ABSTRACT
BACKGROUND: Administering platelets through a rapid infuser is proven to be safe. However, the clinical significance of infusing ABO-incompatible platelets with red blood cells (RBCs) in a rapid infuser remains unclear. There is a theoretical risk that isoagglutinin in the plasma of a platelet unit can interact with RBCs and induce hemolysis. MATERIALS AND METHODS: Seven in vitro studies were performed including five cases (type A RBCs and type O platelets) and two controls (type A RBCs and platelets). Anti-A titers were measured in platelet units. An RBC unit and a platelet unit were mixed in the rapid infuser reservoir and incubated for 30 min. The primary outcome was the presence of hemolysis based on the following parameters: free hemoglobin concentration, hemolysis check, direct antiglobulin test (DAT), and direct agglutination. RESULTS: The post-mix DAT was positive for IgG in all test samples (5/5), and weakly positive for complement in 3/5. The changes in free Hb in test cases between measured and calculated post-mix spanned -2.2 to +3.4 mg/dL. Post-mix hemolysis check was negative in 3/5 and slightly positive in 2/5 cases, with no significant differences compared to the control case. Anti-A titers ranged from 16 to 512 and were not associated with hemolysis. All samples were negative for direct agglutination. CONCLUSION: Our study suggested that mixing ABO-incompatible platelets with RBCs in a rapid infuser does not induce in vitro hemolysis. These findings support the use of rapid infusers regardless of platelet compatibility in support of hemostatic resuscitation.
Subject(s)
ABO Blood-Group System , Hemolysis , Humans , Platelet Transfusion/adverse effects , Blood Group Incompatibility , Blood Platelets , AntibodiesABSTRACT
Granulocyte transfusion (GT) may be used to treat and prevent infections in patients with severe neutropenia or nonfunctioning granulocytes. For pediatric patients, the volume of granulocyte unit transfused is a crucial consideration given smaller blood volume and increased risk of volume overload compared to adults. There is limited literature on the optimal dosing or the maximum amount of granulocytes that can be tolerated, especially in pediatric patients. Additionally, no consensus exists regarding granulocyte collection method, frequency, or timing of GT initiation. Previous studies have described splitting or limiting collection volume for GT in pediatric patients, but these methods yield lower absolute neutrophil count (ANC) increment. Our blood supplier provides high-volume (0.5-1 L/unit), high-dose apheresis-collected granulocytes from donors stimulated with both granulocyte colony-stimulating factor and steroids. Here, we report cases of two pediatric patients with active infection undergoing bone marrow transplant with dramatic ANC increments (median one-hour ANC increment 5524/µL, interquartile range (IQR) 4417-10087; median 24-hour ANC increment 3880/µL, IQR 2550-5263) after infusing 100 mL plasma-reduced, apheresis collected GT. Our cases indicate that pediatric patients can tolerate 4-6 × 109/kg plasma-reduced GT and have detectable ANC with GT every 3 days.
Subject(s)
Blood Component Removal , Granulocytes , Adult , Humans , Child , Neutrophils , Leukocyte Transfusion , Blood Donors , Granulocyte Colony-Stimulating Factor/therapeutic useABSTRACT
INTRODUCTION: Lipoprotein X (Lp-X) is an abnormal lipoprotein found in multiple disease conditions, including liver dysfunction and cholestasis. High Lp-X concentrations can interfere with some laboratory testing that may result in spurious results. The detection of Lp-X can be challenging, and there is currently a lack of consensus regarding the management of Lp-X other than treating the underlying disease. CASE PRESENTATION: A 42-year-old female with Hodgkin's lymphoma treated with dexamethasone, high dose cytarabine and cisplatin and vanishing bile duct syndrome confirmed by liver biopsy presented with cholestasis, pseudohyponatremia (sodium, 113 mmol/L; reference range 136-146 mmL/L; serum osmolality, 303 mOsm/kg), and hypercholesterolemia (> 2800 mg/dL, reference range < 200 mg/dL). Lp-X was confirmed by lipoprotein electrophoresis (EP). Although she did not manifest any specific signs or symptoms, therapeutic plasma exchange (TPE) was initiated based on laboratory findings of extreme hypercholesterolemia, spuriously abnormal serum sodium, and HDL values, and the potential for short- and long-term sequelae such as hyperviscosity syndrome, xanthoma, and neuropathy. During the hospitalization, she was treated with four 1.0 plasma volume TPE over 6 days using 5% albumin for replacement fluid. After the first TPE, total cholesterol (TC) decreased to 383 mg/dL and sodium was measured at 131 mmol/L. The patient was transitioned into outpatient maintenance TPE to eliminate the potential of Lp-X reappearance while the underlying disease was treated. Serial follow-up laboratory testing with lipoprotein EP showed the disappearance of Lp-X after nine TPEs over a 10-week period. LITERATURE REVIEW: There are seven and four case reports of Lp-X treated with TPE and lipoprotein apheresis (LA), respectively. While all previous case reports showed a reduction in TC levels, none had monitored the disappearance of Lp-X after completing a course of therapeutic apheresis. CONCLUSION: Clinicians should have a heightened suspicion for the presence of abnormal Lp-X in patients with cholestasis, hypercholesterolemia, and pseudohyponatremia. Once Lp-X is confirmed by lipoprotein EP, TPE should be initiated to reduce TC level and remove abnormal Lp-X. Most LA techniques are not expected to be beneficial since Lp-X lacks apolipoprotein B. Therefore, we suggest that inpatient course of TPE be performed every other day until serum sodium, TC and HDL levels become normalized. Outpatient maintenance TPE may also be considered to keep Lp-X levels low while the underlying disease is treated. Serum sodium, TC, and HDL levels should be monitored while on maintenance TPE.
