RESUMO
BACKGROUND: Microsatellite instability (MSI)/mismatch repair deficiency (dMMR) is a key genetic feature which should be tested in every patient with colorectal cancer (CRC) according to medical guidelines. Artificial intelligence (AI) methods can detect MSI/dMMR directly in routine pathology slides, but the test performance has not been systematically investigated with predefined test thresholds. METHOD: We trained and validated AI-based MSI/dMMR detectors and evaluated predefined performance metrics using nine patient cohorts of 8343 patients across different countries and ethnicities. RESULTS: Classifiers achieved clinical-grade performance, yielding an area under the receiver operating curve (AUROC) of up to 0.96 without using any manual annotations. Subsequently, we show that the AI system can be applied as a rule-out test: by using cohort-specific thresholds, on average 52.73% of tumors in each surgical cohort [total number of MSI/dMMR = 1020, microsatellite stable (MSS)/ proficient mismatch repair (pMMR) = 7323 patients] could be identified as MSS/pMMR with a fixed sensitivity at 95%. In an additional cohort of N = 1530 (MSI/dMMR = 211, MSS/pMMR = 1319) endoscopy biopsy samples, the system achieved an AUROC of 0.89, and the cohort-specific threshold ruled out 44.12% of tumors with a fixed sensitivity at 95%. As a more robust alternative to cohort-specific thresholds, we showed that with a fixed threshold of 0.25 for all the cohorts, we can rule-out 25.51% in surgical specimens and 6.10% in biopsies. INTERPRETATION: When applied in a clinical setting, this means that the AI system can rule out MSI/dMMR in a quarter (with global thresholds) or half of all CRC patients (with local fine-tuning), thereby reducing cost and turnaround time for molecular profiling.
Assuntos
Neoplasias Colorretais , Instabilidade de Microssatélites , Inteligência Artificial , Neoplasias Colorretais/diagnóstico , Neoplasias Colorretais/genética , Neoplasias Colorretais/patologia , Reparo de Erro de Pareamento de DNA/genética , Detecção Precoce de Câncer , HumanosRESUMO
Epidemiology of bacteria isolated from pyogenic liver abscesses change, and an increase in enterococci has been reported in European hospitals. The aim of this study was to investigate the clinical characteristics and outcome of enterococcal PLA. We performed a retrospective analysis of patients with microbiologically confirmed PLA at three German university centers. Indicators of enterococcal PLA were determined using binary logistic regression, and survival analysis was performed using Kaplan-Meier statistics and Cox regression analysis. Enterococci were isolated in 51/133 (38%) patients with PLA. Patients with enterococcal PLA had smaller abscess diameter (4.8 vs. 6.7 cm, p = 0.03) than patients with non-enterococcal PLA, but had more frequent polymicrobial culture results. In univariate logistic regression analysis, alcohol abuse (OR 3.94, 95% CI 1.24-12.49, p = 0.02), hepatobiliary malignancies (OR 3.90, 95% CI 1.86-8.18, p < 0.001) and cirrhosis (OR 6.36, 95% CI 1.27-31.96, p = 0.02) were associated with enterococcal PLA. Patients with enterococcal PLA had a higher mortality than patients with non-enterococcal PLA (hazard ratio 2.92; 95% confidence interval 1.09-7.80; p = 0.03), which remained elevated even after excluding patients with hepatobiliary malignancies, cirrhosis, and transplant recipients in a sensitivity analysis. The increased mortality was associated with non-fecal enterococci but not in patients with Enterococcus faecalis. In this retrospective, multicenter study, enterococcal PLA was common and indicated an increased risk of mortality, underscoring the need for close clinical monitoring and appropriate treatment protocols in these patients.
