RESUMO
BACKGROUND: Children with medical complexity (CMC) have been defined (Cohen et al., Pediatrics 127: 529-538, 2011.) as an emerging population potentially eligible for PPC. The current study investigated the prevalence of children with medical complexities eligible for a local palliative care network, including a paediatric hospice. METHODS: A retrospective cross-sectional survey has been conducted using children clinical charts from 14 local health authorities of our region (Emilia Romagna, Italy). RESULTS: The total number of children with life-limiting conditions was 601, with a mean age of 7.4 ± 4.8 years, a prevalence of 8.4/10.000 residents < 19 years of age and a heterogeneous presentation among the provinces in the region. Neurological diseases affect 51% of patients, followed by congenital diseases (21%) and pathologies originating in the perinatal period (6%), while only 4% of the patients had a cancer diagnosis. Patients are dependent from many devices and supports: 32% had a gastrostomy, 22% a respiratory support and 15% of patients had both of them. CONCLUSIONS: Observed regional prevalence of complex needs is lower than that published from other European countries. More research is needed to raise awareness of palliative care for children with medical complexities in order to address specific needs.
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Doença Crônica/epidemiologia , Doença Crônica/terapia , Necessidades e Demandas de Serviços de Saúde , Cuidados Paliativos , Criança , Estudos Transversais , Feminino , Cuidados Paliativos na Terminalidade da Vida , Humanos , Itália/epidemiologia , Masculino , Estudos Retrospectivos , Inquéritos e QuestionáriosRESUMO
Polyposis of the gallbladder is rare during childhood. This condition can be associated with three other conditions: metachromatic leukodystrophy, Peutz-Jeghers' syndrome, and pancreaticobiliary maljunction. We report the case of a child with hemobilia in metachromatic leukodystrophy, which rendered cholecystectomy necessary. Macroscopically, the gallbladder measured 4.6 cm in length and showed an opaque serous surface and focal brown petechiae. Moreover, a yellow polypoid lesion of 2 cm in diameter and a diffuse thickening of the fundus wall were observed. Many reports describe the importance of the association of gallbladder papillomatosis with metachromatic leukodystrophy, but only three cases presented with massive intestinal bleeding, such as our young patient had. It is thus imperative that this life-threatening condition should be well known.
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Neoplasias da Vesícula Biliar/complicações , Hemobilia/complicações , Leucodistrofia Metacromática/complicações , Leucodistrofia Metacromática/patologia , Papiloma/complicações , Pré-Escolar , Neoplasias da Vesícula Biliar/patologia , Hemobilia/patologia , Humanos , Masculino , Papiloma/patologiaRESUMO
OBJECTIVE: To gain an insight into parental perceptions of infant feeding practices in five European countries. DESIGN: An exploratory investigation using focus group discussions. Various aspects addressed included social and cultural setting for the consumption of food, infant feeding practice and behaviour, consumer health awareness and sources of information, and attitudes towards a healthy infant diet. SETTING: Focus group participants were recruited from centres in five countries, Germany, Italy, Scotland, Spain and Sweden, with three focus groups being conducted in each centre. SUBJECTS: A total of 108 parents with infants up to the age of 12 months participated in focus group discussions across these centres. METHODS: Focus groups were conducted with participants from centres in five countries. RESULTS: The majority of parents in this study chose to initiate breastfeeding and prepare infant food at home. Parents did not strictly adhere to infant feeding guidelines when introducing complementary foods into their infant's diets. There were cross-cultural differences in sources of information on infant feeding practice with the paediatrician in Germany, Italy and Spain. The health visitor in Scotland and the child welfare clinics in Sweden were the most popular sources. CONCLUSIONS: A number of cultural differences and similarities in attitudes towards infant feeding practice were revealed. This makes European wide approaches to promoting healthy infant feeding difficult as different infant feeding practices are influenced not only by parental perceptions but also by advice from health professionals and feeding guidelines. Further data need to be available on parents' attitudes and beliefs towards infant feeding practice to investigate further the rationale for differing beliefs and attitudes towards infant feeding practice. SPONSORSHIP: EU Fifth Framework QLRT 2002 02606.
