ABSTRACT
Fructose-1,6-bisphosphate (FBP) has been reported to have a protective effect on liver injury following ischemic/reperfusion periods. FBP maintains ATP levels and thereby cellular energy metabolism, which is important to the liver during cold preservation. In the present study, we evaluated the effects of FBP on the composition of storage solutions for cold liver preservation. Adult male Wistar rats were randomly divided into three experimental groups. Hepatic perfusion and preservation were performed with UW, UW plus 10 mmol/L FBP (UWM), and FBP 10 mmol/L (FBPS) alone solutions. Biochemical measurements of AST, ALT, and TBARS were performed on samples of the cold storage solution at 0, 12, 18, and 24 hours preservation. FBPS and UW solutions showed similar preservation grades during 18 hours. Addition of 10 mmol/L of FBP to UW solution induced liver injury and a poor preservation grade. FBP appears to protect the liver from injury caused by free radicals when the preservation time is less than 18 hours. Therefore, FBP may exert a protective effect for the preservation of livers during cold storage, and could represent an important component of new cold storage solutions.
Subject(s)
Fructosediphosphates , Liver Transplantation/physiology , Liver , Organ Preservation Solutions , Adenosine , Alanine Transaminase/analysis , Allopurinol , Animals , Glutathione , Insulin , Liver/physiology , Male , Raffinose , Rats , Rats, Wistar , Thiobarbituric Acid Reactive Substances/analysisABSTRACT
BACKGROUND: Alcoholic hepatic disease is a severe and frequent disease and its diagnosis is not always an easy task. AIM: To assess the contribution of immunoglobulin A (IgA) in the hepatic sinusoids for diagnosis of alcoholic hepatopathy. PATIENTS AND METHODS: The presence of IgA was studied through direct immunofluorescence in 59 patients submitted to hepatic needle biopsy, indicated by clinical or in vitro changes suggestive of chronic hepatopathy. RESULTS: A significant deposition of IgA was found in alcoholic patients as compared to non-alcoholic patients, with 76% sensitivity (95% CI: 54.5-89.8) and 73.5% specificity (95% CI: 55.3-86.5). In individuals who present only alcohol as the etiological agent of hepatopathy, compared with the subgroup of B or C virus carriers, the results were even more significant, with 85.7% sensitivity (95% CI: 56.2-97.5) and 89.5% specificity (95% CI: 65.5-98.2). CONCLUSION: The deposition of IgA in the hepatic sinusoids present sensitivity and specificity for the diagnosis of an alcohol-induced hepatic lesion. This resource can be particularly useful when conventional histology can not be define a specific cause for the change found.
Subject(s)
Immunoglobulin A/analysis , Liver Diseases, Alcoholic/pathology , Liver/chemistry , Adult , Aged , Biomarkers/analysis , Cross-Sectional Studies , Female , Humans , Liver/pathology , Liver Diseases, Alcoholic/metabolism , Male , Middle Aged , Prospective Studies , Sensitivity and SpecificityABSTRACT
Development of a recombinant insulin like growth factor I (IGF-I) that is distinguishable from its endogenous counterpart would provide a powerful tool for delineating the role of IGF in myogenesis. Therefore, the objective of this study was to create an epitope-tagged IGF-I that retains biological activity and determine whether expression of this construct is possible in muscle tissue following direct DNA injection. Expression vectors were created that encoded porcine IGF-I containing a T7 (11-amino acid) epitope-tag (TIGF). Immunoreactivity of the purified recombinant TIGF was confirmed using monoclonal antibodies. Biological activity was evaluated by examining differentiation of myoblasts cultured with TIGF or transfected with TIGF plasmid DNA. Addition of purified TIGF to myoblast cultures stimulated (P < 0.05) muscle creatine kinase levels similar to insulin (10(-5) M). Likewise, transfection of L6A1 with TIGF DNA hastened (P < 0.01) differentiation compared to control pcDNA-transfected myoblasts. The integrity of the recombinant protein was confirmed using a sandwich-configured enzyme linked immunosorbent assay. Finally, recombinant TIGF DNA was injected in porcine muscle and the ability to detect TIGF protein was evaluated. TIGF expression was detected in muscle fibers of injected porcine muscle. These data show that a T7 amino acid tag placed on the amino terminus of the IGF-I protein remains intact during processing and does not interfere with the biological activity of the molecule. Use of this DNA construct is an excellent tool for investigating the role of IGFs in control muscle development and provides a model to investigate other regulators of animal growth.
