Koichi Shinohara

Effects of iron-unsaturated human lactoferrin on hydrogen peroxide-induced oxidative damage in intestinal epithelial cells.

Human milk (HM) contains various bioactive antioxidants. Lactoferrin (Lf) has been assumed to be one of the major antioxidants in HM. We examined the antioxidative properties of iron-unsaturated human Lf (apo-hLf, the major form of Lf in HM) in two intestinal epithelial cell lines: (1) An intestinal epithelial cell line (IEC-6) were preincubated for 24 h with either 50 μg/mL of apo-hLf, iron-saturated human Lf (holo-hLf), iron-unsaturated bovine transferrin (apo-bTf), or 800 ng/mL of the iron-chelating compound deferoxamine (DFX), followed by hydrogen peroxide (H2O2) challenge to induce oxidative stress. Survival rates were significantly higher in the cells preincubated with apo-hLf and DFX than those preincubated with holo-hLf. (2) Caco-2 cells were preincubated with or without apo-hLf for 24 h, followed by an H2O2 challenge. Intracellular oxidative stress was assessed by a fluorescent probe, 2′,7′-dichlorodihydrofluorescein diacetate (DCF-DA). Fluorescent intensity of cell images and cell homogenates was significantly lower in the cells preincubated with apo-hLF than those preincubated without apo-hLF. Our study indicates that apo-hLf alleviates H2O2-induced oxidative damage in intestinal cells due to the iron-chelating capacity. Therefore, Lf in HM may act as an antioxidant in the gastrointestinal tract (GIT).

Suppressive effects of breast milk on oxidative DNA damage in very low birthweight infants

Background: Human milk contains many kinds of antioxidant and is considered to prevent diseases mediated by oxygen free radicals in very low birthweight (VLBW) infants. Aims: To examine the antioxidant effects of breast milk in VLBW infants by determining urinary 8-hydroxydeoxyguanosine (8-OHdG) excretion, which is known to be a non-invasive marker for in vivo oxidative DNA damage. Methods: Urinary 8-OHdG concentrations were measured in 15 breast fed and 14 formula fed VLBW infants at 2, 7, 14, and 28 days of age. Results: Urinary 8-OHdG excretion at 14 and 28 days of age was significantly lower than at 2 and 7 days of age in the breast fed group, and significantly lower than in the formula fed group. Conclusion: This is the first direct evidence of the antioxidant action of human milk in VLBW infants.

Effects of human milk and spermine on hydrogen peroxide-induced oxidative damage in IEC-6 cells

Objective: Oxidative stress is intimately involved in the pathologic processes of serious diseases in the perinatal period. Human milk (HM) contains various bioactive substances, some of which are known as antioxidants, including polyamines such as spermine (SPM). We examined the antioxidative properties of HM and SPM in an intestinal epithelial cell line. Method: Confluent Intestinal Epithelial Cells-6 (IEC-6) cells were preincubated with 100-fold dilutions of defatted HM, bovine milk, or three artificial milks for 24 hours, followed by hydrogen peroxide (H2O2) challenge (0.5 mM, 30 min) for oxidative stress. Cells were preincubated with either HM or increasing concentrations (within the range of HM) of SPM for 24 hours followed by an H2O2 challenge (0.25 mM, 30 min). Results: HM-treated cells showed the highest survival rate (50%) compared with no pretreatment (27%), bovine milk-treated (6%), or artificial formula-treated (13-16%) cells. Significantly higher survival rates were observed in the cells treated with HM (44.0%) and in those treated with 0.5, 1, or 5 μM of SPM (12.6, 13.1, or 22.2%, respectively) in comparison with the nontreated cells (7.0%). Conclusion: Our results demonstrated that HM and SPM alleviated H2O2-induced oxidative damage in IEC-6 cells, whereas bovine milk and artificial formula did not show any antioxidative capacity. These results suggest that HM acts as an antioxidant in the gastrointestinal tract of infants and that SPM plays an important role in the antioxidative properties of HM.

