RESUMO
Aerosol-generating procedures are medical interventions considered high risk for transmission of airborne pathogens. Tracheal intubation of anaesthetised patients is not high risk for aerosol generation; however, patients often perform respiratory manoeuvres during awake tracheal intubation which may generate aerosol. To assess the risk, we undertook aerosol monitoring during a series of awake tracheal intubations and nasendoscopies in healthy participants. Sampling was undertaken within an ultraclean operating theatre. Procedures were performed and received by 12 anaesthetic trainees. The upper airway was topically anaesthetised with lidocaine and participants were not sedated. An optical particle sizer continuously sampled aerosol. Passage of the bronchoscope through the vocal cords generated similar peak median (IQR [range]) aerosol concentrations to coughing, 1020 (645-1245 [120-48,948]) vs. 1460 (390-2506 [40-12,280]) particles.l-1 respectively, p = 0.266. Coughs evoked when lidocaine was sprayed on the vocal cords generated 91,700 (41,907-166,774 [390-557,817]) particles.l-1 which was significantly greater than volitional coughs (p < 0.001). For 38 nasendoscopies in 12 participants, the aerosol concentrations were relatively low, 180 (120-525 [0-9552]) particles.l-1 , however, five nasendoscopies generated peak aerosol concentrations greater than a volitional cough. Awake tracheal intubation and nasendoscopy can generate high concentrations of respiratory aerosol. Specific risks are associated with lidocaine spray of the larynx, instrumentation of the vocal cords, procedural coughing and deep breaths. Given the proximity of practitioners to patient-generated aerosol, airborne infection control precautions are appropriate when undertaking awake upper airway endoscopy (including awake tracheal intubation, nasendoscopy and bronchoscopy) if respirable pathogens cannot be confidently excluded.
Assuntos
Tosse , Vigília , Humanos , Tosse/etiologia , Aerossóis e Gotículas Respiratórios , Intubação Intratraqueal/métodos , LidocaínaRESUMO
A simple, compact, and high-sensitivity optical sensor for salinity measurement is reported based on an optical microfiber coil resonator (MCR). The MCR is manufactured by initially wrapping microfiber on a polymethylmethacrylate (PMMA) rod, which is dissolved to leave a hollow cylindrical fluidic channel within the coil for measurement. Based on the light propagation through the MCR, the device's spectrum moves to long wavelengths with increased salinity in the fluid. The MCR device's sensitivity can reach up to 15.587 nm/% with a resolution of 1.28 × 10-3%. It is also confirmed that the temperature dependence is 79.87 pm/°C, which results from the strong thermal-expansion coefficient of the low refractive index epoxy. The experimental results indicate that the device can be widely used as a high sensitivity salinity sensor in water and other liquids due to its stability, compactness, electromagnetic immunity, and high sensitivity.
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BACKGROUND: Entecavir and tenofovir potently suppress hepatitis B virus (HBV) replication so that serum HBV DNA levels <20 IU/mL can be achieved after 2 years. Despite this, inadequate suppression is reported in >20% of cases for unclear reasons. AIM: We tested whether 4-week viral load (VL) assessment could improve 96-week treatment outcome. METHODS: Data on all chronic hepatitis B patients treated with entecavir or tenofovir between 2005 and 2014 were entered prospectively. Full data capture included pre-treatment, weeks 4, 24, 48 and 96 HBV DNA titre, HBeAg, age, gender, antiviral agent and dose escalation. Compliance data were compiled from pharmacy records, doctors' letters and clinic bookings/attendance. Time to achieve complete viral suppression (HBV DNA < 20 IU/mL) was graphed using Kaplan-Meier curves. Factors affecting this were examined using a multivariate Cox Proportional Hazard model. RESULTS: Among 156 patients treated, 72 received entecavir and 84 tenofovir. Pre-treatment HBV DNA titre, 4-week assessment and compliance impacted significantly on time to complete viral suppression. At 96 weeks, 90% of those assessed as compliant by 4-week HBV DNA had complete viral suppression versus 50% followed by 6-month VL estimation. Continuing care by the same physician was related to 4-week VL testing and optimal compliance. CONCLUSIONS: Medium-term outcomes of HBV antiviral therapy are improved by early on-treatment VL testing, facilitating patient engagement and improved compliance. The observation that 90% complete viral suppression after 2 years monotherapy is achievable in a routine clinic setting questions the need for combination therapy in HBV cases with suboptimal response.
