Urinary (1)H-NMR-based metabolic profiling of children with NAFLD undergoing VSL#3 treatment.

BACKGROUND: Nowadays, non-alcoholic fatty liver disease (NAFLD) is one of the most common chronic liver diseases in children. Our recent clinical trial demonstrated that dietary and VSL#3-based interventions may improve fatty liver by ultrasound and body mass index (BMI) after 4 months. OBJECTIVES: As in this short-term trial, as in others, it is impracticable to monitor response to therapy or treatment by liver biopsy, we aimed to identify a panel of potential non-invasive metabolic biomarkers by a urinary metabolic profiling. METHODS: Urine samples from a group of 31 pediatric NAFLD patients, enrolled in a VSL#3 clinical trial, were analyzed by high-resolution proton nuclear magnetic resonance spectroscopy in combination with analysis of variance-Simultaneous Component Analysis model and multivariate data analyses. Urinary metabolic profiles were interpreted in terms of clinical patient feature, treatment and chronology pattern correlations. RESULTS: VSL#3 treatment induced changes in NAFLD urinary metabolic phenotype mainly at level of host amino-acid metabolism (that is, valine, tyrosine, 3-amino-isobutyrate or ß-aminoisobutyric acid (BAIBA)), nucleic acid degradation (pseudouridine), creatinine metabolism (methylguanidine) and secondarily at the level of gut microbial amino-acid metabolism (that is, 2-hydroxyisobutyrate from valine degradation). Furthermore, some of these metabolites correlated with clinical primary and secondary trial end points after VSL#3 treatment: tyrosine and the organic acid U4 positively with alanine aminotransferase (R=0.399, P=0.026) and BMI (R=0.36, P=0.045); BAIBA and tyrosine negatively with active glucagon-like-peptide 1 (R=-0.51, P=0.003; R=-0.41, P=0.021, respectively). CONCLUSIONS: VSL#3 treatment-dependent urinary metabotypes of NAFLD children may be considered as non-invasive effective biomarkers to evaluate the response to treatment.

Biomarkers of intake for tropical fruits.

Consumption of fruit and vegetable is a key component of a healthy and sustainable diet. However, their accurate dietary assessment remains a challenge. Due to errors in self-reporting methods, the available dietary information is usually biased. Biomarkers of intake constitute objective tools to better reflect the usual or recent consumption of different foods, including fruits and vegetables. Partners of The Food Biomarker Alliance (FoodBall) Project have undertaken the task of reviewing the available literature on putative biomarkers of tropical fruit intake. The identified candidate biomarkers were subject to validation evaluation using eight biological and chemical criteria. This publication presents the current knowledge on intake biomarkers for 17 tropical fruits including banana, mango, and avocado as the most widely consumed ones. Candidate biomarkers were found only for banana, avocado, and watermelon. An array of banana-derived metabolites has been reported in human biofluids, among which 5-hydroxyindole-acetic acid, dopamine sulfate, methoxyeugenol glucuronide, salsolinol sulfate, 6-hydroxy-1-methyl-1,2,3,4-tetrahydro-ß-carboline-sulfate, and other catecholamine metabolites. Their validation is still at an early stage, with insufficient data on dose-response relationship. Perseitol and mannoheptulose have recently been reported as candidate biomarkers for avocado intake, while the amino acid citrulline has been associated with watermelon intake. Additionally, the examination of food composition data revealed some highly specific phytochemicals, which metabolites after absorption may be further studied as putative BFI for one or several tropical fruits. To make the field move forward, untargeted metabolomics, as a data-driven explorative approach, will have to be applied in both intervention and observational studies to discover putative BFIs, while their full validation and the establishment of dose-response calibration curves will require quantification methods at a later stage.

Validation of biomarkers of food intake-critical assessment of candidate biomarkers.

