The Journal of Allergy and Clinical Immunology, Feb 1, 2016
Autophagy has been investigated to be involved in inflammatory diseases, but its implications in ... more Autophagy has been investigated to be involved in inflammatory diseases, but its implications in asthma have rarely been studied. This study aimed to explore the possible role of autophagy and its therapeutic potentials in severe allergic asthma. METHODS: BALB/c mice were sensitized by ovalbumin (OVA) on days 0 and 14, followed by primary OVA challenges on day 28-30. Mice received a secondary 1% or 2% OVA challenges on days 44-46. After the final OVA challenge, mice were assessed for airway responsiveness (AHR), cell composition and cytokine levels in bronchoalveolar lavage fluid (BALF). LC3 expression in lung tissue was measured by Western blot and immunofluorescence staining. Autophagosomes were detected by electron microscopy. 3-methyladenine (3-MA), Atg5 knockdown and anti-IL5 antibody were applied to investigate the potential role of autophagy in allergic asthma mice. RESULTS: The AHR, inflammation in BALF and LC3 expression in lung tissue were significantly increased in 2% OVA challenged mice compared to those of 1% OVA challenged mice (P < 0.05). Additionally, eosinophils showed prominently formation of autophagosomes and increase LC3 expression compared with other inflammatory cells in BALF and lung tissue. After inhibiting autophagy by 3-MA and Atg5 shRNA, the AHR, eosinophilia, IL-5 level in BALF, and inflammatory findings in histology were significantly improved. Finally, treatment of anti-IL-5 antibody considerably reduced LC3 II expression in lung homogenate. CONCLUSIONS: Our findings suggest that autophagy is closely correlated with the severity of asthma through eosinophilic inflammation, and its modulation may provide novel therapeutic approaches for severe allergic asthma.
This article is an open access article distributed under the terms and conditions of the Creative... more This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY
The Journal of Allergy and Clinical Immunology, Feb 1, 2016
The purpose of this study is to investigate the effects of anti-IL-9 antibody on oral tolerance i... more The purpose of this study is to investigate the effects of anti-IL-9 antibody on oral tolerance in a mouse model of allergic rhinitis. METHODS: BALB/c mice were divided into 4 groups: control, allergic rhinitis (AR), oral tolerance (OT) , and OT with anti-IL-9 antibody (OT + IL9AB) group. All mice except control group were sensitized with ovalbumin (OVA) three times for two weeks consecutively. After two weeks, mice in OT and OT + IL9AB group underwent immunotherapy by feeding of OVA. During the immunotherapy, mice in OT + IL9AB group were injected with purified anti-mouse IL-9 antibody. All sensitized mice were challenged intranasally with OVA. Allergic symptoms, eosinophil counts in nasal mucosa and serum OVA-specific IgE were measured. Interferone-g, IL-4, IL-17, IL-10, TGF-ß, T-bet, GATA-3, ROR-g t, PU.1 and Foxp3 mRNA expression in nasal mucosa determined by real-time polymerase chain reaction. Flow cytometry of CD4 + CD25 + Foxp3 + T cells in spleen was analyzed. RESULTS: In OT and OT + IL9AB group, symptom score, serum OVAspecific IgE, eosinophils, IL-4, PU.1 and GATA-3 mRNA were decreased (p<0.05), and IL-10, Foxp3 mRNA, and CD4 + CD25 + Foxp3 + T cells were increased compared with those in AR group (p<0.05). In OT + IL9AB group, symptom score and serum OVA-specific IgE were lower than those in OT group (p<0.05). Foxp3 mRNA and CD4 + CD25 + Foxp3 + T cells were higher than those in OT group (p<0.05). CONCLUSIONS: Administration of anti-IL-9 antibody increased the induction of tolerance in a mouse model of allergic rhinitis. These results indicate that anti-IL-9 antibody have immunomodulatory effect on immune tolerance in allergic rhinitis model.
