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Introduction: Managing breast cancer in female-to-male (FtM) transgender patients is complicated and challenging. Androgens play a crucial role in the development of secondary sexual identity in FtM transgender patients, but their effectiveness in breast cancer remains unclear. Furthermore, the considerations for adjuvant endocrine therapy in this population are highly intricate and warrant thorough discussion. Case Presentation: We describe the case of a 44-year-old FtM transgender diagnosed with breast cancer 3 years after initiating androgen receptor agonist therapy as part of his gender identity transition. After mastectomy, adjuvant endocrine therapy was initiated, consisting of a combination of an aromatase inhibitor and a gonadotropin-releasing hormone agonist, along with a cross-sex hormone. Conclusion: Estradiol levels were significantly reduced, and male-typical levels of sex hormones were attained.
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Extranodal natural killer (NK)/T-cell lymphoma, nasal type (ENKL), is characterized by Epstein-Barr virus infection and poor prognosis. We established a novel cell line, ENKL-J1, from bone marrow cells of an ENKL patient. We found that ENKL-J1 cells express the ganglioside GD2 (GD2) and that GD2-directed chimeric antigen receptor T cells exhibit cytotoxicity against ENKL-J1 cells, indicating that GD2 would be a suitable target of GD2-expressing ENKL cells. Targeted next-generation sequencing revealed TP53 and TET2 variants in ENKL-J1 cells. Furthermore, single-cell RNA sequencing in ENKL-J1 cells showed high gene-expression levels in the oncogenic signaling pathways JAK-STAT, NF-κB, and MAPK. Genes related to multidrug resistance (ABCC1), tumor suppression (ATG5, CRYBG1, FOXO3, TP53, MGA), anti-apoptosis (BCL2, BCL2L1), immune checkpoints (CD274, CD47), and epigenetic regulation (DDX3X, EZH2, HDAC2/3) also were expressed at high levels. The molecular targeting agents eprenetapopt, tazemetostat, and vorinostat efficiently induced apoptosis in ENKL-J1 cells in vitro. Furthermore, GD2-directed chimeric antigen receptor T cells showed cytotoxicity against ENKL-J1 cells in vivo. These findings not only contribute to understanding the molecular and genomic characteristics of ENKL; they also suggest new treatment options for patients with advanced or relapsed ENKL.
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Infecções por Vírus Epstein-Barr , Linfoma Extranodal de Células T-NK , Receptores de Antígenos Quiméricos , Humanos , Infecções por Vírus Epstein-Barr/genética , Herpesvirus Humano 4/genética , Herpesvirus Humano 4/metabolismo , Epigênese Genética , Receptores de Antígenos Quiméricos/genética , Análise da Expressão Gênica de Célula Única , Linfoma Extranodal de Células T-NK/genética , Linfoma Extranodal de Células T-NK/terapia , Linfoma Extranodal de Células T-NK/patologia , Linhagem CelularRESUMO
Primary breast angiosarcoma is an extremely rare disease with a poor prognosis. Primary angiosarcoma is distinct from secondary angiosarcoma, which usually occurs in patients who have been previously treated for breast cancer. The low incidence of primary breast angiosarcoma has hindered the elucidation of its etiology and potential therapies. Here, we report a case of a patient with primary breast angiosarcoma who experienced recurrence after surgery. The tumor was refractory to systemic treatments, and the patient died 18 months after the surgery. We used RNA sequencing for gene expression profiling of the tumor. A high tumor inflammation signature score indicated enrichment in immune-related signaling. CIBERSORTx, a tool used to characterize the cellular composition of complex tissues based on gene expression, indicated that the immune cells in the tumor were predominantly macrophages, and this was confirmed using immunohistochemical analysis. These findings indicate the possible use of checkpoint immunotherapy for the treatment of primary breast angiosarcoma.
