Antibiotic resistance is one of the greatest crises in human medicine. Increased incidents of ant... more Antibiotic resistance is one of the greatest crises in human medicine. Increased incidents of antibiotic resistance are linked to clinical overuse and overreliance on antibiotics. Among the ESKAPE pathogens, Acinetobacter baumannii, especially carbapenem-resistant isolates, has emerged as a significant threat in the context of blood, urinary tract, lung, and wound infections. Therefore, new approaches that limit the emergence of antibiotic resistant A. baumannii are urgently needed. Recently, we have shown that random peptide mixtures (RPMs) are an attractive alternative class of drugs to antibiotics with strong safety and pharmacokinetic profiles. RPMs are antimicrobial peptide mixtures produced by incorporating two amino acids at each coupling step, rendering them extremely diverse but still defined in their overall composition, chain length, and stereochemistry. The extreme diversity of RPMs may prevent bacteria from evolving resistance rapidly. Here, we demonstrated that RPMs ra...
A polarized macrophage response into inflammatory (M1) or regenerative/anti-inflammatory (M2) phe... more A polarized macrophage response into inflammatory (M1) or regenerative/anti-inflammatory (M2) phenotypes is critical in host response to multiple intracellular bacterial infections. Ehrlichia is an obligate Gram-negative intracellular bacterium that causes human monocytic ehrlichiosis (HME): a febrile illness that may progress to fatal sepsis with multi-organ failure. We have shown that liver injury and Ehrlichia-induced sepsis occur due to dysregulated inflammation. Here, we investigated the contribution of macrophages to Ehrlichia-induced sepsis using murine models of mild and fatal ehrlichiosis. Lethally-infected mice showed accumulation of M1 macrophages (iNOS-positive) in the liver. In contrast, non-lethally infected mice showed polarization of M2 macrophages and their accumulation in peritoneum, but not in the liver. Predominance of M1 macrophages in lethally-infected mice was associated with expansion of IL-17-producing T, NK, and NKT cells. Consistent with the in vivo data, ...
Human monocytic ehrlichiosis, a tick transmitted infection, ranges in severity from apparently su... more Human monocytic ehrlichiosis, a tick transmitted infection, ranges in severity from apparently subclinical to fatal toxic shock-like disease. Models in immunocompetent mice range from abortive to uniformly lethal infection, depending on the Ehrlichia species, inoculum dose, and inoculation route. Effective immunity is mediated by CD4+ T lymphocytes and gamma interferon. Lethal infection occurs with early overproduction of proinflammatory cytokines and overproduction of TNF alpha and IL-10 by CD8+ T lymphocytes. Furthermore, fatal ehrlichiosis is associated with TLR 9/MyD88 signaling, upregulation of several inflammasome complexes, and secretion of IL-1 beta, IL-1 alpha, and IL-18 by hepatic mononuclear cells, thus suggesting activation of canonical and noncanonical inflammasome pathways, a deleterious role of IL-18, and a protective role of caspase 1. Autophagy promotes ehrlichial infection, whereas MyD88 signaling hinders ehrlichial infection by inhibiting autophagy induction and f...
Uterine fibroids (UF; aka leiomyoma, myomas) are the most common benign tumors of female reproduc... more Uterine fibroids (UF; aka leiomyoma, myomas) are the most common benign tumors of female reproductive tract. They are highly prevalent, with 70 to 80% of women burdened by the end of their reproductive years. Fibroids are a leading cause of pelvic pain, abnormal vaginal bleeding, pelvic bulk symptoms, miscarriage, and infertility. They are the leading indication for hysterectomy, and costs exceed 34 billion dollars annually in the United States alone. Recently, somatic mutations in exons 1 and 2 of Med12 gene emerged as common UF driver mutations. Unfortunately, the detailed etiology of UF is not fully realized. Particularly, very little is known about possible dysregulation of inflammatory and immune processes and their possible contribution to UF pathogenesis. The notion on possible impact of altered estrogen and progesterone signaling in UF on inflammatory responses and DNA repair machinery that can conceivably lead to tumor-specific somatic mutation is indeed an intriguing conce...
Our murine models of human monocytic ehrlichiosis (HME) have shown that severe and fatal ehrlichi... more Our murine models of human monocytic ehrlichiosis (HME) have shown that severe and fatal ehrlichiosis is due to generation of pathogenic T cell responses causing immunopathology and multi-organ failure. However, the early events in the liver, the main site of infection, are not well understood. In this study, we examined the liver transcriptome during the course of lethal and nonlethal infections caused by Ixodes ovatus Ehrlichia and Ehrlichia muris, respectively. On day 3 post-infection (p.i.), although most host genes were down regulated in the two groups of infected mice compared to naı̈ve counterparts, lethal infection induced significantly higher expression of caspase 1, caspase 4, nucleotide binding oligomerization domain-containing proteins (Nod1), tumor necrosis factor-alpha, interleukin 10, and CCL7 compared to nonlethal infection. On day 7 p.i., lethal infection induced highly significant upregulation of type-1 interferon, several inflammatory cytokines and chemokines, whi...