Compromised DNA repair capacity of individuals could play a critical role in the severity of SARS... more Compromised DNA repair capacity of individuals could play a critical role in the severity of SARS-CoV-2 infection-induced COVID-19. We therefore analyzed the expression of DNA repair genes in publicly available transcriptomic datasets of COVID-19 patients and found that the level of NEIL2, an oxidized base specific mammalian DNA glycosylase, is particularly low in the lungs of COVID-19 patients displaying severe symptoms. Downregulation of pulmonary NEIL2 in CoV-2-permissive animals and postmortem COVID-19 patients validated these results. To investigate the potential roles of NEIL2 in CoV-2 pathogenesis, we infected Neil2-null (Neil2−/−) mice with a mouse-adapted CoV-2 strain and found that Neil2−/− mice suffered more severe viral infection concomitant with increased expression of proinflammatory genes, which resulted in an enhanced mortality rate of 80%, up from 20% for the age matched Neil2+/+ cohorts. We also found that infected animals accumulated a significant amount of damage...
Background and aim: Allergic asthma is a complex inflammatory disease involving type 2 innate lym... more Background and aim: Allergic asthma is a complex inflammatory disease involving type 2 innate lymphoid cells, type 2 T helper cells, macrophages, and eosinophils. The disease is characterized by wheezing, dyspnea, coughing, chest tightness and variable airflow limitation for which there is no cure and is symptomatically treated with inhaled corticosteroids and β2-agonists. Molecular mechanisms underlying its complex pathogenesis are not fully understood. However, 8-oxoguanine DNA glycosylase-1 (OGG1), a DNA repair protein may play a central role, as OGG1 deficiency decreases both innate and allergic inflammation.Methods: Using a murine ovalbumin (OVA) model of allergic airway inflammation we assessed the utility of an inhibitor of OGG1 (TH5487) in this disease context. Cytokines and chemokines, promoting immune cell recruitment were measured using a 23-multiplex assay and Western blotting. Additionally, immune cell recruitment to bronchi was measured using flow cytometry. Histologic...
Proinflammatory cytokines/chemokines are commonly regulated by RNA-binding proteins at posttransc... more Proinflammatory cytokines/chemokines are commonly regulated by RNA-binding proteins at posttranscriptional levels. Human Ag R (HuR)/embryonic lethal abnormal vision-like 1 (ELAVL1) is one of the well-characterized RNA-binding proteins that increases the stability of short-lived mRNAs, which encode proinflammatory mediators. HuR employs its nucleocytoplasmic shuttling sequence (HNS) domain, interacting with poly(ADP-ribose) polymerase 1 (PARP1), which accounts for the enhanced poly-ADP-ribosylation and cytoplasmic shuttling of HuR. Also by using its HNS domain, HuR undergoes dimerization/oligomerization, underlying the increased binding of HuR with proinflammatory cytokine/chemokine mRNAs and the disassociation of the miRNA-induced silencing complex from the targets. Therefore, competitively blocking the interactions of HuR with its partners may suppress proinflammatory mediator production. In this study, peptides derived from the sequence of the HuR-HNS domain were synthesized, and ...
Poly(ADP-ribosyl)ation (PARylation) is an important post-translational modification mainly cataly... more Poly(ADP-ribosyl)ation (PARylation) is an important post-translational modification mainly catalyzed by poly-ADP-ribose polymerase 1 (PARP1). In addition to having important roles in DNA damage detection and repair, it functions in gene expression regulation, especially at the posttranscriptional level. Embryonic lethal abnormal vision-like 1/human antigen R (ELAVL/HuR), a canonical 3′ untranslated region AU-rich element-binding protein, is a crucial mRNA-stabilizing protein that protects target mRNAs from RNA-destabilizing protein- or microRNA-induced silencing complex (miRISC)-mediated degradation. Additionally, in some cases, HuR itself either promotes or suppresses translation. Here, we demonstrated that in response to inflammatory stimuli, the PARylation of HuR, mostly at the conserved D226 site, by PARP1 increased the formation of the HuR oligomer/multimer, and HuR oligomerization promoted the disassociation of miRISC and stabilized the pro-inflammatory gene mRNAs. The prevent...
