Human milk components cross-reacting with antibodies against bovine β-lactoglobulin

The human whey components cross-reacting with antibodies raised against bovine and/or equine beta-lactoglobulin were screened systematically. The milk of six women on a normal diet was collected within 72 h of confinement and whey components were fractionated by high-speed size exclusion chromatography and reversed-phase techniques. The fractions which were immunoreactive in double diffusion experiments with antisera anti-bovine and/or equine beta-lactoglobulin were subsequently purified by native PAGE and then electroblotted on Pro-blott membrane (Western blotting). Pro-blot membranes were stained in parallel with Coomassie and by immunostaining using antibodies against bovine and/or equine beta-lactoglobulin as first antibody solution. The immunoreactive bands were cut out from the membrane and N-terminally sequenced; all the immunoreactive components were clearly identified as human beta-casein or its (mainly tryptic) fragments. The strong antigenic similarity between human beta-casein and beta-lactoglobulin (bovine and equine) might be of immunological importance; it could mean that breast-fed neonates risk being sensitized to beta-lactoglobulin irrespective of the presence of cow's milk in the mother's diet.     

Human milk proteins may interfere in ELISA measurements of bovine β-lactoglobulin in human milk

It is widely believed that cow's milk proteins ingested by the mother, in particular β-lactoglobulin ([β-LG), can pass into breast milk and thus sensitize predisposed infants. However, studies to evaluate bovine β-LG in human milk have given conflicting results. The aim of this study was to analyse the correlation between the amount of cow's milk in the mother's diet and the presence of bovine β-LG in breast milk. Human milk samples from 14 healthy non-atopic women on diets with different cow's milk contents were examined. The total concentration of bovine β-LG or β-LG immuno-like proteins (β-LGIP) was determined by enzyme-linked immunosorbent assay (ELISA). Two separation procedures utilizing ELISA plates and an affinity chromatography column were set up to identify the human whey components recognized by the anti-β-LG antibodies. β-LGIP reactivities of milk from three groups on different diets were not significantly different. After splitting the antigen antibody complexes, three main protein components, human lactoferrin, human β-casein and human α-lactalbumin, were identified. This study would suggest that, at least in healthy subjects, false-positive results in ELISA determinations of bovine β-LG in human milk might be due to cross-reactions between polyclonal antibodies and different protein antigens.     

Breast milk jaundice; the role of lipoprotein lipase and the free fatty acids

Lipoprotein lipase activity and free fatty acid concentrations were measured in samples of milk collected from mothers of infants without and with prolonged neonatal jaundice. The lipoprotein lipase and free fatty acid values in the milk from mothers of infants without jaundice were found to increase with the duration of breast-feeding until the 12th postpartum day, and then to fall to the original levels. In the group of mothers with jaundiced infants both lipoprotein lipase and free fatty acid values were found within normal limits when measured between 15th and 37th days post-partum.These findings indicate that increased values of lipoprotein lipase and free fatty acids in the milk are not responsible for the development of breast-milk jaundice.     

新生儿母乳性黄疸的临床研究

目的　了解母乳性黄疸 (BMJ)患儿胆红素均值及峰值水平 ,探讨BMJ患儿胆红素增高对肝脏及心肌酶的影响 ,寻求最佳干预方案。方法　以 160例确诊为BMJ患儿为研究对象 ,观察黄疸出现及消退时间 ,并行经皮胆红素、血清胆红素测定及心肌酶谱、肝功能等检查 ,并将胆红素 >2 91μmol/L 80例患儿随机分成 4组治疗 ,并评价其疗效。结果　 1.BMJ胆红素高峰期在生后 2～ 4周 ,血清胆红素峰值和均值分别为 3 16μmol/L和 2 3 1μmol/L。黄疸消退时间为 6～ 12周 ;2 .经皮测胆红素与血清总胆红素呈正相关　r =0 .92 ;3 .BMJ患儿心肌酶、肝功能、总蛋白、清蛋白、球蛋白、碱性磷酸酶均在正常范围 ;4.继续母乳组、停止母乳组、光疗暂停母乳组、光疗继续母乳组患儿黄疸消退时间以光疗暂停母乳组与光疗继续母乳组患儿黄疸消退时间快 ,但两组间无显著性差异 (P >0 .0 5)。结论　 1.经皮测胆仪不仅可作为筛查、随访工具 ,且其结果可代替血清总胆红素作为指导治疗的依据 ;2 .BMJ患儿胆红素对肝功能、心肌酶无影响 ;3 .各组中以光疗不停母乳组黄疸消退时间最快 ,疗效最显著 ,故光疗时不必停母乳     