Subject(s)
Cholestasis , Hypercholesterolemia , Female , Humans , Adult , Hypercholesterolemia/complications , Hypercholesterolemia/therapy , Lipoprotein-X , Plasma Exchange , Cholestasis/etiology , Cholestasis/therapy , Lipoproteins , Sodium , Bile DuctsABSTRACT
Previous reports have shown that PPARß/δ agonists ameliorate insulin resistance associated with type 2 diabetes mellitus (T2DM). To determine the role of PPARß/δ in tumor necrosis factor α (TNFα)-mediated insulin resistance, we investigated expression levels of adiponectin and insulin receptor (IR) in response to treatment with the PPARß/δ agonist GW501516 with or without TNFα, a proinflammatory cytokine, in differentiated 3T3-L1 adipocytes. GW501516 induced adipocyte differentiation and the expression of adiponectin in a dose-dependent manner in differentiated adipocytes. TNFα treatment reduced adiponectin expression at the end of differentiation. This effect was reversed by GW501516 co-treatment with TNFα. TNFα treatment decreased adipogenic marker genes such as PPARγ, aP2, resistin, and GLUT4, and GW501516 reversed the effects of TNFα. GW501516 treatment increased the expression of insulin receptor and inhibited TNFα-mediated repression of insulin receptor. Our results showed that GW501516 abrogated TNFα-induced insulin resistance. In summary, our study demonstrated that the PPARß/δ agonist, GW501516 reversed TNFα-induced decreases in adipocyte differentiation and adiponectin expression, and improved insulin sensitivity by increasing the expression of insulin receptor. Therefore, PPARδ may be a promising therapeutic target for treatment of insulin resistance in patients with T2DM.
Subject(s)
Adiponectin/metabolism , PPAR delta/agonists , PPAR-beta/agonists , Receptor, Insulin/metabolism , Thiazoles/pharmacology , Tumor Necrosis Factor-alpha/metabolism , 3T3-L1 Cells , Adipocytes/drug effects , Adipocytes/metabolism , Animals , Insulin Resistance , Mice , PPAR delta/metabolism , PPAR-beta/metabolismABSTRACT
Eupatilin (5,7-dihydroxy-3',4',6-trimethoxyflavone) is the main lipophilic flavonoid obtained from the Artemisia species. Eupatilin has been reported to have anti-apoptotic, anti-oxidative and anti-inflammatory activities. Previously, we found that eupatilin increases transcriptional activity and expression of peroxisome proliferator-activated receptor α (PPARα) in a keratinocyte cell line and acts as an agonist of PPARα. PPARα agonists ameliorate atopic dermatitis (AD) and restore the skin barrier function. In this study, we confirmed that the effects of eupatilin improved AD-like symptoms in an oxazolone-induced AD-like mouse model. Furthermore, we found that eupatilin suppressed the levels of serum immunoglobulin E (IgE), interleukin-4 (IL-4), and AD involved cytokines, such as tumor necrosis factor α (TNFα), interferon-γ (IFN-γ), IL-1ß, and thymic stromal lymphopoietin (TSLP), IL-33, IL-25 and increased the levels of filaggrin and loricrin in the oxazolone-induced AD-like mouse model. Taken together, our data suggest that eupatilin is a potential candidate for the treatment of AD.