Inflammasomes are an important innate immune host defense against intracellular microbial infecti... more Inflammasomes are an important innate immune host defense against intracellular microbial infection. Activation of inflammasomes by microbial or host ligands results in cleavage of caspase‐1 (canonical pathway) or caspase‐11 (noncanonical pathway), release of interleukin (IL)‐1β, IL‐18, high mobility group box 1 (HMGB1), and inflammatory cell death known as pyroptosis. Ehrlichia are obligate, intracellular, gram‐negative bacteria that lack lipopolysaccharide but cause potentially life‐threatening monocytic ehrlichiosis in humans and mice that is characterized by liver injury followed by sepsis and multiorgan failure. Employing murine models of mild and fatal ehrlichiosis caused by infection with mildly and highly virulent Ehrlichia muris (EM) and Ixodes ovatus Ehrlichia (IOE), respectively, we have previously shown that IOE infection triggers type I interferon (IFN‐I) response and deleterious caspase‐11 activation in liver tissues, which promotes liver injury and sepsis. In this study, we examined the contribution of IFN‐I signaling in hepatocytes (HCs) to Ehrlichia‐induced liver injury. Compared to EM infection, we found that IOE enter and replicate in vitro cultured primary murine HCs and induce secretion of IFNβ and several chemokines, including regulated upon activation, normal T‐cell expressed, and secreted (RANTES), monocyte chemoattractant protein 1 (MCP1), monokine induced by gamma (MIG)/chemokine (C‐X‐C motif) ligand 9 (CXCL9), macrophage inflammatory protein 1 alpha (MIP1α), keratinocyte‐derived chemokine (KC), and granulocyte‐macrophage colony‐stimulating factor (GM‐CSF). Notably, in vitro stimulation of uninfected and Ehrlichia‐infected HCs with recombinant IFNβ triggered activation of caspase‐1/11, cytosolic translocation of HMGB1, and enhanced autophagy and intracellular bacterial replication. Secretion of HMGB1 by IOE‐infected HCs was dependent on caspase‐11. Primary HCs from IOE‐ but not EM‐infected mice also expressed active caspase‐1/11. Conclusion: HC‐specific IFN‐I signaling may exacerbate liver pathology during infection with obligate intracellular Ehrlichia by promoting bacterial replication and detrimental caspase‐11‐mediated inflammasome activation.
The performance of a fully automated, random access, enhanced chemiluminescence immunoassay (Orth... more The performance of a fully automated, random access, enhanced chemiluminescence immunoassay (Ortho/ ECi) for the detection of antibody to hepatitis C virus (HCV) (anti-HCV), HBsAg, and antibody to HBsAg (anti-HBsAg), in human serum was compared to a Abbott second-generation enzyme immunoassay (EIA 2.0). The Ortho/ECi assays employ an immunometric technique with enhanced chemiluminescence for optimal assay performance. With regard to the study of clinical laboratory performance, six groups of sera prescreened with Abbott EIAs were assayed: anti-HCV-negative samples (n ؍ 318), anti-HCV-positive samples (n ؍ 177), anti-HBsAg-negative samples (n ؍ 241), anti-HBsAg-positive samples (n ؍ 239), HBsAg-positive samples (n ؍ 158), and HBsAg-negative samples (n ؍ 312). Sera with discrepant results in the two serological assays were resolved by confirmatory tests. Sera with indeterminate results by one or more confirmatory tests were evaluated by reviewing medical records. The overall concordance between the Ortho/ECi assay and the Abbott EIA were 97.78, 93.54, and 97.66% for anti-HCV antibodies, anti-HBsAg antibodies, and HBsAg, respectively. After resolving the discrepancies, the specificities of the new assay for anti-HCV and anti-HBsAg antibodies and HBsAg were 98.1, 92.8, and 100%, respectively. The sensitivities of the new assay for anti-HCV, anti-HBsAg, and HBsAg were 100, 98.8, and 97.4%, respectively. In conclusion, The Ortho/ECi assays for diagnosis of HCV and hepatitis B virus (HBV) infections are highly specific and sensitive assays. The rapid turnaround time, random access, full automation, and high throughput make it an effective assay system for clinical laboratory diagnosis of HCV and HBV infections.
Purpose. The number of individuals affected by job stress is growing day by day in almost every i... more Purpose. The number of individuals affected by job stress is growing day by day in almost every industry. According to Health and Safety Executive (2006) workplace stress is now the fastest growing cause of absence from work. The current study aims to verify the effectiveness of management stress in performance level of manager's productivity of youth centers in Cairo. Methods. The sample contains 40 mangers and 80 sports specialists, to collect the research data the researchers have built a questionnaire to measure the administrative empowerment which contains 5 factors, the initial questionnaire consists of 106 items. Results. statistical analyses showed that the stress natural was connected of planning and make decision , organize, control and guidance ( according its importance). Conclusions. Finally, the leaders in youth centers face many of stress which affected of their performance.
