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1.
Clin Radiol ; 74(3): 220-227, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30554806

RESUMO

AIM: To identify, in vitro, the best fruit juice to use as oral contrast agent in magnetic resonance cholangiopancreatography (MRCP) and to test, in vivo, the best natural juice and the new parameters in MRCP sequences identified in vitro. MATERIALS AND METHODS: The in vitro evaluations consisted of measuring the T2 values of a pure solution of manganese (Mn) and iron (Fe) at different concentrations, measuring the content of Mn and Fe in five commercial juices and their T2 relaxation times, and identifying the optimal juice dilution for suppressing the gastrointestinal fluid signal. The new parameters of MRCP sequences were tested in vivo. RESULTS: Manganese alone strongly influenced the shortening of the T2 values (p=0.004). The T2 value with an echo time (TE) of ≥1,000 ms enabled sufficient intestinal fluid suppression in the case of high juice dilution. A flip angle of 90° maximised the differences between the high signal from static fluids, such as the bile and the fluid in the gastrointestinal tract, using fast imaging employing steady-state acquisition (FIESTA) sequences (p<0.001). CONCLUSION: The shortening of the T2 relaxation time depended only on the Mn concentration. All the commercial juices had an Mn concentration sufficient to suppress the gastrointestinal fluid signal using long TE sequences. The oral ingestion of commercial juice before MRCP was enough to suppress the signal from the gastrointestinal fluids, regardless of its dilution after ingestion. When using FIESTA sequences, a flip angle of 90° allowed the best suppression of gastrointestinal fluid signals.


Assuntos
Colangiopancreatografia por Ressonância Magnética/métodos , Meios de Contraste/administração & dosagem , Sucos de Frutas e Vegetais , Administração Oral , Meios de Contraste/química , Humanos , Técnicas In Vitro , Ferro/química , Manganês/química
2.
Br J Cancer ; 112(1): 69-76, 2015 Jan 06.
Artigo em Inglês | MEDLINE | ID: mdl-25314061

RESUMO

BACKGROUND: Surveillance for hepatocellular carcinoma (HCC) is recommended in patients with cirrhosis. As α-fetoprotein (AFP) is considered a poor surveillance test, we tested the performance of its changes over time. METHODS: Eighty patients were diagnosed with HCC (cases) during semiannual surveillance with ultrasonography and AFP measurement were recruited and matched for age, gender, etiology and Child-Pugh class with 160 contemporary cancer-free controls undergoing the same surveillance training group (TG). As a validation group (VG) we considered 36 subsequent patients diagnosed with HCC, matched 1 : 3 with contemporary cancer-free controls. α-Fetoprotein values at the time of HCC diagnosis (T0) and its changes over the 12 (Δ12) and 6 months (Δ6) before cancer detection were considered. RESULTS: In both TG and VG, >80% of HCCs were found at an early stage. In TG, AFP significantly increased over time only in cases. T0 AFP and a positive Δ6 were independently associated with HCC diagnosis (odds ratio: 1.031 and 2.402, respectively). The area under the curve of T0 AFP was 0.76 and its best cutoff (BC) was 10 ng ml(-1) (sensitivity 66.3%, specificity 80.6%). The combination of AFP >10 ng ml(-1) or a positive Δ6 composite α-fetoprotein index (CAI) increased the sensitivity to 80% with a negative predictive value (NPV) of 86.2%. Negative predictive value rose to 99%, considering a cancer prevalence of 3%. In the VG, the AFP-BC was again 10 ng ml(-1) (sensitivity 66.7%, specificity 88.9%), and CAI sensitivity was 80.6% with a NPV value of 90.5%. CONCLUSIONS: CAI achieves adequate sensitivity and NPV as a surveillance test for the early detection of HCC in cirrhosis.