Assuntos
Enterococcus , Infecções por Bactérias Gram-Positivas/diagnóstico , Infecções por Bactérias Gram-Positivas/microbiologia , Abscesso Hepático Piogênico/diagnóstico , Abscesso Hepático Piogênico/microbiologia , Idoso , Idoso de 80 Anos ou mais , Comorbidade , Estudos Transversais , Gerenciamento Clínico , Suscetibilidade a Doenças , Feminino , Alemanha/epidemiologia , Infecções por Bactérias Gram-Positivas/epidemiologia , Humanos , Abscesso Hepático Piogênico/epidemiologia , Masculino , Pessoa de Meia-Idade , Avaliação de Resultados da Assistência ao Paciente , Prognóstico , Estudos Retrospectivos , Avaliação de SintomasRESUMO
The COVID-19 pandemic represents a huge burden on global health systems. Although far-reaching prevention measures such as the increase of intensive care capacities and drastic restrictions of public life have so far been able to avert an overload of the German health care system, the current situation implies an exceptionally high burden on medical professionals. The current study presents the results of an opinion evaluation among 513 pneumology specialists in Germany in the period from March 27th to April 11th, 2020. While the majority of respondents stated that Germany was "well" prepared for the pandemic, this assessment was significantly worse among participants from the outpatient sector compared to the hospital sector (pâ<â0.001). Furthermore, a lack of medical protective equipment was reported significantly more frequently by respondents from the outpatient sector (pâ<â0.001). The importance of telemedicine approaches during the COVID-19 pandemic was rated "high" (35.2â%) or "very high" (17.2â%) by most pneumology professionals, with participants from the hospital sector giving a higher rating (pâ<â0.001). Finally, 45.8â% of the respondents expressed a "negative" influence of the COVID-19 pandemic on their personal mood and 58.3â% expressed "strong" or "very strong" concerns about the health of their fellow human beings. This assessment was significantly stronger among female participants and participants from the nursing sector (pâ<â0.001). In summary, the current study analyses for the first time the professional and personal impact of the COVID-19 pandemic on pneumology professionals in Germany. The results could help to identify first starting points to better support health professionals during the current and future challenges.
Assuntos
COVID-19 , Pneumologia , Feminino , Alemanha/epidemiologia , Humanos , Pandemias/prevenção & controle , Percepção , SARS-CoV-2 , Inquéritos e QuestionáriosRESUMO
Cholangiocarcinoma (CCC) is the second most common primary malignancy of the liver and is typically diagnosed at advanced disease stages. Among curative treatment options for CCC, radical surgical resection with extrahepatic bile duct resection, hepatectomy, and en-bloc lymphadenectomy are considered the mainstay of curative therapy. The assessment of the functional liver reserve by dynamic liver function tests and the estimation of the remaining future liver volume (future liver remnant, FLR) are of paramount importance. The introduction of novel interventional and surgical techniques, such as portal vein embolization, associating liver partition, and portal vein ligation for staged hepatectomy (ALPPS), have enabled clinicians to achieve resectability even in patients previously deemed unresectable. Radiofrequency ablation (RFA) shows acceptable results in small intrahepatic cholangiocarcinoma (IHCC) in liver cirrhosis and should be evaluated if cirrhosis precludes surgical treatment. Transarterial chemoembolization (TACE) or transarterial radioembolization (TARE) alone or in combination with systemic therapy may be applied in cases of surgical irresectability. According to recent results of the British BILCAP trial, adjuvant therapy may be considered after surgical resection in curative intent.
Assuntos
Algoritmos , Neoplasias dos Ductos Biliares , Quimioembolização Terapêutica , Colangiocarcinoma , Neoplasias Hepáticas , Adulto , Neoplasias dos Ductos Biliares/terapia , Colangiocarcinoma/terapia , Hepatectomia , Humanos , Ligadura , Neoplasias Hepáticas/terapia , Veia Porta , Resultado do TratamentoRESUMO
Acute pancreatitis is a potentially life-threatening disease, which is morphologically classified into interstitial edematous or necrotizing pancreatitis. According to the revised Atlanta classification, mild, moderate and severe clinical courses are differentiated regarding local and systemic complications as well as concomitant organ failure. In the initial disease phase, the therapeutic measures are focused on (aggressive) volume replacement, early enteral nutrition and adequate analgesia. Characteristic in the course of severe acute pancreatitis are abdominal necroses, which require individualized and interdisciplinary treatment with antibiotic therapy, drainage and definitive necrosectomy. Necrosectomy should be planned as a "step-up approach" using interventional-radiological, endoscopic and minimally invasive surgical procedures.