Background Polycystic ovary syndrome (PCOS) is the most common endocrine and metabolic disorder i... more Background Polycystic ovary syndrome (PCOS) is the most common endocrine and metabolic disorder in reproductive-age women. Excessive inflammation and elevated androgen production from ovarian theca cells are key features of PCOS. Human bone marrow mesenchymal stem cells (BM-hMSC) and their secreted factors (secretome) exhibit robust anti-inflammatory capabilities in various biological systems. We evaluated the therapeutic efficacy of BM-hMSC and its secretome in both in vitro and in vivo PCOS models. Methods For in vitro experiment, we treated conditioned media from BM-hMSC to androgen-producing H293R cells and analyzed androgen-producing gene expression. For in vivo experiment, BM-hMSC were implanted into letrozole (LTZ)-induced PCOS mouse model. BM-hMSC effect in androgen-producing cells or PCOS model mice was assessed by monitoring cell proliferation (immunohistochemistry), steroidogenic gene expression (quantitative real-time polymerase chain reaction [qRT-PCR] and Western blot,...
M1-M2 Macrophages Polarization Macrophages are innate immune cells that play a key role in regula... more M1-M2 Macrophages Polarization Macrophages are innate immune cells that play a key role in regulation of innate and adaptive immune responses against infections with several pathogens as they respond to pathogens and tissue injury, serve as antigen presenting cells priming the adaptive immune response, drive inflammation and host defense as well as repairing damaged tissue [5-7]. Macrophages reside in almost all of the lymphoid and non-lymphoid tissues [8]. Two major lineages are currently known: cells that are derived from myeloid progenitor cells in the bone marrow (BM) that give rise to blood-circulating monocytes that migrate to sites of host insult, and macrophages that are derived from the yolk sac and present in the tissues as resident macrophages [9-11]. Examples of tissue-resident macrophages are alveolar macrophages in the lung and Kupffer cells in the liver. Several macrophage subsets with distinct functions have been described. Classically activated macrophages (M1 macrophages) mediates host defense against several bacterial, viral and protozoal pathogens and they play a role in antitumor immunity [12,13]. These M1 cells are characterized by MHC class II, CD86 and CD40 upregulation [14,15], downregulation of mannose receptor (CD206), their ability to engulf and digest microbes, secrete Pro-inflammatory/T helper 1 (Th1)-promoting cytokines such as IL-12, IL-1β, IL-6, TNF-α production of microbicidal molecules; nitric oxide (NO), and reactive oxygen species (ROS), secretion of chemokines for recruitment of immune cells such as CCL2/MCP-1, and present antigens to T cells for the initiation of adaptive immunity [16-19]. Alternatively activated macrophages (M2 macrophages), on the other hand, are characterized by upregulation of markers such as CD206, arginase-1, IL-10, and TGF-β [20,21] (Figure 1). M2 cells play a role in down regulating the inflammatory mediators, up regulating the antiinflammatory mediators, expressing scavenging receptors (e.g. mannose receptor), phagocytizing apoptotic bodies and cellular debris, and promoting wound healing and tissue repair [17]. These M2 macrophages are a strong inducer of T helper 2 (Th2) cells and/or regulatory T cells. M1-macrophage polarization is enhanced by both IFN-γ and Lipopolysaccharide (LPS), while M2 polarization promoted via IL-4, IL-10 and IL-13 [22,23].
Background. Despite the high prevalence of type 2 diabetes mellitus in Gulf countries, standards ... more Background. Despite the high prevalence of type 2 diabetes mellitus in Gulf countries, standards of diabetes care at the primary care level have not been widely studied. Aim. To compare the results of diabetes clinical indicators from the American Diabetes Association (ADA) 2017 guidelines to the reference benchmarks in the Behavioral Risk Factor Surveillance System. Materials and Methods. A cross-sectional analysis of electronic medical records in 643 randomly selected adult patients with type 2 diabetes was undertaken. A checklist enabled the collection of sociodemographic, clinical, biochemical, and quality measurement data. Data were analyzed using Stata 9.0. The chi-squared test was used to compare two or more proportions. Results. There were 643 patients (male = 60.3%; female = 39.7%), and the majority (71.7%) aged between 40 and 64 years. Common comorbidities were dyslipidemia (72.3%), hypertension (70%), obesity (50.1%), and preobesity (overweight) (37.9%). Over 15% were smo...