Subject(s)
Dermatitis, Atopic/drug therapy , Dermatologic Agents/pharmacology , Drugs, Chinese Herbal/pharmacology , Flavonoids/pharmacology , PPAR alpha/genetics , Animals , Cell Line, Tumor , Cytokines/genetics , Cytokines/immunology , Dermatitis, Atopic/chemically induced , Dermatitis, Atopic/immunology , Dermatitis, Atopic/pathology , Dose-Response Relationship, Drug , Female , Filaggrin Proteins , Gene Expression Regulation , Immunoglobulin E/blood , Immunoglobulin E/genetics , Interferon-gamma/genetics , Interferon-gamma/immunology , Interleukin-1beta/genetics , Interleukin-1beta/immunology , Interleukin-33/genetics , Interleukin-33/immunology , Interleukin-4/genetics , Interleukin-4/immunology , Interleukins/genetics , Interleukins/immunology , Intermediate Filament Proteins/genetics , Intermediate Filament Proteins/immunology , Membrane Proteins/genetics , Membrane Proteins/immunology , Mice , Mice, Inbred BALB C , Oxazolone , PPAR alpha/immunology , Rats , Signal Transduction , Tumor Necrosis Factor-alpha/genetics , Tumor Necrosis Factor-alpha/immunology , Thymic Stromal LymphopoietinABSTRACT
Eupatilin (5,7-dihydroxy-3,4,6-trimethoxyflavone) is a flavonoid compound exhibiting several beneficial biological activities, including neuroprotection, anti-cancer, antinociception, chondroprotection, anti-oxidation, and anti-inflammation. Our previous study demonstrated that eupatilin specifically activates peroxisome proliferator-activated receptor alpha (PPARα) through direct binding. The PPAR subfamily includes ligand-dependent transcription factors that consist of three isotypes: PPARα, PPARß/δ, and PPARγ. All isotypes are involved in inflammation, epidermal proliferation/differentiation and skin barrier function. Among them, PPARα regulates lipid and glucose metabolism and skin homeostasis. In this study, we confirm that the ability of eupatilin as a PPARα activator significantly inhibited tumor necrosis factor-alpha (TNFα)-induced matrix metalloproteinase (MMP)-2/-9 expression and proteolytic activity in HaCaT human epidermal keratinocytes. Furthermore, we found that eupatilin subsequently suppressed IκBα phosphorylation, blocked NF-κB p65 nuclear translocation and down-regulated MAPK/AP-1 signaling via PPARα activation. Taken together, our data suggest that eupatilin inhibits TNFα-induced MMP-2/-9 expression by suppressing NF-κB and MAPK/AP-1 pathways via PPARα. Our findings suggest the usefulness of eupatilin for preventing skin aging.
Subject(s)
Flavonoids/administration & dosage , Keratinocytes/drug effects , Keratinocytes/metabolism , Matrix Metalloproteinase 2/metabolism , NF-kappa B/metabolism , PPAR alpha/agonists , Tumor Necrosis Factor-alpha/administration & dosage , Cell Line , Dose-Response Relationship, Drug , Drug Therapy, Combination , Drugs, Chinese Herbal/administration & dosage , Drugs, Chinese Herbal/pharmacology , Humans , MAP Kinase Signaling System/drug effects , MAP Kinase Signaling System/physiology , PPAR alpha/metabolism , Tumor Necrosis Factor-alpha/pharmacologyABSTRACT
Peroxisome proliferator-activated receptors (PPARs) are members of the nuclear hormone receptor superfamily of ligand-activated transcription factors and consist of three isotypes: PPARα, PPARß/δ and PPARγ. PPARs are expressed in various cell types in the skin, including keratinocytes, fibroblasts and infiltrating immune cells. Thus, these receptors are highly studied in dermato-endocrine research, and their ligands are targets for the treatment of various skin disorders, such as photoageing and chronological ageing of skin. Intensive studies have revealed that PPARα/γ functions in photoageing and age-related inflammation by regulating matrix metalloproteinases (MMPs) via nuclear factor-kappa B (NF-κB) and activator protein-1 (AP-1). However, the detailed mechanism of PPARα/γ's role in photoageing has not yet been elucidated. In this study, we confirmed that abietic acid (AA) is a PPARα/γ dual ligand and significantly decreased UVB-induced MMP-1 expression by downregulating UVB-induced MAPK signalling and downstream transcription factors, subsequently reducing IκBα degradation and blocking NF-κB p65 nuclear translocation in Hs68 human dermal fibroblast cells. Treatment of cells with AA and GW6471 or bisphenol A diglycidyl ether (BADGE), PPARα or PPARγ antagonists, respectively, reversed the effect on UVB-induced MMP-1 expression and inflammatory signalling pathway activation. Taken together, our data suggest that AA acts as a PPARα/γ dual activator to inhibit UVB-induced MMP-1 expression and age-related inflammation by suppressing NF-κB and the MAPK/AP-1 pathway and can be a useful agent for improving skin photoageing.
