Clinical reactivity to beef in children allergic to cow's milk

BACKGROUND: Cow's milk is one of the most common food allergens in children. Limited information is available on the prevalence of reactivity to a related food source, beef. The purposes of this study were to examine the prevalence of symptomatic sensitivity to beef in a selected pediatric population and to determine the frequency of concomitant reactivity to cow's milk and beef. METHODS: Children referred for assessment of atopic dermatitis and possible food hypersensitivity were evaluated for symptomatic reactivity to beef by double-blind placebo-controlled food challenges (DBPCFCs) and subsequent open feedings of beef. Sodium dodecyl-sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), immunoblot, and immunodot blot analyses were performed with patients' sera on preparations of beef extracts subjected to different cooking conditions: raw (no heating), medium, and well-cooked. RESULTS: Eleven of 335 children referred for evaluation of atopic dermatitis and possible food hypersensitivity were found to have symptomatic sensitivity to beef; eight were also sensitive to milk, as demonstrated in previous DBPCFCs. Eight patients reacted to beef during DBPCFC, and three tolerated beef in a DBPCFC and well-cooked beef in an open challenge but reacted to ingestion of less well-cooked beef. SDS-PAGE of raw beef revealed at least 24 protein fractions. Several protein bands in raw beef appeared to denature with heating. Bovine serum albumin and bovine gamma globulin were heat-labile in the beef extract, but six protein fractions persisted even after heating the beef extract for 2 hours at 85 degrees C. IgE from patients reacting to rare and well-cooked beef bound up to six of these heat-resistant fractions, but IgE from patients reacting only to rare beef failed to bind any of these fractions with one exception. In addition, patients reacting to rare and well-cooked beef had specific IgE to a 17.8 kd fraction, which was only weakly recognized by one patient reacting only to rare beef. CONCLUSIONS: Specific IgE antibodies to heat-labile beef proteins might explain why some patients can tolerate well-cooked beef but not medium-rare and rare beef. Patients reacting only to rare beef may not need to maintain a complete beef elimination diet.     

Cisapride in neurologically impaired children with chronic constipation

The efficacy of cisapride as a treatment for chronic constipation in children with severe brain damage was studied in 20 children. Each subject was randomly assigned to double-blind treatment with either cisapride (N = 10) or placebo (N = 10) for 12 weeks. Stool habits, total gastrointestinal transit time, colonic segmental transit times, and anorectal motility were evaluated in all children before and at the end of the treatment period. Although cisapride significantly (P < 0.05) increased stool frequency from baseline to week 12 and no significant change was documented in the placebo group, the mean change in stool frequency per week from baseline to 12 week was not significantly different between the two treatment groups. The use of laxatives or suppositories was significantly (P < 0.05) decreased by cisapride, but remained unchanged in the placebo group. Furthermore, cisapride significantly (P < 0.05) reduced rectal compliance but had no effect on total gastrointestinal transit time and colonic segmental transit times. In summary, in neurologically impaired children with chronic constipation, cisapride increased bowel frequency but did not alter the delay in total and segmental gastrointestinal transit times.     

Auxological evaluation of children with cow's milk protein intolerance treated an elimination diet

A somatic development assesment, including the nutritive condition at the moment of diagnosis and after at least a year's dietary treatment was carried out in a group of 55 children with primary cow's milk protein intolerance. The influence of the applied elimination diet and/or anti-allergic supporting treatment on the clinical picture was evaluated as well as calcium--phosphorus--magnesium balance parameters.     

Intestinal barrier function and cow's milk sensitization in guinea pigs fed milk or fermented milk

