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1.
Transl Med UniSa ; 22: 24-27, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32523904

RESUMO

During the pediatric age range, one the most frequent deformities of the chest wall are Pectus Excavatum (PE). Currently the treatment of choice for PE is surgical intervention following the Nuss procedure. In this study, we present a description of the complications associated with surgical treatment of PE with the Nuss technique, in patients with symmetrical or asymmetrical deformity, in different stages of disease severity. The study was conducted in collaboration with the Pediatric Hospital "Istituto G. Gaslini" of Genoa. We analyzed a cohort of 402 patients (334 males and 68 females), who underwent corrective surgery between 2005 and 2018. Within this group of patients, we observed 82 cases with complications (20.39%), 20 of which were intraoperative (4.98%) and 62 post-operative (15.42%). For the evaluation of complications, this group was arbitrarily divided into patients with symmetric and asymmetric Pectus and in patients with mild, moderate and severe Pectus using Haller's index. Although a small group of patients presented complications, overall results from data analysis show that the Nuss technique represents the preferred surgical procedure for the treatment of PE, in agreement with literature. Furthermore, in our results show no correlation between asymmetry or severity of PE with complication related to the surgery.

2.
Transl Med UniSa ; 22: 28-32, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32523905

RESUMO

In children, differentiated thyroid carcinoma is a rare condition. Early diagnosis is not always easy, because of the lack of clinical symptoms, but it has a pivotal role in performing a correct therapeutic process. The study describes three cases of papillary thyroid carcinoma. None of the three patients had a positive familiarity or exposure to risk factors. In two cases, the tumor occurred as a non-injurious swelling in the anterior cervical region, in the other case it occurred with a latero-cervical lymphadenopathy that had been persistent for a year. In the first two patients we made a certain diagnosis by the needle aspiration of the thyroid nodule; in the other case the diagnosis was made by surgical exeresis and histological analysis of the lymph nodes. We also performed blood chemistry and hormonal tests, neck ultrasound, chest x-ray. The three children underwent total thyroidectomy and two of them also underwent right-sided cervical lymph node exeresis because there was the presence of metastasis. In our experience, the best therapeutic strategy for children with differentiated thyroid carcinoma is the total thyroidectomy, followed or not by latero-cervical lymph node exeresis and radioiodiotherapy. The removal of the whole gland reduces the risk of relapse.

3.
Transl Med UniSa ; 22: 33-37, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32523906

RESUMO

In paediatric age, indirect inguinal hernia represents more than 95% of the hernial disease. It is a congenital type, in contrast with adulthood in which acquired forms are more frequently found (1). The laparoscopic correction of indirect inguinal hernia is one of the most common surgeries performed in paediatric age. In recent years, various techniques have been introduced for the videolaparoscopic correction of this disease. The aim of this study is to provide an assessment of the efficacy and safety of the execution of a modified Flip-Flap technique, using tissue glue for filling of Peritoneal-vaginal duct (DPV), performed in order to ensure greater suture tightness and reduce the incidence of postoperative hydrocele. author1, author2, author 2, author1, author1, author 2 The Authors present a retrospective review of their record of cases, considering a total of 187 patients aged between 18 months and 14 years. For the correction of the hernial defect, the modified VLS Flip-Flap technique was carried out. The evaluation of safety, efficacy, operating time, relapse rate and development of short-term complications (such as postoperative hydrocele, scrotal hematoma or ecchymosis, atrophy or iatrogenic testicle ascension) was considered in a mean follow-up of 6 months. The Authors suggest that this variant of the peritoneal Flip-Flap technique is simple to perform; its safety, reproducibility and effectiveness is proven and has a percentage of relapses and complications overlapping with the "open" approach and superior to other laparoscopic techniques.