Dietary Bovine β Mactoglobulin is Transferred to Human Milk

ABSTRACT. Human milk from 38 mothers was analysed by radioimmunological method for content of bovine (5-Iactoglobulin. Detectable amounts (5-33 μg/1) of immunoactive β-lactoglobulin were found in 18 human milk samples. Milk from 3 mothers, whose infants suffered from infantile colic contained high amounts of fi-lactoglobulin (32, 18 and 14 μg/1 respectively). With the mothers on a cow's milk free diet the contents fell to non-detectable amounts in two mothers and to 6 μg/1 in the third. All three infants became free from colic,     

Successful pregnancy in β-thalassaemia major

ABSTRACT. Successful pregnancy is described in a patient with β-thalassaemia major, transfusion-dependent from four months of age and treated with desferrioxamine from 13 years of age. The pregnancy was concealed. No antenatal care was given nor any planned alteration in management of her thalassaemic state.     

Breast feeding practices and severe hyperbilirubinaemia

This study establishes the association between early onset severe hyperbilirubinaemia (serum bilirubin (SBR) level greater than or equal to 272 mumol/L) with no assigned cause and breast feeding. The rates of breast feeding at hospital discharge increased from 46.4 to 75.4% in non-insured (public) women and 58.6 to 89.2% in insured (private) women between 1975 and 1987, and were accompanied by an increase in severe hyperbilirubinaemia from 1.6 to 3.1% in public and 0.9 to 3.6% in private babies. The case control study involved 125 term breast feeding infants born between 1 July 1985 and 1 July 1986 with severe hyperbilirubinaemia (SBR level 272 mumol/L) with no assigned cause who were compared with 125 matched controls who had peak SBR levels less than or equal to 272 mumol/L. Severe hyperbilirubinaemia was associated with primiparous and non-Caucasian mothers, non-smoking and oxytocin usage. Univariate analysis of feeding practice variables revealed that less frequent breast feeds, greater weight loss and less frequent stools over the first 3 days related to severe hyperbilirubinaemia (P less than 0.05). Multivariate analysis of the eight significant univariate factors revealed that maternal non-smoking, less frequent breast feeding, less frequent stooling and excessive infant weight loss were the best predictors of severe hyperbilirubinaemia. With the wide promotion of breast feeding, the contribution of individual feeding practices to severe hyperbilirubinaemia demands ongoing analysis and review.     

Cytokines in human milk and late‐onset breast milk jaundice

Background: Maternal milk plays an important role in the development of late‐onset breast milk jaundice (BMJ), possibly due to the unique characteristics of breast milk. The aim of this study was to investigate whether there is a relation between cytokine concentrations in the milk of nursing mothers and BMJ. Methods: Breast milk samples were collected from breast‐feeding mothers of healthy full‐term neonates, 40 with BMJ and 40 without jaundice. Milk samples were taken between the second and the fourth postpartum week. The concentrations of interleukin (IL)‐1 β, IL‐6, IL‐8, IL‐10, and tumor necrosis factor‐α were measured by flow cytometric bead array. Results: There were significant differences between the study groups in terms of IL‐1 β concentrations (P= 0.013). Not statistically significant but similar trends were also seen for IL‐10 (P= 0.067) and tumor necrosis factor‐α (P= 0.053) concentrations. However, no significant differences were noted in IL‐6 (P= 0.174) and IL‐8 (P= 0.285) concentrations. Conclusions: IL‐1 β concentration seems to be increased in milk of mothers whose infants had BMJ. Although the effect of these cytokines on BMJ is unknown, it may cause prolonged jaundice via hepatic uptake, hepatic excretion, conjugation and intestinal absorption.     

