Louisville, KY, United States
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Xiang X.,University of Louisville | Liu Y.,University of Alabama at Birmingham | Zhuang X.,University of Louisville | Zhang S.,University of Alabama at Birmingham | And 5 more authors.
American Journal of Pathology | Year: 2010

Exosomes released from tumor cells having been shown to induce interleukin-6 release from myeloid-derived suppressor cells in a Toll-like receptor 2/Stat3-dependent manner. In this study, we show that exosomes released from tumor cells re-isolated from syngeneic mice are capable of inducing interleukin-6 in a Toll-like receptor 2-independent manner, whereas the data generated from exosomes of tumor cells having undergone numerous in vitro passages induce interleukin-6 in a Toll-like receptor 2-dependent manner. This discrepancy may be due to the source of tumor cells used to generate the exosomes for this study. These results suggest that exosomes released from tumor cells that are not within a tumor microenvironment may not realistically represent the role of tumor exosomes in vivo. This is an important consideration since frequently passing tumor cells in vivo is an accepted practice for studying tumor exosome-mediated inflammatory responses. Copyright © American Society for Investigative Pathology.


Lee L.-Y.,University of Kentucky | Yu J.,University of Louisville | Yu J.,Robley Rex Medical Center | Yu J.,Fudan University
Comprehensive Physiology | Year: 2014

Sensory nerves innervating the lung and airways play an important role in regulating various cardiopulmonary functions and maintaining homeostasis under both healthy and disease conditions. Their activities conducted by both vagal and sympathetic afferents are also responsible for eliciting important defense reflexes that protect the lung and body from potential health-hazardous effects of airborne particulates and chemical irritants. This article reviews the morphology, transduction properties, reflex functions, and respiratory sensations of these receptors, focusing primarily on recent findings derived from using new technologies such as neural immunochemistry, isolated airway-nerve preparation, cultured airway neurons, patch-clamp electrophysiology, transgenic mice, and other cellular and molecular approaches. Studies of the signal transduction of mechanosensitive afferents have revealed a new concept of sensory unit and cellular mechanism of activation, and identified additional types of sensory receptors in the lung. Chemosensitive properties of these lung afferents are further characterized by the expression of specific ligandgated ion channels on nerve terminals, ganglion origin, and responses to the action of various inflammatory cells, mediators, and cytokines during acute and chronic airway inflammation and injuries. Increasing interest and extensive investigations have been focused on uncovering the mechanisms underlying hypersensitivity of these airway afferents, and their role in the manifestation of various symptoms under pathophysiological conditions. Several important and challenging questions regarding these sensory nerves are discussed. Searching for these answers will be a critical step in developing the translational research and effective treatments of airway diseases. © 2014 American Physiological Society.


Garlie J.B.,University of Louisville | Hamid T.,University of Louisville | Gu Y.,University of Louisville | Ismahil M.A.,University of Louisville | And 3 more authors.
Basic Research in Cardiology | Year: 2011

The in vivo role of TNF signaling in the genesis of β-adrenergic receptor (β-AR)-mediated cardiac hypertrophy is unknown. Wild-type (WT), TNF receptor 1 (TNFR1)-/-and TNFR2-/-mice were given isoproterenol (ISO, 12.5 μg/kg/h) or saline (SAL) for 1 or 7 days. In WT mice, 7 days of ISO yielded chamber/myocyte hypertrophy and hyperdynamic function without hypertension or fibrosis. WT ISO hearts exhibited an early (1 day) pro-inflammatory response with significant (p <0.05) activation of nuclear factor (NF)-κB and activator protein 1 (AP-1) and upregulation of TNF, interleukin (IL)-1β and IL-6, inducible nitric oxide synthase (iNOS) and monocyte chemotactic protein-1 (MCP-1), together with increased anti-inflammatory IL-10. This response diminished markedly by 7 days. As compared with WT ISO mice, TNFR1-/-ISO mice exhibited significantly (p< 0.05) less NF-κB and AP-1 activation, less IL-1β, TNF, iNOS and MCP-1 upregulation, but greater IL-10 at 1 day. However, there were no differences in hypertrophy or contractility at 7 days. In contrast, TNFR2-/-ISO mice exhibited augmented NF-κB and AP-1 activation, increased IL-1β and diminished IL-10 expression at 1 day, and significant exaggeration of hypertrophy and less contractile augmentation at 7 days. Moreover, TNFR2-/-mice exposed to tenfold higher ISO doses displayed significant mortality. TNF signaling contributes to β-AR-mediated cardiac remodeling in vivo in a receptor-specific manner. Unopposed TNFR1 activation is pro-inflammatory, pro-hypertrophic and promotes functional decline. However, co-activation of TNFR2 during β-AR stress is antiinflammatory and counterbalances these deleterious effects. TNF modulatory strategies that maintain TNFR2 signaling may help prevent the detrimental long-term effects of β-AR stimulation in the heart. © Springer-Verlag 2011.


