Stephen E. Welty

Docosahexaenoic Acid and Amino Acid Contents in Pasteurized Donor Milk are Low for Preterm Infants

OBJECTIVEnTo evaluate whether pasteurized donor human milk meets the nutritional needs of preterm infants in terms of free fatty acid and amino acid contents.nnnSTUDY DESIGNnMilk samples were prospectively collected from 39 donors to the Mothers Milk Bank of Ohio. The fatty acid and amino acid compositions in donor milk samples were measured before and after pasteurization, and values were compared with previously published findings and preterm infant nutrition guidelines. The nutritional adequacy of donor milk for preterm infants was based on estimated daily intake of 150 mL/kg. Statistical significance was adjusted to account for multiple comparisons.nnnRESULTSnPasteurization did not appreciably affect donor milk composition. Docosahexaenoic acid level (0.1 mol wt %), and concentrations of glycine, aspartate, valine, phenylalanine, proline, lysine, arginine, serine, and histidine in donor milk were all significantly lower than previously reported concentrations in milk.nnnCONCLUSIONSnDonor milk is not substantially affected by pasteurization, but has low concentrations of docosahexaenoic acid and amino acids. Targeted nutritional supplementation of human donor milk for feeding preterm infants might be warranted.

The role of MAP kinase phosphatase-1 in the protective mechanism of dexamethasone against endotoxemia

AIMSnWe have previously shown that glucocorticoids induce the expression of MAP kinase phosphatase (Mkp)(a)-1 in innate immune cells. Since Mkp-1 is a critical negative regulator of the innate immune response, we hypothesize that Mkp-1 plays a significant role in the anti-inflammatory action of glucocorticoids. The specific aim of the present study is to understand the role of Mkp-1 in the anti-inflammatory function of glucocorticoids.nnnMAIN METHODSnWild-type and Mkp-1(-/-) mice were treated with different doses of dexamethasone and then challenged with different doses of lipopolysaccharide (LPS). The survival and blood cytokines were assessed. The effects of dexamethasone on cytokine production in wild-type and Mkp-1(-/-) primary macrophages ex vivo were also examined.nnnKEY FINDINGSnWe found that dexamethasone induced the expression of Mkp-1 in vivo. Dexamethasone treatment completely protected wild-type mice from the mortality caused by a relatively high dose of LPS. However, dexamethasone treatment offered only a partial protection to Mkp-1(-/-) mice. Dexamethasone attenuated TNF-alpha production in both wild-type and Mkp-1(-/-) mice challenged with LPS, although TNF-alpha production in Mkp-1(-/-) mice was significantly more robust than that in wild-type mice. Dexamethasone pretreatment shortened the duration of p38 and JNK activation in LPS-stimulated wild-type macrophages, but had little effect on p38 or JNK activation in similarly treated Mkp-1(-/-) macrophages.nnnSIGNIFICANCEnOur results indicate that the inhibition of p38 and JNK activities by glucocorticoids is mediated by enhanced Mkp-1 expression. These results demonstrate that dexamethasone exerts its anti-inflammatory effects through both Mkp-1-dependent and Mkp-1-indepent mechanisms.

Myocardial tissue Doppler changes in patients with bronchopulmonary dysplasia.

OBJECTIVEnTo determine whether tissue Doppler measurements provide sensitive measures of right ventricular function that correlate with the severity of bronchopulmonary dysplasia (BPD).nnnSTUDY DESIGNn21 subjects (6 control subjects with no/mild BPD, 7 patients with moderate BPD, and 8 patients with severe BPD) underwent limited echocardiograms with standard M-mode, mitral and tricuspid inflow velocities, and tissue Doppler features measured. BPD severity was scored by using the NICHD/NHLBI/ORD workshop rating scale by physicians blinded to the echocardiogram results. Groups were compared by using 1-way analysis of variance with post-hoc testing and linear regression.nnnRESULTSnPatients with moderate BPD had a higher early Doppler inflow velocity to the early tissue Doppler velocity ratio (E/E ratio; P = .03), corresponding to increased end diastolic pressure, compared with patients with no/mild BPD. Patients with severe BPD had a higher E/E ratio (P = .004) than patients with no/mild BPD. Linear regression demonstrated a correlation between BPD category and right ventricle E/E (P = .007, R(2) = 0.33) and left ventricular myocardial performance index (P = .02, R(2) = 0.28).nnnCONCLUSIONnIncreasing right ventricle E/E ratio correlates with clinical severity of BPD. Abnormal left ventricular myocardial performance index was noted to correlate with the grade of BPD. Further longitudinal studies of tissue Doppler echocardiographic assessment of cardiac function in premature infants with BPD are needed.