BACKGROUND: The respective effect of milk and fermented milks on intestinal barrier capacity and on sensitization to beta-lactoglobulin was studied using a guinea pig model of cow's milk allergy. METHODS: Guinea pigs were fed a control diet or the same diet supplemented with milk, fermented milk (Streptococcus thermophilus and Bifidobacterium breve), or dehydrated fermented milk. Intestinal barrier capacity to macromolecules was assessed in an Ussing chamber, and sensitization to cow's milk proteins was measured by systemic anti-beta-lactoglobulin immunoglobulin G1 titers and by intestinal anaphylaxis, the latter assessed by the beta-lactoglobulin-induced increase in short-circuit current of jejunal fragments (deltaIsc(beta-LG)). RESULTS: The electrical resistance of jejunum was similar in the four groups (approximately 80 omega/cm2) suggesting the same paracellular permeability. The transport of 14C-beta-lactoglobulin from mucosa to serosa was significantly decreased in the animals fed dehydrated fermented milk (403+/-131 ng / hr x cm2) compared with that in control animals or animals fed milk (767+/-250 ng / hr x cm2 and 749+/-475 ng / hr x cm2, respectively; p < 0.05). Milk fermentation did not modify native beta-lactoglobulin concentration but anti-beta-lactoglobulin immunoglobulin G1 titers were higher in fermented milk and dehydrated fermented milk (log10 titer = 2.86 and 2.79, respectively) than in guinea pigs fed milk (log10 titer = 2.5; p < 0.007). However, beta-lactoglobulin-induced intestinal anaphylaxis remained the same in the three groups (deltaIsc(beta-LG), 9.6+/-4.1 microA/cm2, 8.5+/-4.3 microA/cm2, and 8.5+/-3.4 microA/cm2 in milk-fed, fermented milk-fed, and dehydrated fermented milk-fed guinea pigs, respectively). CONCLUSIONS: The intestinal barrier capacity to milk proteins seems to be reinforced by dehydrated fermented milk, but milk and fermented milks are equally efficient in inducing cow's milk allergy in guinea pigs.     

Assay of prolactin after freezing cow's milk

Concentrations of prolactin were similar in aliquots of the same milk sample stored for 2 days at 4 C or --23 C but averaged lower if prepared for assay at 30 C than if prepared at 40 or 50 C. Average deviations in prolactin between duplicate measurements relative to respective treatment averages were lowest generally when technique of mixing included vortexing for 5 s immediately prior to pipetting volumes of milk for assay. The average deviation of differences in prolactin among duplicate measurements relative to average concentrations of prolactin were about the same for colostrum, milk, and blood plasma (11, 17, and 14%) stored frozen. Prolactin can be measured reliably in frozen cow's milk provided samples are warmed to 40 C to 50 C and mixed thoroughly when prepared for assay.     

Ultrasonographic evaluation of urinary tract in children with chronic functional constipation

Urinary tract anomalies were prospectively investigated with ultrasound in 31 children with chronic functional constipation. These children were compared with 29 healthy controls without constipation both before and after treatment. Bladder residue and upper renal tract dilatation after micturition were significantly more common in the group with constipation than in the improved after-treatment and control groups.     

Duphalac (lactulose) in the treatment of chronic constipation in children

The aim of this study was to estimate the effectiveness of DUPHALAC preparation, produced by Solvay-Pharma, in treatment of functional chronic constipation. 112 children, aged 10 months-15.5 years were examined. DUPHALAC appeared to be very effective, were examined. Only two patients did not accept its taste. Rapidly (approximately after 2.5 weeks) improvement was achieved such as of restoration normal peristalsis and of previously disturbed defecation rhythm, softening of a stool, regression of pain and/or dyspeptic ailments, encopresis and improvement of appetite. In no case undesirable side effects were observed.     

Mucosal enterokinase activity in cow's milk protein sensitive enteropathy

Enterokinase has a critical role in initiating proteolytic digestion by hydrolysing the conversion of pancreatic trypsinogen into trypsin. The enzyme is synthesised by enterocytes of the proximal small intestine and initially incorporated into the brush border from where it is released into the intestinal lumen by the action of pancreatic secretions. The aim of the study was to analyse enterokinase activity in the duodenal mucosa of infants with diarrhoeal disease including cow's milk protein-sensitive enteropathy. Our observations show that the mean depletion of enterokinase was only 17% compared to 60-80% for other brush border enzymes like disaccharidases, peptidases and alkaline phosphatases in infants with diarrhoea. This suggests that enterokinase activity in the small bowel enteropathies may be dependent not only on the degree of mucosal damage specifically but also on the extent of damage to the goblet cell population where the enzyme is synthesised. Thus the enterokinase activity was reduced in acute and chronic diarrhoea with marked mucosal damage where significant reduction of goblet cell population was evident but the enzyme was relatively little affected when the mucosa was damaged mildly.     

