... IN ACUTE VENOUS THROMBOEMBOLISM Julio A. Chirinos MD* Gustavo A. Heresi MD Hermes Velasquez M... more ... IN ACUTE VENOUS THROMBOEMBOLISM Julio A. Chirinos MD* Gustavo A. Heresi MD Hermes Velasquez MD Lawrence Horstman BS Joaquin J ... RISK Steven A. Baroletti PharmD* John Fanikos RPh, MBA Jennifer Cina PharmD Karen Fiumara PharmD Kristin Munz PharmD ...
Heme is a complex of iron and the tetrapyrrole protoporphyrin IX with essential functions in aero... more Heme is a complex of iron and the tetrapyrrole protoporphyrin IX with essential functions in aerobic organisms. Heme is the prosthetic group of hemoproteins such as hemoglobin and myoglobin, which are crucial for reversible oxygen binding and transport. By contrast, high levels of free heme, which may occur in various pathophysiological conditions, are toxic via pro-oxidant, pro-inflammatory and cytotoxic effects. The toxicity of heme plays a major role for the pathogenesis of prototypical hemolytic disorders including sickle cell disease and malaria. Moreover, there is increasing appreciation that detrimental effects of heme may also be critically involved in diseases, which usually are not associated with hemolysis such as severe sepsis and atherosclerosis. In mammalians homeostasis of heme and its potential toxicity are primarily controlled by two physiological systems. First, the scavenger protein hemopexin (Hx) non-covalently binds extracellular free heme with high affinity and...
Heme is a ubiquitous compound of human tissues, and it is involved in cellular physiology and met... more Heme is a ubiquitous compound of human tissues, and it is involved in cellular physiology and metabolism. Once released from the cell, free heme oxidizes to the ferric state (hemin). High levels of hemin can cause oxidative stress and inflammation if not neutralized immediately by specialized scavenger proteins. Human alpha1-antitrypsin (A1AT), an acute-phase glycoprotein and important inhibitor of neutrophil proteases, is also a hemin-binding protein. A short-term exposure of freshly isolated human blood neutrophils to 4 µM hemin results in cell spreading, surface expression of filament protein, vimentin, free radical production, expression of heme oxygenase-1 (HO-1), release of IL-8, and enhanced neutrophil adhesion to human endothelial cells. Consequently, the phosphorylation of protein kinase C (PKC) occurs after 25 min. Under the same experimental conditions, addition of 1 mg/ml A1AT markedly reduces or abolishes neutrophil-activating effects of hemin and prevents PKC phosphory...
Journal of immunology (Baltimore, Md. : 1950), Jan 15, 2017
Peroxisomes are proposed to play an important role in the regulation of systemic inflammation; ho... more Peroxisomes are proposed to play an important role in the regulation of systemic inflammation; however, the functional role of these organelles in inflammatory responses of myeloid immune cells is largely unknown. In this article, we demonstrate that the nonclassical peroxisome proliferator 4-phenyl butyric acid is an efficient inducer of peroxisomes in various models of murine macrophages, such as primary alveolar and peritoneal macrophages and the macrophage cell line RAW264.7, but not in primary bone marrow-derived macrophages. Further, proliferation of peroxisomes blocked the TLR4 ligand LPS-induced proinflammatory response, as detected by the reduced induction of the proinflammatory protein cyclooxygenase (COX)-2 and the proinflammatory cytokines TNF-α, IL-6, and IL-12. In contrast, disturbing peroxisome function by knockdown of peroxisomal gene Pex14 or Mfp2 markedly increased the LPS-dependent upregulation of the proinflammatory proteins COX-2 and TNF-α. Specifically, inducti...