Subject(s)
Abietanes/chemistry , Fibroblasts/cytology , Matrix Metalloproteinase 1/metabolism , PPAR alpha/metabolism , PPAR gamma/metabolism , Skin/metabolism , Transcription Factor AP-1/metabolism , Active Transport, Cell Nucleus , Cell Line , Fibroblasts/metabolism , Humans , Inflammation , Ligands , Light , Signal Transduction , Skin/drug effects , Skin Aging , Skin Physiological Phenomena , Transcription Factor RelA/metabolism , Ultraviolet RaysABSTRACT
Peroxisome proliferator-activated receptors (PPARs) are key nuclear receptors and therapeutic targets for the treatment of metabolic diseases through the regulation of insulin resistance, diabetes, and dyslipidemia. Although a few drugs that target PPARs have been approved, more diverse and novel PPAR ligands are necessary to improve the safety and efficacy of available drugs. To expedite the search for new natural agonists of PPARs, we developed a screening assay based on ultrafiltration liquid chromatography-mass spectrometry (LC-MS) that is compatible with complex samples such as dietary foods or botanical extracts. The known PPARα and/or PPARγ ligands resveratrol and rosiglitazone were used as positive controls to validate the developed method. When applied to the screening of an Artemisia argyi extract, eupatilin was identified as a selective PPARα ligand. A PPAR competitive binding assay based on FRET detection also confirmed eupatilin as a selective PPARα agonist exhibiting a binding affinity of 1.18 µM (IC50). Furthermore, eupatilin activation of the transcriptional activity of PPARα was confirmed using a cell-based transactivation assay. Thus, ultrafiltration LC-MS is a suitable assay for the identification of PPAR ligands in complex matrixes such as extracts of dietary foods and botanicals.
Subject(s)
Artemisia/chemistry , Flavonoids , PPAR alpha , Transcriptional Activation/drug effects , Cell Line , Flavonoids/chemistry , Flavonoids/isolation & purification , Flavonoids/pharmacokinetics , Flavonoids/pharmacology , Humans , PPAR alpha/agonists , PPAR alpha/chemistry , PPAR alpha/metabolism , Protein BindingABSTRACT
Peroxisome proliferator-activated receptors (PPARs) are members of the nuclear hormone receptor superfamily of ligand-activated transcription factors and expressed in various cell types in the skin, including keratinocytes, fibroblasts and infiltrating immune cells. Thus, their ligands are targets for the treatment of various skin disorders, such as photo-aging and chronological aging of skin. Intensive studies have revealed that PPARα/γ functions in photo-aging and age-related inflammation by regulating matrix metalloproteinases (MMPs) via activator protein-1 (AP-1) and nuclear factor kappa B (NF-κB). However, the detailed mechanism of PPARα/γ's role in skin aging has not yet been elucidated. In this study, we confirmed that sargahydroquinoic acid (SHQA) as a PPARα/γ ligand significantly decreased Tumor Necrosis Factor-alpha (TNFα)-induced MMP-2/-9 expression by downregulating TNFα-induced transcription factors, subsequently reducing IκBα degradation and blocking NF-κB p65 nuclear translocation in HaCaT human epidermal keratinocyte cells. Treatment of cells with SHQA and GW6471 (PPARα antagonist) not bisphenol A diglycidyl ether (PPARγ antagonists), reversed the effect on TNFα-induced inflammatory signaling pathway activation. Taken together, our data suggest that SHQA inhibit TNFα-induced MMP-2/-9 expression and age-related inflammation by suppressing AP-1 and NF-κB pathway via PPARα.
Subject(s)
Alkenes/pharmacology , Benzoquinones/pharmacology , NF-kappa B/metabolism , PPAR alpha/agonists , Signal Transduction/drug effects , Transcription Factor AP-1/metabolism , Tumor Necrosis Factor-alpha/physiology , Base Sequence , Cell Line , DNA Primers , Humans , PhosphorylationABSTRACT
OBJECTIVE: Independent assessment of SARS-CoV-2 antigen (COV2Ag) tests remains important as varying performance between assays is common. We assessed the performance of a new high-throughput COV2Ag test compared to SARS-CoV-2 nucleic acid amplification tests (NAAT). METHODS: A total of 347 nasopharyngeal samples collected from January to October 2021 were assessed by NAAT as part of standard-of-care testing (CDC LDT or GeneXpert System, Cepheid) and COV2Ag using the ADVIA Centaur CoV2Ag assay (Siemens Healthineers). RESULTS: Among NAAT positive specimens we found 82.4% agreement and in NAAT negative specimens we found 97.3% agreement (overall agreement 85.6%). In symptomatic persons, COV2Ag agreed with NAAT 90.0% (nâ =â 291), and in asymptomatic persons, 62.5% (nâ =â 56). Agreement between positive NAAT and COV2Ag increased at lower cycle threshold (Ct) values. CONCLUSION: The COV2Ag assay exceeded the World Health Organization minimum performance requirements ofâ ≥â 80% sensitivity andâ ≥â 97% specificity. The COV2Ag assay is helpful for large scale screening efforts due to high-throughput and reduced wait times.