Biomarkers of food intake (BFIs) are a promising tool for limiting misclassification in nutrition research where more subjective dietary assessment instruments are used. They may also be used to assess compliance to dietary guidelines or to a dietary intervention. Biomarkers therefore hold promise for direct and objective measurement of food intake. However, the number of comprehensively validated biomarkers of food intake is limited to just a few. Many new candidate biomarkers emerge from metabolic profiling studies and from advances in food chemistry. Furthermore, candidate food intake biomarkers may also be identified based on extensive literature reviews such as described in the guidelines for Biomarker of Food Intake Reviews (BFIRev). To systematically and critically assess the validity of candidate biomarkers of food intake, it is necessary to outline and streamline an optimal and reproducible validation process. A consensus-based procedure was used to provide and evaluate a set of the most important criteria for systematic validation of BFIs. As a result, a validation procedure was developed including eight criteria, plausibility, dose-response, time-response, robustness, reliability, stability, analytical performance, and inter-laboratory reproducibility. The validation has a dual purpose: (1) to estimate the current level of validation of candidate biomarkers of food intake based on an objective and systematic approach and (2) to pinpoint which additional studies are needed to provide full validation of each candidate biomarker of food intake. This position paper on biomarker of food intake validation outlines the second step of the BFIRev procedure but may also be used as such for validation of new candidate biomarkers identified, e.g., in food metabolomic studies.

[Neonatal hypoxia and hemocoagulative changes]. / Ipossia neonatale e modificazioni emocoagulative.

Coagulation abnormalities with and without haemorrhagic manifestations have been frequently reported in newborn-infants affected by hypoxia. Particularly in postmature-infants and in those ones with acute asphyxia at birth, respiratory distress syndrome (RDS), intra-uterine growth retardation (IUGR) and cyanotic congenital heart disease (CCHD). A reduction of synthesis or a consumption of blood coagulation factors are the main causes of these abnormalities. The anomalies of platelet number and of their function, of haemostasis global tests, of coagulation factors and physiologic inhibitors levels, of fibrinogenesis and fibrinolysis are examined, including authors' studies and a review of literature too. The authors think platelet count, PT, PTT, fibrinogen, factor V and VIII, and PDF determinations are necessary laboratory investigations for newborn-infants with RDS or acute asphyxia for about the first week of life, because of the risk of consumption coagulopathy. In the other hypoxic newborns (IUGR, CCHD, postmature infants) platelets count, PT, PTT and serum PDF determinations could be enough in order to value any coagulation abnormalities presence.

[Transverse hemimelia of the lower right limb associated with sysmetrical malformations of the hands in a newborn infant]. / Emimelia trasversa dell'arto inferiore destro associata a malformazioni simmetriche delle mani in un neonato.

The authors report a newborn infant affected by terminal transverse hemimelia of the right lower limb, associated with symmetric malformations of the hands, simulating the "amniotic bands syndrome". The symmetric hand anomalies and the presence of slight malformations of the left lower limb lead us to think that the etiology of this case is genetic, probably related to a new mutation.

[Platelet factor 4 levels in full-term newborns undergoing phototherapy]. / Livelli di fattore piastrinico 4 (FP4) in neonati a termine sottoposti a fototerapia.

PF4 levels and platelets counts were studied in a group of 15 term newborn infants before treatment and after 24-48-72 and 96 hours of phototherapy and in a control group of 10 babies. Unlike data found by other AA. in vitro and in preterm infants, our values show only minimal, not statistically significant, differences in PF4 levels and platelets counts between the two groups. The AA. believe that in term infants, with adequate weight for gestational age, proper phototherapy treatment does not cause any change in platelet function, owing to the thicker and more mature skin and to the better bone marrow compensation typical of term versus preterm infants.

[A case of late neonatal hemorrhagic disease associated with intolerance for cow's milk proteins]. / Su un caso di malattia emorragica neonatale tardiva associata ad intolleranza alle proteine del latte vaccino.

The authors describe a two months aged patient affected by cow's milk protein intolerance (CMPI) with serious haemorrhagic manifestations. As blood coagulative laboratory findings demonstrated a prolongation of P.T. and P.T.T. with a marked reduction of vitamin K-dependent factors only, the authors believe these bleeding manifestations secondary to a case of late haemorrhagic disease of the newborn. Vitamin K treatment determined a rapid normalization of haemorrhagic symptoms and laboratory clotting tests, without any alteration of these ones during the patient's follow-up too. The authors suggest that blood coagulative pattern must be investigated in all CMPI cases, especially in the ones with a precocious onset of clinical symptoms. In the cases with vitamin K-dependent factors deficiency the treatment is immediately necessary, while in other cases a daily dietary supplementation or a vitamin K weekly or monthly injection could be enough in order to prevent any further vitamin K-dependent factors deficiency.