Assuntos
Antivirais/uso terapêutico , Guanina/análogos & derivados , Hepatite B Crônica/tratamento farmacológico , Tenofovir/uso terapêutico , Carga Viral , Adulto , Idoso , Idoso de 80 Anos ou mais , Austrália , DNA Viral/sangue , Quimioterapia Combinada , Feminino , Guanina/uso terapêutico , Vírus da Hepatite B/efeitos dos fármacos , Humanos , Estimativa de Kaplan-Meier , Masculino , Pessoa de Meia-Idade , Modelos de Riscos Proporcionais , Resultado do TratamentoRESUMO
BACKGROUND: We have previously demonstrated that peroxisome proliferator-activated receptor (PPARγ) activation inhibits hepatocarcinogenesis. We aim to investigate the effect of PPARγ on hepatocellular carcinoma (HCC) metastatic potential and explore its underlying mechanisms. METHODS: Human HCC cells (MHCC97L, BEL-7404) were infected with adenovirus-expressing PPARγ (Ad-PPARγ) or Ad-lacZ and treated with or without PPARγ agonist (rosiglitazone). The effects of PPARγ on cell migration and invasive activity were determined by wound healing assay and Matrigel invasive model in vitro, and in an orthotopic liver tumour metastatic model in mice. RESULTS: Pronounced expression of PPARγ was demonstrated in HCC cells (MHCC97L, BEL-7404) treated with Ad-PPARγ, rosiglitazone or Ad-PPARγ plus rosiglitazone, compared with control (Ad-LacZ). Such induction markedly suppressed HCC cell migration. Moreover, the invasiveness of MHCC97L and BEL-7404 cells infected with Ad-PPARγ, or treated with rosiglitazone was significantly diminished up to 60%. Combination of Ad-PPARγ and rosiglitazone showed an additive effect. Activation of PPARγ by rosiglitazone significantly reduced the incidence and severity of lung metastasis in an orthotopic HCC mouse model. Key mechanisms underlying the effect of PPARγ in HCC include upregulation of cell adhesion genes, E-cadherin and SYK (spleen tyrosine kinase), extracellular matrix regulator tissue inhibitors of metalloproteinase (TIMP) 3, tumour suppressor gene retinoblastoma 1, and downregulation of pro-metastatic genes MMP9 (matrix metallopeptidase 9), MMP13, HPSE (heparanase), and Hepatocyte growth factor (HGF). Direct transcriptional regulation of TIMP3, MMP9, MMP13, and HPSE by PPARγ was shown by ChIP-PCR. CONCLUSION: Peroxisome proliferator-activated receptor-gamma exerts an inhibitory effect on the invasive and metastatic potential of HCC in vitro and in vivo, and is thus, a target for the prevention and treatment of HCC metastases.
Assuntos
Carcinoma Hepatocelular/tratamento farmacológico , Neoplasias Hepáticas/tratamento farmacológico , Neoplasias Pulmonares/tratamento farmacológico , PPAR gama/agonistas , PPAR gama/metabolismo , Tiazolidinedionas/farmacologia , Animais , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Western Blotting , Caderinas , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/patologia , Adesão Celular/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Imunoprecipitação da Cromatina , Perfilação da Expressão Gênica , Humanos , Hipoglicemiantes/farmacologia , Técnicas In Vitro , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/patologia , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/secundário , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Análise de Sequência com Séries de Oligonucleotídeos , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Rosiglitazona , Células Tumorais CultivadasRESUMO
OBJECTIVE: Fat Aussie mice (foz/foz) are morbidly obese, glucose intolerant and have liver steatosis that develops into steatohepatitis on a high-fat diet. The cannabinoid 1 receptor (CB1) antagonist SR141716 has been shown to improve obesity-associated metabolic complications in humans and rodent models. The aim of this study was to assess the effect of SR141716 in foz/foz mice. DESIGN: Male wildtype (WT) and foz/foz mice were fed a chow or high-fat diet (45% saturated fat). Vehicle or SR141716 (10 mg kg(-1) per day) was administered in jelly once daily for 4 weeks from 4 months of age. RESULTS: Foz/foz mice were obese but had less epididymal adipose tissue mass than fat-fed WT mice despite being significantly heavier. Liver weight was increased by twofold in foz/foz compared with WT mice and showed significant steatogenesis associated with impaired liver function. Foz/foz and fat-fed WT mice were glucose intolerant as determined by oral glucose tolerance test. In chow-fed foz/foz mice, SR141716 reduced body weight, liver weight, reversed hepatosteatosis and glucose intolerance. Subcutaneous white adipose tissue gene expression of the macrophage-specific marker Cd68 reflected the improvements in the metabolic status by SR141716 in these mice. CONCLUSION: The results are consistent with the hypothesis that foz/foz mice have defective lipid metabolism, are unable to adequately store fat in adipose tissue but instead sequester fat ectopically in other metabolic tissues (liver) leading to insulin resistance and hepatic steatosis associated with inflammation. Our findings suggest that SR141716 can improve liver lipid metabolism in foz/foz mice in line with improved insulin sensitivity and adipose tissue inflammation.