Subject(s)
COVID-19 , Nucleic Acids , Humans , SARS-CoV-2/genetics , COVID-19/diagnosis , COVID-19 Serological Testing , Nucleic Acid Amplification TechniquesABSTRACT
The aim of the present study was to verify the important role of Maillard reaction in the protective effect of heat-processed ginsenoside Re-serine mixture against oxidative stress-induced nephrotoxicity. The free radical-scavenging activity of ginsenoside Re-serine mixture was increased by heat-processing. Ginsenoside Re was transformed into less-polar ginsenosides such as Rg(2), Rg(6) and F(4) by heat-processing, and the glucose molecule at carbon-20 was separated. The improved-free radical-scavenging activity by heat-processing was mediated by the generation of antioxidant Maillard reaction products (MRPs) from the reaction of glucose with serine. Moreover, MRPs from ginsenoside Re-serine mixture showed protective effect against cisplatin-induced renal epithelial cell damage.
Subject(s)
Antineoplastic Agents/toxicity , Antioxidants/pharmacology , Cisplatin/toxicity , Cytoprotection/drug effects , Ginsenosides/pharmacology , Serine/pharmacology , Animals , Antioxidants/chemistry , Cell Line , Cell Survival/drug effects , Cytotoxins/toxicity , Ginsenosides/chemistry , Hot Temperature , Kidney/cytology , Kidney/drug effects , LLC-PK1 Cells , Maillard Reaction/drug effects , Oxidative Stress/drug effects , Panax/chemistry , Serine/chemistry , SwineABSTRACT
OBJECTIVES: Acute viral infections and some vaccines have been shown to increase false positivity in serologic assays. We assessed if the messenger RNA coronavirus disease 2019 (COVID-19) vaccines could cause false reactivity in common serologic assays in a pilot longitudinal cohort. METHODS: Thirty-eight participants with sera available prevaccination, 2 weeks after each vaccine dose, and monthly thereafter for up to 5 months were tested for common infectious disease serologies and antiphospholipid syndrome (APS) serology markers on the BioPlex 2200, Sure-Vue rapid plasma reagin (RPR), and Macro-Vue RPR. Twenty-two participants received the Moderna vaccine and 16 received the Pfizer vaccine. RESULTS: Most assays had no change in reactivity over the course of the sample draws, including APS markers. Epstein-Barr virus immunoglobulin G (IgG), measles IgG, and rubella immunoglobulin M all had possible false reactivity in one to two participants. RPR tests demonstrated false reactivity, with baseline nonreactive participant samples becoming reactive following vaccination. There were more false reactive participants (7/38) in the BioPlex RPR than in the Sure-Vue (2/38) and Macro-Vue (1/38) tests. All falsely reactive RPR tests were in participants who received the Moderna vaccine. CONCLUSIONS: Serologic assays with results that do not fit the clinical picture following COVID-19 vaccination should be repeated. Effects of false reactivity can last more than 5 months in some assays. In particular, RPR is susceptible to false reactivity, and there is variability among assays. Larger longitudinal studies are needed to determine the incidence and window of false reactivity.
Subject(s)
COVID-19 , Epstein-Barr Virus Infections , COVID-19/diagnosis , COVID-19/prevention & control , COVID-19 Testing , COVID-19 Vaccines , Herpesvirus 4, Human , Humans , Immunoglobulin G , RNA, Messenger , Reagins , Serologic Tests , Syphilis Serodiagnosis/methods , Vaccines, Synthetic , mRNA VaccinesABSTRACT
OBJECTIVE: Neutralizing antibodies are among the factors used to measure an individual's immune status for the control of infectious diseases. We aimed to confirm the persistence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) neutralizing antibody levels in patients who had recovered from coronavirus disease 2019 (COVID-19). METHODS: Plasma donors in South Korea who had completely recovered from SARS-CoV-2 infection had follow-up testing to determine the persistence of neutralizing antibodies using a plaque-reduction neutralization test and ELISA. RESULTS: Of the 111 participants-aged 20-29 years, 37/111 (33.3%); 30-39 years, 17/111 (15.3%); 40-49 years, 23/111 (20.7%); 50-59 years, 21/111 (18.9%); 60-65 years, 13/111 (11.7%); male, 43/111 (38.7%); female, 68/111 (61.3%)-66.1% still had neutralizing antibodies approximately 9 months (range 255-302 days) after confirmation of the diagnosis. CONCLUSIONS: In this study we analysed the titre of neutralizing antibodies associated with predicting immune status in individuals with natural infection. Information about the persistence and change in levels of neutralizing antibodies against SARS-CoV-2 can be utilized to provide evidence for developing vaccination schedules for individuals with previous infection.