4.
Transl Med UniSa ; 22: 46-49, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32523909

RESUMO

Urolithiasis is a multifactorial disease; in recent years, its incidence has gradually increased in pediatric age. Among the factors involved in urolithiasis pathophysiology, urinary tract anomalies and metabolic diseases are the most relevant, although ethnicity and environmental factors may have an important role. The advances in technology and miniaturization of endoscopic devices have permitted the use of Retrograde Intrarenal Surgery (RIRS) to treat kidney and ureteral stones. Nowadays, flexible ureterorenoscopy and laser lithotripsy, which are techniques that have been applied in the management of adult upper urinary tract disorders, are also used in children as a minimally invasive treatment of urolithiasis with encouraging, effective and safe results. The Authors report a retrospective review of their record of cases considering 21 pediatric urolithiasis treatment procedures performed between October 2017 and April 2019 in a total of 17 patients (10 males and 7 females). Six procedures involved the use of the flexible ureterorenoscope (FURS) while in 15 procedures the application of the laser fiber was used (FURSL). A case of laser lithotripsy for bladder stone was included. The average age of patients was 10.5 years (2-18 years). The renal pelvis dilatation pretreatment was evaluated in post-operative follow-up. From the evaluation of the sample in analysis, the use of RIRS has good results in the treatment of paediatric urolithiasis, emerging as a valid option in the management of the paediatric population in terms of efficacy and safety, with an improvement in patient outcomes.

5.
Transl Med UniSa ; 20: 9-12, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31850246

RESUMO

In paediatric population, the laparoscopic splenectomy has been preferred to the open surgery during the last years. Due to the improvement of the technique and the devices, the indications to the laparoscopic splenectomy have been increased, even though there is still a variety of conditions in which the execution of this technique is arduous. During the preoperative consult there is the need to carefully evaluate the existence of cholecystic lithiasis, the haemoglobin level in patients with SCA, platelet count in children with ITP and the vaccination status. An anterior and a lateral or hanging spleen approach are primarily used for laparoscopic splenectomy. In the last four years, near the Section of Pediatric Surgery of the Department of Pediatrics, Obstetrics and Medicine of the Reproduction of Siena University, 8 cases of splenomegaly have been treated, 7 by lateral videolaparoscopic splenectomy (5 males and 2 females, with medium age of 10,5 years) and 1 by anterior approach (10 years). The advantages shown by these techniques allow the laparoscopic splenectomy to be considered as a valid alternative to the open surgery. In children's laparoscopic splenectomy, the rate of complications is considerably low and the the major problem is the intraoperative hemorrhage. With increasing surgical experience, the minimally invasive approach appears to be superior in terms of faster postoperative recovery, shorter hospital stay, perioperative and postoperative advantages. Therefore, the laparoscopic technique may soon be accepted as the standard method in patients requiring splenectomy.

6.
Transl Med UniSa ; 20: 19-21, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31850248

RESUMO

Gastro-oesophageal reflux is common in children, especially in the first year of life, and it may be regarded as physiological. Good functioning of the lower oesophageal sphincter depends largely on the anatomical relationships between oesophagus, stomach and diaphragm hiatus. Relative immaturity of these structures in newborn babies and young children is a risk factor in reflux disease, which may result in a wide variety of typical and/or atypical symptoms and, sometimes, serious complications such as oesophagitis and stenosis. Reflux disease may be diagnosed and studied, basing on morphological and functional aspects and, since the advent of pH-metry, it is possible to personalise the therapeutic approach to children with reflux. Surgical treatment of reflux disease in children has recently been improved due to a mini-invasive surgical approach. Absolute indications are recurrent pneumonia, intractable pain due to oesophagitis and retarded growth, often in association with neurological impairment. In the last three years, 18 children with reflux disease underwent videolaparoscopic surgery in our department, 14 by the Nissen and 4 by the Toupet technique. Post-operative pH-metry always showed a reduction in exposure of the distal oesophagus to acid (integral of H+) and an improvement in oesophageal clearance (short refluxes percentage) indicative of good functioning of the gastro-oesophageal junction. PH-metry proved to be an invaluable technique for planning therapeutic strategy. In follow-up evaluations, it enabled us to monitor functioning of the gastro-oesophageal junction and to avoid other more difficult and invasive tests in patients with severe neurological impairment.