Assuntos
Carcinoma Hepatocelular/química , Cirrose Hepática/diagnóstico , Neoplasias Hepáticas/química , alfa-Fetoproteínas/análise , Adulto , Idoso , Idoso de 80 Anos ou mais , Carcinoma Hepatocelular/diagnóstico , Estudos de Casos e Controles , Feminino , Humanos , Neoplasias Hepáticas/diagnóstico , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Estudos Retrospectivos
3.
Aliment Pharmacol Ther ; 24(8): 1151-61, 2006 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-17014574

RESUMO

BACKGROUND: Alcoholic liver disease has a known aetiology but a complex and incompletely known pathogenesis. It is an extremely common disease with significant morbidity and mortality, but the reason why only a relatively small proportion of heavy drinkers progress to advanced disease remains elusive. AIM: To recognize the factors responsible for the development and progression of alcoholic liver disease, in the light of current knowledge on this matter. METHODS: We performed a structured literature review identifying studies focusing on the complex pathogenetic pathway and risk factors of alcoholic liver disease. Results In addition to the cumulative amount of alcohol intake and alcohol consumption patterns, factors such as gender and ethnicity, genetic background, nutritional factors, energy metabolism abnormalities, oxidative stress, immunological mechanisms and hepatic co-morbid conditions play a key role in the genesis and progression of alcoholic liver injury. CONCLUSIONS: Understanding the pathogenesis and risk factors of alcoholic liver disease should provide insight into the development of therapeutic strategies.


Assuntos
Hepatopatias Alcoólicas/etiologia , Consumo de Bebidas Alcoólicas , Citocromo P-450 CYP2E1/genética , Metabolismo Energético/fisiologia , Feminino , Predisposição Genética para Doença/genética , Humanos , Ferro/metabolismo , Hepatopatias Alcoólicas/genética , Hepatopatias Alcoólicas/fisiopatologia , Masculino , Distúrbios Nutricionais/complicações , Distúrbios Nutricionais/fisiopatologia , Estresse Oxidativo/fisiologia , Fatores de Risco , Fatores Sexuais
4.
Dig Liver Dis ; 38(1): 27-32, 2006 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-16311084

RESUMO

BACKGROUND: Hepatitis C virus recurrence after liver transplantation is universal, leading to chronic hepatitis and cirrhosis. AIMS AND PATIENTS: We evaluated the efficacy and safety of pegylated interferon and ribavirin in 20 patients with recurrent Hepatitis C virus after liver transplantation (10 naïve and 10 non-responders to a previous interferon course). METHODS: Treatment consisted of pegylated interferon alfa-2b (1.0 microg/kg once weekly) and ribavirin (600 mg/daily) for at least 6 months. Therapy continued for an additional 6 months only in patients with undetectable serum Hepatitis C virus-RNA or >2 log drop from baseline levels. RESULTS: Eleven out of 20 patients (55%) completed 1 year of treatment. Nine patients (45%) had undetectable Hepatitis C virus-RNA at the end of treatment, six of them were naïves and three non-responders. In all of them, virological response persisted 6 months after discontinuation of therapy, so the sustained virological response rate was 60% in naïve patients and 30% in non-responders. CONCLUSIONS: Our results suggest that pegylated interferon plus ribavirin combination therapy may be effective in patients with post-liver transplantation recurrent chronic Hepatitis C, even in those previously non-responders to interferon plus ribavirin. These results need to be confirmed by large studies.


Assuntos
Antivirais/administração & dosagem , Hepatite C Crônica/terapia , Terapia de Imunossupressão , Interferon-alfa/administração & dosagem , Transplante de Fígado , Ribavirina/administração & dosagem , Idoso , Antivirais/efeitos adversos , Feminino , Hepacivirus/genética , Humanos , Interferon alfa-2 , Interferon-alfa/efeitos adversos , Masculino , Pessoa de Meia-Idade , Polietilenoglicóis , RNA Viral/sangue , Proteínas Recombinantes , Recidiva , Ribavirina/efeitos adversos
5.
Trends Biotechnol ; 14(12): 471-7, 1996 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-8987916

RESUMO

On-line analysis of one component in a complex media used for bioprocesses requires the application of selective tests such as enzymes assays. Because these assays are susceptible to interference by other medium components and have a limited detection range, automatic sample pretreatment is a prerequisite. The progress made with automatic sample pretreatment in flow-injection analysis makes this technique particularly suitable for on-line monitoring of bioprocesses. Moreover, newly developed software control systems may improve the necessary robustness of flow-infection analysis systems.