RATIONALE: Lipoxins(LXs) are a group of lipoxygenase derived eicosanoids which in contrast to oth... more RATIONALE: Lipoxins(LXs) are a group of lipoxygenase derived eicosanoids which in contrast to other eicosanoids including leukotrienes, are recognized to have potent anti-inflammatory and pro-resolution properties. Formyl peptide receptor-like receptor 1(FPRL1) also called also called LXA4 receptor, functions as a component of the inflammatory response. To date, only a few reports have shown the existence of this receptor in the airway. The purpose of this study was to investigate the expression and the localization of FPRL1 protein in human nasal mucosa by western blotting and immunohistochemical analysis. METHODS: Human turbinates were obtained after turbinectomy from 6 patients with nasal obstruction refractory to medical therapy. The expressionof FPRL1 protein was evaluated by western blotting. To identify the cells expressing FPRL1 protein, immunostaining was performed using anti-human FPRL1 antibody. RESULTS: A single band of approximately 38kDa was detected in human turbinates and primary cultured nasal epithelial cells by western blot analysis using anti-FPRL1 antibody. The immunohistochemical studies revealed that anti-FPRL1 antibody mainly labeled epithelial cells, submucosal glands and some inflammatory leukocytes in nasal mucosa. CONCLUSIONS: These results may have an important clinical implication for understanding the role of LXA4 receptor on upper airway diseases such as allergic rhinitis and non-allergic rhinitis.
Ten per cent foetal bovine serum had been added to human embryonic fibroblasts which had previous... more Ten per cent foetal bovine serum had been added to human embryonic fibroblasts which had previously been incubated in serum free cultures. The effect has been studied on quantity of synthetized DNA in next 30 hours and on inositol-1,4,5-phosphate contents of cells. On the effect of foetal bovine serum synthesis of DNA has increased four times and 1 minute after its addition inositol-1,4,5-phosphate level has doubled. Papaverine given together with FBS has suspended inductive effect of FBS on DNA synthesis and its increasing effect on level of inositol-1,4,5-phosphate.
Biochemical and Biophysical Research Communications, Dec 1, 1993
Increased levels of active sequence-specific DNA-binding proteins, AP-1, CRE/B, and NF kappa B we... more Increased levels of active sequence-specific DNA-binding proteins, AP-1, CRE/B, and NF kappa B were observed in nuclear lysates of human cytomegalovirus-infected cells from 15 min postinfection. The activation of these cellular factors did not require infectious virus or de novo viral protein synthesis, but their abundance was significantly reduced by inhibitors of protein kinase C and/or A. These data suggest that formation of these transcription factors resulted from the virus-cell membrane interaction and/or through the action of virion structural proteins on cytoplasmic forms of these cellular factors.
Abstract This is the first study documenting the induction of γ-interferon (IFN–γ) in human embry... more Abstract This is the first study documenting the induction of γ-interferon (IFN–γ) in human embryonic fibroblasts during human cytomegalovirus (HCMV) replication. Infection of cells with HCMV resulted in the consistent production of IFN-γ RNA, as determined by RT-PCR and Northern blot analysis. Western blot analysis of cell lysates and immunoprecipitates from the cultural fluids of infected cells demonstrated the presence of IFN-γ at the protein level. Induction of IFN-γ required infectious HCMV, since high-dose ultraviolet inactivation of the virus stock eliminated IFN-γ production. Further, IFN-γ induction appears to be a late event in the virus replication cycle, since inhibition of HCMV DNA synthesis (e.g., phosphonoacetic acid) blocked the increase in IFN-γ. Soluble factor(s) released from HCMV-infected cells apparently did not contribute to the induction of IFN-γ, since virus stocks from which virus had been removed by sedimentation did not induce production of IFN-γ. The appearance of IFN-γ at late stages of HCMV infection and its elimination in the presence of an inhibitor (Actinomycin D) of RNA synthesis indicate a true transcriptional induction of this lymphokine at the RNA and protein levels. The significance of IFN-y production with regard to the replication and pathogenesis of HCMV in vitro and In vivo will require further investigation.