Assuntos
Pancreatite Necrosante Aguda , Drenagem , Endoscopia , Nutrição Enteral , Humanos , Pancreatite Necrosante Aguda/diagnóstico , Pancreatite Necrosante Aguda/terapiaRESUMO
BACKGROUND: Elderly patients are vulnerable to adverse drug reactions (ADRs). Drug-related readmissions (DRRs) can be a major consequence of ADR. Therefore, this study aimed to investigate the effects of a ward-based, comprehensive pharmaceutical care service on the occurrence of DRRs as the endpoint in dependent-living elderly patients. METHODS: A randomized, controlled trial was performed at a German University Hospital. Patients fulfilling the following criteria were eligible: admission to a cooperating ward, existing drug therapy at admission, 65 years of age and older, home-care or nursing home residents in ambulatory care, and a minimum hospital stay of three days. Patients received either standard care (control group) or pharmaceutical care (intervention group). Follow-up consultations were conducted for each patient at 1, 8, 26, and 52 weeks after discharge. The time to DRR was defined as the primary outcome measure and was analysed using the log-rank test. The Cox-proportional hazard model was used for risk factor analysis. RESULTS: Sixty patients (n = 31 intervention group, n = 29 control group) participated in the study. For patients in the intervention group, the median time to DRR was prolonged; however, the level of statistical significance was not reached (log-rank test P = 0.068; HR = 3.28, P = 0.086). When the risk factors 'age' or 'length of stay on the ward' were added to the Cox proportional hazard model, patients in the control group exhibited a significantly higher risk of experiencing a DRR than patients of the intervention group (HR = 4.62; P = 0.028 including age and HR = 5.76; P = 0.033 including length of stay on the ward). CONCLUSIONS: Our findings demonstrate the successful implementation of ward-based, comprehensive pharmaceutical care for dependent-living elderly. Despite a low participation rate, which led to an underpowered study, the results provide a preliminary efficacy signal and effect size estimates to power a definitive trial. TRIAL REGISTRATION: Clinicaltrials.gov identifier: NCT01578525 , prospectively registered April 13, 2012.
Assuntos
Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos/prevenção & controle , Serviços de Assistência Domiciliar/tendências , Casas de Saúde/tendências , Readmissão do Paciente/tendências , Assistência Farmacêutica/tendências , Idoso , Idoso de 80 Anos ou mais , Assistência Ambulatorial/normas , Assistência Ambulatorial/tendências , Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos/diagnóstico , Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos/epidemiologia , Feminino , Seguimentos , Serviços de Assistência Domiciliar/normas , Hospitalização/tendências , Humanos , Tempo de Internação/tendências , Masculino , Casas de Saúde/normas , Alta do Paciente/tendências , Assistência Farmacêutica/normasRESUMO
Opposing activities of Notch and Wnt signaling regulate mucosal barrier homeostasis and differentiation of intestinal epithelial cells. Specifically, Wnt activity is essential for differentiation of secretory cells including Wnt3-producing Paneth cells, whereas Notch signaling strongly promotes generation of absorptive cells. Loss of caspase-8 in intestinal epithelium (casp8∆int) is associated with fulminant epithelial necroptosis, severe Paneth cell death, secondary intestinal inflammation, and an increase in Notch activity. Here, we found that pharmacological Notch inhibition with dibenzazepine (DBZ) is able to essentially rescue the loss of Paneth cells, deescalate the inflammatory phenotype, and reduce intestinal permeability in casp8∆int mice. The secretory cell metaplasia in DBZ-treated casp8∆int animals is proliferative, indicating for Notch activities partially insensitive to gamma-secretase inhibition in a casp8∆int background. Our data suggest that casp8 acts in the intestinal Notch network.