Gram-negative bacterial infections are a significant public health concern, and the lack of new d... more Gram-negative bacterial infections are a significant public health concern, and the lack of new drug classes for these pathogens is linked to the inability of most drug leads to accumulate inside Gram-negative bacteria 1 – 7 . Here, we report the development of a web application—eNTRyway—that predicts compound accumulation (in Escherichia coli ) from its structure. In conjunction with structure–activity relationships and X-ray data, eNTRyway was utilized to re-design Debio-1452—a Gram-positive-only antibiotic 8 —into versions that accumulate in E. coli and possess antibacterial activity against high-priority Gram-negative pathogens. The lead compound Debio-1452-NH3 operates as an antibiotic via the same mechanism as Debio-1452, namely potent inhibition of the enoyl-acyl carrier protein reductase FabI, as validated by in vitro enzyme assays and the generation of bacterial isolates with spontaneous target mutations. Debio-1452-NH3 is well tolerated in vivo, reduces bacterial burden in mice and rescues mice from lethal infections with clinical isolates of Acinetobacter baumannii , Klebsiella pneumoniae and E. coli . This work provides tools for the facile discovery and development of high-accumulating compounds in E. coli , and a general blueprint for the conversion of Gram-positive-only compounds into broad-spectrum antibiotics. Hergenrother and colleagues develop a web portal to help predict key permeation aspects of query compounds using the eNTRy rules they recently developed. They use this approach to screen antibiotics that are effective against Gram-positive bacteria and engineer a modified version of a FabI inhibitor that is an effective Gram-negative antibiotic.
Human monocytic ehrlichiosis (HME) is a potentially life-threatening tick-borne rickettsial disea... more Human monocytic ehrlichiosis (HME) is a potentially life-threatening tick-borne rickettsial disease (TBRD) caused by the obligate intracellular Gram-negative bacteria, Ehrlichia. Fatal HME presents with acute ailments of sepsis and toxic shock-like symptoms that can evolve to multi-organ failure and death. Early clinical and laboratory diagnosis of HME are problematic due to non-specific flu-like symptoms and limitations in the current diagnostic testing. Several studies in murine models showed that cell-mediated immunity acts as a "double-edged sword" in fatal ehrlichiosis. Protective components are mainly formed by CD4 Th1 and NKT cells, in contrast to deleterious effects originated from neutrophils and TNF-α-producing CD8 T cells. Recent research has highlighted the central role of the inflammasome and autophagy as part of innate immune responses also leading to protective or pathogenic scenarios. Recognition of pathogen-associated molecular patterns (PAMPS) or damage-associated molecular patterns (DAMPS) triggers the assembly of the inflammasome complex that leads to multiple outcomes. Recognition of PAMPs or DAMPs by such complexes can result in activation of caspase-1 and-11, secretion of the pro-inflammatory cytokines IL-1β and IL-18 culminating into dysregulated inflammation, and inflammatory cell death known as pyroptosis. The precise functions of inflammasomes and autophagy remain unexplored in infections with obligate intracellular rickettsial pathogens, such as Ehrlichia. In this review, we discuss the intracellular innate immune surveillance in ehrlichiosis involving the regulation of inflammasome and autophagy, and how this response influences the innate and adaptive immune responses against Ehrlichia. Understanding such mechanisms would pave the way in research for novel diagnostic, preventative and therapeutic approaches against Ehrlichia and other rickettsial diseases.
Previously, we reported a significantly higher prevalence of uterine fibroids (UFs) in African Am... more Previously, we reported a significantly higher prevalence of uterine fibroids (UFs) in African American women. This minority group also commonly suffers from vitamin D deficiency. We have demonstrated that 1,25(OH)D attains a fibroid growth inhibitory impact through its ability to block the G1/S (gap 1/synthesis) phase of the cell cycle. Vitamin D is involved in DNA damage as well as in immune response regulation, anti-inflammation, autoimmunity and cancer. Since most of the prior data on vitamin D and UF were generated in vitro via established cell lines, it was necessary to verify and validate this observation in vivo using a diet-induced vitamin D-deficient mouse model. Our model of vitamin D lacking function was established using 8-week exposure of C57/BL6 mice to vitamin D-deficient diet provides evidence of different functions accomplished by vitamin D in the regulation of myometrium homeostasis disrupted in the context of uterine fibroid. We found that vitamin D deficiency wa...
American journal of clinical pathology, Jan 13, 2018
To determine a quantitative herpes simplex virus (HSV) DNA threshold in lower respiratory tract s... more To determine a quantitative herpes simplex virus (HSV) DNA threshold in lower respiratory tract specimens that correlates with positive viral culture and clinical outcomes. Bronchoalveolar lavage and bronchial wash samples from 53 HSV culture-positive and 61 culture-negative matched controls were tested using HSV-1 and HSV-2 quantitative polymerase chain reaction (qPCR). Median viral culture turnaround time was 21.8 days and 9.9 days for culture-negative and culture-positive specimens, respectively. Using an HSV-1 viral load threshold of 1.62 × 103 copies/mL, there was 93% agreement with viral culture. An HSV-1 viral load ≥1.3 × 104 copies/mL was associated with worse clinical outcome compared to a viral load <1.3 × 104 copies/mL (hazard ratio [HR] = 4.27, P = .017), and there was a trend of worse outcome compared to patients with undetectable HSV-1 DNA (HR = 1.60, P = .056). qPCR has clinical utility for rapid accurate identification of HSV-1 in lower respiratory tract specimens.