Antibody-mediated rejection (AMR) is a key limiting factor for long-term graft survival in solid ... more Antibody-mediated rejection (AMR) is a key limiting factor for long-term graft survival in solid organ transplantation. Human leukocyte antigen (HLA) class I (HLA I) antibodies (Abs) play a major role in the pathogenesis of AMR via their interactions with HLA molecules on vascular endothelial cells (ECs). The antioxidant enzyme heme oxygenase (HO)-1 has anti-inflammatory functions in the endothelium. As complement-independent effects of HLA I Abs can activate ECs, it was the goal of the current study to investigate the role of HO-1 on activation of human ECs by HLA I Abs. In cell cultures of various primary human macro- and microvascular ECs treatment with monoclonal pan- and allele-specific HLA I Abs up-regulated the expression of inducible proinflammatory adhesion molecules and chemokines (vascular cell adhesion molecule-1 [VCAM-1], intercellular cell adhesion molecule-1 [ICAM-1], interleukin-8 [IL-8] and monocyte chemotactic protein 1 [MCP-1]). Pharmacological induction of HO-1 with cobalt-protoporphyrin IX reduced, whereas inhibition of HO-1 with either zinc-protoporphyrin IX or siRNA-mediated knockdown increased HLA I Ab-dependent up-regulation of VCAM-1. Treatment with two carbon monoxide (CO)-releasing molecules, which liberate the gaseous HO product CO, blocked HLA I Ab-dependent EC activation. Finally, in an in vitro adhesion assay exposure of ECs to HLA I Abs led to increased monocyte binding, which was counteracted by up-regulation of HO-1. In conclusion, HLA I Ab-dependent EC activation is modulated by endothelial HO-1 and targeted induction of this enzyme may be a novel therapeutic approach for the treatment of AMR in solid organ transplantation.
Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive, lung disease of unknown cause. Rea... more Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive, lung disease of unknown cause. Reactive oxygen species (ROS), accumulation and proinflammatory mediators are considered as pathogenic mechanisms of IPF. In this respect, it is of interest that peroxisomes degrade ROS and that patients with peroxisomal biogenesis disorders develop liver fibrosis during their first year of life. We hypothesized that peroxisomal enzymes or biogenesis proteins might be affected in IPF, leading to aggravation of the disease. Methods and Results: Human donor (DO) and IPF fibroblasts were obtained from University of Giessen Lung Center. They were analyzed by different methods: Immunofluorescence, Immunohistochemistry, RT-PCR, western blotting, CM-H2DCFDA-FACS for ROS measurement and TUNEL assay for apoptosis. Our results revealed significant downregulation of peroxisomal metabolism, (peroxisomal acyl-coenzyme A oxidase 1-ACOX1) and biogenesis (peroxisomal membrane protein-PEX13), both in lung t...
Journal of immunology (Baltimore, Md. : 1950), 2015
Spermatogenic cells express cell-specific molecules with the potential to be seen as "foreig... more Spermatogenic cells express cell-specific molecules with the potential to be seen as "foreign" by the immune system. Owing to the time difference between their appearance in puberty and the editing of the lymphocyte repertoire around birth, local adaptations of the immune system coined immune privilege are required to confer protection from autoattack. Testicular macrophages (TM) play an important role in maintaining testicular immune privilege and display reduced proinflammatory capacity compared with other macrophages. However, the molecular mechanism underlying this macrophage phenotype remained elusive. We demonstrate that TM have a lower constitutive expression of TLR pathway-specific genes compared with peritoneal macrophages. Moreover, in TM stimulated with LPS, the NF-κB signaling pathway is blocked due to lack of IκBα ubiquitination and, hence, degradation. Instead, challenge of TM with LPS or polyinosinic-polycytidylic acid induces MAPK, AP-1, and CREB signaling ...
Proceedings of the National Academy of Sciences of the United States of America, Jan 6, 2015
Idiopathic pulmonary fibrosis (IPF) is a devastating disease, and its pathogenic mechanisms remai... more Idiopathic pulmonary fibrosis (IPF) is a devastating disease, and its pathogenic mechanisms remain incompletely understood. Peroxisomes are known to be important in ROS and proinflammatory lipid degradation, and their deficiency induces liver fibrosis. However, altered peroxisome functions in IPF pathogenesis have never been investigated. By comparing peroxisome-related protein and gene expression in lung tissue and isolated lung fibroblasts between human control and IPF patients, we found that IPF lungs exhibited a significant down-regulation of peroxisomal biogenesis and metabolism (e.g., PEX13p and acyl-CoA oxidase 1). Moreover, in vivo the bleomycin-induced down-regulation of peroxisomes was abrogated in transforming growth factor beta (TGF-β) receptor II knockout mice indicating a role for TGF-β signaling in the regulation of peroxisomes. Furthermore, in vitro treatment of IPF fibroblasts with the profibrotic factors TGF-β1 or tumor necrosis factor alpha (TNF-α) was found to do...