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BACKGROUND: Postoperative prolonged air leakage is a frequent complication after lung resection. We have developed a new sealant based on a hydrophobically modified Alaska pollock-derived gelatin (ApGltn) sealant. The purpose of this study was to evaluate the adhesive strength of the ApGltn sealant in comparison with a fibrin sealant using a new spray system in ex vivo and in vivo models. METHODS: Pleural defects in ex vivo and in vivo porcine models were created, to which the ApGltn sealant or the fibrin sealant was applied. The pressure resistance was assessed with a stepwise increase in airway pressure to confirm air leakage from the sealing site. Tissue samples covered with each sealant were obtained for histologic assessment. RESULTS: In the ex vivo experiment, the leak pressures of the ApGltn sealant were significantly greater than those of the fibrin sealant (102.94 ± 15.6 cm H2O and 28.37 ± 5.1 cm H2O, respectively) (P < .01). In the in vivo experiment, the leak pressures of the ApGltn sealant were also significantly greater than those of the fibrin sealant (68.82 ± 18.04 cm H2O and 43.33 ± 7.13 cm H2O, respectively) (P = .043). The histologic examination confirmed that the ApGltn sealant adhered tightly to both the pleura and the surface of the pleural defect. CONCLUSIONS: The ApGltn sealant has sufficiently high adhesive quality in ex vivo and in vivo porcine lungs, which could be considered suitable and effective for use in the prevention of air leakage from the lungs.
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Pneumopatias , Adesivos Teciduais , Alaska , Animais , Adesivo Tecidual de Fibrina/uso terapêutico , Gelatina/uso terapêutico , Humanos , Suínos , Adesivos Teciduais/farmacologia , Adesivos Teciduais/uso terapêuticoRESUMO
The pregnane X receptor (PXR) is the key regulator of our defense mechanism against foreign substances such as drugs, dietary nutrients, or environmental pollutants. Because of increased health consciousness, the use of dietary supplements has gradually increased, and most of them can activate PXR. Therefore, an analysis of the interaction between drugs and nutrients is important because altered levels of drug-metabolizing enzymes or transporters can remarkably affect the efficiency of a co-administered drug. In the present study, we analyzed the effect of vitamin K-mediated PXR activation on drug metabolism-related gene expression in intestine-derived LS180 cells via gene expression studies and western blotting analyses. We demonstrated that menaquinone 4 (MK-4), along with other vitamin Ks, including vitamin K1, has the potential to induce MDR1 and CYP3A4 gene expression. We showed that PXR knockdown reversed MK-4-mediated stimulation of these genes, indicating the involvement of PXR in this effect. In addition, we showed that the expression of MDR1 and CYP3A4 genes increased synergistically after 24 h of rifampicin and MK-4 co-treatment. Our study thus elucidates the importance of drug-nutrient interaction mediated via PXR.
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Citocromo P-450 CYP3A/efeitos dos fármacos , Expressão Gênica/efeitos dos fármacos , Receptor de Pregnano X/efeitos dos fármacos , Vitamina K/farmacologia , Subfamília B de Transportador de Cassetes de Ligação de ATP/efeitos dos fármacos , Carcinoma/tratamento farmacológico , Carcinoma/metabolismo , Linhagem Celular Tumoral , Humanos , Neoplasias Intestinais/tratamento farmacológico , Neoplasias Intestinais/metabolismo , Fenômenos Fisiológicos da Nutrição/genética , Rifampina/administração & dosagem , Vitamina K 1/farmacologia , Vitamina K 2/análogos & derivados , Vitamina K 2/farmacologiaRESUMO
Cell identity is determined by the selective activation or silencing of specific genes via transcription factor binding and epigenetic modifications on the genome. Chromatin immunoprecipitation (ChIP) has been the standard technique for mapping the sites of transcription factor binding and histone modification. Recently, alternative methods to ChIP have been developed for addressing the increasing demands for low-input epigenomic profiling. Chromatin integration labeling (ChIL) followed by sequencing (ChIL-seq) has been demonstrated to be particularly useful for epigenomic profiling of low-input samples or even single cells because the technique amplifies the target genomic sequence before cell lysis. After labeling the target protein or modification in situ with an oligonucleotide-conjugated antibody (ChIL probe), the nearby genome sequence is amplified by Tn5 transposase-mediated transposition followed by T7 RNA polymerase-mediated transcription. ChIL-seq enables the detection of the antibody target localization under a fluorescence microscope and at the genomic level. Here we describe the detailed protocol of ChIL-seq with assessment methods for the key steps, including ChIL probe reaction, transposition, in situ transcription and sequencing library preparation. The protocol usually takes 3 d to prepare the sequencing library, including overnight incubations for the ChIL probe reaction and in situ transcription. The ChIL probe can be separately prepared and stored for several months, and its preparation and evaluation protocols are also documented in detail. An optional analysis for multiple targets (multitarget ChIL-seq) is also described. We anticipate that the protocol presented here will make the ChIL technique more widely accessible for analyzing precious samples and facilitate further applications.