BCG vaccine modulates intestinal and systemic response to β-lactoglobulin

Beta-Lactoglobulin (BLG) is a clinically important antigen in cow's milk and one of the major allergens causing cow's milk allergy. Bacillus Calmette-Guerin (BCG) vaccination has been suggested to modify immune response possibly decreasing the risk of allergy to some antigens in both human and experimental animals. In the present study, we have analyzed whether the early BCG vaccination has any effect on the markers of systemic and gastrointestinal (GI) sensitization to BLG. We immunized two groups of Hooded-Lister rat puppets with intraperitoneal injections of native BLG at 43 and 62 days with pertussis vaccine as adjuvant, one group receiving additionally BCG. The animals were then fed native and denatured milk products twice weekly from 73 to 131 days of age, when they were killed. Control group was not vaccinated and received normal rat forage. Total immunoglobulin E (IgE) levels and BLG-specific IgG(1) and IgG(2a) concentrations were determined in serum samples. Spontaneous interleukin (IL)-4 and interferon (IFN)-gamma production from duodenal specimens were measured, and the inflammatory cells were quantitated in specimens from different sections of the GI tract. Administration of BCG simultaneously with BLG resulted in reduced IgE concentration in serum, while the specific IgG(1) and IgG(2a) antibody responses and the spontaneous secretion of IL-4 and IFN-gamma were not affected. Furthermore, BCG-induced eosinophilic infiltration and increase of intraepithelial lymphocytes (IEL) in the GI mucosa, and a trend toward increased number of lamina propria mononuclear inflammatory cells in the colon (BCG compared with BLG, p = 0.09; BCG compared with controls, p = 0.02). Controls showed increment of IgG(1) response in comparison with the BLG group (p = 0.04) and increase of mucosal eosinophilic infiltration. The BCG modified the response to BLG both at the systemic level as shown by decrease of total IgE and at GI mucosa where increase of eosinophilic infiltration and increased number of IEL were seen. Increment of IgG(1) level and eosinophils in the controls might be related with the lack of modulatory effect of pertussis vaccination. A shift of response toward the lower GI tract after BCG immunization as shown by a trend for increase of mononuclear inflammatory cells in colon lamina propria mimics disease development in some cases of clinical food allergy, and emphasizes the need for evaluation of the changes in the whole GI tract in food allergy models.     

Coexisting Hereditary Methaemoglobinaemia and Heterozygous β-Tialassaemia

ABSTRACT. Cyanosis was noted within a few weeks after birth in two sisters. On investigation the cause of cyanosis was found to be congenital methaemoglobinaemia due to NADH diaphorase deficiency. Heterozygous β-thaiassaemia was present as an additional incidental finding in one of the sisters, but did not contribute to the symptoms, thus showing that the two diseases, when coexistent, do not pose additional haematological problems. However, it is possible that the β-thalassaemia counteracts the tendency to compensatory erythrocytosis induced by methaemoglobinaemia     

Values for urinary β-microglobulin and N-acetyl-β-D-glucosaminidase in normal healthy infants

Abstract Background : Measuring urinary β₂ microglobin (B2M) and N -acetyl-β-D-glucosaminidase (NAG) excretion is widely used as a valuable clinical tool in assessing renal tubular lesions. However, few data are available on normal values for urinary excretion of B2M and NAG in infancy.
Methods : Urinary B2M and NAG were measured in healthy infants. The logarithmic values of urinary B2M, NAG. B2M/creatinine ratio and NAG/creatinine ratio were distributed almost normally and reference ranges were calculated from the logarithms of the observed values.
Results : The levels of urinary B2M and B2M/creatinine ratio were highest in the 1-month-old group, followed by a decrease during the first 3 months. Urinary B2M excretions in the 3-monfh-old group showed rather lower levels than those of the 12-month-old and 36-month-old groups. Although urinary NAG excretions were almost constant throughout all groups, urinary NAG/creatinine ratio decreased gradually until 3 years of age.
Conclusions : We suggest that these reference ranges are of importance in evaluating tubular damage due to a variety of renal diseases in infancy.     

Breast mild jaundice: Natural history,familial incidence and late neurodevelopmental outcome of the infant

Jaundice associated with breast feeding is a frequent problem facing the paediatrician. Despite numerous reports on this subject, the natural history, familial occurrence and late neurodevelopment of children with breast milk jaundice remain unclear. The follow up of 60 infants with breast milk jaundice showed that there are two bilirubin peaks, on the 4th and 5th day and on the 14th–15th day of life. In the infants with uninterrupted breast feeding, the hyperbilirubinaemia disappeared slowly and could still be detected 12 weeks after birth. The familial incidence was 13.9%, indicating that in some cases a unique genetic factor is expressed. Late neurodevelopment or hearing defects were not observed, thus enabling the paediatrician to encourage continuation of breast feeding in most cases of healthy infants with breast milk jaundice.     