Assuntos
Biotecnologia , Análise de Injeção de Fluxo , Animais , Células Cultivadas , Enzimas Imobilizadas , Sistemas On-Line
6.
J Nucl Med ; 41(3): 556-64, 2000 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-10716332

RESUMO

UNLABELLED: A fluidized-bed cell-culture reactor with on-line radioactivity detection was developed for the in vitro evaluation of radiopharmaceuticals. The technique was applied to measure the dependency of the lumped constant (LC) of FDG on the glucose concentration in the culture medium in a human glioma cell line. METHODS: Human glioblastoma cells (86HG39) immobilized in open porous microcarriers were cultivated in a continuously operating fluidized-bed bioreactor. At different glucose concentrations in the culture medium, step inputs (0.1 MBq/mL) of FDG were performed and the cellular uptake of FDG was measured on-line and compared with analyzed samples. From these results, the LC of FDG and its dependency on the glucose concentration were calculated. RESULTS: This fluidized-bed technique enabled precise and reproducible adjustment of all relevant experimental parameters, including radiotracer time-concentration course, medium composition, pH, dissolved oxygen and temperature under steady-state conditions, and an on-line determination of the intracellular radiotracer uptake. The immobilized glioma cells formed stable, 3-dimensional, tumor-like spheroids and were continuously proliferating, as proven by an S-phase portion of 25%-40%. For further examination of the cells, an enzymatic method for detachment from the carriers without cellular destruction was introduced. In the FDG experiments, a significant dependency of the LC on the glucose level was found. For normoglycemic glucose concentrations, the LC was determined to be in the range of 0.7+/-0.1, whereas in hypoglycemia LC increased progressively up to a value of 1.22+/-0.01 at a glucose concentration of 3 mmol/L. CONCLUSION: The bioreactor represents an improved in vitro model for the on-line evaluation of radiotracers and combines a wide range of experimental setups and 3-dimensional, tissue-like cell cultivation with a technique for on-line radioactivity detection.


Assuntos
Reatores Biológicos , Fluordesoxiglucose F18 , Glioblastoma/metabolismo , Compostos Radiofarmacêuticos , Meios de Cultura , Glucose/metabolismo , Humanos , Técnicas In Vitro , Microscopia Eletrônica , Células Tumorais Cultivadas/metabolismo
7.
Antiviral Res ; 49(2): 75-81, 2001 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-11248360

RESUMO

BACKGROUND AND AIMS: Interferon-alpha treatment has been the treatment of choice for chronic hepatitis with unpredictable results. Recently, Lamivudine has been licensed for use against HBV infection with good results. Unfortunately, recurrence of viremia after lamivudine withdrawal is common and prolonged treatment can induce the emergence of resistant mutant strains. It has been shown that vitamin E can increase the host immune response, and this may provide protection against infectious diseases. METHODS: We evaluated vitamin E supplementation as therapy for chronic hepatitis B in a pilot study including 32 patients. Patients were randomly allocated to receive vitamin E at the dose of 300 mg twice daily for 3 months (15 patients) or no treatment (17 patients). They were seen monthly during the first 3 months and thereafter quarterly for additional 12 months. RESULTS: The two groups were comparable at enrollment. At the end of the study period, alanine aminotransferase (ALT) normalization was observed in 7 (47%) patients in vitamin E group and only in 1 (6%) of the controls (P=0.011); HBV-DNA negativization was observed in 8 (53%) patients in the vitamin E group as compared to 3 (18%) in the control group, respectively (P=0.039). A complete response (normal ALT and negative HBV-DNA) was obtained in 7 (47%) patients taking vitamin E and in none of the controls (P=0.0019). CONCLUSION: Vitamin E supplementation might be effective in the treatment of chronic hepatitis B.


Assuntos
Hepatite B Crônica/tratamento farmacológico , Vitamina E/uso terapêutico , Adulto , Alanina Transaminase/sangue , DNA Viral/sangue , Feminino , Vírus da Hepatite B/isolamento & purificação , Vírus da Hepatite B/fisiologia , Hepatite B Crônica/virologia , Humanos , Masculino , Pessoa de Meia-Idade , Projetos Piloto , Resultado do Tratamento
8.
J Gerontol A Biol Sci Med Sci ; 52(5): M305-9, 1997 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-9310085