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Sequenciamento de Cromatina por Imunoprecipitação/métodos , Mapeamento Cromossômico/métodos , Proteínas de Ligação a DNA/análise , Animais , Linhagem Celular , Linhagem Celular Tumoral , Cromatina/metabolismo , Imunoprecipitação da Cromatina/métodos , Epigênese Genética/genética , Epigenômica/métodos , Biblioteca Gênica , Genoma , Genômica , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Histonas/metabolismo , Humanos , Camundongos , Processamento de Proteína Pós-Traducional/genética , Análise de Sequência de DNA/métodos , Fatores de Transcrição/metabolismo , Transposases/metabolismoRESUMO
Geranylgeraniol (GGOH), a natural isoprenoid found in plants, has anti-inflammatory effects via inhibiting the activation of nuclear factor-kappa B (NFκB). However, its detailed mechanism has not yet been elucidated. Recent studies have revealed that isoprenoids can modulate signaling molecules in innate immune responses. We found that GGOH decreased the expression of lipopolysaccharide (LPS)-induced inflammatory genes in human macrophage-like THP-1 cells. Furthermore, we observed that the suppression of NFκB signaling proteins, in particular interleukin-1 receptor-associated kinase 1 (IRAK1) and tumor necrosis factor receptor-associated factor 6 (TRAF6), occurred in GGOH-treated cells prior to LPS stimulation, suggesting an immunomodulatory effect. These results indicate that GGOH may modulate and help prevent excessive NFκB activation that can lead to numerous diseases.
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Diterpenos/farmacologia , Inflamação/patologia , Quinases Associadas a Receptores de Interleucina-1/metabolismo , Macrófagos/metabolismo , Macrófagos/patologia , NF-kappa B/metabolismo , Fator 6 Associado a Receptor de TNF/metabolismo , Animais , Linhagem Celular , Humanos , Inflamação/genética , Lipopolissacarídeos , Macrófagos/efeitos dos fármacos , Camundongos , Modelos Biológicos , Fosforilação/efeitos dos fármacos , Células RAW 264.7 , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Transdução de Sinais/efeitos dos fármacos , Células THP-1RESUMO
BACKGROUND: Air leakage still remains a major problem in lung resection despite the introduction of surgical sealants. We have developed a novel sealant based on hydrophobically modified Alaska pollock-derived gelatin (ApGltn), which showed high adhesive quality in vitro. In this study, we evaluated the adhesive quality and conformability of our ApGltn sealant compared with a fibrin sealant. METHODS: The adhesive quality of the sealants was evaluated using excised porcine lungs with ventilation. Pleural defects were created, to which the ApGltn sealant or fibrin sealant was applied. Pressure resistance was assessed using a stepwise increase of airway pressure. Conformability was evaluated by measuring the area of the sealant for its maximum conformity on the gradually inflated lung surface. RESULTS: Leak and burst pressures of the ApGltn sealant were significantly higher than those of the fibrin sealant (47.1 ± 10.5 cm H2O and 52.3 ± 9.4 cm H2O versus 33.9 ± 6.0 cm H2O and 37.5 ± 5.9 cm H2O, respectively). Maximum expansion areas of the ApGltn sealant and fibrin sealant were 2652.4 ± 324.6 mm2 and 1276.6 ± 323.5 mm2, respectively. The ApGltn sealant also showed higher adhesive quality and conformability compared with the fibrin sealant. Histological examination confirmed that the ApGltn sealant showed tight adhesion to the pleural surface, while a gap was observed with the fibrin sealant. CONCLUSIONS: The ApGltn sealant showed higher adhesive quality and conformability than the fibrin sealant in an excised porcine lung model. We expect that, in the future, the ApGltn sealant will be used for lung resections in clinical settings.