Smart phones are one of the most troublesome technologies that exist today and steadily gaining p... more Smart phones are one of the most troublesome technologies that exist today and steadily gaining popularity with better connectivity, functionality and productivity. The ever-increasing complexity of these smart phones gives a new level of threats. Android is one of the newer operating systems based on smart phones, is emerging as a significant force within the highly competitive smart phone market. Android phones stores an enormous amount of data that can be stored locally or remotely and allows forensic analysts to acquire these data and evidence, collecting this valuable information with regard to the investigation. There are number of forensic tools both open source and closed source are available in the market for the purpose of extracting data from the Android devices.
The key focus of this paper is to evaluate the performances of extracting tools that support the Android phones. This research project reviews the literature in the field of digital forensic and methods of retrieving data and discusses the criteria and methodology to be achieved for a meaningful evaluation. Three different software tools are evaluated along with the manual method of extraction on two Android phones. The actual forensic data within the mobile devices were identified and counted. All the three software tools were tested on both phones, to find out how much data can be extracted and these results were compared and evaluated based on the actual data on the phones.
... IN ACUTE VENOUS THROMBOEMBOLISM Julio A. Chirinos MD* Gustavo A. Heresi MD Hermes Velasquez M... more ... IN ACUTE VENOUS THROMBOEMBOLISM Julio A. Chirinos MD* Gustavo A. Heresi MD Hermes Velasquez MD Lawrence Horstman BS Joaquin J ... RISK Steven A. Baroletti PharmD* John Fanikos RPh, MBA Jennifer Cina PharmD Karen Fiumara PharmD Kristin Munz PharmD ...
Heme is a complex of iron and the tetrapyrrole protoporphyrin IX with essential functions in aero... more Heme is a complex of iron and the tetrapyrrole protoporphyrin IX with essential functions in aerobic organisms. Heme is the prosthetic group of hemoproteins such as hemoglobin and myoglobin, which are crucial for reversible oxygen binding and transport. By contrast, high levels of free heme, which may occur in various pathophysiological conditions, are toxic via pro-oxidant, pro-inflammatory and cytotoxic effects. The toxicity of heme plays a major role for the pathogenesis of prototypical hemolytic disorders including sickle cell disease and malaria. Moreover, there is increasing appreciation that detrimental effects of heme may also be critically involved in diseases, which usually are not associated with hemolysis such as severe sepsis and atherosclerosis. In mammalians homeostasis of heme and its potential toxicity are primarily controlled by two physiological systems. First, the scavenger protein hemopexin (Hx) non-covalently binds extracellular free heme with high affinity and...
Heme is a ubiquitous compound of human tissues, and it is involved in cellular physiology and met... more Heme is a ubiquitous compound of human tissues, and it is involved in cellular physiology and metabolism. Once released from the cell, free heme oxidizes to the ferric state (hemin). High levels of hemin can cause oxidative stress and inflammation if not neutralized immediately by specialized scavenger proteins. Human alpha1-antitrypsin (A1AT), an acute-phase glycoprotein and important inhibitor of neutrophil proteases, is also a hemin-binding protein. A short-term exposure of freshly isolated human blood neutrophils to 4 µM hemin results in cell spreading, surface expression of filament protein, vimentin, free radical production, expression of heme oxygenase-1 (HO-1), release of IL-8, and enhanced neutrophil adhesion to human endothelial cells. Consequently, the phosphorylation of protein kinase C (PKC) occurs after 25 min. Under the same experimental conditions, addition of 1 mg/ml A1AT markedly reduces or abolishes neutrophil-activating effects of hemin and prevents PKC phosphory...