RESUMO

BACKGROUND: Age is considered one of the important contraindications to surgery for hepatocellular carcinoma (HCC) in cirrhosis patients. We therefore evaluated the safety and prevalence of side effects in endoarterial therapy (EAT) in subjects aged over 65 years compared with younger treated patients. METHODS: Thirty-eight patients with HCC aged 65 years and over underwent transcatheter arterial chemoembolization (TACE) (n = 28) or intraarterial chemotherapy (IAC) (n = 10). The survival rate was calculated using Kaplan-Meier's method with respect to a control group consisting of younger treated subjects (44 TACE; 21 IAC) comparable for stage of HCC and severity of the underlying cirrhosis. RESULTS: The comparison between the two groups regarding side effects, procedure-related death, and survival did not show any difference considering the whole EAT procedure. TACE in elderly subjects reached a statistically lower outcome with respect to younger patients (p < .025) but remained statistically superior in survival versus both older and younger patients treated with IAC (p < .05, respectively). Stratifying the patients following the degree of Lipiodol uptake of tumor mass in the three groups (Group I, > 75%; Group II, 50-75%; Group III, < 50%), in the young subjects a higher probability of survival was strictly correlated to a degree of uptake over 75%, while in the elderly patients an impregnation over 50% was sufficient to obtain a satisfactory survival curve. CONCLUSIONS: EAT is a reliable and safe therapeutic option for the geriatric patient with HCC, with TACE showing a better efficacy than IAC, requiring a lesser degree of Lipiodol uptake to achieve an improvement of outcome.


Assuntos
Carcinoma Hepatocelular/terapia , Quimioembolização Terapêutica , Meios de Contraste/farmacocinética , Óleo Iodado/farmacocinética , Neoplasias Hepáticas/terapia , Adulto , Idoso , Idoso de 80 Anos ou mais , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/mortalidade , Feminino , Humanos , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/mortalidade , Masculino , Pessoa de Meia-Idade , Prognóstico , Taxa de Sobrevida
9.
J Biotechnol ; 63(3): 187-98, 1998 Aug 27.
Artigo em Inglês | MEDLINE | ID: mdl-9803533

RESUMO

Dielectric spectroscopy, also referred to as capacitance measurement, was evaluated as a tool for on-line and real-time monitoring of hybridoma cell growth in suspension batch culture and in immobilized cell culture in porous glass carriers. The capacitance signal proved to be a lumped parameter influenced by cell concentration, cell size and culture conditions. During a batch culture the cell specific capacitance signal changes by about 45% having a maximum value at the maximum growth rate. An excellent correlation between the specific capacitance and the specific amount of nucleotidetriphosphates in the cells could be shown. Dielectric spectroscopy proved to be a useful tool for on-line monitoring of cell attachment and growth in open porous microcarriers in fluidized bed fermenters. Also, in this system only an approximate correlation with viable cell concentration appeared, whereas an exact correlation with the glutamine consumption rate, a measure of the metabolic activity of the cells, could be shown. This allowed a closed loop control of the medium feed rate, which was directly linked to the capacitance signal during the entire course of a continuous fermentation.


Assuntos
Hibridomas/citologia , Análise Espectral/métodos , Animais , Reatores Biológicos , Biotecnologia , Contagem de Células , Divisão Celular , Tamanho Celular , Células Imobilizadas , Condutividade Elétrica , Fermentação , Glutamina/metabolismo , Hibridomas/metabolismo , Líquido Intracelular/metabolismo , Camundongos , Nucleotídeos/metabolismo , Sistemas On-Line
10.
J Biotechnol ; 37(3): 253-64, 1994 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-7765574

RESUMO

A multi-channel flow injection analysis system was used for on-line monitoring of a continuous animal cell culture with high cell density. With this system, the glucose, lactate and glutamine concentration were determined using immobilized dehydrogenases, ammonium using an aqueous o-phthaldialdehyde solution. Glutamine concentration was determined on the basis of the difference between a glutamine and a glutamate measurement. To prevent disturbance of the measurement and pollution of the system, the analytes in the sample were separated from high molecular compounds by on-line dialysis. On-line gas dialysis was used to avoid interference of other amino groups with the ammonium determination. In addition, dialysis was used as a dilution step. The measurement time for all four components was 42 min. This time included a final washing period after the analysis cycle. The system was calibrated once a day. Two continuous cultivations of a hybridoma cell line immobilized in open-porous glass carriers were monitored, using a fluidized bed reactor as cultivation system. The concentration of glutamine, glucose and ammonium determined with the on-line FIA system were in good agreement with the off-line data determined once a day. Only the lactate data showed some deviation. The immobilized enzyme reactors could be used for up to 3000-5000 injections. During the first cultivation, lasting 200 h, the start up period of the reactor was monitored. The on-line measurements described much better the time-course of the concentrations than the off-line data. It was possible to estimate the growth rate of the cells in the micro-carriers by the on-line data. In the course of the second cultivation, which lasted almost 1000 h, the influence of the dissolved oxygen concentration on the cell metabolism was monitored. It was noted that a sudden change of the glutamine concentration in the feed caused a fast change of the consumption and production rate of the measured metabolites.