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Adesivo Tecidual de Fibrina , Pulmão/cirurgia , Adesivos Teciduais , Animais , Gelatina , Teste de Materiais , SuínosRESUMO
Cardiopulmonary bypass (CPB) recovery is complicated by lung inflammation from bone marrow (BM)-derived polymorphonuclear leukocytes (PMNs) and monocytes (MO). Although Sivelestat reduces inflammatory mediators and Rolipram inhibits PMN and MO activation, any kinetic effects to improve CPB recovery in vivo are unknown. We hypothesized that intraoperative co-administration of these compounds would reduce CPB-induced lung inflammation through downregulation of PMN and MO recruitment. A 2-h CPB was surgically established in cynomolgus monkeys (n = 13), and BM leukocyte release and lung recruitment were monitored postoperatively by flow cytometry with 5'-bromo-2'-deoxyuridine (BrdU) and cytokine ELISA. Either Sivelestat, Sivelestat plus Rolipram, or saline (control) was administered intraoperatively and both peripheral and perfusion sampling courses revealed BrdU-labeled cells representative of activated leukocyte infiltration. Levels of cytokines CD11b and CD18 were leukocytic activation markers. Sivelestat plus Rolipram attenuated increases in CPB-associated circulating band cells, prolonged BM-transit time (PMN: 121.0 ± 3.7 to 96.2 ± 4.3 h [control], p = 0.012; MO: 84.4 ± 4.1 to 61.4 ± 3.0 h [control], p = 0.003), and reduced their alveolar appearance. CD11b-mediated PMN and MO changes during CPB and the post-surgical increases of Interleukin (IL)-6 and IL-8 in the bronchoalveolar lavage fluid were suppressed. Sivelestat alone increased PMN transit time to 115.8 ± 6.6 h, but monocytes were unaffected. Therefore, Rolipram has additive inhibitory effects with Sivelestat on the CPB-induced activation and release of BM-derived PMNs and MO and their recruitment to the lungs. Co-administration of these compounds could, therefore, hold value for preventing CPB-induced lung injury.
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Células da Medula Óssea/citologia , Ponte Cardiopulmonar/efeitos adversos , Glicina/análogos & derivados , Pulmão/patologia , Neutrófilos/citologia , Rolipram/farmacologia , Sulfonamidas/farmacologia , Animais , Células da Medula Óssea/efeitos dos fármacos , Quimiotaxia de Leucócito/efeitos dos fármacos , Modelos Animais de Doenças , Glicina/farmacologia , Macaca fascicularis , Masculino , Neutrófilos/efeitos dos fármacos , Inibidores da Fosfodiesterase 4/farmacologia , Inibidores de Serina Proteinase/farmacologiaRESUMO
Sialic acid-binding lectin from Rana catesbeiana eggs (cSBL) is a multifunctional protein that has lectin and ribonuclease activity. In this study, the anti-tumor activities of cSBL were assessed using a panel of breast cancer cell lines. cSBL suppressed the cell growth of all cancer cell lines tested here at a concentration that is less toxic, or not toxic at all, to normal cells. The growth suppressive effect was attributed to the cancer-selective induction of apoptosis. We assessed the expressions of several key molecules associated with the breast cancer phenotype after cSBL treatment by western blotting. cSBL decreased the expression level of estrogen receptor (ER) α, while it increased the phosphorylation level of p38 mitogen-activated protein kinase (MAPK). cSBL also suppressed the expression of the progesterone receptor (PgR) and human epidermal growth factor receptor type 2 (HER2). Furthermore, it was revealed that cSBL decreases the expression of the epidermal growth factor receptor (EGFR/HER1) in triple-negative breast cancer cells. These results indicate that cSBL induces apoptosis with decreasing ErbB family proteins and may have great potential for breast cancer chemotherapy, particularly in triple-negative phenotype cells.