αβ and γδ T cell subsets in chronic renal failure in children on dialysis treatment

BACKGROUND: Impaired immunity, particularly cell-mediated, is one of the features of chronic renal failure. This also concerns impaired T cell dependent responsiveness. METHODS: The expression of T cell surface antigens (CD3, CD25, TCRalphabeta, TCRgammadelta) was evaluated on peripheral blood (PB) mononuclear cells using two-color flow cytometry in 10 children on continuous ambulatory peritoneal dialysis (CAPD) and in 13 children on maintenance hemodialysis (HD) with polysulfone and cuprophane dialysers. RESULTS: In HD children absolute numbers of leukocytes, lymphocytes, CD3+, alphabeta, gammadelta T cells and a percentage of gammadelta T cells were decreased versus healthy children. Also, we observed a relative increase of CD3+, CD3+/CD25+ and alphabeta T cells after sessions with cuprophane membranes, and an increase of CD3+/CD25+, alphabeta T cell percentages after sessions with the polysulfone membranes. Additionally we found a decrease of both relative and absolute numbers of gammadelta T cells after HD with polysulfone. In CAPD children we found declined absolute numbers of total lymphocytes, CD3+ and alphabeta T cells and higher relative values of CD3+ and alphabeta T cells versus controls. CONCLUSIONS: The T cell depletion in chronic renal failure (CRF) patients primarily results from uremic-related toxicities, rather than from CAPD or HD-related incompatibilities. We showed a significant decrease of gammadelta T cells in CRF patients on HD, that may be partly responsible for impaired T-dependent responsiveness in that group. The intradialytic changes of gammadelta Tcells may result from a different degree of biocompatibility during the application of various dialysis membranes.     

Serum bile acids and their conjugates in breast-fed infants with prolonged jaundice

Serum bile acids and their conjugates were analysed in 20 breast-fed infants with prolonged jaundice. The mean total bile acid levels in serum were increased in the breast-fed infants with jaundice, as compared with those in either breastor bottle-fed infants without jaundice. However, there were no significant differences between the groups. All the breast-fed infants examined, regardless of association with jaundice, had a bile acid pattern dominated by taurine conjugates (the ratio of glycine- to taurine-conjugated bile acid, G/T ratio, less than 1.00). In contrast, the bottle-fed infants without jaundice had a pattern dominated by glycine conjugates (G/T ratio, more than 1.00). Among the breast-fed infants with jaundice, the mean G/T ratio in those who had serum bilirubin levels over 10 mg/100 ml was significantly lower than that in those who had serum bilirubin levels of less than 10 mg/100 ml. The altered bile acid metabolism might be associated with the pathology of breast milk jaundice.Abbreviation LP-X lipoprotein-X     

PLASMA AND URINARY β-HEXOSAMINIDASE IN JUVENILE DIABETES MELLITUS

ABSTRACT. The activity of β-hexosaminidase (E C 3.2.1.30) in plasma and urine was determined in 99 patients with juvenile diabetes mellitus and in 40 age-matched controls. Plasma enzyme activities were above normal in plasma from diabetics and showed significant correlations with blood glucose and glycosylated HbA₁ (GHbA₁) but not with the presence of retinopathy. The urinary excretion of β-hexosaminidase, which is a sensitive indicator of renal injury, was also excessive in the diabetics and showed a significant correlation with blood GHbA₁. The results show that plasma and urinary β-hexosaminidase correlate with parameters of diabetic control and that urinary β-hexosaminidase may be more sensitive than urinary albumin as an indicator of early renal damage.     

Low breast milk TGF-β2 is induced by Lactobacillus reuteri supplementation and associates with reduced risk of sensitization during infancy

