Leche materna deslactosada con la enzima beta galactosidasa para lactantes intolerantes / Leite materno sem lactose com a enzima beta-galactosidase para lactentes intolerantes / Lactose-free breast milk with the enzyme beta galactosidase for intolerant infants

Introducción: los bebés deben beneficiarse de la leche materna, incluso cuando presentan intolerancia a la lactosa. Por esto, se debe recurrir a la obtención de leche materna deslactosada. Objetivo: analizar el efecto de la enzima beta galactosidasa en la hidrólisis de la lactosa de leche materna madura para bebés clínicamente diagnosticados con intolerancia a la lactosa. Materiales y método: estudio exploratorio, descriptivo y explicativo. El contenido de lactosa se cuantificó desde el inicio hasta el final del tratamiento, controlando temperatura, tiempos y cantidad de enzima ß-galactosidasa adicionada en la leche materna. Se recolectaron 1000 ml de leche materna, obtenidos del Banco de Leche del Hospital General de Medellín (Antioquia, Colombia). Resultados: las muestras donadas se encontraban pasteurizadas y posteriormente fueron sometidas a la acción de la enzima lactasa. Se cuantificó el contenido de lactosa sin la enzima, reportando en promedio 6,34 mg/100 ml ± 0,23. El mayor aporte de lactosa obtenido posterior a la exposición a la enzima (30 minutos) fue de 6,07 mg/ml ± 0,35 (correspondiente a 95 % del contenido inicial), finalizando con un aporte de 0,35 % a una concentración de 0,4 % tras 24 horas, porcentaje que representa 95 % de la hidrólisis total en la leche materna. Conclusiones: en todas las muestras analizadas de diferentes madres se pudo obtener leche materna con bajas concentraciones de lactosa tras 24 horas de haber sido sometidas a la acción de ß-galactosidasa. Lo anterior se establece como una alternativa para los bebés intolerantes a la lactosa, que permitiría no privarlos de todos los beneficios que ofrece este alimento.

Introduction: Babies should benefit from breast milk, even when they are lactose intolerant. For this reason, parents should resort to obtaining lactose-free breast milk. Objective: To examine the effect of the enzyme ß-galactosidase on the hydrolysis of lactose in mature breast milk for babies clinically diagnosed with lactose intolerance. Materials and method: Exploratory, descriptive, and explanatory study. The lactose content was quantified from the beginning to the end of the treatment, controlling variables such as temperature, times, and the amount of ß-galactosidase enzyme added in breast milk. A total of 1000 ml of breast milk were obtained from the milk bank at Hospital General de Medellín (Antioquia, Colombia). Results: Donated samples were first pasteurized and subsequently subjected to the action of the enzyme lactase. The lactose content without the enzyme was quantified, reporting an average of 6.34 mg/100 mL±0.23. The highest contribution of lactose obtained after exposure to the enzyme was 6.07 mg/mL±0.35 (corresponding to 95% of the initial content), at 30 minutes, ending with a contribution of 0.35% at a concentration of 0.4% in 24 hours, percentage that represents 95% of total hydrolysis in breast milk. Conclusions: In all the examined samples from different mothers, it was possible to obtain breast milk with low concentrations of lactose 24 hours after these were exposed to the action of ß-galactosidase. This becomes an alternative for feeding lactose intolerant babies and not deprive them from all the benefits offered by breast milk.