Assuntos
Análise de Injeção de Fluxo , Hibridomas , Animais , Meios de Cultura/química , Enzimas Imobilizadas/metabolismo , Glucose/análise , Glutaminase/metabolismo , Glutamina/análise , L-Lactato Desidrogenase/metabolismo , Lactatos/análise , Ácido Láctico , Compostos de Amônio Quaternário/análise
11.
J Biotechnol ; 43(3): 229-42, 1995 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-8590649

RESUMO

An immobilized hybridoma cell line was cultivated at controlled glucose and glutamine concentrations. On-line analysis of the substrates was carried out with a multi-channel flow injection analysis system. The analysis system also determined on-line the lactate and ammonium concentration. The substrate concentrations were controlled using an adaptive-control strategy. This strategy consisted of the estimation of the real-time concentrations and volumetric substrate consumption rates by an Extended Kalman Filter, and a minimum variance controller, which used the estimated parameters to set the feed rates of the substrates. The closed-loop control was used to start-up two cultures with either glucose or glutamine as control-substrate for the medium feed rate. The controller kept the concentration of the control-substrate constant by enhancing the medium feed rate simultaneously to the increasing volumetric consumption rate of the substrate. When glutamine was used as control-substrate, the glucose concentration remained relatively constant, whereas the glutamine concentration decreased during the start-up at a constant glucose concentration. This indicates that glutamine is consumed faster than glucose and will be a better control-substrate to avoid limitation during the start-up of a culture with the applied hybridoma cell line. During the colonization of the microcarriers, the yield of ammonium on glutamine decreased from 0.80 to 0.55 (mol mol-1), indicating a change in the glutamine metabolism. The yield of lactate on glucose stayed constant for both experiments. During long-term culture of more than 800 h, the controller kept both the glucose and glutamine concentrations constant at perfusion rates between 0.50 h-1 and 0.15 h-1. The medium, glucose and glutamine feed rate were independently controlled. Both the specific glutamine and glucose consumption rates remained constant for all perfusion rates, which was probably as a result of the constant concentrations. The specific monoclonal antibody production rate decreased with the perfusion rate decreasing from 0.40 h-1 to 0.20 h-1. The immobilized-cell concentration decreased only at the lowest perfusion rate. Both effects could not be explained directly by the increasing ammonium and lactate concentrations nor by the decreasing amino-acid concentrations.


Assuntos
Biotecnologia/instrumentação , Hibridomas/metabolismo , Animais , Anticorpos Monoclonais/biossíntese , Glucose/administração & dosagem , Glucose/metabolismo , Glutamina/administração & dosagem , Glutamina/metabolismo , Hibridomas/citologia , Hibridomas/imunologia , Cinética , Lactatos/metabolismo , Ácido Láctico , Modelos Biológicos , Compostos de Amônio Quaternário/metabolismo
12.
Dig Liver Dis ; 33(8): 693-7, 2001 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-11785716

RESUMO

BACKGROUND: Recurrence of hepatitis C after liver transplantation is almost constant and may lead to graft loss. The results of treatment with interferon and/or other agents have been controversial. AIMS: To evaluate the efficacy and safety of combination therapy with interferon-alpha2b (3 MU, 3 times weekly), ribavirin (600 mg daily) and amantadine (100 mg daily) in post-transplant hepatitis C. PATIENTS AND METHODS: Enrolled in the study were 9 liver transplant recipients with histologically proven recurrent hepatitis C. Patients were treated for 12 months and followed up for 6 months after treatment. RESULTS: Treatment was not tolerated: only one patient completed the planned course, two stopped therapy within the first 3 months and 6 needed a change. However, mean alanine aminotransferase levels significantly decreased during treatment and were significantly lower than baseline at the end of follow-up. One patient out of 9 (11%) achieved a biochemical and virological sustained response. Control liver biopsy showed improvement in 2/7 patients, no change in 3 and worsening in 2. CONCLUSIONS: In recurrent post-transplant hepatitis C, antiviral treatment with interferon, ribavirin and amantadine seems to be poorly tolerated. However further studies are needed before expressing any conclusion on this potentially important option.