Evidence for the presence of cytomegaloviruses (CMV) as pathogenic agents in man was first obtain... more Evidence for the presence of cytomegaloviruses (CMV) as pathogenic agents in man was first obtained by examination of tissues derived from stillborn infants that demonstrated severe developmental abnormalities (Jesionek and Kiolemeno
During exercise, muscle contraction increases the level of reactive oxygen species (ROS), which h... more During exercise, muscle contraction increases the level of reactive oxygen species (ROS), which has been acknowledged as key signaling entities in muscle remodeling and to underlie the healthy adaptation of skeletal muscle. ROS inevitably endows damage to genomic DNA of the myo bers, and DNA damage ought to be repaired to ensure the integrity and delity of genetic materials; yet, how DNA repair metabolites affect muscle remodeling remains elusive. Here, we demonstrated that exercise induces 8oxo-7,8-dihydroguanine (8-oxoG) generation in nuclei of myo ber, which subsequently is repaired. In myoblasts, 8-oxoG functions as a signal messenger, triggering myogenic differentiation via activating RAS-MEK-MyoD signal axis. 8-OxoG glycosylae 1 (OGG1) silencing from cells or depletion from mice decreased Ras activation, ERK phosphorylation, MyoD transcriptional activation, myogenic regulatory factors gene (MRFs) expression and consequently myogenic differentiation of myoblasts. Microscale Thermophoresis (MST) assay showed high a nity of 8-OxoG binding with OGG1, the complex functions as guanine nucleotide exchange factor (GEF) to activate Ras. In reconstruction experiments, exogenously added 8-oxoG base increases in the expression of MRFs and accelerates recovery of the injured skeletal muscle. Collectively, these data not only suggest that DNA repair metabolite 8-oxoG function as a signal entity for muscle remodeling and contribute to exercise-induced adaptation of skeletal muscle, but also pave the way for utilizing 8-oxoG in clinical treatment to skeletal muscle damage-related disorders.
<p>The ciliated monolayer of airway cells at ALI was treated with AR inhibitor for 24 h and... more <p>The ciliated monolayer of airway cells at ALI was treated with AR inhibitor for 24 h and stimulated with IL-13 for different time periods as indicated. At the end of incubation, cells were lysed and cell lysate was subjected to western blotting using antibodies against phosphorylated and non-phosphorylated JAK-1, ERK1/2 and STAT-6 to analyze the activation of these signaling proteins. A representative blot is shown (n = 4). #p<0.001 vs Control; *p<0.01 and **p<0.001 vs IL-13.</p
This is a PDF file of an article that has undergone enhancements after acceptance, such as the ad... more This is a PDF file of an article that has undergone enhancements after acceptance, such as the addition of a cover page and metadata, and formatting for readability, but it is not yet the definitive version of record. This version will undergo additional copyediting, typesetting and review before it is published in its final form, but we are providing this version to give early visibility of the article. Please note that, during the production process, errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
Macrophage disappearance reaction was induced by intraperitoneal injections of specific antigens ... more Macrophage disappearance reaction was induced by intraperitoneal injections of specific antigens in Rana esculenta sensitized by bovine gamma globulin and Mycobacterium bovis BCG. Rabbit lymphokines derived from concanavalin A stimulated blood lymphocytes injected intraperitoneally were able to induce macrophage disappearance reaction in normal Rana esculenta. This suggests that mammalian lymphokines are capable of acting in amphibia. Mammalian lymphokines have not an exclusive class-specificity in vertebrates.