Assuntos
Caspase 8/metabolismo , Dibenzazepinas/farmacologia , Celulas de Paneth/efeitos dos fármacos , Receptor Notch1/antagonistas & inibidores , Animais , Caspase 8/genética , Morte Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Masculino , Metaplasia , Camundongos Endogâmicos C57BL , Camundongos Knockout , Celulas de Paneth/enzimologia , Celulas de Paneth/patologia , Permeabilidade , Fenótipo , Receptor Notch1/metabolismo , Via Secretória , Via de Sinalização Wnt/efeitos dos fármacosRESUMO
Acute liver failure (ALF) is a rare, but life-threatening disease that is characterized by the acute onset of jaundice, coagulopathy, and hepatic encephalopathy (HE) in patients without pre-existing liver disease. Main causes in Germany are drug toxicity, acetaminophen overdose, and viral hepatitis (A, B, E). For the initial assessment of patients with ALF and the diagnostic algorithm, the early detection of HE, exclusion of liver cirrhosis, immediate diagnosis of the underlying etiology, and evaluation for the necessity of liver transplantation (LT) are critical. Intensive care therapeutic measures aim at preventing or treating complications of ALF. Potentially, plasmapheresis (full plasma exchange) offers a survival benefit for ALF patients who do not undergo LT. The King's College criteria and the Clichy criteria are used as prognostic tools for the indication for LT.
Assuntos
Cuidados Críticos/métodos , Falência Hepática Aguda/terapia , Algoritmos , Diagnóstico Diferencial , Falência Hepática Aguda/complicações , Falência Hepática Aguda/diagnóstico , Falência Hepática Aguda/etiologia , Transplante de Fígado , Troca Plasmática , Prognóstico , Fatores de Risco , Taxa de SobrevidaRESUMO
WHAT IS KNOWN AND OBJECTIVE: In kidney transplant patients, clinically relevant drug-drug interactions (DDIs) with immunosuppressants potentially lead to serious adverse drug events (ADEs). The aim of this study was (i) to show that five clinical decision support systems (CDSSs) differ in their ability to identify clinically relevant potential DDIs (pDDIs) of immunosuppressants in kidney transplant patients and (ii) to compare CDSSs in terms of their ability to identify clinically relevant pDDIs in this context. METHODS: All pDDIs being possible between nine immunosuppressants and 234 comedication drugs were identified for 264 intensive care unit (ICU) kidney transplant patients from 1999 to 2010. For pDDI identification, five CDSSs were used: DRUG-REAX® , ID PHARMA CHECK® , Lexi-Interact, mediQ and Meona. PDDIs from high severity categories were defined as clinically relevant. Classification of pDDIs as clinically relevant/non-clinically relevant by a clinical pharmacist using Stockley's Drug Interactions was employed as benchmark. We analysed inter-rater agreement, sensitivity, specificity, positive predictive value and negative predictive value. RESULTS AND DISCUSSION: Clinical decision support systems generated a total of 759 pDDI alerts. A total of 240 pDDI alerts were in high severity categories. A total of 391 different pDDIs were identified. Only 5% (n = 35) of different pDDIs were identified by all CDSSs. A total of 49 pDDIs were classified as clinically relevant by clinical pharmacists' rating using Stockley's Drug Interactions. Meona (0·72) has the highest inter-rater agreement with the benchmark for clinically relevant pDDIs. ID PHARMA CHECK® and mediQ show highest sensitivities (0·74, respectively). Meona has the highest specificity (0·99) and positive predictive value (0·89). WHAT IS NEW AND CONCLUSION: Five CDSSs differ in their ability to identify clinically relevant pDDIs of immunosuppressants in kidney transplant patients. Data may assist in selecting CDSSs for kidney transplant patients in the ICU. Using CDSSs to identify clinically relevant pDDIs could prevent ADEs and contribute to the overall goal of avoiding patient harm and increasing patient safety.
Assuntos
Sistemas de Apoio a Decisões Clínicas , Imunossupressores/efeitos adversos , Transplante de Rim/métodos , Interações Medicamentosas , Humanos , Imunossupressores/administração & dosagem , Unidades de Terapia Intensiva , Farmacêuticos , Valor Preditivo dos Testes , Estudos Retrospectivos , Sensibilidade e EspecificidadeRESUMO
In this manuscript, we report an exceptional observation study of a young woman suffering from an autoimmune syndrome induced by adjuvants (ASIA). Until today only seven cases of adult-onset Still's disease (AOSD) induced by breast implants have been published. This may be due to the fact that this illness itself is very rare; however, the reason might also be that the community is not sensitized to the case-specific symptoms. Within this article, we show for the first time highly detailed diagnostic test procedures such as PET-CT scans and specific histological staining of the breast tissue, displaying proinflammatory macrophages that are a well-known activator and booster of autoimmune diseases. We hope to give new insights into the clinical picture and pathogenesis of AOSD in order to improve the challenging diagnosis.