Assuntos
Amantadina/uso terapêutico , Antivirais/uso terapêutico , Hepatite C/tratamento farmacológico , Interferon-alfa/uso terapêutico , Transplante de Fígado , Ribavirina/uso terapêutico , Idoso , Quimioterapia Combinada , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Projetos Piloto , Recidiva , Resultado do Tratamento
13.
Hepatogastroenterology ; 45(19): 209-15, 1998.
Artigo em Inglês | MEDLINE | ID: mdl-9496515

RESUMO

BACKGROUND/AIMS: To evaluate the tolerability and therapeutic potential of the immunostimulating adjuvant alpha-1-thymosin in patients with hepatocellular carcinoma. METHODOLOGY: Twelve patients with hepatocellular carcinoma were treated with alpha-1-thymosin (900 micrograms/m2 subcutaneously twice per week for 6 months) and transcatheter arterial chemoembolization and compared to a historical control group (matched for gender, age, Okuda staging, Child's score, alpha-fetoprotein serum levels and viral infection) treated with transcatheter arterial chemoembolization alone. RESULTS: No severe side effects were recorded in the 2 treatment groups. The combination of alpha-1-thymosin plus transcatheter arterial chemoembolization resulted in a longer survival that reached statistical significance 7 months after the end of treatment (p < 0.05). Patients receiving combined treatment demonstrated a significant increase in peripheral blood mononuclear cells expressing CD3 (p < 0.05) and CD8 (p < 0.025) 3 months after beginning treatment. They also had a significant increase (p < 0.05) in CD16+ and CD56+ cells after 1 month, and a slight reduction in mononuclear cells expressing CD25, a marker for cell activation. No alterations in the response to phytohemagglutinin stimulation were seen during the alpha-1-thymosin treatment. CONCLUSIONS: The absence of toxicity and the favourable effects observed in this open study call for a double blind control study to confirm the efficacy of the combined treatment.


Assuntos
Adjuvantes Imunológicos/uso terapêutico , Carcinoma Hepatocelular/terapia , Quimioembolização Terapêutica , Neoplasias Hepáticas/terapia , Timosina/análogos & derivados , Adulto , Idoso , Carcinoma Hepatocelular/imunologia , Terapia Combinada , Feminino , Humanos , Neoplasias Hepáticas/imunologia , Subpopulações de Linfócitos , Masculino , Pessoa de Meia-Idade , Timalfasina , Timosina/uso terapêutico
14.
Hepatogastroenterology ; 44(17): 1356-9, 1997.
Artigo em Inglês | MEDLINE | ID: mdl-9356855

RESUMO

BACKGROUND/AIMS: Liver transplantation for alcoholic end-stage liver disease remains controversial at many transplant centers. The aim of the present study was to evaluate the outcome of patients with alcoholic liver disease who underwent liver transplantation at Centro Trapianti, Policlinico S. Orsola, Bologna. METHODOLOGY: We describe the outcomes of 18 alcoholic patients with end-stage liver disease who received orthotopic liver transplants at our center from April, 1986 to February, 1996. The data obtained was compared with that of 114 patients with virus-related cirrhosis selected as transplant controls. An absolute period of abstinence from alcohol consumption for at least six months was required. RESULTS: Regarding the actuarial survival rate and non-fatal post-transplant complications, no significant differences were noted in comparing the non-alcoholic with the alcoholic recipients, except for a higher incidence of Cytomegalovirus infection in the alcoholic group followed-up for more than four months. The alcoholic relapse rate was 27.2%, but only one patient returned to harmful drinking. Seventy-three percent of subjects who were followed-up for at least six months were occupied in gainful employment. Alcoholic relapse did not affect employment status. CONCLUSION: This data demonstrates that liver transplantation for selected patients with end-stage alcohol-related cirrhosis achieves good results in survival, complications and employment status, and it appears difficult to defend an inflexible claim to have demonstrated an absolute long-term abstinence before transplantation when a severe deterioration of liver function is present.