Assuntos
Tecido Adiposo/metabolismo , Fígado Gorduroso/tratamento farmacológico , Inflamação/tratamento farmacológico , Metabolismo dos Lipídeos/efeitos dos fármacos , Fígado/metabolismo , Obesidade Mórbida/tratamento farmacológico , Obesidade Mórbida/metabolismo , Piperidinas/farmacologia , Pirazóis/farmacologia , Receptor CB1 de Canabinoide/antagonistas & inibidores , Tecido Adiposo/efeitos dos fármacos , Tecido Adiposo/patologia , Animais , Dieta Hiperlipídica , Modelos Animais de Doenças , Fígado Gorduroso/metabolismo , Fígado Gorduroso/patologia , Intolerância à Glucose/metabolismo , Teste de Tolerância a Glucose , Inflamação/etiologia , Inflamação/metabolismo , Inflamação/patologia , Resistência à Insulina , Fígado/efeitos dos fármacos , Fígado/patologia , Masculino , Camundongos , Obesidade Mórbida/complicações , RimonabantoRESUMO
Despite advances in antiviral therapy, molecular drivers of Hepatitis C Virus (HCV)-related liver disease remain poorly characterised. Chronic infection with HCV genotypes (1 and 3) differ in presentation of liver steatosis and virological response to therapies, both to interferon and direct acting antivirals. To understand what drives these clinically important differences, liver expression profiles of patients with HCV Genotype 1 or 3 infection (n = 26 and 33), alcoholic liver disease (n = 8), and no liver disease (n = 10) were analysed using transcriptome-wide microarrays. In progressive liver disease, HCV genotype was the major contributor to altered liver gene expression with 2151 genes differentially expressed >1.5-fold between HCV Genotype 1 and 3. In contrast, only 6 genes were altered between the HCV genotypes in advanced liver disease. Induction of lipogenic, lipolytic, and interferon stimulated gene pathways were enriched in Genotype 1 injury whilst a broad range of immune-associated pathways were associated with Genotype 3 injury. The results are consistent with greater lipid turnover in HCV Genotype 1 patients. Moreover, the lower activity in inflammatory pathways associated with HCV genotype 1 is consistent with relative resistance to interferon-based therapy. This data provides a molecular framework to explain the clinical manifestations of HCV-associated liver disease.
Assuntos
Regulação Viral da Expressão Gênica/genética , Hepacivirus/genética , Hepatite C/metabolismo , Inflamação/metabolismo , Metabolismo dos Lipídeos , Fígado/virologia , Adolescente , Adulto , Feminino , Perfilação da Expressão Gênica , Genótipo , Humanos , Fígado/metabolismo , Cirrose Hepática/metabolismo , Hepatopatias Alcoólicas/metabolismo , Masculino , Redes e Vias Metabólicas , Pessoa de Meia-Idade , Transcrição Gênica , Adulto JovemRESUMO
The objective of the study was to evaluate the role of rapid virological response (RVR) in predicting sustained virological response (SVR) rates to hepatitis C virus (HCV) therapy. 65 HIV / HCV co-infected patients commenced HCV treatment per protocol. HIV / HCV patients with a mean CD4 count of 502 were treated for 24-48 weeks depending on genotype. Virological response was assessed at weeks 4 (RVR), 12 [early virological response (EVR)], 24, at end of treatment (EOTR) and 24 weeks post-completion of treatment (SVR). Primary end-point was defined as undetectable HCV RNA at 24 weeks post-treatment completion. Fifty-five per cent of co-infected patients were on highly active anti-retroviral therapy. A majority of patient group were male. 60% of HIV / HCV patients achieved SVR (35% genotype 1 / 4; 77% genotype 2 / 3). 24 HIV / HCV patients achieved undetectable HCV levels compared with baseline by week 4. The positive predictive value (PPV) of RVR at week 4 for subsequent SVR in HIV-HCV co-infected patients was 100%; the negative predictive value (NPV) was 57%. Significant variables associated with SVR were: (i) lower median pre-treatment HCV viral load, (ii) genotype 2 / 3 disease and (iii) achievement of RVR. Independent variables associated with RVR were low pre-treatment HCV viral load and genotype 2 / 3 disease. Achievement of RVR, a negative HCV-PCR, at week 4 of treatment is predictive of SVR in this cohort of patients. This may be used to guide optimal treatment duration in patient groups. More significantly, the data serve to highlight the subgroup of patients who, on achieving RVR, should be actively supported to complete HCV treatment with full dose therapy, especially patients co-infected with G2 / 3 disease for whom 6 months' full dose therapy may be sufficient to obtain a SVR.