Antibiotic resistance is one of the greatest crises in human medicine. Increased incidents of ant... more Antibiotic resistance is one of the greatest crises in human medicine. Increased incidents of antibiotic resistance are linked to clinical overuse and overreliance on antibiotics. Among the ESKAPE pathogens, Acinetobacter baumannii, especially carbapenem-resistant isolates, has emerged as a significant threat in the context of blood, urinary tract, lung, and wound infections. Therefore, new approaches that limit the emergence of antibiotic resistant A. baumannii are urgently needed. Recently, we have shown that random peptide mixtures (RPMs) are an attractive alternative class of drugs to antibiotics with strong safety and pharmacokinetic profiles. RPMs are antimicrobial peptide mixtures produced by incorporating two amino acids at each coupling step, rendering them extremely diverse but still defined in their overall composition, chain length, and stereochemistry. The extreme diversity of RPMs may prevent bacteria from evolving resistance rapidly. Here, we demonstrated that RPMs ra...
A polarized macrophage response into inflammatory (M1) or regenerative/anti-inflammatory (M2) phe... more A polarized macrophage response into inflammatory (M1) or regenerative/anti-inflammatory (M2) phenotypes is critical in host response to multiple intracellular bacterial infections. Ehrlichia is an obligate Gram-negative intracellular bacterium that causes human monocytic ehrlichiosis (HME): a febrile illness that may progress to fatal sepsis with multi-organ failure. We have shown that liver injury and Ehrlichia-induced sepsis occur due to dysregulated inflammation. Here, we investigated the contribution of macrophages to Ehrlichia-induced sepsis using murine models of mild and fatal ehrlichiosis. Lethally-infected mice showed accumulation of M1 macrophages (iNOS-positive) in the liver. In contrast, non-lethally infected mice showed polarization of M2 macrophages and their accumulation in peritoneum, but not in the liver. Predominance of M1 macrophages in lethally-infected mice was associated with expansion of IL-17-producing T, NK, and NKT cells. Consistent with the in vivo data, ...
Human monocytic ehrlichiosis, a tick transmitted infection, ranges in severity from apparently su... more Human monocytic ehrlichiosis, a tick transmitted infection, ranges in severity from apparently subclinical to fatal toxic shock-like disease. Models in immunocompetent mice range from abortive to uniformly lethal infection, depending on the Ehrlichia species, inoculum dose, and inoculation route. Effective immunity is mediated by CD4+ T lymphocytes and gamma interferon. Lethal infection occurs with early overproduction of proinflammatory cytokines and overproduction of TNF alpha and IL-10 by CD8+ T lymphocytes. Furthermore, fatal ehrlichiosis is associated with TLR 9/MyD88 signaling, upregulation of several inflammasome complexes, and secretion of IL-1 beta, IL-1 alpha, and IL-18 by hepatic mononuclear cells, thus suggesting activation of canonical and noncanonical inflammasome pathways, a deleterious role of IL-18, and a protective role of caspase 1. Autophagy promotes ehrlichial infection, whereas MyD88 signaling hinders ehrlichial infection by inhibiting autophagy induction and f...
Uterine fibroids (UF; aka leiomyoma, myomas) are the most common benign tumors of female reproduc... more Uterine fibroids (UF; aka leiomyoma, myomas) are the most common benign tumors of female reproductive tract. They are highly prevalent, with 70 to 80% of women burdened by the end of their reproductive years. Fibroids are a leading cause of pelvic pain, abnormal vaginal bleeding, pelvic bulk symptoms, miscarriage, and infertility. They are the leading indication for hysterectomy, and costs exceed 34 billion dollars annually in the United States alone. Recently, somatic mutations in exons 1 and 2 of Med12 gene emerged as common UF driver mutations. Unfortunately, the detailed etiology of UF is not fully realized. Particularly, very little is known about possible dysregulation of inflammatory and immune processes and their possible contribution to UF pathogenesis. The notion on possible impact of altered estrogen and progesterone signaling in UF on inflammatory responses and DNA repair machinery that can conceivably lead to tumor-specific somatic mutation is indeed an intriguing conce...
Our murine models of human monocytic ehrlichiosis (HME) have shown that severe and fatal ehrlichi... more Our murine models of human monocytic ehrlichiosis (HME) have shown that severe and fatal ehrlichiosis is due to generation of pathogenic T cell responses causing immunopathology and multi-organ failure. However, the early events in the liver, the main site of infection, are not well understood. In this study, we examined the liver transcriptome during the course of lethal and nonlethal infections caused by Ixodes ovatus Ehrlichia and Ehrlichia muris, respectively. On day 3 post-infection (p.i.), although most host genes were down regulated in the two groups of infected mice compared to naı̈ve counterparts, lethal infection induced significantly higher expression of caspase 1, caspase 4, nucleotide binding oligomerization domain-containing proteins (Nod1), tumor necrosis factor-alpha, interleukin 10, and CCL7 compared to nonlethal infection. On day 7 p.i., lethal infection induced highly significant upregulation of type-1 interferon, several inflammatory cytokines and chemokines, whi...