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Proteínas de Anfíbios/farmacologia , Apoptose/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Lectinas/farmacologia , Ribonucleases/farmacologia , Neoplasias de Mama Triplo Negativas/tratamento farmacológico , Proteínas de Anfíbios/química , Animais , Receptores ErbB/genética , Receptor alfa de Estrogênio/genética , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Lectinas/química , Células MCF-7 , Fenótipo , Rana catesbeiana , Receptor ErbB-2/genética , Ribonucleases/química , Neoplasias de Mama Triplo Negativas/patologia , Proteínas Quinases p38 Ativadas por Mitógeno/genéticaRESUMO
Serum endonucleases are essential for degrading the chromatin released from dead cells and preventing autoimmune diseases such as systemic lupus erythematosus. Serum DNase I is known as the major endonuclease, but recently, another endonuclease, DNase γ/DNase I-like 3, gained attention. However, the precise role of each endonuclease, especially that of DNase γ, remains unclear. In this study, we distinguished the activities of DNase γ from those of DNase I in mouse serum and concluded that both cooperated in degrading DNA during necrosis: DNase γ functions as the primary chromatolytic activity, causing internucleosomal DNA fragmentation, and DNase I as the secondary one, causing random DNA digestion for its complete degradation. These results were confirmed by two in vivo experimental mouse models, in which necrosis was induced, acetaminophen-induced hepatic injury and streptozotocin-induced ß-cell necrosis models. We also determined that DNase γ functions as a backup endonuclease for caspase-activated DNase (CAD) in the secondary necrosis phase after γ-ray-induced apoptosis in vivo.
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Degradação Necrótica do DNA , Desoxirribonucleases de Sítio Específico do Tipo I/sangue , Desoxirribonucleases/sangue , Endodesoxirribonucleases/sangue , Animais , Apoptose , Linhagem Celular Tumoral , Fragmentação do DNA , Feminino , Humanos , Fígado/metabolismo , Fígado/ultraestrutura , Masculino , Camundongos , Camundongos Knockout , Complexos Multienzimáticos , Necrose/sangue , Proteínas de Ligação a Poli-ADP-Ribose , Baço/metabolismoRESUMO
A 30-year-old woman with corrected transposition of great arteries (c-TGA) was scheduled for elective cesarean section at 37 weeks of gestation. At previous cesarean section, she received general anesthesia for dyspnea and lower cardiac function by severe mitral regurgitation, with a pulmonary catheter inserted. In the current pregnancy, she had tricuspid regurgitation, but she had no signs of heart failure. Cardiac index (CI) and stroke volume variation (SVV) were monitored by the FloTrack, before induction of anesthesia. Because the CI was 3.6 l x min(-1) x m(-2), and the SVV was 18%, we decided to perform combined spinal epidural anesthesia. Epidural anesthesia was performed at L1-2, and spinal anesthesia was performed at L3-4. Hyperbaric 0.5% bupivacaine 2.0 ml with fentanyl 10 µg was given to the subarachnoid space. The total dose of phenylephrine administered was 150 µg, and the CI as well as the SVV were stable during surgery. Her postpartum couse was uneventful. Anesthetic management of c-TGA is discussed, and we should select anesthetic method carefully.
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Anestesia Obstétrica , Cesárea , Complicações Cardiovasculares na Gravidez , Transposição dos Grandes Vasos/complicações , Adulto , Transposição das Grandes Artérias Corrigida Congenitamente , Feminino , Humanos , Gravidez , Resultado da Gravidez , Radiografia , Transposição dos Grandes Vasos/diagnóstico por imagemRESUMO
BACKGROUND AND AIM: Infliximab is an established therapy for ulcerative colitis (UC). The aim of this study was to examine various serum cytokine levels and to identify possible markers predictive of therapeutic efficacy of infliximab for UC patients. METHODS: Twenty-one patients with moderately active UC were given intravenous infliximab (5 mg/kg) at 0, 2, and 6 weeks as induction therapy. The serum levels of 17 cytokines were determined using a Bio-Plex suspension array system before and 8 weeks after induction therapy. Partial Mayo score (PMS) and serum C-reactive protein levels were used for the determination of clinical activities at 0 and 8 weeks after the treatment. The overall therapeutic effect was determined at 26 weeks according to the PMS. RESULTS: The median value of the PMS decreased significantly 8 weeks after the treatment (from 6 to 1.5, P < 0.05). However, C-reactive protein levels did not change significantly. Levels of serum interleukin (IL)-8 (P < 0.05) and macrophage inflammatory protein-1ß (P < 0.005) significantly decreased 8 weeks after the induction. Serum levels of the other 15 cytokines did not change significantly. At 26 weeks, 13 of 20 patients (65%) were responders while 7 patients were non-responders. Levels of serum IL-6 at 8 weeks were significantly lower in responders than in non-responders (P < 0.05). CONCLUSIONS: Serum IL-8 and macrophage inflammatory protein-1ß seem to be sensitive markers for UC patients treated with infliximab, while IL-6 at 8 weeks after induction therapy may be predictive of subsequent response to infliximab.