The Journal of Allergy and Clinical Immunology, Feb 1, 2016
Autophagy has been investigated to be involved in inflammatory diseases, but its implications in ... more Autophagy has been investigated to be involved in inflammatory diseases, but its implications in asthma have rarely been studied. This study aimed to explore the possible role of autophagy and its therapeutic potentials in severe allergic asthma. METHODS: BALB/c mice were sensitized by ovalbumin (OVA) on days 0 and 14, followed by primary OVA challenges on day 28-30. Mice received a secondary 1% or 2% OVA challenges on days 44-46. After the final OVA challenge, mice were assessed for airway responsiveness (AHR), cell composition and cytokine levels in bronchoalveolar lavage fluid (BALF). LC3 expression in lung tissue was measured by Western blot and immunofluorescence staining. Autophagosomes were detected by electron microscopy. 3-methyladenine (3-MA), Atg5 knockdown and anti-IL5 antibody were applied to investigate the potential role of autophagy in allergic asthma mice. RESULTS: The AHR, inflammation in BALF and LC3 expression in lung tissue were significantly increased in 2% OVA challenged mice compared to those of 1% OVA challenged mice (P < 0.05). Additionally, eosinophils showed prominently formation of autophagosomes and increase LC3 expression compared with other inflammatory cells in BALF and lung tissue. After inhibiting autophagy by 3-MA and Atg5 shRNA, the AHR, eosinophilia, IL-5 level in BALF, and inflammatory findings in histology were significantly improved. Finally, treatment of anti-IL-5 antibody considerably reduced LC3 II expression in lung homogenate. CONCLUSIONS: Our findings suggest that autophagy is closely correlated with the severity of asthma through eosinophilic inflammation, and its modulation may provide novel therapeutic approaches for severe allergic asthma.
This article is an open access article distributed under the terms and conditions of the Creative... more This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY
The Journal of Allergy and Clinical Immunology, Feb 1, 2016
The purpose of this study is to investigate the effects of anti-IL-9 antibody on oral tolerance i... more The purpose of this study is to investigate the effects of anti-IL-9 antibody on oral tolerance in a mouse model of allergic rhinitis. METHODS: BALB/c mice were divided into 4 groups: control, allergic rhinitis (AR), oral tolerance (OT) , and OT with anti-IL-9 antibody (OT + IL9AB) group. All mice except control group were sensitized with ovalbumin (OVA) three times for two weeks consecutively. After two weeks, mice in OT and OT + IL9AB group underwent immunotherapy by feeding of OVA. During the immunotherapy, mice in OT + IL9AB group were injected with purified anti-mouse IL-9 antibody. All sensitized mice were challenged intranasally with OVA. Allergic symptoms, eosinophil counts in nasal mucosa and serum OVA-specific IgE were measured. Interferone-g, IL-4, IL-17, IL-10, TGF-ß, T-bet, GATA-3, ROR-g t, PU.1 and Foxp3 mRNA expression in nasal mucosa determined by real-time polymerase chain reaction. Flow cytometry of CD4 + CD25 + Foxp3 + T cells in spleen was analyzed. RESULTS: In OT and OT + IL9AB group, symptom score, serum OVAspecific IgE, eosinophils, IL-4, PU.1 and GATA-3 mRNA were decreased (p<0.05), and IL-10, Foxp3 mRNA, and CD4 + CD25 + Foxp3 + T cells were increased compared with those in AR group (p<0.05). In OT + IL9AB group, symptom score and serum OVA-specific IgE were lower than those in OT group (p<0.05). Foxp3 mRNA and CD4 + CD25 + Foxp3 + T cells were higher than those in OT group (p<0.05). CONCLUSIONS: Administration of anti-IL-9 antibody increased the induction of tolerance in a mouse model of allergic rhinitis. These results indicate that anti-IL-9 antibody have immunomodulatory effect on immune tolerance in allergic rhinitis model.