Timar C.I.,Semmelweis University | Lorincz A.M.,Semmelweis University | Csepanyi-Komi R.,Semmelweis University | Valyi-Nagy A.,Semmelweis University | And 8 more authors.
Blood | Year: 2013

Cell-derived vesicles represent a recently discovered mechanism for intercellular communication. We investigated their potential role in interaction of microbes with host organisms. We provide evidence that different stimuli induced isolated neutrophilic granulocytes to release microvesicles with different biologic properties. Only opsonized particles initiated the formation of microvesicles that were able to impair bacterial growth. The antibacterial effect of neutrophil-derived microvesicles was independent of production of toxic oxygen metabolites and opsonization or engulfment of the microbes, but depended on 2 integrin function, continuous actin remodeling, and on the glucose supply. Neutrophil-derived microvesicles were detected in the serum of healthy donors, and their number was significantly increased in the serum of bacteremic patients. We propose a new extracellular mechanism to restrict bacterial growth and dissemination. Copyright 2011 by The American Society of Hematology; all rights reserved. Copyright 2011 by The American Society of Hematology; all rights reserved.


Sethi I.,University of Louisville | Brier M.,University of Louisville | Brier M.,Robley Rex Medical Center | Dwyer A.,University of Louisville
Seminars in Dialysis | Year: 2013

At our institution, kidney biopsies are performed by an interventional nephrologist with standardized guidelines using real-time ultrasound. We hypothesized that patient factors could predict post biopsy complications. We did a retrospective review of 100 patients who underwent renal biopsy. Prebiopsy data obtained included demographics, blood pressure, laboratory studies, and kidney size. Biopsy procedure information was also recorded. Complications and post biopsy imaging was noted. A minor complication was defined as one not requiring intervention while a major complication required interventions like readmission or blood transfusion. The average age was 47years, 41 were men, 51 were black, 30 had diabetes, 42 were obese, and 81 had hypertension. Twenty-six patients had a complication; 14 minor and 12 major including 1 nephrectomy. Factors predictive of a complication were thrombocytopenia (p=0.002) and inpatient status (p=0.04). Drop in hemoglobin at 6hours was moderately sensitive and specific for a bleeding complication with an ROC of 0.723. Thrombocytopenia and inpatient status are risk factors for complications after renal biopsy. Serum creatinine, obesity, blood pressure, kidney size, needle size, and number of passes were not predictive of a major complication in our study. © 2013 Wiley Periodicals, Inc.


McLeish K.R.,University of Louisville | McLeish K.R.,Robley Rex Medical Center | Uriarte S.M.,University of Louisville | Tandon S.,University of Louisville | And 3 more authors.
Journal of Innate Immunity | Year: 2013

This study tested the hypothesis that priming the neutrophil respiratory burst requires both granule exocytosis and activation of the prolyl isomerase Pin1. Fusion proteins containing the TAT cell permeability sequence and either the SNARE domain of syntaxin-4 or the N-terminal SNARE domain of SNAP-23 were used to examine the role of granule subsets in TNF-mediated respiratory burst priming using human neutrophils. Concentration-inhibition curves for exocytosis of individual granule subsets and for priming of fMLF-stimulated superoxide release and phagocytosis-stimulated H2O2 production were generated. Maximal inhibition of priming ranged from 72 to 88%. Linear regression lines for inhibition of priming versus inhibition of exocytosis did not differ from the line of identity for secretory vesicles and gelatinase granules, while the slopes or the y-intercepts were different from the line of identity for specific and azurophilic granules. Inhibition of Pin1 reduced priming by 56%, while exocytosis of secretory vesicles and specific granules was not affected. These findings indicate that exocytosis of secretory vesicles and gelatinase granules and activation of Pin1 are independent events required for TNF-mediated priming of neutrophil respiratory burst. Copyright © 2013 S. Karger AG, Basel.


Uriarte S.M.,University of Louisville | Rane M.J.,University of Louisville | Merchant M.L.,University of Louisville | Jin S.,University of Louisville | And 4 more authors.
Shock | Year: 2013