Assuntos
Doenças Autoimunes , Implantes de Mama , Doença de Still de Início Tardio , Adjuvantes Imunológicos , Adulto , Implantes de Mama/efeitos adversos , Feminino , Humanos , Tomografia por Emissão de Pósitrons combinada à Tomografia Computadorizada , Doença de Still de Início Tardio/diagnóstico , Doença de Still de Início Tardio/etiologiaRESUMO
c-Jun N-terminal kinases (JNKs) contribute to immune signaling but their functional role during intestinal mucosal inflammation has remained ill defined. Using genetic mouse models, we characterized the role of JNK1 and JNK2 during homeostasis and acute colitis. Epithelial apoptosis, regeneration, differentiation, and barrier function were analyzed in intestinal epithelium-specific (ΔIEC) or complete JNK1 and bone marrow chimeric or complete JNK2 deficient mice as well as double-knockout animals (JNK1ΔIECJNK2-/-) during homeostasis and acute dextran sulfate sodium (DSS)-induced colitis. Results were confirmed using human HT-29 cells and wild-type or JNK2-deficient mouse intestinal organoid cultures. We show that nonhematopoietic JNK2 but not JNK1 expression confers protection from DSS-induced intestinal inflammation reducing epithelial barrier dysfunction and enterocyte apoptosis. JNK2 additionally enhanced Atonal homolog 1 expression, goblet cell and enteroendocrine cell differentiation, and mucus production under inflammatory conditions. Our results identify a protective role of epithelial JNK2 signaling to maintain mucosal barrier function, epithelial cell integrity, and mucus layer production in the event of inflammatory tissue damage.
Assuntos
Colite/imunologia , Enterócitos/fisiologia , Células Caliciformes/fisiologia , Intestinos/imunologia , Proteína Quinase 9 Ativada por Mitógeno/metabolismo , Doença Aguda , Animais , Apoptose , Diferenciação Celular , Sobrevivência Celular , Sulfato de Dextrana , Modelos Animais de Doenças , Células HT29 , Humanos , Camundongos , Camundongos Knockout , Proteína Quinase 9 Ativada por Mitógeno/genética , Transdução de SinaisRESUMO
Acute liver failure (ALF) is a rare, but life-threatening disease that is characterized by the acute onset of jaundice, coagulopathy, and hepatic encephalopathy (HE) in patients without pre-existing liver disease. Main causes in Germany are drug toxicity, acetaminophen overdose, and viral hepatitis (A, B, E). For the initial assessment of patients with ALF and the diagnostic algorithm, the early detection of HE, exclusion of liver cirrhosis, immediate diagnosis of the underlying etiology, and evaluation for the necessity of liver transplantation (LT) are critical. Intensive care therapeutic measures aim at preventing or treating complications of ALF. Potentially, plasmapheresis (full plasma exchange) offers a survival benefit for ALF patients who do not undergo LT. The King's College criteria and the Clichy criteria are used as prognostic tools for the indication for LT.
RESUMO
To identify molecular interactions between hepatitis B virus (HBV) and hepatitis delta virus (HDV), HBV sequences were analysed in HBV/HDV-infected patients. Characteristic amino acid substitutions were found in cytosolic domains of hepatitis B surface antigen (HBsAg), in contrast to HBV-mono-infected controls. The functional impact of HDV on the replication of wild-type and mutant HBV was assessed in vitro. HDV co-transfection significantly reduced the replication of HBV strains containing precore or basal core promoter mutations, and HBV polymerase or surface antigen mutants affected HDV replication in vitro. Conclusively, our study revealed distinct HBsAg mutational patterns in HBV/HDV-infected patients and novel functional interactions between HBV and HDV.