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Colite Ulcerativa/diagnóstico , Colite Ulcerativa/tratamento farmacológico , Citocinas/sangue , Fármacos Gastrointestinais/uso terapêutico , Infliximab/uso terapêutico , Biomarcadores/sangue , Proteína C-Reativa , Quimiocina CCL4/sangue , Previsões , Fármacos Gastrointestinais/administração & dosagem , Quimioterapia de Indução , Infliximab/administração & dosagem , Interleucina-6/sangue , Interleucina-8/sangue , Fatores de TempoRESUMO
Since bioprosthetic valve dysfunction may arise due to histological calcification in the crosslinking process by glutaraldehyde (GA), non-GA crosslinking reagents have been investigated. We compared the efficacy of triglycidylamine (TGA), a newly synthesized epoxy compound, and GA as crosslinking reagents for the treatment of autologous tissues. We assessed the strength of crosslinked tissues using shrinkage temperature (Ts) measured by differential scanning calorimetry. We also conducted subdermal allografting of the crosslinked pericardium and thoracic aorta in rats, and verified the anti-calcification efficacy of TGA by histological evaluations with von Kossa stain, and immunological evaluations using tenascin-C (TN-C) or matrix metalloproteinase-9 (MMP-9). TGA treatment resulted in slower increases in Ts of the pericardium, and it required 9-12 h to reach Ts achieved by GA. In subdermal implantation of rat tissues, calcium content was lower in the TGA group than in the GA groups (p < 0.005). The expression site of TN-C and MMP-9 differed from the primary location of calcium deposition in the thoracic aorta treated with TGA suggesting a different underlying mechanism in calcification between GA and TGA crosslinking. In conclusion, TGA crosslinking in the allograft showed superior anti-calcification effect as compared to brief treatment by GA, although TGA crosslinking process was slow.
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Aorta Torácica/transplante , Bioprótese , Calcinose/prevenção & controle , Compostos de Epóxi/farmacologia , Glutaral/farmacologia , Próteses Valvulares Cardíacas , Pericárdio/transplante , Animais , Aorta Torácica/efeitos dos fármacos , Calcinose/metabolismo , Reagentes de Ligações Cruzadas/farmacologia , Modelos Animais de Doenças , Humanos , Masculino , Pericárdio/efeitos dos fármacos , Ratos , Ratos Wistar , Temperatura , Transplante AutólogoRESUMO
BACKGROUND: A bone marrow (BM) response induced by cardiopulmonary bypass (CPB) as a systemic inflammatory reaction has previously been postulated but not clarified. Newly released polymorphonuclear leukocytes (PMNs) and monocytes from the BM are known to be immature, indicating their greater potential to damage tissue. The present study aimed to examine the kinetics of BM-derived leukocytes associated with CPB in a nonhuman primate model. METHODS: Normothermic CPB was performed in cynomolgus monkeys for 2 hours through a median sternotomy. Leukocyte precursors were labeled in the BM of the monkeys in vivo by an intravenous injection of 5-bromo-2'-deoxyuridine (BrdU), and their release into the circulation and recruitment to the lungs after operation with or without CPB (control group) were monitored over time by flow cytometry. RESULTS: In normal-state monkeys, the calculated transit time of BrdU-labeled PMNs (PMNBrdU) through the BM was 143.6±4.5 hours and that of monocytes was 100.9±7.6 hours. CPB caused a rapid release of PMNs and monocytes from the BM, shortened their transit through the BM to 92.0±4.1 and 60.3±2.9 hours, respectively, and further induced their increased appearance in the alveolar spaces, with a significant increase in both interleukin (IL)-6 and IL-8 levels in the bronchoalveolar lavage fluid (BALF) 24 hours after CPB. CONCLUSIONS: CPB accelerated the release of PMNs and monocytes from the BM and their recruitment to the lungs in our monkey model, indicating that this model is relevant for monitoring the kinetics of BM-derived leukocytes in humans.