Subject(s)
COVID-19 , Adult , Antibodies, Neutralizing , Antibodies, Viral , Antibody Formation , Female , Humans , Male , SARS-CoV-2 , Young AdultABSTRACT
BACKGROUND: The skin acts as a barrier to protect organisms against harmful exogenous agents. Compound K (CK) is an active metabolite of ginsenoside Rb1, Rb2 and Rc, and researchers have focused on its skin protective efficacy. In this study, we hypothesized that increased expression of the serine protease inhibitor Kazal type-5 (SPINK5) may improve skin barrier function. METHODS: We screened several ginsenosides to increase SPINK5 gene promoter activity using a transactivation assay and found that CK can increase SPINK5 expression. To investigate the protective effect of CK on the skin barrier, RT-PCR and Western blotting were performed to investigate the expression levels of SPINK5, kallikrein 5 (KLK5), KLK7 and PAR2 in UVB-irradiated HaCaT cells. Measurement of transepidermal water loss (TEWL) and histological changes associated with the skin barrier were performed in a UVB-irradiated mouse model and a 1-chloro-2,4-dinitrobenzene (DNCB)-induced atopic dermatitis-like model. RESULTS: CK treatment increased the expression of SPINK5 and decreased the expression of its downstream genes, such as KLKs and PAR2. In the UVB-irradiated mouse model and the DNCB-induced atopic dermatitis model, CK restored increased TEWL and decreased hydration and epidermal hyperplasia. In addition, CK normalized the reduced SPINK5 expression caused by UVB or DNCB, thereby restoring the expression of the proteins involved in desquamation to a level similar to normal. CONCLUSIONS: Our data showed that CK contributes to improving skin-barrier function in UVB-irradiated and DNCB-induced atopic dermatitis-like models through SPINK5. These results suggest that therapeutic attempts with CK might be useful in treating barrier-disrupted diseases.
ABSTRACT
Miniature pigs are worth notice as candidate donors for xenotransplantation. However, donor organs are inevitably subjected to hypoxia, which causes vascular endothelial dysfunction. In this respect, hypoxia-inducible factor 1alpha (HIF-1alpha), the key factor for hypoxic adaptation, should be expressed in grafts. However, some immunosuppressive drugs have been reported to suppress HIF-1alpha in rat cells. Here, we first identified the cDNA and protein structures of miniature pig HIF-1alpha, and next investigated the effects of cyclosporine and FK506 on HIF-1alpha expression in endothelial cells of miniature pig. Thus, we conclude that FK506, rather than cyclosporine, may be recommended for xenotransplantation using miniature pig organs.
Subject(s)
Hypoxia-Inducible Factor 1, alpha Subunit/metabolism , Immunosuppressive Agents/pharmacology , Swine, Miniature/metabolism , Swine/metabolism , Tacrolimus/pharmacology , Transplantation, Heterologous , Amino Acid Sequence , Anaerobiosis/drug effects , Anaerobiosis/genetics , Animals , Basic Helix-Loop-Helix Transcription Factors/antagonists & inhibitors , Basic Helix-Loop-Helix Transcription Factors/metabolism , Cloning, Molecular , Cyclosporine/pharmacology , DNA, Complementary , Endothelial Cells/drug effects , Endothelial Cells/metabolism , Hypoxia-Inducible Factor 1, alpha Subunit/antagonists & inhibitors , Hypoxia-Inducible Factor 1, alpha Subunit/genetics , Molecular Sequence Data , Protein Biosynthesis/drug effects , Protein Conformation , Swine/genetics , Swine/immunology , Swine, Miniature/genetics , Swine, Miniature/immunologyABSTRACT
Hypoxia-inducible factor-1alpha (HIF-1alpha) is believed to promote tumor growth, and thus, is viewed as one of the most compelling cancer therapy targets. YC-1 is widely used as a potent inhibitor of HIF-1alpha both in vitro and in vivo, and is also being developed as a novel anticancer drug. However, little is known about the effects of YC-1 on tumor invasion or metastasis. In the present study, we found that the Hep3B cell migration-stimulatory effect of hypoxia was abolished by HIF-1alpha siRNA or YC-1. YC-1 also significantly inhibited the migrations of other cancer cells. Furthermore, YC-1 effectively inhibited cell invasion through Matrigel. In nude mice, GFP-expressing stable cell-lines of Hep3B or H1299 were inoculated into spleens to induce liver metastasis or into the pleural cavity to induce lung invasion. In untreated mice, many tumor lesions emitting strong fluorescence were found in livers or lungs, and fluorescence intensities and tumor lesion numbers were markedly reduced in YC-1-treated mice. These results suggest that YC-1 effectively inhibits tumor invasion and metastasis, and imply that YC-1 is worth while to further develop as a multipurpose anticancer drug.