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Aleitamento Materno/psicologia , Conhecimentos, Atitudes e Prática em Saúde , Alimentos Infantis , Fenômenos Fisiológicos da Nutrição do Lactente/fisiologia , Pais/psicologia , Desmame , Adulto , Atitude , Comparação Transcultural , Feminino , Grupos Focais , Alemanha , Humanos , Lactente , Alimentos Infantis/normas , Recém-Nascido , Itália , Masculino , Escócia , Espanha , SuéciaRESUMO
INTRODUCTION: the prevalence of malnutrition in children and its impact on clinical outcomes is underrecognized by clinicians in Italy as well as worldwide. A novel definition of pediatric malnutrition has been recently proposed by a working group of the Academy of Nutrition and Dietetics and American Society for Parenteral and Enteral Nutrition (A.S.P.E.N.), based on the correlation between illness and the use of zscores of anthropometric measurements. AIM: to investigate the prevalence of malnutrition and related nutritional support among hospitalized children in Italy, in a nationwide survey performed in a single day (16/4/2015). METHODS: an open access website (http://nday.biomedia.net) was used to collected data from 73 hospitals and 101 wards in 14 Italian regions (1994 patients). Anonymous information was collected on hospitals' characteristics, patient's anthropometry, admission diagnosis, presence of chronic diseases and use of nutritional support: oral nutritional supplements (ONS), enteral nutrition (EN) or parenteral nutrition (PN). Z-scores of anthropometric measurements, calculated with Epi Info 7.1.5, defined nutritional status: wasting was identified by BMI or Weight-for-Length z-score (<-1 mild, <-2 moderate, <-3 severe), stunting by Height-for-Age Z-score <-2. WHO 2006 and CDC 2000 growth charts were used respectively for children younger and older than 2 years old. RESULTS: 1790 complete records were obtained for hospitalized patients aged 0-20 years, with median age 6.16 (0.1-20 years and 53.3% males). 52.9% were aged 0-6 years and 58.8% of children suffered from chronic diseases. Wasting was detected in 28.7% of the total sample with higher occurrence observed in age ranges 0-6 and 14-20 years, while 17.3% of patients showed stunting; surprisingly almost 27% of them were aged 0-2. A ranking of the admission diagnosis with the highest rate of malnutrition was complied. The prevalence of wasting was significantly (p < 0.005) higher amongst children with chronic diseases (34.1% vs. 27.1%); stunting prevalence tripled in patients with chronic disease (24.5% vs. 8.3%). Only 23.5% of malnourished children (17%, 25.6% and 36.7%, respectively mild, moderate and severe malnutrition) received nutritional support: 11.7% received oral nutrition supplements (ONS, modular or complete), 11.5% enteral nutrition (EN, 6.4% via nasogastric tube, 5.1% via gastrostomy) and 6.8 % received parenteral nutrition (PN); in some patients a combination of two. Nutritional support is more commonly used among stunting patients, 39.5% of children under treatment. CONCLUSION: Malnutrition of any grade was observed in nearly 1/3 and stunting in 17% of the reported hospitalized children, and it is likely to be underrecognized as the nutritional support reached only a small part of the malnourished children.
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Transtornos do Crescimento/epidemiologia , Desnutrição/epidemiologia , Inquéritos Nutricionais , Adolescente , Criança , Desenvolvimento Infantil , Criança Hospitalizada , Pré-Escolar , Doença Crônica , Feminino , Gráficos de Crescimento , Transtornos do Crescimento/terapia , Humanos , Lactente , Itália/epidemiologia , Masculino , Desnutrição/diagnóstico , Desnutrição/terapia , Estado Nutricional , Apoio Nutricional , Prevalência , Adulto JovemRESUMO
INTRODUCTION: Little is known about changes in intestinal microbiota during the important period of complementary feeding (weaning). This descriptive study investigated changes of selected gut microbiota and markers of gut permeability and the immune system in breast fed infants during the complementary feeding period. METHODS: 22 healthy, exclusively breast fed infants (from birth to 4 months) with no antibiotic intake during the month prior to the study, were followed from 4 to 9 months of age. Faecal and saliva samples were collected at the start of the study (V0) and at monthly intervals (V1-V5) for measurement of selective gut microbiota (bifidobacteria, lactobacilli, vancomycin-insensitive lactobacilli, enterobacteria, enterococci, Clostridium perfringens) using semi-selective media. Immune markers (alpha-1-antitrypsin, eosinophil cationic protein (ECP), secretory IgA and TNF-alpha were measured in saliva and secretory IgA and TNF-alpha in faecal samples. RESULTS: High stool bifidobacteria counts at the start of the study (7.99 1 1.95 log10 CFU/g faeces) remained stable throughout the 5 months of complementary feeding while counts of enterobacteria and enterococci increased with age (P < 0.05 and P = 0.02 respectively). Vancomycin-insensitive lactobacilli increased significantly during weaning for V0 to V3 (P < 0.01), and then decreased slightly (V4). Faecal Clostridium perfringens remained below the detection limit during the study and parameters measured in saliva did not change. Faecal ECP decreased significantly from 1.011.4 (V0) to 0.510.9 mg/mg protein (V5) P = 0.03. CONCLUSION: Age and/or diet modifications during complementary feeding had no impact on faecal bifidobacteria counts but increased those of enterobacteria and enterococci. Transient increases in faecal lactobacilli and vancomycin-insensitive lactobacilli counts were observed. The reduction in faecal ECP may indicate a decrease in gut permeability (reinforcement of gut mucosa integrity) during the weaning period with age [corrected]
Assuntos
Aleitamento Materno , Intestinos/imunologia , Intestinos/microbiologia , Biomarcadores , Fezes/microbiologia , Humanos , Lactente , Fenômenos Fisiológicos da Nutrição do Lactente , Recém-Nascido , Mucosa Intestinal/fisiologia , Saliva/microbiologia , DesmameRESUMO
BACKGROUND/PURPOSE: Scintigraphy is regarded as the "gold standard" procedure in measuring gastric emptying (GE) rates. 13C-acetate breath test (ABT), which already has been validated in adults, is a noninvasive and nonradioactive alternative method. The aim of the current study was to validate ABT against technetium Tc 99m scintigraphy in children affected by delayed GE. METHODS: Sixty children were recruited and divided into 2 groups: group A, 30 healthy controls; group B, 30 patients with gastroesophageal reflux, and scintigraphy-documented DGE (15 neurologically impaired). After an overnight fast, all of them underwent ABT using 25 to 150 mg 13C-acetate. Breath samples were obtained at baseline and then every 10 minutes for 2 hours. The 13CO2 to 12CO2 ratio in breath samples was analysed by isotope ratio mass spectrometry. Data are expressed as follows: time of peak 13C exhalation (tP13CO2b) and half emptying time in ABT (t(1/2b)), and scintigraphy half emptying time (t(1/2s)). RESULTS: In controls tP13CO2b was 37 +/- 13 minutes and t(1/2b) 74 +/- 12 minutes. In patients tP13CO2b and t(1/2b) were, respectively, 65 +/- 26 minutes and 104 +/- 18 minutes t(1/2s) was 91 +/- 21 minutes. In group B tP13CO2b and t(1/2b) were delayed significantly compared with controls, respectively, P < .03 and P < .01. In group B significant correlation between t(1/2s) and t(1/2b) was noted (r1 = 0.97). A close correlation was also observed between t(1/2s) and tP13CO2b (r2 = 0.95). CONCLUSION: The 13C ABT is an easy, reliable, and less expensive procedure for measuring GE, and its results closely correlate with those of scintigraphy in a paediatric population.
Assuntos
Testes Respiratórios , Esvaziamento Gástrico , Acetatos , Isótopos de Carbono , Criança , Pré-Escolar , Feminino , Refluxo Gastroesofágico/diagnóstico , Refluxo Gastroesofágico/diagnóstico por imagem , Humanos , Masculino , Cintilografia , Estômago/diagnóstico por imagem , TecnécioRESUMO
Sedation for children doing diagnostic or operative pediatric gastrointestinal endoscopy (PE) procedures is performed differently over the world and no consensus is yet agreed on the best paediatric endoscopy sedation (PES). Some centres do not use any sedation, especially in infants, most centre use some form of sedation: conscious sedation, deep sedation and general anaesthesia. We review sedation drugs and describe our centre protocol on 188 consecutive PE: oral premedication with flunitrazepam (0.05 mg/kg/dose) at least 30 min before procedure, petidine (1 mg/kg) followed by increasing boluses of midazolam (0.05 mg/kg up to a maximal 0.2 mg/kg or 5 mg) were given i.v. to obtain a conscious sedation. All PE could be performed and ended safely, PES resulted satisfactory in approximately 65% of patient having conscious sedation. SaO2 < 90% was observed in 2% of cases, one child had a respiratory depression after PE that resolved with flumanezil. Endoscopy and sedation was always performed by the PE team in the immediate vicinity of anaesthesiologists at work. PE can be safely performed with conscious sedation. Basic and advanced resuscitation skills are needed for the PE team who wish to perform both endoscopic and sedation procedures.