Assuntos
Vírus da Hepatite B/fisiologia , Hepatite B/virologia , Hepatite D/virologia , Vírus Delta da Hepatite/fisiologia , Interações Microbianas , Seleção Genética , Replicação Viral , Adolescente , Adulto , Coinfecção/virologia , Feminino , Hepatite B/complicações , Antígenos de Superfície da Hepatite B/genética , Hepatite D/complicações , Humanos , Masculino , Pessoa de Meia-Idade , Mutação , Virulência , Adulto JovemRESUMO
BACKGROUND AND AIM: Accurate quantification of progressive liver disease is essential for therapeutic decisions and follow-up for patients who underwent liver transplantation. To evaluate the quality of noninvasive assessment of liver fibrosis in these patients, we compared Doppler ultrasound of the hepatic blood vessels as well as transient elastography (TE, FibroScan(®)) with liver biopsy following transplantation. METHODS: We performed Doppler ultrasound of the hepatic veins, hepatic artery, and portal vein as well as a TE in 48 patients who underwent liver transplantation 12 months ago. Hepatic venous flow was evaluated by determination of the resistance index (HVRI) of the right hepatic vein. Doppler and TE results were compared with histopathologic workup of a 12-month protocol liver biopsy after transplantation. RESULTS: HVRI showed a high reliability in predicting liver fibrosis stage FII or higher (AUROC of 0.99 ± 0.001 for FII or higher, the HVRI < 1.05 with a sensitivity and specificity of 100 and 91.43 %) compared to histopathologic workup (Desmet's score) and was comparable to TE analysis. Both HVRI and TE differed significantly in no or minimal fibrosis versus FII or higher (p < 0.001). In contrast, portal vein and hepatic artery did not show significant changes in blood flow in our study population. CONCLUSIONS: Hepatic vein flow resistance index is a valuable tool in noninvasive evaluation of liver fibrosis in liver transplantation follow-up predicting FII or higher and might help reducing the number of protocol biopsies needed.
Assuntos
Técnicas de Imagem por Elasticidade , Cirrose Hepática/diagnóstico por imagem , Cirrose Hepática/patologia , Fígado/patologia , Ultrassonografia Doppler , Resistência Vascular , Adulto , Idoso , Área Sob a Curva , Biópsia , Feminino , Artéria Hepática/diagnóstico por imagem , Artéria Hepática/fisiopatologia , Veias Hepáticas/diagnóstico por imagem , Veias Hepáticas/fisiopatologia , Humanos , Cirrose Hepática/fisiopatologia , Transplante de Fígado , Masculino , Pessoa de Meia-Idade , Veia Porta/diagnóstico por imagem , Veia Porta/fisiopatologia , Valor Preditivo dos Testes , Estudos Prospectivos , Curva ROCRESUMO
The surgical procedure of two-thirds partial hepatectomy (PH) in rodents was first described more than 80 years ago by Higgins and Anderson. Nevertheless, this technique is still a state-of-the-art method for the community of liver researchers as it allows the in-depth analysis of signalling pathways involved in liver regeneration and hepatocarcinogenesis. The importance of PH as a key method in experimental hepatology has even increased in the last decade due to the increasing availability of genetically-modified mouse strains. Here, we propose a standard operating procedure (SOP) for the implementation of PH in mice, which is based on our experience of more than 10 years. In particular, the SOP offers all relevant background information on the PH model and provides comprehensive guidelines for planning and performing PH experiments. We provide established recommendations regarding optimal age and gender of animals, use of appropriate anaesthesia and biometric calculation of the experiments. We finally present an easy-to-follow step-by-step description of the complete surgical procedure including required materials, critical steps and postoperative management. This SOP especially takes into account the latest changes in animal welfare rules in the European Union but is still in agreement with current international regulations. In summary, this article provides comprehensive information for the legal application, design and implementation of PH experiments.