Subject(s)
Drug Screening Assays, Antitumor , Hypoxia-Inducible Factor 1, alpha Subunit/antagonists & inhibitors , Indazoles/pharmacology , Liver Neoplasms/drug therapy , Neoplasm Invasiveness/prevention & control , Animals , Antineoplastic Agents/pharmacology , Cell Line, Tumor , Cell Movement/drug effects , Collagen/chemistry , Drug Combinations , Green Fluorescent Proteins/metabolism , Humans , Laminin/chemistry , Liver Neoplasms/pathology , Mice , Mice, Inbred BALB C , Mice, Nude , Neoplasm Metastasis , Proteoglycans/chemistryABSTRACT
In this study, we aimed to demonstrate the antidiabetic potential of (E)-N-(4-(3-(5-bromo-4-hydroxy-2-methoxyphenyl)acryloyl) phenyl)-4-tert-butylbenzamide (SN158) through peroxisome proliferator-activated receptor (PPAR)-α/γ dual activation. SN158 interacted with both PPARα and PPARγ, and increased their transcriptional activities. Simultaneously, SN158 treatment led to an increase in adipogenic differentiation of 3T3-L1 preadipocytes and fatty acid oxidation in hepatocytes. In addition, glucose uptake in myotubes was significantly increased by SN158 treatment. Finally, SN158 significantly lowered the plasma levels of glucose, triglycerides, and free fatty acids in ob/ob mice without severe weight gain and hepatomegaly. These results suggest that SN158 can be useful as a potential therapeutic agent against type 2 diabetes and related metabolic disorders by alleviating glucose and lipid abnormalities.
Subject(s)
Benzamides/pharmacology , Chalcones/pharmacology , Hypoglycemic Agents/pharmacology , PPAR alpha/agonists , PPAR delta/agonists , Adipocytes/cytology , Adipocytes/drug effects , Adiponectin/genetics , Animals , Benzamides/administration & dosage , Blood Glucose/metabolism , CCAAT-Enhancer-Binding Protein-alpha/genetics , Carnitine O-Palmitoyltransferase/metabolism , Cell Differentiation/drug effects , Chalcones/administration & dosage , Fatty Acids/metabolism , Fatty Acids, Nonesterified/blood , Hypoglycemic Agents/administration & dosage , Male , Mice, Inbred C57BL , Oxidoreductases/metabolism , Pioglitazone , RNA, Messenger/metabolism , Stereoisomerism , Thiazolidinediones/pharmacology , Transcription, Genetic , Triglycerides/bloodABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: Jawoongo is a traditional drug ointment (with a traditional botanic formula) used for the treatment of burns and wounds in Korea. One of the components of Jawoongo is Lithospermi Radix (LR, the dried root of Lithospermum erythrorhizon Siebold & Zucc., also known as Zicao or Gromwell), which contains deoxyshikonin and its derivatives. OBJECTIVE: The aim of the present study was to investigate the effects of deoxyshikonin on wound healing. MATERIALS AND METHODS: The effects of LR extract and deoxyshikonin on tube formation and migration were measured in human umbilical vein vascular endothelial cells (HUVEC) and HaCaT cells, respectively. We evaluated protein expression of mitogen-activated protein kinase (MAPK) activation by Western blotting. The wound healing effects of deoxyshikonin was assessed in a mouse model of cutaneous wounds. RESULTS: The results showed that deoxyshikonin enhanced tube formation in HUVEC and migration in HaCaT cells. From the western blot analysis, we found that deoxyshikonin stimulated the phosphorylation of p38 and extracellular signal-regulated kinase (ERK) in HaCaT cells. Moreover, 20µm deoxyshikonin-treated groups showed accelerated wound closure compared with the controls in a mouse model of cutaneous wounds. CONCLUSION: In conclusion, the current data indicate that deoxyshikonin treatment elevated tube formation in HUVECs, and that deoxyshikonin-induced proliferation and migration in HaCaT cells were mediated by the activation of ERK and p38 MAPKs, respectively. Collectively, these data suggest that deoxyshikonin in Jawoongo must be an active compound for may be wound healing.
