enhanced wound healing activity in Wistar Albino rats.