Compromised DNA repair capacity of individuals could play a critical role in the severity of SARS... more Compromised DNA repair capacity of individuals could play a critical role in the severity of SARS-CoV-2 infection-induced COVID-19. We therefore analyzed the expression of DNA repair genes in publicly available transcriptomic datasets of COVID-19 patients and found that the level of NEIL2, an oxidized base specific mammalian DNA glycosylase, is particularly low in the lungs of COVID-19 patients displaying severe symptoms. Downregulation of pulmonary NEIL2 in CoV-2-permissive animals and postmortem COVID-19 patients validated these results. To investigate the potential roles of NEIL2 in CoV-2 pathogenesis, we infected Neil2-null (Neil2−/−) mice with a mouse-adapted CoV-2 strain and found that Neil2−/− mice suffered more severe viral infection concomitant with increased expression of proinflammatory genes, which resulted in an enhanced mortality rate of 80%, up from 20% for the age matched Neil2+/+ cohorts. We also found that infected animals accumulated a significant amount of damage...
Background and aim: Allergic asthma is a complex inflammatory disease involving type 2 innate lym... more Background and aim: Allergic asthma is a complex inflammatory disease involving type 2 innate lymphoid cells, type 2 T helper cells, macrophages, and eosinophils. The disease is characterized by wheezing, dyspnea, coughing, chest tightness and variable airflow limitation for which there is no cure and is symptomatically treated with inhaled corticosteroids and β2-agonists. Molecular mechanisms underlying its complex pathogenesis are not fully understood. However, 8-oxoguanine DNA glycosylase-1 (OGG1), a DNA repair protein may play a central role, as OGG1 deficiency decreases both innate and allergic inflammation.Methods: Using a murine ovalbumin (OVA) model of allergic airway inflammation we assessed the utility of an inhibitor of OGG1 (TH5487) in this disease context. Cytokines and chemokines, promoting immune cell recruitment were measured using a 23-multiplex assay and Western blotting. Additionally, immune cell recruitment to bronchi was measured using flow cytometry. Histologic...
Proinflammatory cytokines/chemokines are commonly regulated by RNA-binding proteins at posttransc... more Proinflammatory cytokines/chemokines are commonly regulated by RNA-binding proteins at posttranscriptional levels. Human Ag R (HuR)/embryonic lethal abnormal vision-like 1 (ELAVL1) is one of the well-characterized RNA-binding proteins that increases the stability of short-lived mRNAs, which encode proinflammatory mediators. HuR employs its nucleocytoplasmic shuttling sequence (HNS) domain, interacting with poly(ADP-ribose) polymerase 1 (PARP1), which accounts for the enhanced poly-ADP-ribosylation and cytoplasmic shuttling of HuR. Also by using its HNS domain, HuR undergoes dimerization/oligomerization, underlying the increased binding of HuR with proinflammatory cytokine/chemokine mRNAs and the disassociation of the miRNA-induced silencing complex from the targets. Therefore, competitively blocking the interactions of HuR with its partners may suppress proinflammatory mediator production. In this study, peptides derived from the sequence of the HuR-HNS domain were synthesized, and ...
Poly(ADP-ribosyl)ation (PARylation) is an important post-translational modification mainly cataly... more Poly(ADP-ribosyl)ation (PARylation) is an important post-translational modification mainly catalyzed by poly-ADP-ribose polymerase 1 (PARP1). In addition to having important roles in DNA damage detection and repair, it functions in gene expression regulation, especially at the posttranscriptional level. Embryonic lethal abnormal vision-like 1/human antigen R (ELAVL/HuR), a canonical 3′ untranslated region AU-rich element-binding protein, is a crucial mRNA-stabilizing protein that protects target mRNAs from RNA-destabilizing protein- or microRNA-induced silencing complex (miRISC)-mediated degradation. Additionally, in some cases, HuR itself either promotes or suppresses translation. Here, we demonstrated that in response to inflammatory stimuli, the PARylation of HuR, mostly at the conserved D226 site, by PARP1 increased the formation of the HuR oligomer/multimer, and HuR oligomerization promoted the disassociation of miRISC and stabilized the pro-inflammatory gene mRNAs. The prevent...