7.
Transl Med UniSa ; 20: 4-8, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31850245

RESUMO

Congenital cystic adenomatoid malformation (CCAM) is a rare congenital lung lesion. It may appear since birth (30-35%) with difficulty breathing or may have a late onset (60-65%) with recurring pulmonary infections or growth failure; in a small percentage of cases, the lesion can be completely asymptomatic. Fetal or post-natal surgery can be used as surgical treatment of these lesions. Postnatal surgery consists of a lobectomy, bilobectomy or pneumonectomy, based on the size of the lesion. The best age to undergo this surgery is around 2 years, but only if the injury is stable and the child has no complications. The study describes three cases of CCAM, observed at the Pediatric Surgery Section of the University of Siena. We analyzed those 3 cases whose approach was defined by the onset of symptoms, age and clinical condition of patients. In the first case the surgery was performed a few hours after birth due to the worsening of the clinical conditions; in the other two cases it was delayed because the patients were asymptomatic. The purpose of this study is to review the management of patients with CCAM in relation to clinical onset and the type of injury.

8.
Pharmacogenomics J ; 18(3): 480-486, 2018 05 22.
Artigo em Inglês | MEDLINE | ID: mdl-28786423

RESUMO

HLA-DRB1*07:01 allele carriage was characterised as a risk biomarker for lapatinib-induced liver injury in a large global study evaluating lapatinib, alone and in combination with trastuzumab and taxanes, as adjuvant therapy for advanced breast cancer (adjuvant lapatinib and/or trastuzumab treatment optimisation). HLA-DRB1*07:01 carriage was associated with serum alanine aminotransferase (ALT) elevations in lapatinib-treated patients (odds ratio 6.5, P=3 × 10-26, n=4482) and the risk and severity of ALT elevation for lapatinib-treated patients was higher in homozygous than heterozygous HLA-DRB1*07:01 genotype carriers. A higher ALT case incidence plus weaker HLA association observed during concurrent administration of lapatinib and taxane suggested a subset of liver injury in this combination group that was HLA-DRB1*07:01 independent. Furthermore, the incidence of ALT elevation demonstrated an expected correlation with geographic HLA-DRB1*07:01 carriage frequency. Robust ALT elevation risk estimates for HLA-DRB1*07:01 may support causality discrimination and safety risk management during the use of lapatinib combination therapy for the treatment of metastatic breast cancer.


Assuntos
Neoplasias da Mama/tratamento farmacológico , Doença Hepática Induzida por Substâncias e Drogas/genética , Cadeias HLA-DRB1/genética , Lapatinib/efeitos adversos , Alelos , Protocolos de Quimioterapia Combinada Antineoplásica/administração & dosagem , Protocolos de Quimioterapia Combinada Antineoplásica/efeitos adversos , Neoplasias da Mama/complicações , Neoplasias da Mama/genética , Neoplasias da Mama/patologia , Doença Hepática Induzida por Substâncias e Drogas/patologia , Feminino , Predisposição Genética para Doença , Genótipo , Humanos , Lapatinib/administração & dosagem , Fígado/efeitos dos fármacos , Fígado/patologia , Estadiamento de Neoplasias , Fatores de Risco , Taxoides/administração & dosagem , Taxoides/efeitos adversos , Trastuzumab/administração & dosagem , Trastuzumab/efeitos adversos
9.
Benef Microbes ; 8(5): 823-831, 2017 Oct 13.
Artigo em Inglês | MEDLINE | ID: mdl-28969443

RESUMO

A well-established rat model of diet-induced metabolic syndrome was used to evaluate the effects of the oral administration of spores or cells of HU16, a carotenoid-producing strain of Bacillus indicus. Symptoms of metabolic syndrome were induced in 90-days old, male Sprague-Dawley rats maintained for eight weeks on a high-fat diet, as previously reported. Parallel groups of animals under the same diet regimen also received a daily dose of 1×1010 cells or spores of B. indicus HU16. Cells of strain HU16 were able to reduce symptoms of metabolic syndrome, plasma markers of inflammation and oxidative markers in plasma and liver to levels similar to those observed in rats under a standard diet. HU16 cells did not affect obesity markers or the accumulation of triglycerides in the liver of treated animals. Denaturing gradient gel electrophoresis analysis showed that the oral administration of HU16 cells did not significantly affect the gut microbiota of high fat-fed rats, suggesting that the observed beneficial effects are not due to a reshaping of the gut microbiota but rather to metabolites produced by HU16 cells.