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Ponte Cardiopulmonar/efeitos adversos , Quimiotaxia de Leucócito , Pulmão/imunologia , Pulmão/patologia , Neutrófilos , Animais , Células da Medula Óssea , Haplorrinos , MasculinoRESUMO
BACKGROUND: We tested the hypothesis that heparin administration prior to the emergence of tissue factor (TF) would increase plasma TF pathway inhibitor (TFPI) and attenuate TF-mediated thrombin generation during simulated cardiopulmonary bypass (CPB). METHODS: Human blood was recirculated for 120 minutes using an oxygenator and roller pump. Four groups were examined: control group (heparin 3.75 U/mL, in donor blood, n = 7), rTF group (heparin + recombinant TF 1000 pg/mL, in donor blood, n = 7), TFPI boost group (heparin, in preheparinized donor blood, n = 8), and rTF + TFPI boost group (heparin + rTF, in preheparinized blood, n = 7). In the two TFPI boost groups, 50 U/kg of heparin was given to the donors intravenously five minutes before donation to boost plasma TFPI levels. Total plasma TFPI, thrombin-antithrombin complex, and prothrombin fragment F1+2 levels were measured before and during CPB. RESULTS: Preheparinization increased total plasma TFPI levels by a factor of 8.0. Administration of rTF significantly enhanced the generation of F1+2 (p = 0.0002). The heparin-induced TFPI elevation reduced both thrombin-antithrombin complex and F1+2 to control levels in rTF + TFPI boost group (p = 0.0158 for thrombin-antithrombin complex, p < 0.0001 for F1+2 ). F1+2 levels were at all times lower than control levels in TFPI boost group (p < 0.0001). CONCLUSIONS: Heparin-induced TFPI elevation attenuates TF-mediated thrombin generation. Early heparin administration prior to the emergence of plasma TF may represent a novel strategy for controlling thrombin generation by the extrinsic coagulation pathway during CPB.
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Ponte Cardiopulmonar , Heparina/administração & dosagem , Heparina/farmacologia , Lipoproteínas/sangue , Trombina/metabolismo , Tromboplastina/metabolismo , Antitrombina III , Coagulação Sanguínea/fisiologia , Humanos , Peptídeo Hidrolases/sangueRESUMO
Although the aryl hydrocarbon receptor (AHR) and glucocorticoid receptor (GR) play essential roles in mammalian development, stress responses, and other physiological events, crosstalk between these receptors has been the subject of much debate. Metallothioneins are classic glucocorticoid-inducible genes that were reported to increase upon treatment with AHR agonists in rodent tissues and cultured human cells. In this study, the mechanism of human metallothionein 2A (MT2A) gene transcription activation by AHR was investigated. Cotreatment with 3-methylcholanthrene and dexamethasone, agonists of AHR and GR respectively, synergistically increased MT2A mRNA levels in HepG2 cells. MT2A induction was suppressed by RNA interference against AHR or GR. Coimmunoprecipitation experiments revealed a physical interaction between AHR and GR proteins. Moreover, chromatin immunoprecipitation assays indicated that AHR was recruited to the glucocorticoid response element in the MT2A promoter. Thus, we provide a novel mechanism whereby AHR modulates expression of human MT2A via the glucocorticoid response element and protein-protein interactions with GR.