Assuntos
Cirrose Hepática Alcoólica/cirurgia , Transplante de Fígado , Análise Atuarial , Adulto , Estudos de Casos e Controles , Feminino , Humanos , Cirrose Hepática/mortalidade , Cirrose Hepática/cirurgia , Cirrose Hepática/virologia , Cirrose Hepática Alcoólica/mortalidade , Masculino , Pessoa de Meia-Idade , Complicações Pós-Operatórias/epidemiologia , Recidiva , Temperança , Resultado do Tratamento
15.
Med Biol Eng Comput ; 41(3): 350-6, 2003 May.
Artigo em Inglês | MEDLINE | ID: mdl-12803302

RESUMO

Human umbilical cord blood (CB) has moved from the status of biological waste to that of a valuable source of haematopoietic stem (HS) cells. There are potentially three major clinical applications for HS cells and ex vivo-expanded HS cells: reconstitution of haematopoiesis in patients undergoing chemotherapy; gene therapy (e.g. in thalassaemia, sickle cell anaemia); and large-scale production of mature blood cells. Erythropoiesis is accomplished by highly complex interactions of haematopoietic progenitor cells, stromal cells and cytokines in the bone marrow. Among them, erythropoietin is the principal regulator. Ex vivo cell culture experiments to obtain mature red blood cells were the focus of this study. Attempts to elucidate appropriate medium components and amounts of haematopoietic growth factors were successful: enucleated and haemoglobin-filled erythroid cells were obtained from primitive HS cells. Dimethylsulphoxide (DMSO) was found to be of particular importance as an efficient differentiation inducer. The differentiation process was followed microscopically and by fluorescence-activated cell sorting (FACS). Using the micropipette aspiration technique, the elastic properties of erythroid cells were evaluated as erythropoiesis progressed. Discocyte-like cells, comprising reticulocytes and finally differentiated red blood cells, showed an about ten-fold higher membrane shear modulus compared with control cells.


Assuntos
Eritrócitos/citologia , Sangue Fetal/citologia , Células-Tronco Hematopoéticas/citologia , Técnicas de Cultura de Células/métodos , Meios de Cultura , Eritrócitos/fisiologia , Eritropoese , Humanos
16.
Biomed Tech (Berl) ; 47 Suppl 1 Pt 1: 106-9, 2002.
Artigo em Inglês | MEDLINE | ID: mdl-12451786

RESUMO

In this study we investigated the mechanical properties of in vitro cultured red blood cells (RBCs) in a liquid system. We used human umbilical cord blood as a highly efficient source of hematopoietic stem cells (HS). Our first goal was to establish an optimal medium composition in order to yield finally differentiated RBCs, i.e. enucleated and hemoglobin-filled cells. Different stages of cell differentiation were distinguished based on morphological observations and flow cytometry measurements. By means of the micropipette aspiration technique we estimated the deformability characteristics of the cultured cells. Up to the stage of oxiphilic normoblasts they readily deformed. Reticulocytes and mature RBCs showed an enhanced stiffness as compared to RBCs obtained from donors.


Assuntos
Deformação Eritrocítica/fisiologia , Membrana Eritrocítica/fisiologia , Adulto , Células Cultivadas , Elasticidade , Envelhecimento Eritrocítico/fisiologia , Sangue Fetal/citologia , Citometria de Fluxo , Humanos , Técnicas In Vitro , Recém-Nascido , Reticulócitos/fisiologia
17.
J Biotechnol ; 163(4): 371-6, 2013 Feb 20.
Artigo em Inglês | MEDLINE | ID: mdl-22465601

RESUMO

A microfluidic chip integrating amperometric enzyme sensors for the detection of glucose, glutamate and glutamine in cell-culture fermentation processes has been developed. The enzymes glucose oxidase, glutamate oxidase and glutaminase were immobilized by means of cross-linking with glutaraldehyde on platinum thin-film electrodes integrated within a microfluidic channel. The biosensor chip was coupled to a flow-injection analysis system for electrochemical characterization of the sensors. The sensors have been characterized in terms of sensitivity, linear working range and detection limit. The sensitivity evaluated from the respective peak areas was 1.47, 3.68 and 0.28 µAs/mM for the glucose, glutamate and glutamine sensor, respectively. The calibration curves were linear up to a concentration of 20 mM glucose and glutamine and up to 10 mM for glutamate. The lower detection limit amounted to be 0.05 mM for the glucose and glutamate sensor, respectively, and 0.1 mM for the glutamine sensor. Experiments in cell-culture medium have demonstrated a good correlation between the glutamate, glutamine and glucose concentrations measured with the chip-based biosensors in a differential-mode and the commercially available instrumentation. The obtained results demonstrate the feasibility of the realized microfluidic biosensor chip for monitoring of bioprocesses.