Assuntos
Antioxidantes/metabolismo , Bacillus/crescimento & desenvolvimento , Bacillus/metabolismo , Carotenoides/metabolismo , Dieta Hiperlipídica/efeitos adversos , Síndrome Metabólica/terapia , Probióticos/administração & dosagem , Administração Oral , Animais , Modelos Animais de Doenças , Inflamação/patologia , Masculino , Síndrome Metabólica/patologia , Estresse Oxidativo , Plasma/química , Ratos Sprague-Dawley , Resultado do Tratamento
10.
Cytopathology ; 28(3): 203-215, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28008668

RESUMO

OBJECTIVES: To evaluate and compare the DNA yield and quality extracted from lymph node fine needle cytology (FNC) samples stored on FTA cards to those cryopreserved, and to assess the immunoglobulin heavy and light chains (IGHK) and T-Cell receptor beta and gamma chains (TCRBG) PCR tests. METHODS: DNA extractions were performed on FNC of 80 non-Hodgkin lymphomas (NHL), four myelomas and 56 benign reactive hyperplasias (BRH) cryopreserved and stored on FTA cards. The JAK2 gene was amplified to assess the DNA integrity and the IGHK/TCRBG clonality status was tested. RESULTS: IGHK monoclonality was found in 99% of B-cell NHL and 100% of myeloma. TCRBG monoclonality was found in 100% of T-cell NHL. TCRBG polyclonality was detected in 97% of B-cell NHL, 100% of myeloma and 96% of BRH. IGHK/TCRBG PCR data were confirmed by histological and/or follow-up controls. No differences were found in the DNA quality between cryopreservation and FTA cards storage methods. CONCLUSIONS: IGHK/TCRBG PCR of the lymphoproliferative process on FTA cards is comparable to those cryopreserved. FTA cards can be used to store lymph node FNC for further molecular investigations.


Assuntos
Cadeias Pesadas de Imunoglobulinas/genética , Cadeias Leves de Imunoglobulina/genética , Linfonodos/patologia , Receptores de Antígenos de Linfócitos T alfa-beta/genética , Receptores de Antígenos de Linfócitos T gama-delta/genética , Adulto , Idoso , Biópsia por Agulha Fina/métodos , Citodiagnóstico/métodos , Feminino , Humanos , Linfoma não Hodgkin/genética , Linfoma não Hodgkin/patologia , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase/métodos , Manejo de Espécimes/métodos
11.
Leukemia ; 29(6): 1390-401, 2015 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-25533804

RESUMO

Although anaplastic large-cell lymphomas (ALCL) carrying anaplastic lymphoma kinase (ALK) have a relatively good prognosis, aggressive forms exist. We have identified a novel translocation, causing the fusion of the TRAF1 and ALK genes, in one patient who presented with a leukemic ALK+ ALCL (ALCL-11). To uncover the mechanisms leading to high-grade ALCL, we developed a human patient-derived tumorgraft (hPDT) line. Molecular characterization of primary and PDT cells demonstrated the activation of ALK and nuclear factor kB (NFkB) pathways. Genomic studies of ALCL-11 showed the TP53 loss and the in vivo subclonal expansion of lymphoma cells, lacking PRDM1/Blimp1 and carrying c-MYC gene amplification. The treatment with proteasome inhibitors of TRAF1-ALK cells led to the downregulation of p50/p52 and lymphoma growth inhibition. Moreover, a NFkB gene set classifier stratified ALCL in distinct subsets with different clinical outcome. Although a selective ALK inhibitor (CEP28122) resulted in a significant clinical response of hPDT mice, nevertheless the disease could not be eradicated. These data indicate that the activation of NFkB signaling contributes to the neoplastic phenotype of TRAF1-ALK ALCL. ALCL hPDTs are invaluable tools to validate the role of druggable molecules, predict therapeutic responses and implement patient specific therapies.