[Normal levels of collagen-type-I telopeptide in the first 90 days of life]. / Livelli normali di telopeptide del collagene di tipo I nei primi 90 giorni di vita.

Serum levels of carboxyterminal telopeptide of type I collagen (ICTP), a marker of matrix degradation, were measured by RIA test, on 184 samples of healthy newborns and children aging from 1 (cord blood) to 90 days of life. We found ICTP values about tenfold higher than the adults', with highly significant variations (P < 0.001) in the whole period studied. During the first three months of life serum levels of the bone marker show a progressive increase from 0 to 7 days, they remain unchanged until the 30th day and then decrease until the 45th day, maintaining similar values from the 45th to the 90th day of life. The authors think that the pattern of ICTP in the first week of life is under the influence of the adapting phenomena following delivery, in which catabolic processes are predominant, while in the second period ICTP modifications are related to growing processes and then to bone turnover.

[The relationships between the degrees of oxygenation and the serum calcitonin levels in the term newborn]. / Relazioni tra gradi di ossigenazione e livelli di calcitonina sierica nel neonato a termine.

The authors have studied the correlations between serum levels of calcitonin and the degree of oxygenation assessed by means of transcutaneous pO2 and pCO2 and capillary pH in 40 term newborns of adequate birth weight. Highly significant correlations (P < 0.001) were found at the 24th hour of life between calcitonin levels and the asphyxia parametres and between the latter measured at the 12th or the 24th hour and calcitonin levels found respectively at the 24th or the 48th hour. Similar correlations were found subdividing the studied newborns with regard to the type of delivery. We conclude that the severity of neonatal asphyxia is indeed the main determining factor of the magnitude of the calcitonin hyperincretion.

[Bone metabolism markers in thalassemia]. / Markers del metabolismo osseo nella talassemia.

The A.A. performed a screening on 113 patients affected by beta-thalassemia major ranging in age between 2 and 40 years, randomized among those which come to the Microcitemic Center of our Institute, and in a control group. In everybody, serum levels of calcium, phosphate, parathyroid hormone (PTH), calcitonin and 25-OH vitamin D were measured. Average serum levels of PTH were significantly (P < 0.001) lower in patients than controls and 12.4% of them were clearly under normal range, especially in the group above 16 years of age. Also serum levels of 25-OH vitamin D were lower in thalassemic subjects than in controls, because of the presence of 32 patients with values under normal limit. Our results are in agreement with current literature that underline the increasing incidence of endocrine complications in thalassemic patients which undergo to high transfusion regimens, owing to the increase of emosiderosis due to the low compliance to iron chelation therapy. Controversial is the pathogenesis of the absence of hypocalcemia in many patients with hypoparathyroidism and the determinism of the deficit of vitamin D.

[Calcium-phosphorus metabolism in celiac disease in children]. / Metabolismo calcio-fosforico nella malattia celiaca del bambino.

The Authors studied the changes of the main parameters of calcium-phosphate metabolism in twenty four untreated celiac children (mean age: 23.7 +/- 14.4 months) and in eleven control subjects (mean age: 28.5 +/- 21.2 months). 16 patients were checked again one and three months after treatment was begun. Compared with controls patients show at diagnosis a significant increase of serum phosphate (P = 0.025) and a decrease of calcitonin levels (P = 0.02), whereas serum calcium is slightly lower and parathyroid hormone higher with serum levels above normal range in 5 of the coeliac patients (20.8%). During the three months of gluten free diet we find a significant increase of calcemia values (ANOVA: P = 0.025) together with a decrease of serum phosphate (ANOVA: P = 0.009) and of parathyroid hormone levels (ANOVA: P = 0.042). No significant change was found in vitamin D metabolites levels. The Authors conclude that coeliac disease affect clearly mineral metabolism. Actually the tendency to hypocalcemia, due to abnormalities of the intestinal mucosa, and the comparative iperphosphatemia, cause in some patients a compensatory increase of PTH levels. This increase seems to be the cause of the osteoporotic lesions described in current literature. Rickets due to the lack of vitamin D does not occur.

[Normal urinary levels of telopeptide aminoterminal (NTx) of type I collagen in healthy term newborns and infants in the first month of life]. / Livelli normali urinari di telopeptide amino terminale (NTx) del collagene di tipo I in neonati a termine e lattanti sani nei primi tre mesi di vita.