Assuntos
Técnicas Biossensoriais , Catalase/metabolismo , Análise de Injeção de Fluxo/métodos , Glucose Oxidase/metabolismo , Glutaminase/metabolismo , Técnicas Biossensoriais/economia , Técnicas Biossensoriais/instrumentação , Técnicas Biossensoriais/métodos , Calibragem , Técnicas Eletroquímicas/economia , Técnicas Eletroquímicas/instrumentação , Eletrodos , Fermentação , Análise de Injeção de Fluxo/economia , Glucose/análise , Ácido Glutâmico/análise , Glutamina/análise
19.
Dig Liver Dis ; 41(10): 700-6, 2009 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-19502118

RESUMO

Given the lack of donors, a correct organ allocation system for candidates to liver transplantation is essential to increase graft and patient survival. The most used organ allocation tools are Child-Turcotte-Pugh and model for end-stage liver disease. It is generally accepted that model for end-stage liver disease score is superior to the Child-Turcotte-Pugh classification in predicting the short-term survival of cirrhotic patients awaiting liver transplantation. Since 2002, model for end-stage liver disease is widely used for liver allocation. In recent years, to overcome limitations of the consolidated scores, some adjustments to the original model for end-stage liver disease formula and new scoring systems have been proposed. Published data suggest that integrating serum sodium and model for end-stage liver disease may improve the score prognostic accuracy but further studies are necessary to confirm this issue. The updated model for end-stage liver disease, obtained through a revision of traditional model for end-stage liver disease parameters and tested in a large cohort of patients, is of great interest at the moment. In conclusion, several scoring systems have been described for organ allocation, but today, none is definitely able to overcome the limitations of the Child-Turcotte-Pugh and model for end-stage liver disease systems.


Assuntos
Falência Hepática/diagnóstico , Transplante de Fígado , Seleção de Pacientes , Obtenção de Tecidos e Órgãos , Humanos , Fígado/cirurgia , Falência Hepática/classificação , Falência Hepática/cirurgia , Modelos Estatísticos , Prognóstico , Medição de Risco/métodos , Fatores de Risco , Índice de Gravidade de Doença , Listas de Espera
20.
Aliment Pharmacol Ther ; 28(4): 450-7, 2008 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-18549463

RESUMO

BACKGROUND: Treatment of hepatitis C virus (HCV) recurrence after liver transplantation (LT) is difficult with low response rates. AIM: To assess the safety and efficacy of pegylated-interferon (PEG-IFN) alfa-2b + ribavirin (RBV) in patients with post-LT recurrent genotype-1 HCV and to establish stopping rules according to response. METHODS: Fifty-three patients with post-LT HCV recurrence were enrolled. Patients received PEG-IFN alfa-2b 1.0 micro/kg/week plus RBV 8-10 mg/kg/day for 24 weeks. Those with 'early virological response at week 24' (EVR24) continued treatment for 24 weeks (group A). Patients without EVR24 were randomized to continue (group B) or to discontinue (group C). RESULTS: Overall sustained virological response (SVR) was 26% (14/53). Alanine aminotransferase, rapid virological response, EVR12, EVR24, undetectable serum HCV-RNA at weeks 12 (cEVR12) and 24 (cEVR24) were related to SVR. cEVR12 and cEVR24 (OR: 14.7; 95% CI: 2.02-106.4) were independent predictors of SVR. All patients with SVR, had cEVR12. No patient in groups B and C achieved end-of-treatment response. One patient in group B had SVR. CONCLUSIONS: Pegylated-interferon alfa-2b was effective in one of four of patients with HCV genotype 1 after LT. Treatment should be discontinued in patients with no virological response at week 12. Further studies are needed to evaluate whether a longer treatment period may be beneficial in patients with > or =2 log10 drop in HCV-RNA at week 24.


Assuntos
Antivirais/uso terapêutico , Hepatite C Crônica/tratamento farmacológico , Interferon-alfa/uso terapêutico , Transplante de Fígado/patologia , Ribavirina/uso terapêutico , Adulto , Idoso , Quimioterapia Combinada , Feminino , Genótipo , Hepacivirus/efeitos dos fármacos , Hepacivirus/genética , Humanos , Interferon alfa-2 , Masculino , Pessoa de Meia-Idade , Seleção de Pacientes , Polietilenoglicóis , RNA Viral/genética , Proteínas Recombinantes , Prevenção Secundária , Resultado do Tratamento
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