RATIONALE: Lipoxins(LXs) are a group of lipoxygenase derived eicosanoids which in contrast to oth... more RATIONALE: Lipoxins(LXs) are a group of lipoxygenase derived eicosanoids which in contrast to other eicosanoids including leukotrienes, are recognized to have potent anti-inflammatory and pro-resolution properties. Formyl peptide receptor-like receptor 1(FPRL1) also called also called LXA4 receptor, functions as a component of the inflammatory response. To date, only a few reports have shown the existence of this receptor in the airway. The purpose of this study was to investigate the expression and the localization of FPRL1 protein in human nasal mucosa by western blotting and immunohistochemical analysis. METHODS: Human turbinates were obtained after turbinectomy from 6 patients with nasal obstruction refractory to medical therapy. The expressionof FPRL1 protein was evaluated by western blotting. To identify the cells expressing FPRL1 protein, immunostaining was performed using anti-human FPRL1 antibody. RESULTS: A single band of approximately 38kDa was detected in human turbinates and primary cultured nasal epithelial cells by western blot analysis using anti-FPRL1 antibody. The immunohistochemical studies revealed that anti-FPRL1 antibody mainly labeled epithelial cells, submucosal glands and some inflammatory leukocytes in nasal mucosa. CONCLUSIONS: These results may have an important clinical implication for understanding the role of LXA4 receptor on upper airway diseases such as allergic rhinitis and non-allergic rhinitis.
Ten per cent foetal bovine serum had been added to human embryonic fibroblasts which had previous... more Ten per cent foetal bovine serum had been added to human embryonic fibroblasts which had previously been incubated in serum free cultures. The effect has been studied on quantity of synthetized DNA in next 30 hours and on inositol-1,4,5-phosphate contents of cells. On the effect of foetal bovine serum synthesis of DNA has increased four times and 1 minute after its addition inositol-1,4,5-phosphate level has doubled. Papaverine given together with FBS has suspended inductive effect of FBS on DNA synthesis and its increasing effect on level of inositol-1,4,5-phosphate.
Biochemical and Biophysical Research Communications, Dec 1, 1993
Increased levels of active sequence-specific DNA-binding proteins, AP-1, CRE/B, and NF kappa B we... more Increased levels of active sequence-specific DNA-binding proteins, AP-1, CRE/B, and NF kappa B were observed in nuclear lysates of human cytomegalovirus-infected cells from 15 min postinfection. The activation of these cellular factors did not require infectious virus or de novo viral protein synthesis, but their abundance was significantly reduced by inhibitors of protein kinase C and/or A. These data suggest that formation of these transcription factors resulted from the virus-cell membrane interaction and/or through the action of virion structural proteins on cytoplasmic forms of these cellular factors.
Abstract This is the first study documenting the induction of γ-interferon (IFN–γ) in human embry... more Abstract This is the first study documenting the induction of γ-interferon (IFN–γ) in human embryonic fibroblasts during human cytomegalovirus (HCMV) replication. Infection of cells with HCMV resulted in the consistent production of IFN-γ RNA, as determined by RT-PCR and Northern blot analysis. Western blot analysis of cell lysates and immunoprecipitates from the cultural fluids of infected cells demonstrated the presence of IFN-γ at the protein level. Induction of IFN-γ required infectious HCMV, since high-dose ultraviolet inactivation of the virus stock eliminated IFN-γ production. Further, IFN-γ induction appears to be a late event in the virus replication cycle, since inhibition of HCMV DNA synthesis (e.g., phosphonoacetic acid) blocked the increase in IFN-γ. Soluble factor(s) released from HCMV-infected cells apparently did not contribute to the induction of IFN-γ, since virus stocks from which virus had been removed by sedimentation did not induce production of IFN-γ. The appearance of IFN-γ at late stages of HCMV infection and its elimination in the presence of an inhibitor (Actinomycin D) of RNA synthesis indicate a true transcriptional induction of this lymphokine at the RNA and protein levels. The significance of IFN-y production with regard to the replication and pathogenesis of HCMV in vitro and In vivo will require further investigation.