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Sedação Consciente , Endoscopia do Sistema Digestório , Criança , Pré-Escolar , Feminino , Humanos , Hipnóticos e Sedativos/administração & dosagem , Lactente , Masculino , Estudos RetrospectivosRESUMO
The Authors evaluate the effectiveness of oral bacteriotherapy using a combination of anaerobe fecal Lactobacilli for chronic non specific diarrhea of infancy. A double blind study was carried out in a total of 40 children treated with low and high doses of bacteria. The results confirm the importance of fecal flora in this disease and support the hypothesis that oral bacteriotherapy can improve clinical and laboratory presentation especially when given at high doses.
Assuntos
Bifidobacterium , Diarreia Infantil/terapia , Lactobacillus acidophilus , Administração Oral , Pré-Escolar , Doença Crônica , Diarreia Infantil/diagnóstico , Método Duplo-Cego , Humanos , Lactente , Intestinos/microbiologiaRESUMO
Authors describe 9 infants aged between 3 and 17 months, affected by recurrent episodes of bronchial asthma (at least one crises even month for three following months) and treated with sustained-release theophylline. During the 6 months of follow-up a total of 8 episodes of bronchial asthma in 7 subjects were observed. Seven out of 8 of these episodes occurred during the period of suspension of the therapy with the sustained-release theophylline. In each subjects, after 6 hours from the drug administration on average of 4-8 determinations of the theophylline blood levels were performed. More than 70 percent of the theophylline blood values has been found within the normal therapeutic range (6-20 micrograms/ml).
Assuntos
Asma/tratamento farmacológico , Teofilina/uso terapêutico , Asma/imunologia , Preparações de Ação Retardada , Feminino , Humanos , Lactente , Masculino , Testes Cutâneos , Teofilina/sangueRESUMO
Cow's milk sensitive enteropathy has been described several times but in spite of that, it is still a problem concerning the pathogenesis. Our study involves the children hospitalized from 1974 to 1984 in the First Department of Pediatrics, University of Modena. Patients were suffering from chronic diarrhea and malabsorption. At the first biopsy each child showed atrophy of the small intestinal mucosa. All patients had been fed fed with gluten. We have followed the protocol for Celiac Disease's diagnosis; we found proved 85% of cases, excluded 15%. These last cases may be considered as cow's milk sensitive enteropathy. We catamnestically considered all the clinical and laboratory data of the two groups in the purpose of selecting significative parameters for a differential diagnosis. The data meaning fully different between the two groups resulted: family history of allergy, recurrent infections, positive occult blood in the stools, eosinophils in blood greater than 400/mm3 serum IgE value greater than 97 degrees P X (p less than 0.01). On the ground of recent studies the involvement of the cell-mediated immunity in cow's milk sensitive enteropathy is supposed.
Assuntos
Duodeno/patologia , Imunoglobulinas/análise , Mucosa Intestinal/patologia , Jejuno/patologia , Síndromes de Malabsorção/patologia , Proteínas do Leite/efeitos adversos , Animais , Atrofia , Bovinos , Doença Celíaca/patologia , Criança , Pré-Escolar , Feminino , Hipersensibilidade Alimentar/etiologia , Humanos , Imunoglobulina A/análise , Imunoglobulina G/análise , Imunoglobulina M/análise , Lactente , Síndromes de Malabsorção/etiologia , Síndromes de Malabsorção/imunologia , MasculinoRESUMO
BACKGROUND: Tissue transglutaminase is the major autoantigen recognized in the sera of coeliac patients. An enzyme-linked immune-adsorbed assay based on tissue transglutaminase was recently used to measure serum tissue transglutaminase immunoglobulins A for coeliac disease diagnosis. OBJECTIVES: To determine the sensitivity, the specificity, the positive and negative predictive values of an immunoenzymatic assay based on guinea pig tissue transglutaminase, to compare antititransglutaminase immunoenzymatic assay to the antiendomysium immunofluorescent assay, and to define a cost-effective sequence to execute serum antibody determination in coeliac patients. METHODS: We assessed for coeliac disease antibodies 91 pediatric patients with symptoms suggestive of coeliac disease, and 23 patients with coeliac disease on a gluten-free diet as controls. RESULTS: Antitransglutaminase immunoglobulins A showed 93.1% sensitivity, 93.6% specificity, 87.1% positive and 96.7% negative predictive values. Antitransglutaminase immunoglobulins A were significantly higher in antiendomysium positive subjects. Correlation between antitransglutaminase immunoglobulins A and antigliadin immunoglobulins A was not significant. DISCUSSION: Our results show that antitransglutaminase immunoenzymatic assay represents a cost-effective strategy for patients' serological evaluation and it could substitute EMA determination, which could be considered a second level evaluation.