Exocytosis of neutrophil granules contributes to acute lung injury (ALI) induced by infection or inflammation, suggesting that inhibition of neutrophil exocytosis in vivo could be a viable therapeutic strategy. This study was conducted to determine the effect of a cell-permeable fusion protein that inhibits neutrophil exocytosis (TAT-SNAP-23) on ALI using an immune complex deposition model in rats. The effect of inhibition of neutrophil exocytosis by intravenous administration of TAT-SNAP-23 on ALI was assessed by albumin leakage, neutrophil infiltration, lung histology, and proteomic analysis of bronchoalveolar lavage fluid (BALF). Administration of TAT-SNAP-23, but not TAT-control, significantly reduced albumin leakage, total protein levels in the BALF, and intra-alveolar edema and hemorrhage. Evidence that TAT-SNAP-23 inhibits neutrophil exocytosis included a reduction in plasma membrane CD18 expression by BALF neutrophils and a decrease in neutrophil granule proteins in BALF. Similar degree of neutrophil accumulation in the lungs and/or BALF suggests that TAT-SNAP-23 did not alter vascular endothelial cell function. Proteomic analysis of BALF revealed that components of the complement and coagulation pathways were significantly reduced in BALF from TAT-SNAP-23-treated animals. Our results indicate that administration of a TAT-fusion protein that inhibits neutrophil exocytosis reduces in vivo ALI. Targeting neutrophil exocytosis is a potential therapeutic strategy to ameliorate ALI. © 2013 by the Shock Society.


Levinson S.S.,Robley Rex Medical Center | Levinson S.S.,University of Louisville
Clinical Chemistry and Laboratory Medicine | Year: 2011

Articles have debated whether or not urine analysis remains valuable for identifying monoclonal gammopathies. A general impression is that the newer serum free light chain (FLC) assay is more analytically sensitive, more quantitative and simpler to perform. Many laboratory directors may have seized on the idea of eliminating urine analysis because it is a tedious procedure and requires expert interpretation while most laboratories can perform automated serum FLC assay. Others have concluded that urine immunofixation electrophoresis (IFE) optimizes the diagnostic sensitivity and should be included when there is a clinical indication. Here, I show that papers faulting urine analysis often used inappropriate urine methodology and this helps explain why there was misinterpretation. Moreover, the literature, shows urine IFE is often more sensitive for identifying low-level monoclonal FLC than the serum assay because urine IFE is as sensitive when performed appropriately and generally more specific. Besides, the reference range for serum FLC assay is unclear which is a great problem in assessing response to treatment and in identifying diseases when there is low concentration monoclonal FLC. I conclude that urine IFE remains important and is complementary to serum FLC assay, although the best algorithms for use remains to be elucidated. © 2011 by Walter de Gruyter Berlin Boston 2011.


Levinson S.S.,Robley Rex Medical Center | Levinson S.S.,University of Louisville
Clinica Chimica Acta | Year: 2011

Background: With the introduction of a new sensitive serum assay for monoclonal free light chains (FLC), there is a question as to whether or not traditional urine immunofixation electrophoresis (IFE) is still necessary. Therapy for B-cell disease has greatly improved in recent time so that noninvasive biomarkers that suggest complete remissions have become increasingly important. A Consensus Opinion stated that because of methodological imperfections, at concentrations of FLC < 100. mg/l, urine IFE should supplement serum FLC ratio. Methods: Examples are presented from a review of laboratory results and medical records from 3 patients on treatment after diagnosis of amyloidosis AL and multiple myeloma. Results: Monoclonal FLC was identified in urine by IFE while the κ/λ ratio from the serum FLC assay was within the reference range. Conclusion: The results from these 3 cases suggest that as the FLC concentration drops, the serum FLC results may become more ambiguous. These results are consistent with guidelines cautioning about conclusions drawn from the serum FLC assay alone. Although more study is needed, the examples presented here illustrate that traditional urine IFE appear to adds important information regarding potential complete remissions and remains an important complementary assay to the serum FLC assay. © 2011 Elsevier B.V.


Levinson S.S.,Robley Rex Medical Center | Levinson S.S.,University of Louisville
Clinica Chimica Acta | Year: 2012

Background: Polyneuropathy organomegaly, endocrinopathy, monoclonal (M) gammopathy, skin syndrome (POEMS) may be difficult to diagnose as a result of methodological and clinical idiosyncrasies. Four of eleven criteria (M-protein, VEGF, endocrinopathy and thrombocytosis/polycythemia) are closely associated with clinical laboratory testing. POEMS has been largely associated with λ-M-gammopathies. Vascular endothelial growth factor (VEGF) is a recent addition to the major diagnostic criteria. VEGF may alter vascular permeability causing some manifestations of POEMS. Methods: Review of the literature that focuses on clinical laboratory issues - endocrinological findings, identification of monoclonal gammopathies by electrophoresis and case demonstration. Results: Based on the criterion of VEGF, POEMS was diagnosed in a patient with a κ-M-gammopathy. Conclusions: Low-level IgA monoclonal proteins that are common in POEMS are often difficult to identify by serum electrophoresis (SPE). Immunofixation electrophoresis is required when polyneuropaties are investigated and an M-protein is not identified by SPE. VEGF may improve the sensitivity for diagnosis of POEMS and findings from capillary leak syndrome which are also associated with elevated VEGF and M-gammopathy suggests that κ-M-gammopathies may be implicated more often. It is demonstrated that due to computerized records, the laboratory practitioner is well suited to help the clinician make this complicated diagnosis. © 2012.

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