The Authors have studied urinary aminoterminal telopeptide of type I collagen (NTx), a bone catabolism marker of recent determination, by an enzyme-linked immunoassorbent assay (OSTEOMARK) in 80 urine samples of term healthy infants in the first 3 months of life. Highly significant variations have been compared in the whole period studied (P = 0.000). Levels of NTx increase significantly from 1 to 7 days of life, reaching a plateau that is kept until 45th day and then significantly decrease until 90th day, when, however, they result higher than the values reported in literature concerning older ages. The Authors conclude that even this bone catabolism marker is influenced, in the first week of life, by the particular phenomenology linked to the neonatal adaptation, and that, subsequently, shows a trend strictly linked to the bone turnover modifications throughout the faster stage of the growth.

[Normal levels of carboxyterminal propeptide of type I procollagen in the first three months of life]. / Livelli normali del propeptide carbossiterminale del procollagene di tipo I nei primi tre mesi di vita.

Serum levels of type I procollagen were measured on 118 samples from cord blood or from healthy infants aging from 1 to 90 days of life. Significant variations (P = 0.001) were noticed in the values of the marker in the whole period under investigation. We observed a decrease of PICP from cord blood to the end of the first day of life with a sharp rise on the 5th day lasting until the 30th day which then became stable till the end of the third month. Our results show a peculiar pattern of PICP levels during the first month of life which has to be taken into account to evaluate normal values of the marker in this period of life.

[Calcium phosphate metabolism in thalassemia]. / Metabolismo calciofosforico nella talassemia.

The AA. performed a screening test on 113 patients affected by beta thalassemia major ranging between 2 and 40 years of age, randomized among those who come to the Microcitemic Center of our Institute, and on a control group. In all of them serum levels of calcium, phosphate, parathyroid hormone (PTH), calcitonin and 25-OH vitamin D were measured. Average serum levels of PTH were significantly (P < 0.001) lower in our patients than in control group and 12.4% of the former were clearly under normal range, especially in the group over 16 years of age. Also serum levels of 25-OH vitamin D were lower in thalassemic patients than in controls, because of the presence of 32 patients with average values under normal limit. Our results are in agreement with current literature and underline the increasing incidence of endocrine complications in thalassemic patients who undergo high transfusion regimens, because of to the increase of hemosiderosis due to the low compliance to iron chelation therapy. Controversial is the pathogenesis of the absence of hypocalcemia in many patients with hypoparathyroidism and the cause of the deficit of vitamin D.

[Effect of the therapy with vitamin K on coagulation factors in celiac disease in children]. / Effetto della terapia con vitamina K sui fattori della coagulazione nella malattia celiaca del bambino.

The Authors carried out a study on 37 untreated coeliac children to investigate the behaviour of K-dependent factors after vitamin K administration. The children were randomized into two groups: 22 children receiving a single dose of 10 mg i.m. of Phytonadione (Konakion, Roche) on the initial day of GFD and 15 children who did not receive vitamin K administration. PT, PTT and clotting activity of Factors II, VII, IX, X were determined before the treatment and/or GFD, and after 24 hours, 7 and 15 days. The results demonstrated that vitamin K administration determined a rapid increase in clotting activity of all K-dependent factors after 24 hours. These values remained normal after 7 and 15 days, except for Factor II, which slightly decreased on the 7th day. On the contrary, the children not treated, had levels similar to those of acute stage. After 7 days these values showed a slight increase and reached normal limits on the 15th day. No significant changes were seen in either PT or PTT in the two groups. They were constantly prolonged, reaching normal limits on the 15th day. These results indicate that the vitamin K deficiency, not only seems constant in children with CD, but also seems responsible for the haemocoagulative deficit of the K-dependent factors. After GFD when intestinal absorption is regained, all parameters returned to normal. The Authors concluded that K-dependent factors can be used as short-term indexes of improved intestinal absorption and that the coeliac children with severely compromised nutritional status can be treated with vitamin K (10 mg bolus).

[Role of dietary prevention in newborns at risk for atopy. Results of a follow-up study]. / Ruolo della prevenzione dietetica nei neonati a rischio per atopia. Risultati del follow-up.