Assuntos
Algoritmos , Doença Celíaca/diagnóstico , Transglutaminases/imunologia , Adolescente , Fatores Etários , Anticorpos/análise , Autoanticorpos/análise , Doença Celíaca/imunologia , Distribuição de Qui-Quadrado , Criança , Pré-Escolar , Interpretação Estatística de Dados , Ensaio de Imunoadsorção Enzimática , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Imunoglobulina A/análise , Imunoglobulina G/análise , Lactente , Masculino , Sensibilidade e EspecificidadeRESUMO
The project "Precursors of Atherosclerosis (ATS) in Children" following the protocol of WHO started in Modena in 1981/82. A total of 527 schoolchildren of 6, 9, 12 and 15, living in the area around Modena, Emilia Romagna Region, were examined, sampled from whole schools or classes. Each group exceeds the minimum of 100 children with the exception of the group including subjects of 15. The population sampled is fairly homogeneous. There are no immigrants to the area and the intra-population differences due to social-classes are minimal. The standard of life is very high: the rate of income is one of the highest in Italy and the incidence of ATS manifestations is very high. The children will be reexamined each year for three years at the same season and the study will be over in 1984. A questionnaire with family story has been filled by the parents and the parents were directly examined about: height, blood pressure, cholesterolemia, triglyceridemia, smoking habit. Following the protocol in each child are determined: height, weight, Quetelet's index (kg/m2), skinfold thickness, blood pressure, cholesterolemia, triglyceridemia, sexual maturity and smoking habit. The statistical elaboration methods of data are chosen in cooperation with the "Centro di Calcolo" of the University of Modena, and elaborated with the Statistical Package for the Social Science (SPSS) method.
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Arteriosclerose/etiologia , Adolescente , Arteriosclerose/patologia , Pressão Sanguínea , Estatura , Peso Corporal , Criança , Colesterol/sangue , Feminino , Humanos , Itália , Masculino , Projetos Piloto , Risco , Maturidade Sexual , Dobras Cutâneas , Fumar , Triglicerídeos/sangueAssuntos
Infecções Bacterianas/diagnóstico , Intestino Delgado/microbiologia , Intestinos/transplante , Vísceras/transplante , Adulto , Antibacterianos/uso terapêutico , Bactérias Aeróbias/classificação , Bactérias Aeróbias/isolamento & purificação , Infecções Bacterianas/tratamento farmacológico , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Complicações Pós-Operatórias/diagnóstico , Complicações Pós-Operatórias/microbiologia , Estômago/transplanteRESUMO
Megalencephaly (MEG), or enlargement of the brain, can either represent a familial variant with normal cerebral structure, or a rare brain malformation associated with developmental delay and neurological problems. MEG has been split into two subtypes: anatomical and metabolic. The latter features a build-up inside the cells owing to metabolic causes. Anatomical MEG has been detected in many different conditions, including many overgrowth syndromes. In 2004 Mirzaa et al. reported five non-consanguineous patients with a new MCA/MR syndrome characterized by severe congenital MEG with polymicrogyria (PMG), postaxial polydactyly (POLY) and hydrocephalus (HYD). The authors argued that these findings identified a new and distinct malformation syndrome, which they named MPPH. We report on a new case of MPPH, the first to be described after the original series (Mirzaa et al., 2004).