Subject(s)
Gene Expression Regulation , Insulin-Like Growth Factor I/physiology , Muscle, Skeletal/physiology , Swine/physiology , Animals , Antibodies, Monoclonal , Blotting, Western/veterinary , Cells, Cultured , Chromatography, Affinity/veterinary , Creatine Kinase/analysis , DNA Primers/chemistry , Enzyme-Linked Immunosorbent Assay/veterinary , Epitopes , Injections, Intramuscular/veterinary , Insulin-Like Growth Factor I/genetics , Insulin-Like Growth Factor I/immunology , Microscopy, Fluorescence/veterinary , Muscle, Skeletal/immunology , Plasmids , Recombinant Proteins/chemistry , Reverse Transcriptase Polymerase Chain Reaction/veterinary , Swine/immunology , Transfection/geneticsABSTRACT
One hundred and sixty seven cases of pituitary adenoma were analysed using the immunocytochemical method of the Avidin-Biotin Complex (ABC), described by Hsu et al. (1981). Six pituitary anti-hormones were utilized: anti-prolactin (aPRL) at a 1:1,500 dilution; anti-growth hormone (aHGH) at a 1:4,000 dilution: anti-adrenocorticotrophic hormone (aACTH) at a 1:3,000 dilution; anti-thyrothrophic hormone (aTSH) at a 1:3,000 dilution; anti-luteinizing hormone (aLH) at a 1:1,000 dilution; and a anti-follicle-stimulating hormone (aFSH) at a 1:300 dilution. Incubation period was 14 to 16 hours at 4 degrees C. The survey of clinical, laboratory and radiological data of cases of pituitary adenomas was performed after reading the stained slides using the immunocytochemical method. Of the 167 cases of pituitary adenomas, 136 (81.4%) disclosed a positive immunoreaction to one or more anti-hormones, and the positivity index of neoplastic cells varied from 1 to 90%. The immunoreaction was positive exclusively to one anti-hormone in 80 cases (58.8%) and to two or more anti-hormones in 56 cases, and the association most frequently found was between both aPRL and aHGH. The positivity to the immunoreaction was distributed as follows: -100 cases were positive for aPRL, exclusively in 4 cases; -65 cases were positive for aHGH, exclusively in 22 cases; -31 cases were positive for aACTH, exclusively in 8 cases; -5 cases were positive for aTSH, exclusively in one case; -one patient presented an adenoma positive to aLH and another patient to aFSH.
Subject(s)
Adenoma/diagnosis , Immunohistochemistry , Pituitary Neoplasms/diagnosis , Adenoma/metabolism , Adolescent , Adult , Aged , Female , Humans , Male , Middle Aged , Pituitary Hormones, Anterior/metabolism , Pituitary Neoplasms/metabolismABSTRACT
Foram analisados 167 casos de adenomas da hipófise pelo método imuno-histoquímico utilizando o Complexo da Avidina Biotina (ABC) descrito por HSU e col. (1981). Foram usados 6 anti-hormônios hipofisários: anti-prolactina (aPRL), na diluiçäo de 1:1.500, anti-hormônio do crescimento (aHGH), na diluiçäo de 1:4.000, anti-hormônio adrenocorticotrófico (aACTH), na diluiçäo de 1:3.000, anti-hormônio tireotrófico (aTSH), na diluiçäo de trófico (aACTH), na diluiçäo de 1:3.000, anti-hormônio tireotrófico (aTSH), na diluiçäo de 1:3.000, anti-hormônio luteinizante (aLH), na diluiçäo de 1:1.000, anti-hormônio folículo estimulante (a(FSH), na diluiçäo de 1;3.000. O período de incubaçäo foi de 14 a 16 horas a 4-C. Foi realizada também a coloraçäo pelo Orange G-PAS. O levantamento dos dados clínicos, laboratoriais, e radiológicos dos casos de adenomas de hipófise foi realizado após leitura das lâminas pelo método imuno-histoquímico. Dos 167 casos de adenomas da hipófise, 136 (81,4%) mostraram imuno-reaçäo positiva a um ou mais anti-hormônios, variando o índice de positividade entre 1 e 90% das células neoplásicas. A imuno-reaçäo foi positiva exclusivamente a um anti-hormônio em 80 casos (58,8%) e para dois ou mais anti-hormônios nos 56 casos restantes (41,2%), sendo a associaçäo mais freqüentemente encontrada aquela em que a positividade ocorreu para o aPRL e o aHGH. A positividade a reaçäo imuno-histoquímica distribuiu-se da seguinte forma: 100 casos foram positivos para o aPRL, em 49 pacientes de forma isolada; 65 casos foram positivos para o aHGH, em 22 pacientes de forma isolada; 31 casos foram positivos para o aACTH, em 8 pacientes de forma isolada; 5 casos foram positivos ao aTSH, em um paciente de forma isolada; um paciente apresentou adenoma positivo ao aLH; um caso foi positivo ao aFSH