Assuntos
Resistencia a Medicamentos Antineoplásicos , Linfoma Anaplásico de Células Grandes/genética , NF-kappa B/metabolismo , Receptores Proteína Tirosina Quinases/genética , Fator 1 Associado a Receptor de TNF/genética , Translocação Genética/genética , Quinase do Linfoma Anaplásico , Animais , Western Blotting , Citometria de Fluxo , Perfilação da Expressão Gênica , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Imunoprecipitação , Hibridização in Situ Fluorescente , Linfoma Anaplásico de Células Grandes/tratamento farmacológico , Linfoma Anaplásico de Células Grandes/mortalidade , Camundongos , Camundongos Endogâmicos NOD , NF-kappa B/genética , Fator 1 de Ligação ao Domínio I Regulador Positivo , Inibidores de Proteassoma/farmacologia , Proteínas Proto-Oncogênicas c-myc/genética , Proteínas Proto-Oncogênicas c-myc/metabolismo , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Receptores Proteína Tirosina Quinases/metabolismo , Proteínas Repressoras/genética , Proteínas Repressoras/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais , Fator 1 Associado a Receptor de TNF/metabolismo , Células Tumorais Cultivadas , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismo , Ensaios Antitumorais Modelo de Xenoenxerto
12.
Horm Metab Res ; 45(12): 874-9, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-23918688

RESUMO

The purpose of the present study was to assess the impact of previous period of caloric restriction on energy balance and skeletal muscle mitochondrial energetics in response to high-fat (HF) diet. To this end, 1 group of rats was subjected to 2 weeks of caloric restriction with nonpurified diet and then fed HF diet (430 kJ metabolizable energy/day) for 1 week, while the second group was fed ad libitum with nonpurified diet for 2 weeks and then fed HF diet (430 kJ metabolizable energy/day) for 1 week. Body composition, energy balance, and glucose homeostasis were measured. Mitochondrial mass, oxidative capacity and efficiency, parameters of oxidative stress, and antioxidant defense were evaluated in subsarcolemmal and intermyofibrillar mitochondria from skeletal muscle. Body energy and lipid content, plasma insulin, and metabolic efficiency were significantly higher, while energy expenditure significantly decreased, in food-restricted rats fed HF diet compared to controls. Mitochondrial efficiency and oxidative damage in skeletal muscle were significantly increased, while antioxidant defence was significantly lower in food-restricted rats fed HF diet, compared with controls. Finally, food-restricted rats fed HF diet exhibited significant reduction in subsarcolemmal mitochondrial mass. In conclusion, caloric restriction elicits higher mitochondrial efficiency and predisposes skeletal muscle to high fat-induced oxidative damage, which in turn could lead to impaired glucose homeostasis in food-restricted rats fed HF diet.


Assuntos
Restrição Calórica , Dieta Hiperlipídica , Comportamento Alimentar , Mitocôndrias Musculares/patologia , Músculo Esquelético/patologia , Estresse Oxidativo , Aconitato Hidratase/metabolismo , Animais , Glicemia/metabolismo , Composição Corporal , Citrato (si)-Sintase/metabolismo , Metabolismo Energético , Insulina/sangue , Cinética , Peroxidação de Lipídeos , Masculino , Mitocôndrias Musculares/enzimologia , Músculo Esquelético/enzimologia , Prótons , Ratos , Ratos Sprague-Dawley , Sarcolema/enzimologia , Superóxido Dismutase/metabolismo
13.
Clin Exp Allergy ; 43(1): 128-40, 2013 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-23278887

RESUMO

BACKGROUND: Among the peach-derived allergens which are already known, the lipid transfer protein (Pru p 3) seems to be the one to exert severe allergic reactions. OBJECTIVE: To identify and characterize a new peach allergen causing a clinical picture similar to that of Pru p 3. METHODS: Patients were selected on the basis of their severe clinical reactivity and negative results to a panel of peach allergens available on the ISAC103 microarray. Several in-house and commercial preparations were compared. Several methods were used to characterize the newly identified molecule. Specific IgE and inhibition assays were performed using the Allergen micro-Beads Array (ABA) assay. RESULTS: Negative ISAC results to Pru p 3 were confirmed by additional testing in contrast with the positive results obtained by commercial Pru p 3-enriched peach peel extracts. The analyses of one of these preparations led to the identification of Peamaclein, a new allergenic protein. It is a small, basic, cysteine-rich, heat-stable, digestion-resistant protein, homologous to a potato antimicrobial peptide. Peamaclein was able to trigger positive skin test reactions and to bind IgE in the ABA assay. It displays an electrophoretic mobility and chromatographic behaviour similar to that of Pru p 3; therefore, it can be hidden in Pru p 3 preparations. In fact, Pru p 3-enriched peach peel extracts were found to contain both Pru p 3 and Peamaclein by means of comparative in vivo testing, and by biochemical and immunochemical assays. Commercially available anti-Pru p 3 polyclonal antibodies were found to have a double specificity for the two molecules. CONCLUSIONS AND CLINICAL RELEVANCE: A new allergen from peach belonging to a new family of allergenic proteins has been identified and characterized. This knowledge on Peamaclein will improve our understanding on the clinical aspects of the peach allergy and the quality of diagnostic reagents.