Assuntos
Encéfalo/anormalidades , Deficiências do Desenvolvimento/complicações , Hidrocefalia/complicações , Malformações do Desenvolvimento Cortical/complicações , Polidactilia/complicações , Feminino , Humanos , Hidrocefalia/patologia , Lactente , Imageamento por Ressonância Magnética/métodos , Malformações do Desenvolvimento Cortical/patologia , Polidactilia/patologia , Tomografia Computadorizada por Raios X/métodosRESUMO
The regulation of different maturational processes in the liver is believed to be influenced by the hormonal system. The aim of this study was to investigate the effect of two hormones, glucagon and dexamethasone, on levels of plasma membrane proteins in rat liver cells during late fetal and early postnatal stages of development. For this purpose, 18-day-old rat fetuses and 1-day-old newborns were treated with glucagon or dexamethasone and killed at 22 days of gestation and 3, 5 and 7 days of age, respectively. Postnuclei liver membranes were isolated using a sucrose gradient method and assessed for levels of specific membrane proteins. Asialoglycoprotein receptor and 110,000 Mr glycoprotein, denoted GP 110, representing the sinusoidal and bile canalicular domains, respectively, were quantitated using the immunoblot method. Membrane enzymes alkaline phosphatase, leucine aminopeptidase and gamma-glutamyl transferase were evaluated using enzymatic methods. The data showed that glucagon and dexamethasone have a differential effect on membrane constituents according to the stage of development. Glucagon increased the levels of membrane enzymes during the late fetal stage but had no effect on liver membrane proteins in the newborn animal. In contrast, although dexamethasone elevated GP 110 in fetal rat livers, none of the other marker proteins was significantly affected. On the other hand, in newborns dexamethasone reduced the amount of asialoglycoprotein receptor and alkaline phosphatase and leucine aminopeptidase enzyme activities but greatly augmented the level of gamma-glutamyl transferase. Thus, glucagon primarily affects plasma membrane proteins in late gestation while dexamethasone does so during the early postnatal period. The roles that these two hormones may play during ontogeny is discussed with respect to liver development.
Assuntos
Dexametasona/farmacologia , Glucagon/farmacologia , Fígado/metabolismo , Proteínas de Membrana/metabolismo , 5'-Nucleotidase , Fosfatase Alcalina/metabolismo , Animais , Animais Recém-Nascidos/metabolismo , Receptor de Asialoglicoproteína , Membrana Celular/enzimologia , Redutases do Citocromo/metabolismo , Complexo IV da Cadeia de Transporte de Elétrons/metabolismo , Feto/metabolismo , Leucil Aminopeptidase/metabolismo , Fígado/enzimologia , Fígado/ultraestrutura , Glicoproteínas de Membrana/metabolismo , Nucleotidases/metabolismo , Ratos , Receptores Imunológicos/metabolismo , gama-Glutamiltransferase/metabolismoRESUMO
Several recent studies have demonstrated the ability of techniques based on immunoadsorption to selectively isolate specialized subregions of membranes, termed domains, which are derived from a larger more complex parent membrane like the plasma membrane. The immunoadsorbent is directed against a specific antigen that resides exclusively or predominantly in the membrane domain to be isolated. Thus, a monospecific antibody to the domain-specific antigen is required. In the present study we developed a method employing a modified immunoblotting strategy which could utilize polyspecific antibodies to isolate membrane vesicles derived from a specific membrane domain of the hepatocyte plasma membrane. We also used specific cell surface labeling of the hepatocyte plasma membrane by lactoperoxidase-catalyzed iodination at 4 degrees C and preparation of different sized vesicles by sonication to facilitate isolation of the specific domain. For this study, polyspecific antisera were raised in goats against a membrane fraction, denoted N2u, which is enriched in bile canalicular proteins. This antiserum recognizes, among other antigens, a 110,000 Mr polypeptide previously shown to be localized in the bile canaliculus (J. Cook et al. (1983) J. Cell. Biol. 97, 1823-1833). A monospecific antiserum was raised in rabbits against the rat hepatocyte asialoglycoprotein receptor, a sinusoidal domain-specific set of glycoproteins whose major form has a Mr of 43,000. These antisera were each coupled indirectly to different pieces of nitrocellulose by the immunoblotting protocol and were used to isolate membrane vesicles from a crude extract of liver plasma membrane prepared by sonication. The ratio of iodinated asialoglycoprotein receptor to the 110,000 Mr polypeptide in vesicles isolated by the affinity nitrocellulose immunoadsorbent method indicate a 10- to 15-fold enrichment of sinusoidal-derived vesicles relative to bile canalicular-derived membrane vesicles. These results show that the affinity nitrocellulose immunoadsorbent method can be used to isolate domain-specific vesicles. Further, the affinity immunoadsorbent method described here for the isolation of domains of the plasma membrane is an integrative one allowing isolation of vesicles present in relatively small concentration in crude cell extracts and it requires minimal ultracentrifugation time.