Introdução: os bebês se devem beneficiar do leite materno, mesmo quando tenham intolerância à lactose, razão pela qual se deve recorrer à obtenção de leite materno sem lactose. Objetivo: analisar o efeito da enzima beta-galactosidase na hidrólise da lactose no leite materno maduro para bebês diagnosticados clinicamente com intolerância à lactose. Materiais e método: estudo exploratório, descritivo, explicativo. O teor de lactose foi quantificado do início ao fim do tratamento; temperatura, tempos e quantidade de enzima beta-galactosidase adicionada no leite materno foram controlados; foram coletados 1000ml de leite materno, obtidos no Banco de Leite do Hospital General de Medellín (Antioquia, Colômbia). VResultados: as amostras doadas foram pasteurizadas e posteriormente submetidas à ação da enzima lactase. O teor de lactose sem a enzima foi quantificado, relatando uma média de 6,34mg/100ml±0,23. A maior contribuição de lactose obtida após a exposição à enzima foi de 6,07mg/ml±0,35 (correspondendo a 95% do conteúdo inicial) em 30 minutos, finalizando com uma contribuição de 0,35% na concentração de 0,4% em 24 horas, percentual que representa 95% da hidrólise total no leite materno. Conclusões: em todas as amostras analisadas de diferentes mães, foi possível obter leite materno com baixas concentrações de lactose 24 horas após ser submetido à ação da beta galactosidase, como alternativa para bebês intolerantes à lactose e não os privar de todos os outros benefícios oferecidos por esse alimento ideal.

Enfermedad de Pompe: reporte de caso / Disease Pompe: report of case

Objetivo. Se describe el caso de un paciente masculino de siete meses de edad, evaluado por cardiología a los quince días de vida por antecedente de muerte súbita cardíaca de hermana a los cuatro meses, no se sospechó nada pese a consanguinidad de padres y diagnóstico temprano del paciente de cardiomiopatía. Evoluciona con un cuadro clínico de infecciones respiratorias a repetición desde los tres meses (bronquiolitis recurrente), falla de medro y cuadro de neumonía reciente y fallece a los ocho días de la consulta con la genetista. Métodos. El abordaje inicial fue la realización de un ecocardiograma a los quince días de nacido por antecedente de muerte súbita de hermana a los cuatro meses por cardiomegalia y consanguinidad de los padres. Continúa con deterioro clínico a través de los meses por lo que se remite a genética, se toman pruebas enzimáticas en gota de sangre seca, el paciente fallece antes de recibir el diagnóstico de Enfermedad de Pompe. Resultados. Se enfocó al paciente con un posible diagnóstico de Enfermedad de Pompe solicitándose enzima lisosomal alfa-glucosidasa (GAA) en muestra de gota de sangre seca y reporte final de la secuenciación genética.

Objective. It is a case of a seven months male patient, evaluated by cardiology fifteen days after he was born secondary, to sudden cardiac death of his sister of four months, nothing suspicious despite consanguinity of parents and early diagnosis of the patient with Cardiomyopathy. Evolved with the following clinical conditions recurrent respiratory infections from three months (recurrent bronchiolitis), widespread malnutrition and recent pneumonia. Died eight days after the consultation with the geneticist. Methods. The initial approach was to perform echocardiogram at fifteen days old, because of history of the sudden death of his sister of four months, cardiomegaly and parental consanguinity. Clinical deterioration continues through months so he is referred to genetic, enzymatic tests, taken in dried blood, the patient dies before receiving the diagnosis of Pompe disease. Results. The patient was focused with a possible diagnosis ofPompe Disease so lysosomal enzyme α-glucosidase (GAA) sample was requested in dried blood and genetic sequencing final report to define diagnosis.

Frequency of the LCT*-13910C>T polymorphism associated with lactase persistence diverges among Euro-descendant groups from Brazil

Objective: The aim of this study was to investigate the frequency of the LCT*-13910C>T polymorphism associated with a high expression of lactase in the small intestine during adulthood, and to infer the lactase persistence and adult-type hypolactasia phenotypes among Euro-Brazilians and Mennonites from South Brazil

Materials and Methods: A sequence-specific PCR method to genotype the LCT*-13910C>T polymorphism in 292 Euro-Brazilians and 151 Mennonites [a group with European ancestry and a long history of endogamy] was developed. Using an exact test of population differentiation, the genotype and allele frequency between these and other Brazilian populations were compared