Assuntos
Alérgenos/imunologia , Antígenos de Plantas/imunologia , Hipersensibilidade Alimentar/imunologia , Proteínas de Plantas/imunologia , Prunus/imunologia , Adolescente , Adulto , Alérgenos/efeitos adversos , Alérgenos/química , Antígenos de Plantas/efeitos adversos , Antígenos de Plantas/química , Criança , Pré-Escolar , Método Duplo-Cego , Feminino , Humanos , Imunoglobulina E/biossíntese , Masculino , Pessoa de Meia-Idade , Extratos Vegetais/química , Extratos Vegetais/imunologia , Proteínas de Plantas/efeitos adversos , Proteínas de Plantas/química , Prunus/efeitos adversos , Prunus/química , Adulto Jovem
14.
Int J Obes (Lond) ; 31(10): 1596-604, 2007 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-17637704

RESUMO

OBJECTIVE: To investigate whether changes in body energy balance induced by long-term high-fat feeding in adult rats could be associated with modifications in energetic behaviour and oxidative stress of skeletal muscle subsarcolemmal (SS) and intermyofibrillar (IMF) mitochondrial populations. DESIGN: Adult rats were fed low-fat or high-fat diet for 7 weeks. MEASUREMENTS: Body energy balance and composition analysis together with plasma insulin and glucose level determination in the whole animal. Oxidative capacity, basal and induced proton leaks as well as aconitase and superoxide dismutase activities in SS and IMF mitochondria from skeletal muscle. RESULTS: High-fat fed rats exhibit increased body lipid content, as well as hyperinsulinemia, hyperglycaemia and higher plasma non-esterified fatty acids. In addition, SS mitochondria display lower respiratory capacity and a different behaviour of SS and IMF mitochondria is found in the prevention from oxidative damage. CONCLUSIONS: A deleterious consequence of decreased oxidative capacity in SS mitochondria from rats fed high-fat diet would be a reduced utilization of energy substrates, especially fatty acids, which may lead to intracellular triglyceride accumulation, lipotoxicity and insulin resistance development. Our results thus reveal a possible role for SS mitochondria in the impairment of glucose homeostasis induced by high-fat diet.


Assuntos
Intolerância à Glucose/metabolismo , Resistência à Insulina/fisiologia , Mitocôndrias Musculares/metabolismo , Músculo Esquelético/metabolismo , Animais , Composição Corporal/fisiologia , Gorduras na Dieta/administração & dosagem , Ingestão de Energia , Metabolismo Energético , Masculino , Obesidade , Estresse Oxidativo/fisiologia , Ratos , Ratos Wistar , Sarcolema/metabolismo
15.
Cell Mol Life Sci ; 63(3): 358-66, 2006 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-16416026

RESUMO

This study was designed to examine energetic behaviour of skeletal muscle subsarcolemmal and intermyofibrillar mitochondrial populations. The data show that subsarcolemmal mitochondria exhibited a lower degree of coupling and efficiency than intermyofibrillar ones, and can therefore be considered less efficient at producing ATP. In addition, subsarcolemmal mitochondria showed an increased sensitivity to palmitate-induced uncoupling, in line with high adenine nucleotide translocator content and decreased oxidative damage. We then determined the effect of 24 h fasting on energetic characteristics of skeletal muscle mitochondrial populations. We found that fasting enhanced proton leak and decreased the degree of coupling and efficiency, both in the absence and in the presence of palmitate only in subsarcolemmal mitochondria. Moreover, this mitochondrial population showed lower oxidative damage, probably due to a counter-regulatory mechanism mediated by uncoupling protein 3. Subsarcolemmal and intermyofibrillar mitochondria appear to exhibit different energetic characteristics and can be differently affected by physiological stimuli.