Assuntos
Antivirais/uso terapêutico , Infecções por HIV/complicações , Hepacivirus/genética , Hepatite C/complicações , Interferon-alfa/uso terapêutico , Ribavirina/uso terapêutico , Viremia , Adulto , Feminino , Genótipo , Hepacivirus/efeitos dos fármacos , Humanos , Interferon alfa-2 , Masculino , Polietilenoglicóis/uso terapêutico , RNA Viral/sangue , Proteínas Recombinantes , Estudos Retrospectivos , Resultado do TratamentoRESUMO
SUMMARY: The impact of mutations in the precore and basal core promoter (BCP) regions of the hepatitis B virus on the course of chronic liver disease is not well established. We sought to examine the relationship of these characteristics to the clinical expression of liver disease in patients infected with genotype D chronic hepatitis B (CHB). BCP and precore mutations in 110 patients with genotype D1 CHB were determined and correlated with clinical phenotype. Of 110 patients, 95 (86.5%) were HBeAg-negative. Compared with HBeAg-positive subjects, HBeAg-negative patients were over a decade older and had lower viral loads (3.70 +/- 0.98 vs 5.77 +/- 0.69 log copies/ml, P < 0.001). The double mutation A1762T-G1764A was more prevalent in patients with advanced liver disease (AdLD) and was associated with higher alanine aminotransferase and viral load. After adjusting for age, there was a more than fourfold increase in the risk of AdLD with this mutation (OR = 4.4; 95% CI: 1.13-16.92, P < 0.03). Conversely, the G1757A substitution was associated with protection, being 90% less frequent among patients with AdLD (P = 0.001). The results indicate that in genotype D CHB, the presence of the A1762T-G1764A mutation was associated with more aggressive liver disease while the G1757A substitution was associated with protection from advanced disease.
Assuntos
Vírus da Hepatite B , Hepatite B Crônica , Mutação , Regiões Promotoras Genéticas/genética , Precursores de Proteínas/genética , Proteínas do Core Viral/genética , Adulto , Idoso , DNA Viral/análise , DNA Viral/isolamento & purificação , Feminino , Genótipo , Antígenos do Núcleo do Vírus da Hepatite B/genética , Vírus da Hepatite B/classificação , Vírus da Hepatite B/genética , Vírus da Hepatite B/patogenicidade , Hepatite B Crônica/genética , Hepatite B Crônica/fisiopatologia , Hepatite B Crônica/virologia , Humanos , Irã (Geográfico) , Masculino , Pessoa de Meia-Idade , FilogeniaRESUMO
Adipose tissue plays a significant role in whole body energy homeostasis. Obesity-associated diabetes, fatty liver and metabolic syndrome are closely linked to adipose stress and dysfunction. Genetic predisposition, overeating and physical inactivity influence the expansion of adipose tissues. Under conditions of constant energy surplus, adipocytes become hypertrophic and adipose tissues undergo hyperplasia so as to increase their lipid storage capacity, thereby keeping circulating blood glucose and fatty acids below toxic levels. Nonetheless, adipocytes have a saturation point where they lose capacity to store more lipids. At this stage, when adipocytes are fully lipid-engorged, they express stress signals. Adipose depots (particularly visceral compartments) from obese individuals with a severe metabolic phenotype are characterized by the high proportion of hypertrophic adipocytes. This review focuses on the mechanisms of adipocyte enlargement in relation to adipose fatty acid and cholesterol metabolism, and considers how this may be related to adipose dysfunction.