Subject(s)
Lithospermum , Naphthoquinones/pharmacology , Plant Extracts/pharmacology , Wound Healing/drug effects , Animals , Cell Line, Transformed , Cell Movement/drug effects , Cell Movement/physiology , Dose-Response Relationship, Drug , Human Umbilical Vein Endothelial Cells/drug effects , Human Umbilical Vein Endothelial Cells/physiology , Humans , Male , Mice , Mice, Inbred ICR , Naphthoquinones/isolation & purification , Plant Extracts/isolation & purification , Random Allocation , Wound Healing/physiologyABSTRACT
The 4-nonylphenol (NP) group is classified as some of the most potent endocrine disrupting chemicals (EDCs) reported to have estrogenic effects on reproductive endocrine system in vertebrates. In the present study, we investigated the effect of NP on expression of gonadotropin (GTH) subunit genes in masu salmon (Oncorhynchus masou) to clarify pituitary-based reproductive impact by EDCs. Female juvenile fish were injected with NP (a mixture of ring and chain isomers; 10 or 50 mg kg(-1) body weight) and maintained for 3 days post-injection. A semi-quantitative reverse transcription-polymerase chain reaction was used to analyze the pituitary GTHalpha, follicle-stimulating hormone beta (FSHbeta), and luteinizing hormone beta (LHbeta), and hepatic vitellogenin (VTG) mRNA levels. A low dose of NP induced the GTHalpha and LHbeta mRNA levels. High dose of NP slightly reduced FSHbeta mRNA levels in contrast to increased VTG mRNA levels. In a promoter study, NP (1-10 nM) increased the activities of luciferase reporter gene located downstream of masu salmon GTHalpha or LHbeta 5'-flanking region depending on the estrogen receptor alpha (ERalpha) in transiently transfected mammalian cells. In contrast, the luciferase activity of FSHbeta was elevated by NP in an ERalpha-independent manner. These results suggest that GTH subunit gene expression of masu salmon may be affected by EDCs at the transcription level and that the genes are useful markers for pituitary effects of xenoestrogens.
Subject(s)
Estrogen Receptor alpha/drug effects , Estrogen Receptor alpha/metabolism , Gonadotropins/genetics , Oncorhynchus/genetics , Phenols/pharmacology , Promoter Regions, Genetic/drug effects , 5' Flanking Region/drug effects , 5' Flanking Region/genetics , Amino Acid Sequence , Animals , Cells, Cultured , Estrogen Receptor alpha/genetics , Female , Follicle Stimulating Hormone/genetics , Follicle Stimulating Hormone/metabolism , Genes, Reporter , Gonadotropins/metabolism , HeLa Cells , Humans , Luteinizing Hormone/drug effects , Luteinizing Hormone/genetics , Luteinizing Hormone/metabolism , Molecular Sequence Data , Oncorhynchus/metabolism , Phenols/metabolism , Protein Subunits/drug effects , Protein Subunits/genetics , Protein Subunits/metabolism , RNA, Messenger/drug effects , RNA, Messenger/metabolism , Sequence Alignment , Transcriptional Activation/drug effects , TransfectionABSTRACT
The present study investigated the presence and mechanism of esculin-mediated renoprotection to assess its therapeutic potential. Esculin was orally administered at 20 mg/kg/day for 2 weeks to streptozotocin-induced diabetic mice, and its effects were compared with those of the vehicle in normal and diabetic mice. After oral administration of esculin to mice, the concentrations of esculin and esculetin in blood were 159.5 ± 29.8 and 9.7 ± 4.9 ng/mL at 30 min, respectively. Food and water intake were significantly increased in the diabetic mice compared to normal mice but attenuated in mice receiving esculin. The elevated blood glucose level and hepatic glucose-6-phosphatase expression were significantly reduced in esculin-treated diabetic mice, supporting the antidiabetic effect of esculin. Esculin also increased the uptake of glucose and induced the insulin-evoked phosphorylation of insulin receptor, Akt, and glycogen synthase kinase 3ß in C2C12 myotubes, indicating a potential for improvement of insulin sensitivity. In addition, esculin lessened the elevated blood creatinine levels in diabetic mice and ameliorated diabetes-induced renal dysfunction by reducing caspase-3 activation in the kidney. Data support the beneficial effect of esculin against diabetes and oxidative stress-related inflammatory processes in the kidney.