Inflammasomes are an important innate immune host defense against intracellular microbial infecti... more Inflammasomes are an important innate immune host defense against intracellular microbial infection. Activation of inflammasomes by microbial or host ligands results in cleavage of caspase‐1 (canonical pathway) or caspase‐11 (noncanonical pathway), release of interleukin (IL)‐1β, IL‐18, high mobility group box 1 (HMGB1), and inflammatory cell death known as pyroptosis. Ehrlichia are obligate, intracellular, gram‐negative bacteria that lack lipopolysaccharide but cause potentially life‐threatening monocytic ehrlichiosis in humans and mice that is characterized by liver injury followed by sepsis and multiorgan failure. Employing murine models of mild and fatal ehrlichiosis caused by infection with mildly and highly virulent Ehrlichia muris (EM) and Ixodes ovatus Ehrlichia (IOE), respectively, we have previously shown that IOE infection triggers type I interferon (IFN‐I) response and deleterious caspase‐11 activation in liver tissues, which promotes liver injury and sepsis. In this study, we examined the contribution of IFN‐I signaling in hepatocytes (HCs) to Ehrlichia‐induced liver injury. Compared to EM infection, we found that IOE enter and replicate in vitro cultured primary murine HCs and induce secretion of IFNβ and several chemokines, including regulated upon activation, normal T‐cell expressed, and secreted (RANTES), monocyte chemoattractant protein 1 (MCP1), monokine induced by gamma (MIG)/chemokine (C‐X‐C motif) ligand 9 (CXCL9), macrophage inflammatory protein 1 alpha (MIP1α), keratinocyte‐derived chemokine (KC), and granulocyte‐macrophage colony‐stimulating factor (GM‐CSF). Notably, in vitro stimulation of uninfected and Ehrlichia‐infected HCs with recombinant IFNβ triggered activation of caspase‐1/11, cytosolic translocation of HMGB1, and enhanced autophagy and intracellular bacterial replication. Secretion of HMGB1 by IOE‐infected HCs was dependent on caspase‐11. Primary HCs from IOE‐ but not EM‐infected mice also expressed active caspase‐1/11. Conclusion: HC‐specific IFN‐I signaling may exacerbate liver pathology during infection with obligate intracellular Ehrlichia by promoting bacterial replication and detrimental caspase‐11‐mediated inflammasome activation.
The performance of a fully automated, random access, enhanced chemiluminescence immunoassay (Orth... more The performance of a fully automated, random access, enhanced chemiluminescence immunoassay (Ortho/ ECi) for the detection of antibody to hepatitis C virus (HCV) (anti-HCV), HBsAg, and antibody to HBsAg (anti-HBsAg), in human serum was compared to a Abbott second-generation enzyme immunoassay (EIA 2.0). The Ortho/ECi assays employ an immunometric technique with enhanced chemiluminescence for optimal assay performance. With regard to the study of clinical laboratory performance, six groups of sera prescreened with Abbott EIAs were assayed: anti-HCV-negative samples (n ؍ 318), anti-HCV-positive samples (n ؍ 177), anti-HBsAg-negative samples (n ؍ 241), anti-HBsAg-positive samples (n ؍ 239), HBsAg-positive samples (n ؍ 158), and HBsAg-negative samples (n ؍ 312). Sera with discrepant results in the two serological assays were resolved by confirmatory tests. Sera with indeterminate results by one or more confirmatory tests were evaluated by reviewing medical records. The overall concordance between the Ortho/ECi assay and the Abbott EIA were 97.78, 93.54, and 97.66% for anti-HCV antibodies, anti-HBsAg antibodies, and HBsAg, respectively. After resolving the discrepancies, the specificities of the new assay for anti-HCV and anti-HBsAg antibodies and HBsAg were 98.1, 92.8, and 100%, respectively. The sensitivities of the new assay for anti-HCV, anti-HBsAg, and HBsAg were 100, 98.8, and 97.4%, respectively. In conclusion, The Ortho/ECi assays for diagnosis of HCV and hepatitis B virus (HBV) infections are highly specific and sensitive assays. The rapid turnaround time, random access, full automation, and high throughput make it an effective assay system for clinical laboratory diagnosis of HCV and HBV infections.
Purpose. The number of individuals affected by job stress is growing day by day in almost every i... more Purpose. The number of individuals affected by job stress is growing day by day in almost every industry. According to Health and Safety Executive (2006) workplace stress is now the fastest growing cause of absence from work. The current study aims to verify the effectiveness of management stress in performance level of manager's productivity of youth centers in Cairo. Methods. The sample contains 40 mangers and 80 sports specialists, to collect the research data the researchers have built a questionnaire to measure the administrative empowerment which contains 5 factors, the initial questionnaire consists of 106 items. Results. statistical analyses showed that the stress natural was connected of planning and make decision , organize, control and guidance ( according its importance). Conclusions. Finally, the leaders in youth centers face many of stress which affected of their performance.