Assuntos
Adipócitos/patologia , Adipócitos/fisiologia , Ácidos Graxos/metabolismo , Metabolismo dos Lipídeos/fisiologia , Obesidade/fisiopatologia , Hipernutrição/fisiopatologia , Adipócitos/citologia , Adiposidade , Humanos , Mediadores da Inflamação/metabolismo , Obesidade/metabolismo , Hipernutrição/complicações , Hipernutrição/metabolismo , Estresse FisiológicoRESUMO
We sought to establish the mechanism for the raised serum estrogen levels that occur in male rats with portal hypertension and resultant portal bypass. Using the portal vein ligated (PVL) rat model, we evaluated plasma steroid hormone concentrations, metabolic clearance rate (MCR) of estradiol, and hepatic metabolism of androstenedione to estrogens and other products. In contrast to serum testosterone levels that were reduced, serum androstenedione levels were normal in the PVL rat. Estradiol MCR was measured by a constant intravenous infusion technique and was found to be similar in PVL and control animals. Androstenedione MCR was determined during constant intravenous infusion of [3H]androstenedione, and the resultant radiolabeled steroids present in plasma were separated by thin layer chromatography. The MCR of androstenedione was not diminished in PVL rats compared with controls. However, there was a sevenfold increase in the plasma estradiol derived from [3H]androstenedione in rats with portal bypass. Examination of radiolabel excreted in bile during infusion of [3H]androstenedione showed that 25-46% of this steroid was converted to estradiol in PVL rats compared with less than 3% in control male rats (P less than 0.001). Moreover, there was a selective reduction in the excretion of 16 alpha-hydroxyandrostenedione, a finding which suggested that the metabolism of androstenedione via this pathway was decreased. Androstenedione 16 alpha-hydroxylation is known to be catalyzed by a male-specific cytochrome P-450 isoform, P-450UT-A. We conclude that raised plasma estradiol levels after portal bypass in male rats are due to increased production rates, resulting in turn from enhanced aromatization of androstenedione to estradiol. On the basis of the observed specific changes in androstenedione hydroxylation pathways, it is proposed that alterations in levels of sex-specific forms of cytochrome P-450 occur in male rats with portal bypass and could account for the enhanced formation of estradiol.
Assuntos
Hidrocarboneto de Aril Hidroxilases , Estrogênios/sangue , Hipertensão Portal/sangue , Derivação Portossistêmica Cirúrgica , Androstenodiona/administração & dosagem , Androstenodiona/farmacocinética , Animais , Família 2 do Citocromo P450 , Estradiol/administração & dosagem , Estradiol/farmacocinética , Hipertensão Portal/fisiopatologia , Masculino , Taxa de Depuração Metabólica , Orquiectomia , Ratos , Ratos Endogâmicos , Esteroide 16-alfa-Hidroxilase , Testosterona/sangueRESUMO
Elevated serum estradiol concentrations and specific changes in the biliary excretion of some androstenedione metabolites have been reported in male rats with portal bypass produced by portal vein ligation (PVL). In this study, the hypothesis that male-specific forms of cytochrome P-450 are altered after PVL was tested by measuring microsomal steroid hydroxylase activities. Consistent with earlier findings in the intact animal, androstenedione 16 alpha-hydroxylase activity was reduced after PVL to 44% of control (P less than 0.05). Other pathways of androstenedione hydroxylation, and total estrogen formation (after androstenedione aromatization) were unchanged. Although total estrogen formation was not different, a sevenfold greater proportion of estradiol was produced in PVL rat microsomes. Additional experiments revealed that PVL selectively reduced the rate of microsomal estradiol 16 alpha-hydroxylation (to 56% of control, P less than 0.02). Levels of cytochrome P-450UT-A, the microsomal steroid 16 alpha-hydroxylase, were lower after PVL (56% of control, P less than 0.05), so that the present observations are consistent with the earlier suggestion that portal bypass is associated with the selective downregulation of this enzyme. Since downregulation of cytochrome P-450UT-A also occurs in experimental hepatic cirrhosis, portal hypertension may well contribute significantly to altered drug metabolism in liver disease. Impaired hepatic elimination of androstenedione by hydroxylation may indirectly enhance extrahepatic aromatization of the androgen. The decreased activity of hepatic estradiol 16 alpha-hydroxylation after PVL would enhance the accumulation of estradiol, the biologically more potent estrogen.
Assuntos
Hidrocarboneto de Aril Hidroxilases , Sistema Enzimático do Citocromo P-450/metabolismo , Estradiol/metabolismo , Cirrose Hepática Experimental/metabolismo , Microssomos Hepáticos/enzimologia , Veia Porta/cirurgia , Esteroide Hidroxilases/metabolismo , Androstenodiona/metabolismo , Animais , Sistema Enzimático do Citocromo P-450/fisiologia , Família 2 do Citocromo P450 , Feminino , Ligadura , Cirrose Hepática Experimental/enzimologia , Masculino , Microssomos Hepáticos/fisiologia , Coelhos , Ratos , Ratos Endogâmicos , Caracteres Sexuais , Esteroide 16-alfa-Hidroxilase , Esteroide Hidroxilases/fisiologiaRESUMO
We tested the hypothesis that ethanol impairs liver regeneration by abrogating receptor-mediated elevation of cytosolic free calcium ([Ca2+]i). In rats fed for 16 weeks with ethanol, hepatocellular proliferation induced by partial hepatectomy was greatly impaired. Similarly, EGF-induced DNA synthesis was reduced in cultured hepatocytes from ethanol-fed rats. There was no change in the number or affinity of EGF receptors on hepatocytes from ethanol-fed rats. Despite this, EGF-mediated production of inositol 1,4,5-trisphosphate (Ins[1,4,5]P3) was lower in hepatocytes from ethanol-fed rats, and the EGF-induced [Ca2+]i transient appeared to be abrogated. When vasopressin or phenylephrine were used as cell surface receptor ligands, hepatocytes cultured from ethanol-fed rats exhibited major reductions in Ins(1,4,5)P3 synthesis. This was associated with greatly truncated [Ca2+]i transients. These changes were not due to an effect on the Ins(1,4,5)P3 receptor on the endoplasmic reticulum or to a decrease in the size of the Ins(1,4,5)P3-mobilizable intracellular Ca+2 store. Further, mobilization of the same Ca2+ store by 2,5-di-tert-butylhydroquinone or thapsigargin restored the ability of hepatocytes from ethanol-fed rats to proliferate when exposed to EGF. It is concluded that chronic ethanol consumption inhibits liver regeneration by a mechanism that is, at least partly, the result of impaired receptor-operated [Ca2+]i signaling due to reduced generation of Ins(1,4,5)P3.