Assuntos
Mitocôndrias Musculares/metabolismo , Músculo Esquelético/metabolismo , Miofibrilas/metabolismo , Sarcolema/metabolismo , Aconitato Hidratase/metabolismo , Animais , Proteínas de Transporte/metabolismo , Metabolismo Energético , Jejum , Ácidos Graxos/metabolismo , Técnicas In Vitro , Canais Iônicos , Masculino , Potenciais da Membrana , Mitocôndrias Musculares/fisiologia , Translocases Mitocondriais de ADP e ATP/metabolismo , Proteínas Mitocondriais , Oxigênio/metabolismo , Ratos , Ratos Wistar , Superóxido Dismutase/metabolismo , Proteína Desacopladora 3
16.
Cell Mol Life Sci ; 61(11): 1366-71, 2004 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-15170514

RESUMO

This study was designed to examine the effect of youth-adulthood transition on hepatic mitochondrial energy efficiency. The changes in basal and palmitate-induced proton leak, which contribute to mitochondrial efficiency, were evaluated in mitochondria isolated from the liver of young and adult rats. Alterations in mitochondrial cytochrome oxidase and aconitase specific activities, and in adenine nucleotide translocator content were also assessed. There was no difference in basal proton leak or thermodynamic coupling and efficiency of oxidative phosphorylation in liver mitochondria between the two rat groups. On the other hand, palmitate-induced proton leak increased significantly in adult rats. The function of this uncoupling could be avoidance of elevated formation of reactive oxygen species, which are known to accelerate ageing.


Assuntos
Envelhecimento/metabolismo , Mitocôndrias Hepáticas/metabolismo , Fosforilação Oxidativa , Aconitato Hidratase/metabolismo , Animais , Complexo IV da Cadeia de Transporte de Elétrons/metabolismo , Metabolismo Energético/fisiologia , Masculino , Potenciais da Membrana , Mitocôndrias Hepáticas/fisiologia , Translocases Mitocondriais de ADP e ATP/análise , Oxigênio/metabolismo , Ácido Palmítico/farmacologia , Ratos , Ratos Wistar
17.
FEBS Lett ; 544(1-3): 138-42, 2003 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-12782304

RESUMO

We investigated, in skeletal muscle mitochondria isolated from semistarved and refed rats, the relation between the protein expression of uncoupling protein 3 (UCP3) and mitochondrial oxidative capacity, assessed as state 4 and state 3 respiration rates in presence of substrates that are either non-lipids (glutamate, succinate) or lipids (palmitoyl CoA, palmitoylcarnitine). During semistarvation, when whole-body thermogenesis is diminished, state 3 respiration was lower than in fed controls by about 30% independently of substrate types, while state 4 respiration was lower by 20% only during succinate oxidation, but UCP3 was unaltered. After 5 days of refeeding, when thermogenesis is still diminished, neither state 4, state 3 nor UCP3 were lower than in controls. Refeeding on a high-fat diet, which exacerbates the suppression of thermogenesis, resulted in a two-fold elevation in UCP3 but no change in state 4 or state 3 respiration. These results during semistarvation and refeeding, in line with those previously reported for fasting, are not in support of the hypothesis that UCP3 is a mediator of adaptive thermogenesis pertaining to weight regulation, and underscore the need for caution in interpreting parallel changes in UCP3 and mitochondrial oxidative capacity as the reflection of mitochondrial uncoupling by UCP3.


Assuntos
Proteínas de Transporte/metabolismo , Músculo Esquelético/metabolismo , Consumo de Oxigênio , Oxigênio/metabolismo , Ração Animal , Animais , Western Blotting , Peso Corporal , Privação de Alimentos , Ácido Glutâmico/química , Canais Iônicos , Metabolismo dos Lipídeos , Masculino , Mitocôndrias/metabolismo , Proteínas Mitocondriais , Músculo Esquelético/citologia , Palmitoil Coenzima A/química , Palmitoilcarnitina/química , Ratos , Ratos Sprague-Dawley , Ácido Succínico/química , Proteína Desacopladora 3
18.
G Ital Med Lav Ergon ; 25 Suppl(3): 32-4, 2003.
Artigo em Italiano | MEDLINE | ID: mdl-14979072