Assuntos
Modelos Animais de Doenças , Ciência dos Animais de Laboratório , Fatores Etários , Bem-Estar do Animal , Animais , União Europeia , Guias como Assunto , Hepatectomia/normas , Humanos , Ciência dos Animais de Laboratório/normas , Camundongos , Fatores SexuaisRESUMO
Chronic liver inflammation is a crucial event in the development and growth of hepatocellular carcinoma (HCC). Compelling evidence has shown that interleukin-6 (IL-6)/gp130-dependent signaling has a fundamental role in liver carcinogenesis. Thus, in the present study we aimed to investigate the role of gp130 in hepatocytes for the initiation and progression of HCC. Hepatocyte-specific gp130 knockout mice (gp130(Δhepa)) and control animals (gp130(f/f)) were treated with diethylnitrosamine (DEN). The role of gp130 for acute injury (0-144 h post treatment), tumor initiation (24 weeks) and progression (40 weeks) was analyzed. After acute DEN-induced liver injury we observed a reduction in the inflammatory response in gp130(Δhepa) animals as reflected by decreased levels of IL-6 and oncostatin M. The loss of gp130 slightly attenuated the initiation of HCC 24 weeks after DEN treatment. In contrast, 40 weeks after DEN treatment, male and female gp130(Δhepa) mice showed smaller tumors and reduced tumor burden, indicating a role for hepatocyte-specific gp130 expression during HCC progression. Oxidative stress and DNA damage were substantially and similarly increased by DEN in both gp130(f/f) and gp130(Δhepa) animals. However, gp130(Δhepa) livers revealed aberrant STAT5 activation and decreased levels of transforming growth factor-ß (TGFß), pSMAD2/3 and SMAD2, whereas phosphorylation of STAT3 at Tyr705 and Ser727 was absent. Our results indicate that gp130 deletion in hepatocytes reduces progression, but not HCC initiation in the DEN model. Gp130 deletion resulted in STAT3 inhibition but increased STAT5 activation and diminished TGF-dependent signaling. Hence, blocking gp130 in hepatocytes might be an interesting therapeutic target to inhibit the growth of HCC.
Assuntos
Carcinoma Hepatocelular/metabolismo , Receptor gp130 de Citocina/metabolismo , Hepatócitos/metabolismo , Animais , Carcinoma Hepatocelular/induzido quimicamente , Receptor gp130 de Citocina/genética , Dano ao DNA/efeitos dos fármacos , Dano ao DNA/genética , Dietilnitrosamina/toxicidade , Feminino , Citometria de Fluxo , Imunofluorescência , Hepatócitos/efeitos dos fármacos , Immunoblotting , Inflamação/genética , Inflamação/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Oncostatina M/toxicidade , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/genética , Fator de Transcrição STAT3/genética , Fator de Transcrição STAT3/metabolismo , Fator de Transcrição STAT5/genética , Fator de Transcrição STAT5/metabolismoRESUMO
Delta hepatitis, caused by co-infection or super-infection of hepatitis D virus (HDV) in hepatitis B virus (HBV) -infected patients, is the most severe form of chronic hepatitis, often progressing to liver cirrhosis and liver failure. Although 15 million individuals are affected worldwide, molecular data on the HDV genome and its proteins, small and large delta antigen (S-/L-HDAg), are limited. We therefore conducted a nationwide study in HBV-HDV-infected patients from Iran and successfully amplified 38 HDV full genomes and 44 L-HDAg sequences from 34 individuals. Phylogenetic analyses of full-length HDV and L-HDAg isolates revealed that all strains clustered with genotype 1 and showed high genotypic distances to HDV genotypes 2 to 8, with a maximal distance to genotype 3. Longitudinal analyses in individual patients indicated a reverse evolutionary trend, especially in L-HDAg amino acid composition, over time. Besides multiple sequence variations in the hypervariable region of HDV, nucleotide substitutions preferentially occurred in the stabilizing P4 domain of the HDV ribozyme. A high rate of single amino acid changes was detected in structural parts of L-HDAg, whereas its post-translational modification sites were highly conserved. Interestingly, several non-synonymous mutations were positively selected that affected immunogenic epitopes of L-HDAg towards CD8 T-cell- and B-cell-driven immune responses. Hence, our comprehensive molecular analysis comprising a nationwide cohort revealed phylogenetic relationships and provided insight into viral evolution within individual hosts. Moreover, preferential areas of frequent mutations in the HDV ribozyme and antigen protein were determined in this study.