Results: The frequency of -13910*T was significantly higher among the Mennonites when compared to the Euro-Brazilian cohort [0.63 vs. 0.33, p < 0.000001]. Accordingly, Mennonites had a higher prevalence of the lactase persistence genotype [88.1 vs. 55.5%, p < 0.000001]. The distribution of -13910*T differed between Mennonites and all other Brazilian groups [p < 0.0001]. The Euro-Brazilians from Curitiba displayed differences when compared to all other Brazilian groups [p < 0.0001], even to Euro-Brazilians from a different geographic region [p = 0.0003], but were similar to those from Porto Alegre [p = 0.2]

Conclusion: Differences in the -13910*T-associated lactase persistence distribution among Euro-Brazilian groups reflect the ancestry and admixture of each particular group and should be considered for adult-type hypolactasia screening

Correlation between lactose absorption and the C/T-13910 and G/A-22018 mutations of the lactase-phlorizin hydrolase (LCT) gene in adult-type hypolactasia

The C/T-13910 mutation is the major factor responsible for the persistence of the lactase-phlorizin hydrolase (LCT) gene expression. Mutation G/A-22018 appears to be only in co-segregation with C/T-13910. The objective of the present study was to assess the presence of these two mutations in Brazilian individuals with and without lactose malabsorption diagnosed by the hydrogen breath test (HBT). Ten milk-tolerant and 10 milk-intolerant individuals underwent the HBT after oral ingestion of 50 g lactose (equivalent to 1 L of milk). Analyses for C/T-13910 and G/A-22018 mutations were performed using a PCR-based method. Primers were designed for this study based on the GenBank sequence. The CT/GA, CT/AA, and TT/AA genotypes (lactase persistence) were found in 10 individuals with negative HBT. The CC/GG genotype (lactase non-persistence) was found in 10 individuals, 9 of them with positive HBT results. There was a significant agreement between the presence of mutations in the LCT gene promoter and HBT results (kappa = -0.9, P < 0.001). The CT/AA genotype has not been described previously and seems to be related to lactase persistence. The present study showed a significant agreement between the occurrence of mutations G/A-22018 and C/T-13910 and lactose absorption in Brazilian subjects, suggesting that the molecular test used here could be proposed for the laboratory diagnosis of adult-type primary hypolactasia.

Hypolactasia as a molecular basis of lactose intolerance.

Lactase-phlorizin hydrolase (LPH), a membrane-bound glycoprotein present in the luminal surface of enterocytes in the intestine is responsible for lactose intolerance, a phenomenon prevalent in humans worldwide. In the rodent intestine, the post-natal development of the LPH follows a specific pattern, such that the enzyme levels are high in the peri-natal period, but declines considerably upon maturation. The observed maturational decline in the LPH activity is very similar to adult-type hypolactasia observed in humans. Majority of the studies have been carried out using animal models or cell lines and a number of hypotheses have been put forward to explain the maturational decline of lactase activity such as: (a) decreased amount of lactase protein, (b) defect in post-translational modification of precursor lactase to the mature enzyme, and (c) synthesis of an inactive, high molecular weight lactase with altered glycosylation, however, the precise underlying mechanism of adult-type hypolactasia remains undefined. The present review describes the recent developments in understanding the regulation of lactase expression and the possible mechanism of adult-type hypolactasia, as a cause of lactose intolerance.

Molecular cloning of a novel cDNA from Mus muscular BALB/c mice encoding glycosyl hydrolase family 1: a homolog of human lactase-phlorizin hydrolase / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To study the mechanism of lactose intolerance (LI) by cloning the mouse lactase cDNA and recombining a vector. METHODS Total murine RNA was isolated from the small intestine of a 4-week-old BALB/c mouse (d). Gene-specific primers were designed and synthesized according to the cDNA sequences of lactase-phlorizin hydrolase (LPH) in human, rat, and rabbit. A coding sequence (CDS) fragment was obtained using RT-PCR, and inserted into a clone vector pNEB-193, then the cDNA was sequenced and analyzed using bioinformatics.</p><p><b>RESULTS</b>The cDNA from the BALB/c mouse with 912 bp encoding 303 amino acid residues. Analysis of the deduced amino acid sequence using bioinformatics revealed that this cDNA shared extensive sequence homology with human LPH containing a conserved glycosyl hydrolase family 1 motif important for regulating lactase intolerance.</p><p><b>CONCLUSION</b>BALB/c mouse LPH cDNA (GenBank accession No: AY751548) provides a necessary foundation for study of the biological function and regulatory mechanism of the lactose intolerance in mice.</p>