Journal of immunology (Baltimore, Md. : 1950), Jan 15, 2017
Peroxisomes are proposed to play an important role in the regulation of systemic inflammation; ho... more Peroxisomes are proposed to play an important role in the regulation of systemic inflammation; however, the functional role of these organelles in inflammatory responses of myeloid immune cells is largely unknown. In this article, we demonstrate that the nonclassical peroxisome proliferator 4-phenyl butyric acid is an efficient inducer of peroxisomes in various models of murine macrophages, such as primary alveolar and peritoneal macrophages and the macrophage cell line RAW264.7, but not in primary bone marrow-derived macrophages. Further, proliferation of peroxisomes blocked the TLR4 ligand LPS-induced proinflammatory response, as detected by the reduced induction of the proinflammatory protein cyclooxygenase (COX)-2 and the proinflammatory cytokines TNF-α, IL-6, and IL-12. In contrast, disturbing peroxisome function by knockdown of peroxisomal gene Pex14 or Mfp2 markedly increased the LPS-dependent upregulation of the proinflammatory proteins COX-2 and TNF-α. Specifically, inducti...
Antibody-mediated rejection (AMR) is a key limiting factor for long-term graft survival in solid ... more Antibody-mediated rejection (AMR) is a key limiting factor for long-term graft survival in solid organ transplantation. Human leukocyte antigen (HLA) class I (HLA I) antibodies (Abs) play a major role in the pathogenesis of AMR via their interactions with HLA molecules on vascular endothelial cells (ECs). The antioxidant enzyme heme oxygenase (HO)-1 has anti-inflammatory functions in the endothelium. As complement-independent effects of HLA I Abs can activate ECs, it was the goal of the current study to investigate the role of HO-1 on activation of human ECs by HLA I Abs. In cell cultures of various primary human macro- and microvascular ECs treatment with monoclonal pan- and allele-specific HLA I Abs up-regulated the expression of inducible proinflammatory adhesion molecules and chemokines (vascular cell adhesion molecule-1 [VCAM-1], intercellular cell adhesion molecule-1 [ICAM-1], interleukin-8 [IL-8] and monocyte chemotactic protein 1 [MCP-1]). Pharmacological induction of HO-1 with cobalt-protoporphyrin IX reduced, whereas inhibition of HO-1 with either zinc-protoporphyrin IX or siRNA-mediated knockdown increased HLA I Ab-dependent up-regulation of VCAM-1. Treatment with two carbon monoxide (CO)-releasing molecules, which liberate the gaseous HO product CO, blocked HLA I Ab-dependent EC activation. Finally, in an in vitro adhesion assay exposure of ECs to HLA I Abs led to increased monocyte binding, which was counteracted by up-regulation of HO-1. In conclusion, HLA I Ab-dependent EC activation is modulated by endothelial HO-1 and targeted induction of this enzyme may be a novel therapeutic approach for the treatment of AMR in solid organ transplantation.
Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive, lung disease of unknown cause. Rea... more Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive, lung disease of unknown cause. Reactive oxygen species (ROS), accumulation and proinflammatory mediators are considered as pathogenic mechanisms of IPF. In this respect, it is of interest that peroxisomes degrade ROS and that patients with peroxisomal biogenesis disorders develop liver fibrosis during their first year of life. We hypothesized that peroxisomal enzymes or biogenesis proteins might be affected in IPF, leading to aggravation of the disease. Methods and Results: Human donor (DO) and IPF fibroblasts were obtained from University of Giessen Lung Center. They were analyzed by different methods: Immunofluorescence, Immunohistochemistry, RT-PCR, western blotting, CM-H2DCFDA-FACS for ROS measurement and TUNEL assay for apoptosis. Our results revealed significant downregulation of peroxisomal metabolism, (peroxisomal acyl-coenzyme A oxidase 1-ACOX1) and biogenesis (peroxisomal membrane protein-PEX13), both in lung t...