RESUMO

This study aims at measuring association between environmental exposure to PAHs and internal or biologically effective dose biomarkers among 212 individuals living at different distance from the industrial area of Taranto, Italy. Environmental PAH exposure was determined by area monitoring. PAH levels was measured by HPLC UV/FL. Urinary 1-hydroxypyrene levels were measured by HPLC/Fluorescence. The levels of DNA adducts were determined by 32P. post-labelling. A questionnaire was administered to gather personal data, residence, occupational history, alcohol, tobacco smoke, and diet, to control for other sources of exposure to PAH. Blood and urinary samples, drawn from the subjects studied were collected and analysed. A multivariate regression model was used to investigate the influence of several confounding factors. Housing was placed within 1 km from smokestacks in Taranto and Statte, whereas they were more far away in Locorotondo and in Alberobello. The prevalence of current smokers was 31.6%.


Assuntos
Adutos de DNA/análise , Exposição Ambiental/efeitos adversos , Exposição Ambiental/análise , Hidrocarbonetos Policíclicos Aromáticos/toxicidade , Pirenos/análise , Feminino , Humanos , Masculino , Inquéritos e Questionários
19.
Int J Obes Relat Metab Disord ; 26(1): 65-72, 2002 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-11791148

RESUMO

OBJECTIVE: To investigate whether young rats respond to high-fat feeding through changes in energy efficiency and fuel partitioning at the level of skeletal muscle, to avoid obesity development. In addition, to establish whether the two mitochondrial subpopulations, which exist in skeletal muscle, ie subsarcolemmal and intermyofibrillar, are differently affected by high-fat feeding. DESIGN: Weaning rats were fed a low-fat or a high-fat diet for 15 days. MEASUREMENTS: Energy balance and lipid partitioning in the whole animal. State 3 and state 4 oxygen consumption rates in whole skeletal muscle homogenate. State 3 and state 4 oxygen consumption rates, membrane potential and uncoupling effect of palmitate in subsarcolemmal and intermyofibrillar mitochondria from skeletal muscle. RESULTS: Rats fed a high-fat diet showed an increased whole body lipid utilization. Skeletal muscle NAD-linked and lipid oxidative capacity significantly increased at the whole-tissue level, due to an increase in lipid oxidative capacity in subsarcolemmal and intermyofibrillar mitochondria and in NAD-linked activity only in intermyofibrillar ones. In addition, rats fed a high-fat diet showed an increase in the uncoupling effect of palmitate in both the mitochondrial populations. CONCLUSIONS: In young rats fed a high-fat diet, skeletal muscle contributes to enhanced whole body lipid oxidation through an increased mitochondrial capacity to use lipids as metabolic fuels, associated with a decrease in energy coupling.


Assuntos
Gorduras na Dieta/administração & dosagem , Músculo Esquelético/metabolismo , Consumo de Oxigênio/efeitos dos fármacos , Ração Animal , Animais , Animais Recém-Nascidos , Composição Corporal , Ingestão de Energia , Metabolismo Energético/efeitos dos fármacos , Metabolismo dos Lipídeos , Masculino , Mitocôndrias Musculares/metabolismo , Músculo Esquelético/enzimologia , Ratos , Ratos Wistar
20.
FEBS Lett ; 505(1): 53-6, 2001 Sep 07.
Artigo em Inglês | MEDLINE | ID: mdl-11557041

RESUMO

We have investigated the effect of 24-h fasting on basal proton leak and uncoupling protein (UCP) 3 expression at the protein level in subsarcolemmal and intermyofibrillar skeletal muscle mitochondria. In fed rats, the two mitochondrial populations displayed different proton leak, but the same protein content of UCP3. In addition, 24-h fasting, both at 24 and 29 degrees C, induced an increase in proton leak only in subsarcolemmal mitochondria, while UCP3 content increased in both the populations. From the present data, it appears that UCP3 does not control the basal proton leak of skeletal muscle mitochondria.


Assuntos
Proteínas de Transporte/metabolismo , Mitocôndrias Musculares/metabolismo , Músculo Esquelético/metabolismo , Miofibrilas/metabolismo , Sarcolema/metabolismo , Animais , Jejum , Canais Iônicos , Cinética , Masculino , Proteínas Mitocondriais , Prótons , Ratos , Ratos Wistar , Temperatura , Proteína Desacopladora 3
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