Lactase mRNA Expression in Small Intestines of Korean Fetuses and Adults / 대한내과학회지

OBJECTIVE: The specific activity of lactase-phlorizin hydrolase (LPH) is very high at birth and sharply declines after weaning, producing lactose intolerance. The prevalence of lactose intolerance is up to 85% in Korean adults. Molecular basis of the regulatory mechanisms responsible for the decline of LPH specific activity is still unknown. In order to elucidate the molecular mechanisms regulating the LPH expression during development, LPH specific activity and mI4NA level of Korean fetal and adult intestines were compared. METHODS: 20 fetal small intestines (16-27 weeks) were obtained during therapeutic abortion and were divided into 3 equal length. 20 adult jejunal tissues were obtained from patients without small intestinal disease during laparotomy. Mucosal homogenates were prepared for dissacharidases specific activities measurement and total RNA was extracted for northern and slot hvbridization. LPH mRNA level was measured by laser densitometer. RESULTS: LPH specific activities of proximal, middle and distal portion of fetal intestines (n=20) were 36.2 +/- 22.5, 38.6 +/- 23.2 and 23.2 +/- 19.9 mu/mg protein, respectively. LPH specific activity of adult jejunum (n=8) was 5.9 +/- 1.8 mu/mg protein and significantly (p<0.05) lower than those of fetal intestines. However, there was no significant difference in sucrase and trehalase specific activities between fetal intestines and adult jejunum. Although LPH specific activity of adult jejunum was lower than those of fetal intestines, LPH mBNA level of adult jejunum was as high as those of fetal intestines. CONCLUSION: These results show that LPH specific activity and mRNA level do not parallel, indicating the posttranscriptional control of fetal development of LPH expression.

Terapia de reemplazo enzimático en la enfermedad de Gaucher Tipo 1 / Enzyme replacement therapy in type 1 Gaucher's disease

La enfermedad de Gaucher es una deficiencia de glucocerebrosidasa (GC) con acumulación de glucocerebrósidos en el sistema macrofágico, y da lugar a tres formas clínicas diferentes. De ellas, el tipo 1 se caracteriza por infiltración del bazo, hígado y médula ósea e incluye además alteraciones esqueléticas, evidenciadas por dolor óseo, osteopenia y fracturas. Se trataron dos niñas con GC disponible comercialmente, usando dosis bimensuales entre 30 y 60 U/Kg. En ambas pudo observarse tendencia a la normalización de la hemoglobina y fosfatasa ácida sérica, y en la más afectada menor hepatomegalia y desaparición de los síntomas esquelético. Un episódio de anafilaxia observado en esta misma enferma cedió rápidamente, y pudo continuar el tratamiento sin que ocurriera disminución de la respuesta. Los resultados coincidentes con la experiencia mundial, confirman que el tratamiento con GC es racional y efectivo en la enfermedad de Gaucher tipo I. La tolerancia es buena y no se describen fenómenos de resistencia. La modalidad de respuesta es variable, pero todos los enfermos responden y al cabo de un tiempo se puede disminuir lentamente la dosis. Aun así, el precio de la enzima impone una precisa definición de las indicaciones en función de una óptima relación conto-beneficio

Doença de Gaucher: Relato de caso e breve revisäo da literatura / Gaucher's disease: a case report and a brief review of the literature

E descrito um caso de doença de Gaucher do adulto e apresentada revisäo sobre aspectos fisiopatológicos, quadro clínico, diagnóstico e terapêutica