Journal of immunology (Baltimore, Md. : 1950), 2015
Spermatogenic cells express cell-specific molecules with the potential to be seen as "foreig... more Spermatogenic cells express cell-specific molecules with the potential to be seen as "foreign" by the immune system. Owing to the time difference between their appearance in puberty and the editing of the lymphocyte repertoire around birth, local adaptations of the immune system coined immune privilege are required to confer protection from autoattack. Testicular macrophages (TM) play an important role in maintaining testicular immune privilege and display reduced proinflammatory capacity compared with other macrophages. However, the molecular mechanism underlying this macrophage phenotype remained elusive. We demonstrate that TM have a lower constitutive expression of TLR pathway-specific genes compared with peritoneal macrophages. Moreover, in TM stimulated with LPS, the NF-κB signaling pathway is blocked due to lack of IκBα ubiquitination and, hence, degradation. Instead, challenge of TM with LPS or polyinosinic-polycytidylic acid induces MAPK, AP-1, and CREB signaling ...
Proceedings of the National Academy of Sciences of the United States of America, Jan 6, 2015
Idiopathic pulmonary fibrosis (IPF) is a devastating disease, and its pathogenic mechanisms remai... more Idiopathic pulmonary fibrosis (IPF) is a devastating disease, and its pathogenic mechanisms remain incompletely understood. Peroxisomes are known to be important in ROS and proinflammatory lipid degradation, and their deficiency induces liver fibrosis. However, altered peroxisome functions in IPF pathogenesis have never been investigated. By comparing peroxisome-related protein and gene expression in lung tissue and isolated lung fibroblasts between human control and IPF patients, we found that IPF lungs exhibited a significant down-regulation of peroxisomal biogenesis and metabolism (e.g., PEX13p and acyl-CoA oxidase 1). Moreover, in vivo the bleomycin-induced down-regulation of peroxisomes was abrogated in transforming growth factor beta (TGF-β) receptor II knockout mice indicating a role for TGF-β signaling in the regulation of peroxisomes. Furthermore, in vitro treatment of IPF fibroblasts with the profibrotic factors TGF-β1 or tumor necrosis factor alpha (TNF-α) was found to do...
Smart phones are one of the most troublesome technologies that exist today and steadily gaining p... more Smart phones are one of the most troublesome technologies that exist today and steadily gaining popularity with better connectivity, functionality and productivity. The ever-increasing complexity of these smart phones gives a new level of threats. Android is one of the newer operating systems based on smart phones, is emerging as a significant force within the highly competitive smart phone market. Android phones stores an enormous amount of data that can be stored locally or remotely and allows forensic analysts to acquire these data and evidence, collecting this valuable information with regard to the investigation. There are number of forensic tools both open source and closed source are available in the market for the purpose of extracting data from the Android devices.
The key focus of this paper is to evaluate the performances of extracting tools that support the Android phones. This research project reviews the literature in the field of digital forensic and methods of retrieving data and discusses the criteria and methodology to be achieved for a meaningful evaluation. Three different software tools are evaluated along with the manual method of extraction on two Android phones. The actual forensic data within the mobile devices were identified and counted. All the three software tools were tested on both phones, to find out how much data can be extracted and these results were compared and evaluated based on the actual data on the phones.
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Papers by Vijith Vijayan
The key focus of this paper is to evaluate the performances of extracting tools that support the Android phones. This research project reviews the literature in the field of digital forensic and methods of retrieving data and discusses the criteria and methodology to be achieved for a meaningful evaluation. Three different software tools are evaluated along with the manual method of extraction on two Android phones. The actual forensic data within the mobile devices were identified and counted. All the three software tools were tested on both phones, to find out how much data can be extracted and these results were compared and evaluated based on the actual data on the phones.
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The key focus of this paper is to evaluate the performances of extracting tools that support the Android phones. This research project reviews the literature in the field of digital forensic and methods of retrieving data and discusses the criteria and methodology to be achieved for a meaningful evaluation. Three different software tools are evaluated along with the manual method of extraction on two Android phones. The actual forensic data within the mobile devices were identified and counted. All the three software tools were tested on both phones, to find out how much data can be extracted and these results were compared and evaluated based on the actual data on the phones.
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