Assuntos
Antígenos de Superfície/isolamento & purificação , Fracionamento Celular , Fígado/citologia , Marcadores de Afinidade , Animais , Receptor de Asialoglicoproteína , Extratos Celulares/análise , Membrana Celular/análise , Colódio , Técnicas de Imunoadsorção , Fígado/análise , Fígado/ultraestrutura , Ratos , Ratos Endogâmicos BUF , Receptores Imunológicos/isolamento & purificação , SonicaçãoRESUMO
Children with cystic fibrosis have variable degrees of exocrine pancreatic insufficiency which, if untreated, is the main cause of fat malabsorption. The impact of pancreatic enzyme supplementation on fat digestion was measured in 41 children with cystic fibrosis, 11 healthy controls, and five children with mucosal diseases by a non-invasive test of intraluminal lipolysis using 13carbon (13C) labelled mixed triglyceride (1,3-distearyl, 2[13C] octanoyl glycerol). The children with cystic fibrosis without pancreatic supplements had a median (range) 13C cumulative percentage dose recovered over six hours (cPDR) of 3.1% (0-31.7), the controls 31.0% (21.8-41.1), and the subjects with mucosal disease 27.8% (19.7-32.5). In 23 subjects with cystic fibrosis the usual dose of pancreatic enzyme supplements increased the cPDR to a median of 23.9% (0-45.6), and twice the usual dose of enteric coated microspheres increased the cPDR to 31.1% (11.1-47.8). There was no significant difference between the median cPDR of normal controls and children with mucosal disease, but there was a highly significant difference between these groups and children with untreated cystic fibrosis. Thirteen children with cystic fibrosis had no 13C recovery in their breath without enzymes and 10 showed marked increases with regular enzymes. In eight children doubling the dose of enzymes caused no or minimal improvement. The mixed triglyceride breath test offers a simple, non-invasive way of assessing the need for pancreatic enzyme supplementation in children with cystic fibrosis and could be used to optimise treatment.
Assuntos
Fibrose Cística/tratamento farmacológico , Lipase/uso terapêutico , Extratos Pancreáticos/uso terapêutico , Testes Respiratórios , Isótopos de Carbono , Criança , Fibrose Cística/diagnóstico , Feminino , Humanos , Masculino , Pancrelipase , Resultado do Tratamento , TriglicerídeosRESUMO
Stable isotope breath tests offer a safe, repeatable non-invasive method of measuring fat digestion. They involve the ingestion of a substrate labelled with 13C followed by serial measurements of the 13C:12C ratio in expired CO2, from which the percentage of the 13C dose recovered (PDR) can be calculated. However PDR depends on the CO2-production rate. Our aim was to compare results obtained using directly measured CO2-production rates with those calculated from two predicted values. Twelve normal healthy children and twenty-four children with cystic fibrosis (CF) (without or with the normal dose of enzyme supplementation) were studied with 1,3-distearyl, 2[carboxyl-13C] octanoyl glycerol. The volume of CO2 produced (litres/min) was measured at rest for 30 min approximately 3 h after substrate ingestion, and the results were converted to mmol/min. For each subject the expected BMR was calculated from the equation of Schofield (1985), based on sex, age, weight and height, and from these values, CO2-production rate was derived. Surface area was calculated and an estimated value of 5 mmol/m2 per min (Shreeve et al. 1970) was used. Using these three CO2-production rates, three different PDR were calculated and compared. In healthy children there was a close concordance between measured and predicted CO2-production rates, but children with CF had a mean measured CO2-production rate 39% higher than normal children. This use of normal data for predicted CO2-production rates in children with CF underestimates cumulative PDR. If direct measurements of CO2-production rate are not available or impossible to perform the VCO2 obtained from the BMR calculated using the equations of Schofield (1985) or Shreeve et al. (1970) can be used in normal children. However, if accurate results for PDR are to be obtained, CO2-production rates should be measured when performing breath tests in conditions where energy expenditure and/or CO2-production rate are not expected to be normal.