Secretory IgA antibodies against cow's milk proteins in human milk and their possible effect in mixed feeding

Antibodies of the secretory IgA type against cow's milk proteins were consistently found in human milk. With the assumption that such antibodies can help to prevent or at least diminish the contact between native cow's milk proteins and the lymphoid system of babies on mixed feeding, the levels of antibodies of various immunoglobulin classes against bovine milk proteins were measured in different groups of babies. Those on artificial feeds who had been on mixed feeding of human milk and cow's milk for less than 1 week had significantly higher levels of IgG antibodies to cow's milk proteins than those who had been on mixed feeding for a period longer than 3 weeks.     

Production of exfoliative toxin A by Staphylococcus aureus isolated from mastitic cow's milk and farm bulk milk

The production of exfoliative toxins A and B (ETA and ETB) by Staphylococcus aureus isolated from mastitic cow's milk and farm bulk milk was examined by the reverse passive latex agglutination method (RPLA). ETA was detected in 2 (1.2%) of 162 isolates from mastitic cow's milk and in 1 (0.6%) of 166 isolates from farm bulk milk. RPLA titers of these isolates were much lower than in human isolates. No ETB was detected in any of the isolates tested. These ETA-positive isolates belonged to bovine ecovar. They were non-typable using the international phage set for human strains. When these ETA-positive isolates were subcutaneously inoculated into neonatal mice, general exfoliation of the epidermis accompanied by the so-called Nikolsky sign was not recognized. By the immunoblotting and PCR methods, however, ETA and eta gene were recognized in the ETA-positive isolates from mastitic cow's milk and farm bulk milk. These data suggest that ETA is also produced by bovine isolates of S. aureus, but in smaller quantities.     

The threshold for immune cell reactivity to milk antigens decreases in cow's milk allergy with intestinal symptoms

BACKGROUND: In cow's milk allergy (CMA) with intestinal symptoms, peripheral blood mononuclear cells (PBMCs) secrete tumor necrosis factor-alpha (TNF-alpha), altering intestinal function. However, the type of cow's milk protein (CMP) that triggers symptoms (intact or intestinally processed) is not known, and neither is the minimal amount required. METHODS: PBMCs were isolated from infants with active CMA or cured infants just before a new challenge and stimulated with intact or intestinally processed CMP. Supernatants were tested for cytokine content and for their ability to perturb intestinal barrier capacity, measured in Using chambers in HT29-19A intestinal cells. RESULTS: PBMCs from infants with active CMA secreted more TNF-alpha, when they were stimulated with intact rather than intestinally processed CMPs, and more TNF-alpha than PBMCs from cured infants. Accordingly, supernatants from PBMCs stimulated with intact but not intestinally processed CMPs significantly increased intestinal permeability. The CMP concentration required to trigger TNF-alpha secretion capable of altering intestinal function was very small in infants with active CMA (approximately 2 micrograms/ml), but about 300 times higher in cured infants. CONCLUSION: Intact rather than intestinally processed proteins stimulate PBMCs to release TNF-alpha and alter intestinal barrier capacity. The threshold for PBMC reactivity to milk antigens drops considerably during active CMA with intestinal symptoms.     

The intestinal ecosystem in chronic functional constipation

Chronic functional constipation is common in infants, and the bacterial composition of stools in this condition is not known. The study aims were to: (i) investigate the composition of the intestinal ecosystem in chronic functional constipation; (ii) establish whether the addition of the water-holding agent calcium polycarbophil to the diet induces an improvement in constipation; and (iii) determine the composition of the intestinal ecosystem after the use of this agent. In total, 42 children (20F, 22M; mean age: 8.6 +/- 2.9 y) were studied. Twenty-eight children with functional chronic constipation without anatomical disorders were treated double-blind in random sequence for 1 month with an oral preparation of calcium polycarbophil (0.62 g/twice daily) or placebo. Intestinal flora composition was evaluated by standard microbiological methods and biochemical assays on faecal samples collected before and after treatment. Fourteen healthy children were studied as controls. The results show that (i) the constipated children presented a significant increase in clostridia and bifidobacteria in faeces compared to healthy subjects--different species of clostridia and enterobacteriaceae were frequently isolated; no generalized overgrowth was observed; Clostridia outnumbered bacteroides and E. coli mean counts by 2-3log, while bacteroides and E. coli counts were similar (5-6 log10/g fresh faeces); these intestinal disturbances could be defined as a dysbiosis, i.e. a quantitative alteration in the relative proportions of certain intestinal bacterial species. (ii) Clinical resolution of constipation was achieved only in 43% of treated children and an improvement in 21% (one bowel movement every 2 d). (iii) Calcium polycarbophil treatment induced no significant changes in the composition of the intestinal ecosystem, nor in blood chemistry parameters.     