Assuntos
Genoma Viral , Hepatite D Crônica/patologia , Hepatite D Crônica/virologia , Vírus Delta da Hepatite/genética , Mutação , Adulto , Criança , Análise por Conglomerados , Progressão da Doença , Evolução Molecular , Feminino , Genótipo , Hepatite B/complicações , Humanos , Irã (Geográfico) , Estudos Longitudinais , Masculino , Pessoa de Meia-Idade , Filogenia , Seleção Genética , Análise de Sequência de DNA , Homologia de Sequência , Adulto JovemRESUMO
The multi-kinase inhibitor Sorafenib increases the survival of patients with advanced hepatocellular carcinoma (HCC). Current data suggest that Sorafenib inhibits cellular proliferation and angiogenesis and promotes apoptosis. However, the underlying pro-apoptotic molecular mechanisms are incompletely understood. Here we compared the pro-apoptotic and anti-proliferative properties of Sorafenib in murine hepatoma cells and syngeneic healthy hepatocytes in vitro and in animal models of HCC and liver regeneration in vivo. In vitro, we demonstrate that cell cycle activity and expression of anti-apoptotic Bcl-2 like proteins are similarly downregulated by Sorafenib in Hepa1-6 hepatoma cells and in syngeneic primary hepatocytes. However, Sorafenib-mediated activation of caspase-3 and induction of apoptosis were exclusively found in hepatoma cells, but not in matching primary hepatocytes. We validated these findings in vivo by applying an isograft HCC transplantation model and partial hepatectomy (PH) in C57BL/6 mice. Sorafenib treatment activated caspase-3 and thus apoptosis selectively in small tumor foci that originated from implanted Hepa1-6 cells but not in surrounding healthy hepatocytes. Similarly, Sorafenib did not induce apoptosis after PH. However, Sorafenib treatment transiently inhibited cell cycle progression and resulted in mitotic catastrophe and enhanced non-apoptotic liver injury during regeneration. Importantly, Sorafenib-mediated apoptosis in hepatoma cells was associated with the expression of p53-upregulated-modulator-of-apoptosis (PUMA). In contrast, regenerating livers after PH revealed downregulation of PUMA and were completely protected from Sorafenib-mediated apoptosis. We conclude that Sorafenib induces apoptosis selectively in hepatoma cells but not in healthy hepatocytes and can additionally increase non-apoptotic hepatocyte injury in the regenerating liver.
Assuntos
Proteínas Reguladoras de Apoptose/genética , Apoptose/efeitos dos fármacos , Carcinoma Hepatocelular/tratamento farmacológico , Carcinoma Hepatocelular/genética , Hepatócitos/citologia , Neoplasias Hepáticas/tratamento farmacológico , Neoplasias Hepáticas/genética , Proteínas Supressoras de Tumor/genética , Animais , Proteínas Reguladoras de Apoptose/metabolismo , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/fisiopatologia , Caspase 3/genética , Caspase 3/metabolismo , Ciclo Celular , Células Cultivadas , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Hepatócitos/efeitos dos fármacos , Hepatócitos/enzimologia , Humanos , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/fisiopatologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Niacinamida/análogos & derivados , Compostos de Fenilureia , Sorafenibe , Proteínas Supressoras de Tumor/metabolismoRESUMO
Acute hepatitis B virus (aHBV) infection can lead to fulminant liver failure, which likely is prevented by early lamivudine therapy. Even nonfulminant but severe acute hepatitis B can lead to significant morbidity and impaired quality of life. Therefore, lamivudine was evaluated in patients with severe aHBV in a placebo-controlled trial. Patients with severe aHBV infection (ALT >10× ULN, bilirubin >85 µm, prothrombin time >50%) were prospectively treated with lamivudine 100 mg/day or with placebo within 8 days after the diagnosis. The primary end point was time to bilirubin <34.2 µm. Secondary end points were time to clear HBsAg and HBV-DNA, development of anti-HBs and normalization of ALT. Eighteen cases were randomized to lamivudine, 17 to placebo. 94% of patients were hospitalized. No individual progressed to hepatic failure; all but one patient achieved the primary end point. Due to smaller than expected patient numbers, all study end points did not become statistically significant between treatment arms. Median time end points [in days] were bilirubin <34.2 µm (26.5 vs 32), ALT normalization (35 vs 48) and HBsAg clearance (48 vs 67) referring to earlier recovery under lamivudine, in contrast to loss of HBV-DNA (62 vs 54) and development of anti-HBs (119 vs 109). In all but two patients (one in every group), HBsAg clearance was reached in the study. Adverse events occurred more frequently during lamivudine therapy, but did not reach statistical significance. Lamivudine may ameliorate severe aHBV infection, but limited patient numbers prevented definite conclusions.