The Authors have studied the role of various preventing diet for a primary prophylaxis of allergy in 125 newborns at risk of atopy: 30 exclusively breast-fed, 50 hypoallergenic milk fed, 30 soy milk fed, and 15 with conventional milk formula. IgE values were determined at 5 days, 6 months, and 12 months of age, IgE values at 5 days were compared to newborns not at atopic risk. The clinical follow-up lasted 4 years. Total IgE values at 5 days were significantly higher in new-born at atopic risk. Only breast-feeding subjects had IgE normal values at six months. Allergic symptoms were observed in 14% of infants with a guided diet and in 53% of infants with a conventional diet. Breast fed subjects had atopic disorders in only 8% of cases, subjects with hypoallergenic formula in 12% while soja milk fed in 25%. The Authors stress the role of breast feeding in preventing allergic disorders in subjects at atopic risk or, when human milk misses, of a hypoallergenic formula, more than soy milk and conventional formula and confirm the possibility of diet and ambiental prophylaxis of allergy.

Microphthalmia due to congenital varicella infection: a case report.

The authors report the clinical case of a newborn affected by congenital varicella syndrome, occurred about the 12th gestational week, with ophthalmic involvement. The ocular anomalies consisted in right microphthalmia, with lens opacities and atrophic chorioretinitis, without any involvement of other pathologies. The ophthalmic lesions and the sierological data confirmed that the infection occurred during the first weeks of gestation. Although the manifestation limited to the eyes is extremely rare, the authors point out the necessity of an appropriate prevention to avoid irreversible involvements of important organs.

Rubinstein-Taybi syndrome associated with Dandy-Walker cyst. Case report in a newborn.

Rubinstein-Taybi is a rare malformative syndrome characterized by dysmorphic features and mental retardation. Early diagnosis in neonatal age can be facilitated by the presence of characteristic broadening of the distal phalanges of thumbs and great toes. Most of the cases are sporadic. Associated malformations such as bone anomalies, heart malformations, cell immunity deficits and metabolic alterations have been observed. This paper reports the first case of Rubinstein-Taybi syndrome associated with Dandy-Walker type cerebral malformation diagnosed in the neonatal period.

[Calcium-phosphate metabolism and bone markers in two patients with Noonan's syndrome treated with growth hormone]. / Metabolismo calcio-fosforico e markers ossei in due pazienti con Sindrome di Noonan trattati con ormone della crescita.

AIM: To evaluate the possible effects of recombinant growth hormone (rhGH) therapy on mineral homeostasis and bone turnover, the authors studied calcium-phosphate metabolism parameters, including some bone markers, in 2 prepubertal subjects with Noonan's syndrome (NS). METHODS: Two prepubertal males suffering from NS, short stature (-3.9 and -5.4 SDS respectively) and low growth velocity (3.9 and 3.3 cm/year), were treated with rhGH (0.85 U/kg/week) for 1 year. Serum levels of total calcium (Ca), inorganic phosphate (P), magnesium (Mg), parathyroid hormone (PTH), calcitonin (CT), 25OH vitamin D, 1.25(OH)(2)D, osteocalcin (BGP), type I procollagen carboxy-terminal propeptide (PICP) and its telopeptide (ICTP) were measured. RESULTS: The baseline values were in the normal range; during the treatment no remarkable difference in the values of every one parameters was detected in the 2 patients studied. In one of them, who responded to GH treatment with significantly improved growth velocity, serum levels of the BGP increased during the first semester, and then progressively declined; conversely, serum levels of the ICTP remained stable during the first 6 months of GH-therapy, whereas increased in the following 6 months. CONCLUSION: The results suggest that in Noonan's syndrome patients responding to GH-therapy, a stimulation of bone turnover, with ensuing increase of height velocity, takes place, at least during the first year of GH-therapy. The authors underline the necessity of confirming their results on a larger group of patients with Noonan's syndrome.

Serum IgE in newborns undergoing exchange transfusion.

Serum IgE was studied in a group of 20 newborns, ten males and ten females, with neonatal non-immunologic jaundice before and after exchange transfusion and in a control group without jaundice or other immunologic problems. Serum IgE was significantly higher in the study group than in the control group (P less than .005). After exchange transfusion there was a further increase (P less than .005). These changes do not seem to be due to the IgE content of the transfused blood. It is suggested that neonatal jaundice and/or exchange transfusion may stimulate the IgE synthesis in the newborn.