Evaluation of an extensively hydrolyzed casein-whey protein formula in immediate cow's milk protein hypersensitivity

BACKGROUND: Hydrolysate formulas are safe for most infants who are allergic to cow's milk. The purpose of this study was to assess the clinical tolerance and safety of an extensively hydrolyzed casein-whey protein formula in a homogeneous group of patients with immediate cow's milk hypersensitivity. METHODS: The study population consisted of 33 infants in whom immediate cow's milk protein allergy had been diagnosed by clinical data, evidence of specific immunoglobulin E antibodies, and positive results in a controlled challenge test with cow's milk proteins. RESULTS: The hydrolysate was well tolerated in 31 of the 33 patients, which represented a 94% effectiveness. Using 95% confidence intervals generated for a binomial outcome (reaction-no reaction), the casein-whey protein formula is tolerated by 84.4% to 99% of infants with immediate immunoglobulinE-mediated cow's milk protein allergy. These figures are in the range of values recommended for this type of product. CONCLUSIONS: This extensively hydrolyzed casein-whey protein formula is generally safe to feed children with immediate hypersensitivity to cow's milk. However. it is advisable that the first intakes be given under direct medical supervision, in that the occurrence of adverse reactions in highly sensitive infants cannot be unequivocally excluded.     

Surgical evaluation and management of refractory constipation in older children

Chronic constipation is a common childhood problem that accounts for 3% to 5% of pediatric visits and 10% to 25% of referrals to pediatric gastroenterologists. The etiology of constipation can be elusive, and extensive investigation often fails to identify a specific cause. The authors conducted a 5-year retrospective review of the patients referred for deep transanal rectal biopsy to determine the usefulness of this procedure in the evaluation and subsequent surgical management of refractory constipation. Specimens obtained by transanal rectal biopsy established a diagnosis for 30 of the 70 patients, and 17 of these 30 had subsequent procedures in the treatment of their constipation. The authors conclude that transanal rectal biopsy identifies a significant number of patients with previously unidentified neuroenteric disorders who may benefit from additional surgery in the treatment of constipation refractory to medical management.     

Detection of Coxiella burnetii in cow's milk using the polymerase chain reaction (PCR)

A PCR approach (transposon PCR) with primers based on repetitive transposon-like sequences, which--depending on the isolate--were found at a minimum frequency of 19 on the C. burnetii genome, was established for the highly sensitive and specific detection of C. burnetii. This study describes the analytical detection of C. burnetii in milk, which requires a special preparation method prior to PCR. Because of the low level of C. burnetii particles in milk samples, template DNA was concentrated by a factor of 200, using cetyltrimethylammonium bromide as the precipitation reagent. Using this particular preparation method, even a single C. burnetii particle could be detected in 1 ml milk.     

In vitro diagnosis of cow's milk protein sensitive enteropathy by organ culture method

The criteria that are used at present to diagnose cow's milk protein sensitive enteropathy (CMPSE) are based on an in vivo milk challenge which can be hazardous and life threatening. We have used an organ culture model to determine the usefulness of this technique in establishing the diagnosis of CMPSE on the basis of a single biopsy with in vitro milk challenge. Fourteen infants with diarrhoea clinically suspected to have CMPSE were studied prospectively. On the basis of milk challenge studies seven infants had CMPSE. They had clinical reaction to cow's milk with associated histological changes and depression of alkaline phosphatase levels in the jejunal mucosa. In all seven cases parallel changes in alkaline phosphatase levels were noted in the organ culture specimens of initial biopsy subjected to in vitro challenge. The seven control infants tolerated cow's milk and did not have histological changes. The alkaline phosphatase levels were moderately increased in the jejunal mucosa in five of the seven infants. The alkaline phosphatase levels in the organ culture specimens of initial biopsy were increased after in vitro challenge in all seven infants. This study suggests that organ culture methods may be useful in the vitro diagnosis of CMPSE, and also obviate the need for in vivo oral milk challenges and repeated biopsies.