Assuntos
Alcoolismo/metabolismo , Cálcio/metabolismo , Inositol 1,4,5-Trifosfato/metabolismo , Regeneração Hepática/efeitos dos fármacos , Fígado/metabolismo , Receptores de Superfície Celular/metabolismo , Animais , Membrana Celular/metabolismo , Permeabilidade da Membrana Celular , Células Cultivadas , Fator de Crescimento Epidérmico/metabolismo , Fator de Crescimento Epidérmico/farmacologia , Hepatectomia , Fígado/citologia , Fígado/crescimento & desenvolvimento , Masculino , Fenilefrina/farmacologia , Ligação Proteica , Ratos , Ratos Wistar , Vasopressinas/farmacologiaRESUMO
The hypothesis that monohydroxy bile acids exert their cholestatic and hepatotoxic effects via a sustained elevation of cytosolic [Ca2+] was tested in the isolated perfused rat liver. Infusion of the specific inhibitor of microsomal Ca2+ sequestration, 2,5-di(tert-butyl)-1,4-benzohydroquinone (tBuBHQ) (25 microM for 10 min) produced efflux of Ca2+ from the liver and a sustained (20 min) increase in cytosolic [Ca2+] as indicated by the threefold increase in hepatic glucose output. Release of the endoplasmic reticular Ca2+ pool was demonstrated by the complete abolition of vasopressin- and phenylephrine-induced Ca2+ exchange between the liver and perfusate. Despite the profound perturbation of intracellular Ca2+ homeostasis produced by tBuBHQ, there was no decrease in bile flow and no evidence of hepatocellular injury (for 60 min), as indicated by lactate dehydrogenase release. In contrast, lithocholic acid (25 microM for 10 or 30 min) or taurolithocholic acid (5 microM for 10 or 30 min) produced an 80-90% inhibition of bile flow and a progressive increase in perfusate lactate dehydrogenase activity. During and after bile acid infusion, there was no change in Ca2+ fluxes between liver and perfusate, no stimulation of glucose output from the liver, and hormone-stimulated Ca2+ responses were preserved. It is concluded that the mechanisms for bile acid-induced cholestasis and hepatotoxicity in the intact liver are not attributable to changes in intracellular Ca2+ homeostasis, and especially not to prolonged release or depletion of Ca2+ sequestered in the endoplasmic reticulum.
Assuntos
Ácidos e Sais Biliares/toxicidade , Cálcio/metabolismo , Colestase/induzido quimicamente , Retículo Endoplasmático/metabolismo , Fígado/efeitos dos fármacos , Bile/efeitos dos fármacos , Glucose/metabolismo , Humanos , Recém-Nascido , L-Lactato Desidrogenase/metabolismo , Fígado/metabolismo , PerfusãoRESUMO
Nonalcoholic steatohepatitis (NASH) and alcoholic liver disease have similar pathological features. Because CYP2E1 plays a key role in alcoholic liver disease with its ability to stimulate lipid peroxidation, we tested the proposal that CYP2E1 could also be a factor in the development of NASH. In a dietary model - mice fed a methionine- and choline-deficient (MCD) diet - liver injury was associated with both induction of CYP2E1 and a 100-fold increase in hepatic content of lipid peroxides. Microsomal NADPH-dependent lipid oxidases contributed to the formation of these lipid peroxides, and in vitro inhibition studies demonstrated that CYP2E1 was the major catalyst. To further define the role of CYP2E1 as an initiator of oxidative stress in NASH, Cyp2e1(-/-)mice were administered the MCD diet. CYP2E1 deficiency neither prevented the development of NASH nor abrogated the increased microsomal NADPH-dependent lipid peroxidation, indicating the operation of a non-CYP2E1 peroxidase pathway. In Cyp2e1(-/-) mice with NASH (but not in wild-type mice), CYP4A10 and CYP4A14 were upregulated. Furthermore, hepatic microsomal lipid peroxidation was substantially inhibited by anti-mouse CYP4A10 antibody in vitro. These results show that experimental NASH is strongly associated with hepatic microsomal lipid peroxidation. CYP2E1, the main enzyme associated with that process in wild-type mice, is not unique among P450 proteins in catalyzing peroxidation of endogenous lipids. We have now identified CYP4A enzymes as alternative initiators of oxidative stress in the liver.