Background Polycystic ovary syndrome (PCOS) is the most common endocrine and metabolic disorder i... more Background Polycystic ovary syndrome (PCOS) is the most common endocrine and metabolic disorder in reproductive-age women. Excessive inflammation and elevated androgen production from ovarian theca cells are key features of PCOS. Human bone marrow mesenchymal stem cells (BM-hMSC) and their secreted factors (secretome) exhibit robust anti-inflammatory capabilities in various biological systems. We evaluated the therapeutic efficacy of BM-hMSC and its secretome in both in vitro and in vivo PCOS models. Methods For in vitro experiment, we treated conditioned media from BM-hMSC to androgen-producing H293R cells and analyzed androgen-producing gene expression. For in vivo experiment, BM-hMSC were implanted into letrozole (LTZ)-induced PCOS mouse model. BM-hMSC effect in androgen-producing cells or PCOS model mice was assessed by monitoring cell proliferation (immunohistochemistry), steroidogenic gene expression (quantitative real-time polymerase chain reaction [qRT-PCR] and Western blot,...
M1-M2 Macrophages Polarization Macrophages are innate immune cells that play a key role in regula... more M1-M2 Macrophages Polarization Macrophages are innate immune cells that play a key role in regulation of innate and adaptive immune responses against infections with several pathogens as they respond to pathogens and tissue injury, serve as antigen presenting cells priming the adaptive immune response, drive inflammation and host defense as well as repairing damaged tissue [5-7]. Macrophages reside in almost all of the lymphoid and non-lymphoid tissues [8]. Two major lineages are currently known: cells that are derived from myeloid progenitor cells in the bone marrow (BM) that give rise to blood-circulating monocytes that migrate to sites of host insult, and macrophages that are derived from the yolk sac and present in the tissues as resident macrophages [9-11]. Examples of tissue-resident macrophages are alveolar macrophages in the lung and Kupffer cells in the liver. Several macrophage subsets with distinct functions have been described. Classically activated macrophages (M1 macrophages) mediates host defense against several bacterial, viral and protozoal pathogens and they play a role in antitumor immunity [12,13]. These M1 cells are characterized by MHC class II, CD86 and CD40 upregulation [14,15], downregulation of mannose receptor (CD206), their ability to engulf and digest microbes, secrete Pro-inflammatory/T helper 1 (Th1)-promoting cytokines such as IL-12, IL-1β, IL-6, TNF-α production of microbicidal molecules; nitric oxide (NO), and reactive oxygen species (ROS), secretion of chemokines for recruitment of immune cells such as CCL2/MCP-1, and present antigens to T cells for the initiation of adaptive immunity [16-19]. Alternatively activated macrophages (M2 macrophages), on the other hand, are characterized by upregulation of markers such as CD206, arginase-1, IL-10, and TGF-β [20,21] (Figure 1). M2 cells play a role in down regulating the inflammatory mediators, up regulating the antiinflammatory mediators, expressing scavenging receptors (e.g. mannose receptor), phagocytizing apoptotic bodies and cellular debris, and promoting wound healing and tissue repair [17]. These M2 macrophages are a strong inducer of T helper 2 (Th2) cells and/or regulatory T cells. M1-macrophage polarization is enhanced by both IFN-γ and Lipopolysaccharide (LPS), while M2 polarization promoted via IL-4, IL-10 and IL-13 [22,23].
Background. Despite the high prevalence of type 2 diabetes mellitus in Gulf countries, standards ... more Background. Despite the high prevalence of type 2 diabetes mellitus in Gulf countries, standards of diabetes care at the primary care level have not been widely studied. Aim. To compare the results of diabetes clinical indicators from the American Diabetes Association (ADA) 2017 guidelines to the reference benchmarks in the Behavioral Risk Factor Surveillance System. Materials and Methods. A cross-sectional analysis of electronic medical records in 643 randomly selected adult patients with type 2 diabetes was undertaken. A checklist enabled the collection of sociodemographic, clinical, biochemical, and quality measurement data. Data were analyzed using Stata 9.0. The chi-squared test was used to compare two or more proportions. Results. There were 643 patients (male = 60.3%; female = 39.7%), and the majority (71.7%) aged between 40 and 64 years. Common comorbidities were dyslipidemia (72.3%), hypertension (70%), obesity (50.1%), and preobesity (overweight) (37.9%). Over 15% were smo...