The immunological composition of breast milk differs between mothers. The reasons for these differences and the consequences for the breast-fed infants are poorly understood. The aim of this study was to evaluate the effect of probiotic Lactobacillus reuteri supplementation on the immunological composition of breast milk in relation to sensitization and eczema in the babies. Total IgA, secretory IgA (SIgA), TGF-β1, TGF-β2, IL-10, TNF, soluble CD14 (sCD14), and Na/K ratios were analyzed in colostrum and mature milk obtained from women treated with L. reuteri (n = 54) or placebo (n = 55) from gestational week 36 until delivery. Bacteriological analyses of L. reuteri were performed in faecal samples of the mothers. The infants were followed prospectively for 2 yr regarding development of eczema and sensitization as defined by a positive skin prick test and/or circulating allergen-specific IgE antibodies at 6, 12, and 24 months of age. Supplementation of L. reuteri during pregnancy was associated with low levels of TGF-β2 and slightly increased levels of IL-10 in colostrum. For TGF-β2, this association was most pronounced in mothers with detectable L. reuteri in faeces. Infants receiving breast milk with low levels of TGF-β2 were less likely to become sensitized during their first 2 yr of life. A similar trend was observed for development of IgE-associated eczema. The levels of total IgA, SIgA, TGF-β1, TNF, sCD14, and Na/K ratios in breast milk were not affected by the intake of L. reuteri . None of these parameters correlated with sensitization or development of eczema in the infant, except for high Na/K ratios that associated with increased risk of sensitization. Supplementation with L. reuteri during late pregnancy reduces breast milk levels of TGF-β2, and low levels of this cytokine are associated with less sensitization and possibly less IgE-associated eczema in breast-fed infants.     

Control of β-catenin/Tcf-directed transcription in medulloblastoma

The β-catenin, glycogen synthase kinase 3β (GSK-3β), and adenomatous polyposis coli (APC) gene products interact to form a network that influences the rate of cell proliferation. Medulloblastoma occurs as part of Turcot's syndrome and patients with Turcot's syndrome, who develop medulloblastomas, have been shown to harbor germline APC mutations. While APC mutations have been investigated and not identified in sporadic medulloblastomas, the status of the β-catenin and GSK-3β genes has not been evaluated in this tumor. This study shows that 3 of 67 medulloblastomas harbor β-catenin mutations, each of which converts a GSK-3β phosphorylation site from serine to cysteine. The β-catenin mutation seen in the tumors was not present in matched constitutional DNA in the 2 cases where matched normal DNA was available. A loss of heterozygosity (LOH) analysis of 32 medulloblastomas with paired normal DNA samples was performed with 4 microsatellite markers flanking the GSK-3β locus; LOH with at least one marker was identified in 7 tumors. Sequencing of the remaining GSK-3β allele in these cases failed to identify any mutations. Taken together, these data suggest that activating mutations in the β-catenin gene may be involved in the development of a subset of medulloblastomas. The GSK-3β gene does not appear to be a target for inactivation in this tumor.     

GLYCOSIDASES IN SKIN AND PLASMA IN HUNTER'S SYNDROME Abnormality of a β-galactosidase in Skin

Two patients with Hunter's syndrome (mucopolysaccharidosis type II) were studied. The exact diagnosis could be settled by the finding of clinical symptoms and signs, typical for this form of mucopolysaccharidosis, and of a greatly increased excretion of glycosaminoglycans (acid mucopolysaccharides) in the urine, as well as by the fact that the patients were half-brothers with unrelated fathers. Analyses of glycosidases in skin demonstrated the existence of low activity of β-galactosidase in both patients. β-acetylglucosaminidase was more active in both patients and β-glucuronidase in one of them than in the controls. In plasma increased activities were noted for β-glucuronidase and β-acetylglucosaminidase in both patients and of β-galactosidase and α-mannosidase activities in one of them and of α-fucosidase activity in the other. The results are very similar to those described in patients with Hurler's syndrome (mucopolysaccharidosis type I).     

Circulating noradrenaline and β-adrenergic receptors in children with congestive heart failure

This study was designed to investigate changes in plasma catecholamine concentrations and the number of β-adrenoceptors (β-AR) of circulating lymphocyte in 94 noncyanotic congenital heart patients, in 43 patients with congestive heart failure, β-AR density was significantly lower ( p < 0.001) and plasma noradrenergic levels were significantly higher ( p < 0.001) compared with corresponding values in 51 patients without heart failure. A significant negative correlation between lymphocyte β-AR density and plasma noradrenergic levels was observed ( r =−0.61, p < 0.001). The degree of left-to-right shunt and pulmonary pressure was correlated directly with noradrenaline level and inversely with lymphocyte β-AR density. Both plasma noradrenaline level and lymphocyte β-AR density return to normal in children with heart failure after surgical repair. Our results support the idea that changes in noradrenaline level and lymphocyte β-adrenoceptor density occur concurrently with the presence and severity of heart failure in children.