Assuntos
Citocromo P-450 CYP2E1/metabolismo , Sistema Enzimático do Citocromo P-450/metabolismo , Fígado Gorduroso/enzimologia , Hepatite/enzimologia , Peroxidação de Lipídeos , Oxigenases de Função Mista/metabolismo , Animais , Catálise , Deficiência de Colina , Citocromo P-450 CYP2E1/genética , Citocromo P-450 CYP4A , Fígado Gorduroso/patologia , Feminino , Expressão Gênica , Hepatite/patologia , Metionina/deficiência , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Microssomos , Redução de PesoRESUMO
Temperature-dependent (TD) Raman measurements at laser excitation 514.5 nm were performed at different concentrations. The spectral profile of the radial breathing modes were investigated up to a polymer concentration of 1 g/L and were found to be dominated by approximately 1.2-1.4 nm diameter tubes at room temperature. Upon heating above the glass transition of the polymer (60 degrees C) the smaller tubes around approximately 0.9 nm increased significantly in relative intensity. This suggests that below the glass transition of the polymer (60 degrees C) RBMs within the composite are damped and spectral changes cannot be interpreted as diameter selective solubilization. The observed RBM damping at room temperature only occurred up to a concentration of approximately 1.2 x 10(-4) g/L and below this no damping was observed. Photoluminescence intensity (PL) measurements were taken for a range of PmPV concentrations, in which HiPco single walled carbon nanotubes (SWNTs) at 100%, 10%, 1%, 0.1%, 0.01%, and 0% mass fractions were added. Fitting of the concentration dependence to a dynamic absorption/desorption model indicates that the polymer interacts with nanotube bundles until a critical concentration of approximately 1.2 x 10(-4) g/L is reached, below which the nanotubes are isolated. The polymer and or solvent has a significant effect on the debundling and aggregation within these systems. Aggregation and/or interaction with the polymer at higher concentrations can effect the RBM profile in the composite at ambient temperatures, providing an incomplete representation of the selection of diameters present within composites at a particular wavelength.
RESUMO
PURPOSE: An exploratory study to test whether body-surface area (BSA) should be used for the calculation of epirubicin dose. PATIENTS AND METHODS: The relationship between pretreatment characteristics and the effects of epirubicin were investigated in 20 chemotherapy-naive patients. Measurements of body size, renal and hepatic function, and other factors were correlated with epirubicin pharmacokinetics (PK) and epirubicin-induced neutropenia. All patients received 150 mg of epirubicin infused continuously over 120 hours, regardless of body size. Factors were analyzed by univariate and multivariate linear regression. RESULTS: There were no correlations between BSA or weight with any PK parameter or with the degree of neutropenia. In multivariate analysis, indicators of liver function were the only factors that correlated with neutropenia and epirubicin PK. Thus, correlations for neutropenia were seen with antipyrine clearance (P = .003), activated partial thromboplastin time (APTT) (P = .005) and serum transferrin (P = .01). Further, the area under the concentration-time curve (AUC) for epirubicin correlated with prothrombin index (P < .01), antipyrine clearance (P < .01), and serum bile salt concentration (P = .03), and there were similar correlations for epirubicin steady-state concentration (CpSS). Epirubicin clearance correlated with antipyrine clearance (P = .02). PK parameters for dihydroepirubicin correlated with prothombin index, serum transferrin, and bile salt concentrations (P < .001 for all correlations). Because of the number of statistical examinations performed, some of these correlations may be spurious. However, some are likely to be real, since the same variables repeatedly correlated with different epirubicin-associated outcomes. There were no correlations between epirubicin PK indices or neutropenia and serum aminotransferase levels or other biochemical liver function tests, creatinine, or any of the clinical factors examined. CONCLUSION: These results led us to question the use of BSA for epirubicin dose calculation. In contrast, quantitative liver function tests may give a better indication of drug handling and toxicity and may be useful to determine more accurate methods for dose calculation of epirubicin.