Gram-negative bacterial infections are a significant public health concern, and the lack of new d... more Gram-negative bacterial infections are a significant public health concern, and the lack of new drug classes for these pathogens is linked to the inability of most drug leads to accumulate inside Gram-negative bacteria 1 – 7 . Here, we report the development of a web application—eNTRyway—that predicts compound accumulation (in Escherichia coli ) from its structure. In conjunction with structure–activity relationships and X-ray data, eNTRyway was utilized to re-design Debio-1452—a Gram-positive-only antibiotic 8 —into versions that accumulate in E. coli and possess antibacterial activity against high-priority Gram-negative pathogens. The lead compound Debio-1452-NH3 operates as an antibiotic via the same mechanism as Debio-1452, namely potent inhibition of the enoyl-acyl carrier protein reductase FabI, as validated by in vitro enzyme assays and the generation of bacterial isolates with spontaneous target mutations. Debio-1452-NH3 is well tolerated in vivo, reduces bacterial burden in mice and rescues mice from lethal infections with clinical isolates of Acinetobacter baumannii , Klebsiella pneumoniae and E. coli . This work provides tools for the facile discovery and development of high-accumulating compounds in E. coli , and a general blueprint for the conversion of Gram-positive-only compounds into broad-spectrum antibiotics. Hergenrother and colleagues develop a web portal to help predict key permeation aspects of query compounds using the eNTRy rules they recently developed. They use this approach to screen antibiotics that are effective against Gram-positive bacteria and engineer a modified version of a FabI inhibitor that is an effective Gram-negative antibiotic.
Human monocytic ehrlichiosis (HME) is a potentially life-threatening tick-borne rickettsial disea... more Human monocytic ehrlichiosis (HME) is a potentially life-threatening tick-borne rickettsial disease (TBRD) caused by the obligate intracellular Gram-negative bacteria, Ehrlichia. Fatal HME presents with acute ailments of sepsis and toxic shock-like symptoms that can evolve to multi-organ failure and death. Early clinical and laboratory diagnosis of HME are problematic due to non-specific flu-like symptoms and limitations in the current diagnostic testing. Several studies in murine models showed that cell-mediated immunity acts as a "double-edged sword" in fatal ehrlichiosis. Protective components are mainly formed by CD4 Th1 and NKT cells, in contrast to deleterious effects originated from neutrophils and TNF-α-producing CD8 T cells. Recent research has highlighted the central role of the inflammasome and autophagy as part of innate immune responses also leading to protective or pathogenic scenarios. Recognition of pathogen-associated molecular patterns (PAMPS) or damage-associated molecular patterns (DAMPS) triggers the assembly of the inflammasome complex that leads to multiple outcomes. Recognition of PAMPs or DAMPs by such complexes can result in activation of caspase-1 and-11, secretion of the pro-inflammatory cytokines IL-1β and IL-18 culminating into dysregulated inflammation, and inflammatory cell death known as pyroptosis. The precise functions of inflammasomes and autophagy remain unexplored in infections with obligate intracellular rickettsial pathogens, such as Ehrlichia. In this review, we discuss the intracellular innate immune surveillance in ehrlichiosis involving the regulation of inflammasome and autophagy, and how this response influences the innate and adaptive immune responses against Ehrlichia. Understanding such mechanisms would pave the way in research for novel diagnostic, preventative and therapeutic approaches against Ehrlichia and other rickettsial diseases.
Previously, we reported a significantly higher prevalence of uterine fibroids (UFs) in African Am... more Previously, we reported a significantly higher prevalence of uterine fibroids (UFs) in African American women. This minority group also commonly suffers from vitamin D deficiency. We have demonstrated that 1,25(OH)D attains a fibroid growth inhibitory impact through its ability to block the G1/S (gap 1/synthesis) phase of the cell cycle. Vitamin D is involved in DNA damage as well as in immune response regulation, anti-inflammation, autoimmunity and cancer. Since most of the prior data on vitamin D and UF were generated in vitro via established cell lines, it was necessary to verify and validate this observation in vivo using a diet-induced vitamin D-deficient mouse model. Our model of vitamin D lacking function was established using 8-week exposure of C57/BL6 mice to vitamin D-deficient diet provides evidence of different functions accomplished by vitamin D in the regulation of myometrium homeostasis disrupted in the context of uterine fibroid. We found that vitamin D deficiency wa...
American journal of clinical pathology, Jan 13, 2018
To determine a quantitative herpes simplex virus (HSV) DNA threshold in lower respiratory tract s... more To determine a quantitative herpes simplex virus (HSV) DNA threshold in lower respiratory tract specimens that correlates with positive viral culture and clinical outcomes. Bronchoalveolar lavage and bronchial wash samples from 53 HSV culture-positive and 61 culture-negative matched controls were tested using HSV-1 and HSV-2 quantitative polymerase chain reaction (qPCR). Median viral culture turnaround time was 21.8 days and 9.9 days for culture-negative and culture-positive specimens, respectively. Using an HSV-1 viral load threshold of 1.62 × 103 copies/mL, there was 93% agreement with viral culture. An HSV-1 viral load ≥1.3 × 104 copies/mL was associated with worse clinical outcome compared to a viral load <1.3 × 104 copies/mL (hazard ratio [HR] = 4.27, P = .017), and there was a trend of worse outcome compared to patients with undetectable HSV-1 DNA (HR = 1.60, P = .056). qPCR has clinical utility for rapid accurate identification of HSV-1 in lower respiratory tract specimens.
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Papers by Nahed Ismail