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Metalotioneína/metabolismo , Receptores de Hidrocarboneto Arílico/metabolismo , Receptores de Glucocorticoides/metabolismo , Animais , Células COS , Chlorocebus aethiops , Imunoprecipitação da Cromatina , Dexametasona/farmacologia , Células Hep G2 , Humanos , Imunoprecipitação , Metalotioneína/genética , Metilcolantreno/farmacologia , Regiões Promotoras Genéticas , Domínios e Motivos de Interação entre Proteínas/efeitos dos fármacos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Receptores de Hidrocarboneto Arílico/agonistas , Receptores de Hidrocarboneto Arílico/genética , Receptores de Glucocorticoides/agonistas , Receptores de Glucocorticoides/genética , Elementos de Resposta/efeitos dos fármacos , Transdução de Sinais , Ativação TranscricionalRESUMO
We describe the clinical use of a new video-laryngoscope (McGRATH MAC, McG) in patients with a difficult airway and morbid obesity. In a patient, case no. 2, with a difficult airway, showing a Cormack-Lehane grade III view with Macintosh direct laryngoscope, the glottis opening (Cormack-Lehane grade I) was visualized with McG. McG also provided a good view of glottis opening in a patient with morbid obesity. McG will have a profound impact on the management of the difficult airway.
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Intubação Intratraqueal/instrumentação , Laringoscópios , Idoso , Humanos , Masculino , Obesidade Mórbida/complicações , Cirurgia Vídeoassistida/instrumentaçãoRESUMO
Paroxysmal nocturnal hemoglobinuria (PNH) is characterized by complement (C')-induced lysis of PNH red blood cells (RBCs), which are deficient in the expression of CD55 and CD59. Surgery is one of the major clinical situations that trigger hemolytic attack and thrombosis in PNH. We describe here a case of 64-year-old man with classic PNH complicated by early-stage gastric cancer requiring distal gastrectomy under general anesthesia. We administered humanized monoclonal anti-C5 antibody (eculizumab; Soliris) for a limited period (600 mg, once a week × four times) perisurgically. Eculizumab effectively inhibited the C' system and the patient underwent a curative distal gastrectomy without significant surgery-triggered hemolytic attack. Although discontinuation of eculizumab induced mild hemolysis 2 weeks after the last administration, it was treated conservatively without thrombotic complication. Limited-term induction of eculizumab could be an option for PNH patients with transient and anticipated high risks, with careful preparation for the discontinuation-related risks afterwards.
Assuntos
Anticorpos Monoclonais Humanizados/administração & dosagem , Hemoglobinúria Paroxística/sangue , Hemoglobinúria Paroxística/cirurgia , Anticorpos Monoclonais Humanizados/efeitos adversos , Hemoglobinúria Paroxística/tratamento farmacológico , Hemólise/efeitos dos fármacos , Humanos , Masculino , Pessoa de Meia-Idade , Cuidados Pré-Operatórios/métodosRESUMO
PURPOSE: The isoprenoid geranylgeraniol (GGOH) inhibits nuclear factor-kappa B (NF-κB) activation in the liver, yet the mechanism remains unclear. We investigated the modulation and inhibition of lipopolysaccharide (LPS)-induced NF-κB signaling in the liver of rats fed a GGOH-supplemented diet. METHODS: Rats were fed a diet supplemented with or without GGOH for 10 days. Rats were then intraperitoneally injected with 0.5 mg/kg LPS or vehicle (sterilized saline) and fasted for 18 h. Plasma levels of the inflammatory cytokines tumor necrosis factor (TNF)-α, interleukin (IL)-1ß, and IL-6, and the liver damage indicators alanine and aspartate aminotransferases (ALT and AST) were assessed. Liver mRNA and proteins were assayed for changes in NF-κB target genes and signal transduction genes. RESULTS: Rats fed a high-dose, GGOH-supplemented diet showed significantly lower levels of plasma inflammatory cytokines and ALT and AST activities. In the liver, GGOH significantly suppressed NF-κB activation and mRNA expression of its pro-inflammatory target genes. Furthermore, GGOH supplementation substantially suppressed mRNA expression of signal transducer genes upstream of the IκB kinase complex. Western blotting of liver extracts further demonstrated the substantial decrease in total IL-1 receptor-associated kinase 1 (IRAK1) and TNF receptor-associated factor 6 (TRAF6), leading to lower signal transduction and inhibition of NF-κB after LPS. CONCLUSION: A 10-day, high-dose, GGOH-supplemented diet was sufficient to inhibit LPS-induced inflammation and activation of NF-κB in rat livers. GGOH significantly modulated NF-κB signaling molecules, inhibiting its signal transduction and activation in the liver, thus protecting against liver damage.