Plasma levels of active ghrelin until 8 weeks after birth in preterm infants: relationship with anthropometric and biochemical measures

This study investigated the relationship between plasma levels of ghrelin and postnatal growth in preterm infants. The levels of active ghrelin in cord blood and in plasma in 25 very low birthweight (VLBW) infants were measured. The results indicate that the levels of circulating active ghrelin markedly increases after birth in VLBW infants, and suggest that the increased levels of ghrelin reflects the maturation of ghrelin production in the stomach and an increased physiological need for ghrelin.

Comparison of the phospholipid classes in human milk in Japanese mothers of term and preterm infants

Background: Phospholipids (PLs) play an essential role in the growth and brain development of infants. Aim: To investigate PL composition in human milk (HM), including lysophosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine, phosphatidylcholine (PC) and sphingomyelin (SM), from healthy Japanese mothers. Analyses were performed on colostrum, transitional milk and mature milk from mothers of preterm and term infants. Methods: HM samples were collected from mothers of 15 term infants (term group) and of 19 preterm infants (preterm group). PL composition was determined by two‐dimensional thin‐layer chromatography in conjunction with phosphorus analysis. Results: In both groups, the PL content (% of total lipid) of mature milk was significantly lower than in colostrum. SM and PC were the main PLs in HM, but in the preterm group, the percentage of SM in mature milk was significantly higher and PC in mature milk was significantly lower than in the term group.

Growth Factors in Breast Milk and Their Effect on Gastrointestinal Development

Breast milk contains various biologically active factors including, hormones, peptide growth factors, and cytokines. Epidermal growth factor (EGF) and insulinlike growth factor-I (IGF-I) are two of the major milk-derived peptide growth factors. Colostrum contains higher levels of these growth factors than mature milk does, and, these factors are relatively resistant to proteolysis and stable in the G-I tract. There are specific receptors found in G-I mucosa. Luminal EGF and IGF-I stimulate growth and development of gastrointestinal tract. Intestinal epithelial Caco-2 cells are a good model for studying physiological roles of exogenous growth factors in the G-I development. Effect of EGF and IGF-I on proliferation, differentiation, and IGF-binding protein (IGFBP) production of intestinal Caco-2 cells were studied. Both EGF and IGF-I increased cell proliferation in dose dependent manner. The number of IGF-I receptors on Caco-2 cells increased after differentiation, in contrast to EGF binding which was reported to decrease. Caco-2 cells produced at least three IGFBPs, namely IGFBP-2, -3, and -4. The profile of these IGFBPs varied with differentiation. Secretion of IGFBP-2 and -3 increased with differentiation, but IGFBP-4 diminished. IGF-I stimulated mainly IGFBP-3 production, while EGF stimulated predominantly IGFBP-4. The effects of IGF-I and EGF on IGFBP secretion diminished with increasing cell differentiation. Thus, the interaction between intestinal epithelial cells and extrinsic growth factors are complex and the stage of differentiation is an important determinant of this phenomenon.

Lack of plasma leptin response to feeding in newborn infants.

Objective:  Previous reports of the postprandial regulation of leptin are controversial, and there have been few studies on the effects of breast‐feeding on postprandial regulation in newborn infants. We examined the response of plasma leptin to breast‐ and formula‐feeding in newborn infants.

Hypertrophic pyloric stenosis in mono-ovular extremely preterm twins after use of erythromycin.

Although male predominance (male : female ratio, 4 – 5:1), familial recurrence and concordance of HPS in twins suggests some polygenetic relationship, 3 the pathogenesis and inheritance patterns underlying HPS remain relatively uncharacterized. There are many indications for administration of erythromycin, a macrolide antibiotic, in the neonatal period. These indications include promotion of gastrointestinal motility, 4 reduction of Ureaplasma urealyticum colonization in the respiratory tract 5 in preterm infants, and prophylaxis against pertussis infection. 6 Since the SanFilippo report in 1976, 7 however, several case series and cohort studies have suggested that early exposure to erythromycin increases the risk of HPS. 8