Evidence for the presence of cytomegaloviruses (CMV) as pathogenic agents in man was first obtain... more Evidence for the presence of cytomegaloviruses (CMV) as pathogenic agents in man was first obtained by examination of tissues derived from stillborn infants that demonstrated severe developmental abnormalities (Jesionek and Kiolemeno
During exercise, muscle contraction increases the level of reactive oxygen species (ROS), which h... more During exercise, muscle contraction increases the level of reactive oxygen species (ROS), which has been acknowledged as key signaling entities in muscle remodeling and to underlie the healthy adaptation of skeletal muscle. ROS inevitably endows damage to genomic DNA of the myo bers, and DNA damage ought to be repaired to ensure the integrity and delity of genetic materials; yet, how DNA repair metabolites affect muscle remodeling remains elusive. Here, we demonstrated that exercise induces 8oxo-7,8-dihydroguanine (8-oxoG) generation in nuclei of myo ber, which subsequently is repaired. In myoblasts, 8-oxoG functions as a signal messenger, triggering myogenic differentiation via activating RAS-MEK-MyoD signal axis. 8-OxoG glycosylae 1 (OGG1) silencing from cells or depletion from mice decreased Ras activation, ERK phosphorylation, MyoD transcriptional activation, myogenic regulatory factors gene (MRFs) expression and consequently myogenic differentiation of myoblasts. Microscale Thermophoresis (MST) assay showed high a nity of 8-OxoG binding with OGG1, the complex functions as guanine nucleotide exchange factor (GEF) to activate Ras. In reconstruction experiments, exogenously added 8-oxoG base increases in the expression of MRFs and accelerates recovery of the injured skeletal muscle. Collectively, these data not only suggest that DNA repair metabolite 8-oxoG function as a signal entity for muscle remodeling and contribute to exercise-induced adaptation of skeletal muscle, but also pave the way for utilizing 8-oxoG in clinical treatment to skeletal muscle damage-related disorders.
<p>The ciliated monolayer of airway cells at ALI was treated with AR inhibitor for 24 h and... more <p>The ciliated monolayer of airway cells at ALI was treated with AR inhibitor for 24 h and stimulated with IL-13 for different time periods as indicated. At the end of incubation, cells were lysed and cell lysate was subjected to western blotting using antibodies against phosphorylated and non-phosphorylated JAK-1, ERK1/2 and STAT-6 to analyze the activation of these signaling proteins. A representative blot is shown (n = 4). #p<0.001 vs Control; *p<0.01 and **p<0.001 vs IL-13.</p
This is a PDF file of an article that has undergone enhancements after acceptance, such as the ad... more This is a PDF file of an article that has undergone enhancements after acceptance, such as the addition of a cover page and metadata, and formatting for readability, but it is not yet the definitive version of record. This version will undergo additional copyediting, typesetting and review before it is published in its final form, but we are providing this version to give early visibility of the article. Please note that, during the production process, errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
Macrophage disappearance reaction was induced by intraperitoneal injections of specific antigens ... more Macrophage disappearance reaction was induced by intraperitoneal injections of specific antigens in Rana esculenta sensitized by bovine gamma globulin and Mycobacterium bovis BCG. Rabbit lymphokines derived from concanavalin A stimulated blood lymphocytes injected intraperitoneally were able to induce macrophage disappearance reaction in normal Rana esculenta. This suggests that mammalian lymphokines are capable of acting in amphibia. Mammalian lymphokines have not an exclusive class-specificity in vertebrates.
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