Assuntos
Antibióticos Antineoplásicos/administração & dosagem , Antibióticos Antineoplásicos/farmacocinética , Superfície Corporal , Epirubicina/administração & dosagem , Epirubicina/farmacocinética , Adulto , Idoso , Idoso de 80 Anos ou mais , Anti-Inflamatórios não Esteroides , Antibióticos Antineoplásicos/efeitos adversos , Antibióticos Antineoplásicos/sangue , Antipirina , Ácidos e Sais Biliares/sangue , Relação Dose-Resposta a Droga , Esquema de Medicação , Epirubicina/efeitos adversos , Epirubicina/sangue , Feminino , Humanos , Testes de Função Hepática , Masculino , Pessoa de Meia-Idade , Neutropenia/induzido quimicamenteRESUMO
Hybrid systems of the conjugated organic polymer poly(p-phenylene vinylene-co-2,5-dioctyloxy-m-phenylene vinylene)(PmPV) and HiPco single-walled carbon nanotubes (SWNTs) are explored using spectroscopic and thermal techniques to determine specific interactions. Vibrational spectroscopy indicates a weak interaction, and this is further elucidated using differential scanning calorimetry (DSC), confocal laser scanning microscopy, temperature-dependent Raman spectroscopy, and temperature-dependent infrared spectroscopy of the raw materials and the composite. An endothermic transition is observed in the DSC of both the polymer and the 0.1% HiPco composite in the region of 50 degrees C. Also observed in the DSC of the composite is a double-peaked endotherm at -39 and -49 degrees C, which does not appear in the polymer. The Raman spectroscopy of the polymer upon increasing the temperature to 60 degrees C shows a diminished cis-vinylene mode at 1575 cm(-1), with an increase in relative intensity of the trans-vinylene mode at 1630 cm(-1). Partially irreversible change in isomerization suggests increased order in the polymer. This change in the polymer is also manifest in the Raman composite spectrum upon increase of the temperature to 60 degrees C, where the spectrum becomes abruptly dominated by nanotubes. Raman spectroscopy of the composite shows no change at -35 degrees C; however, infrared absorption measurements suggest that the transition at -35 degrees C derives from the polymer side chains. Here the composite at -35 degrees C shows a change in the absorbance of the polymer side chain aryl-oxide linkage at 1250 cm(-1) and alkyl-oxide stretch at 1050 cm(-1). Infrared spectra thus suggest that the transitions in the lower temperature region around -35 degrees C are side chain-induced, while Raman spectra suggest that the transition at 60 degrees C is backbone-induced. Furthermore, temperature cycling induces an irreversible decrease in the mean fluorescence intensity of the polymer, coupled with a further reduction in the mean fluorescence intensity of the composite. This suggests that an increase in crystallization of the composite is supported and enhanced by an increase in ordering of the polymer. Implications are discussed.
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Hepatocellular carcinoma (HCC) is a leading cause of cancer death worldwide, and incidence rates in Western countries are on the rise. Despite many options, no ideal treatment yet exists for this highly malignant tumour, and management strategies have varied accordingly. This review summarises current strategies for the diagnosis and management of HCC.
Assuntos
Carcinoma Hepatocelular/diagnóstico , Carcinoma Hepatocelular/terapia , Neoplasias Hepáticas/diagnóstico , Neoplasias Hepáticas/terapia , Terapia Combinada , HumanosRESUMO
CYP3A4, the predominant cytochrome P450 expressed in human liver, is responsible for the metabolism of endogenous steroids and many drugs. On the basis of pharmacokinetic studies in patients with hormonal derangements and the effects of replacement therapy, it has been suggested that iodothyronines decrease CYP3A4-mediated drug metabolism, whereas glucocorticoids and GH enhance CYP3A4 activity. The aim of the present study, using well differentiated human hepatocytes in primary culture, was to examine directly whether hormonal factors regulate CYP3A4 gene expression. Addition of T3 to primary hepatocytes resulted in a marked reduction of CYP3A4-catalyzed testosterone 6 beta-hydroxylase activity and corresponding levels of CYP3A4 protein and messenger ribonucleic acid compared to those in untreated cells. Conversely, both dexamethasone and GH treatment substantially increased CYP3A4 gene expression. None of the hormones studied consistently altered the expression of other human cytochrome P450 genes. We conclude that iodothyronines, glucocorticoids, and GH act directly on human hepatocytes to regulate the expression of CYP3A4, and these effects appear to be exerted at a pretranslational level. Altered regulation of hepatic CYP3A4 is, therefore, likely to account for previous observations concerning the effects of endocrine diseases and hormonal treatments on human cytochrome P450-mediated drug and steroid metabolism.