Maternal Docosahexaenoic Acid Supplementation Decreases Lung Inflammation in Hyperoxia-Exposed Newborn Mice

DHA is a long-chain fatty acid that has potent antiinflammatory properties. Whereas maternal DHA dietary supplementation has been shown to improve cognitive development in infants fed DHA-supplemented milk, the antiinflammatory effects of maternal DHA supplementation on the developing fetus and neonate have not been extensively explored. Pregnant C3H/HeN dams were fed purified control or DHA-supplemented diets (~0.25% of total fat) at embryonic d 16 and consumed these diets throughout the study. At birth, the nursing mouse pups were placed in room air (RA; 21% O(2)) or >95% O(2) (hyperoxia) for up to 7 d. These studies tested the hypothesis that maternal DHA supplementation would decrease inflammation and improve alveolarization in the lungs of newborn mouse pups exposed to hyperoxia. Survival, inflammatory responses, and lung growth were compared among control diet/RA, DHA/RA, control/O(2), and DHA/O(2) pups. There were fewer neutrophils and macrophages in lung tissues from pups nursed by DHA-supplemented dams than in those nursed by dams fed the control diet at 7 d of hyperoxia exposure (P < 0.015). Although differences due to hyperoxia exposure were observed, maternal diet did not affect keratinocyte-derived chemokine, macrophage inflammatory protein-2, IL-1β, or TNFα mRNA levels in pup tissues. Hyperoxia also induced NF-κB activity, but maternal diet did not affect NF-κB or PPARγ activities. In mice, DHA supplementation decreases leukocyte infiltration in the offspring exposed to hyperoxia, suggesting a potential role for DHA supplementation as a therapy to reduce inflammation in preterm infants.

Thioredoxin interacting protein inhibits hypoxia-inducible factor transcriptional activity

Vascular endothelial growth factor (VEGF) is required for proper lung development and is transcriptionally regulated in alveolar epithelial cells by hypoxia-inducible factor (HIF). Previous findings in a newborn mouse model of bronchopulmonary dysplasia (BPD) suggest that thioredoxin-interacting protein (Txnip) is a novel regulator of VEGF expression. The present studies were designed to test the hypothesis that Txnip negatively regulates VEGF through effects on HIF-mediated gene expression. To test this hypothesis, we first examined the levels of VEGF and Txnip protein in the lungs of 1-day-old newborn mice and E19 embryos and detected a significant inverse correlation. To elucidate the mechanisms underlying this relationship, we studied the effects of Txnip overexpression on HIF-mediated transcription using murine lung epithelial (MLE-12) cells. Overexpression of Txnip inhibited HIF-mediated reporter activity in both hypoxia and room air. Suppression of HIF activity by Txnip seemed to be independent of the ability of Txnip to bind to thioredoxin. Thus, our studies support a model in which Txnip is a potentially critical regulator of HIF-mediated gene transcription in the murine lung. Alterations in Txnip expression could alter lung VEGF expression in prematurely born human infants and contribute to the development of BPD.

Mitochondrial thiol status in the liver is altered by exposure to hyperoxia

Patients with poorly functioning lungs often require treatment with high concentrations of supplemental oxygen, which, although often necessary to sustain life, can cause lung injury. The mechanisms responsible for hyperoxic lung injury have been investigated intensely and most probably involve oxidant stress responses, but the details are not well understood. In the present studies, we exposed adult male C57/Bl6 mice to >95% O2 for up to 72 h and obtained lung and liver samples for assessment of lung injury, measurements of tissue concentrations of coenzyme A (CoASH) and the corresponding mixed disulfide with glutathione (CoASSG), as possible biomarkers of intramitochondrial thiol redox status. Subcellular fractions were prepared from both tissues for determination of glutathione reductase (GR) activities. Lung injury in the hyperoxic mice was demonstrated by increases in lung weight to body weight ratios at 48 h and by increases in bronchoalveolar lavage protein concentrations at 72 h. Lung CoASH concentrations declined in the hyperoxic mice, but CoASSG concentrations were not increased nor were CoASH/CoASSG ratios decreased, as would be expected for an oxidant shift in mitochondrial thiol-disulfide status. Interestingly, CoASSG concentrations increased (from 6.72+/-0.54 to 14.10+/-1.10 nmol/g of liver in air-breathing controls and 72 h of hyperoxia, respectively, P<0.05), and CoASH/CoASSG ratios decreased in the livers of mice exposed to hyperoxia. Some apparent effects of duration of hyperoxia on GR activities in lung or liver cytosolic, mitochondrial, or nuclear fractions were observed, but the changes were not consistent or progressive. Yields of isolated hepatic nuclear protein were decreased in the hyperoxic mice within 24 h of exposure, and by 72 h of hyperoxia, protein recoveries in purified nuclear fractions had declined from 41.8 to 14.8 mg of protein/g animal body weight. Concentrations of 10-formyltetrahydrofolate dehydrogenase were diminished in hepatic mitochondria of hyperoxic mice. A second protein in hepatic mitochondria of approximately 25 kDa showed apparent decreases in thiol content, as determined by fluorescence intensities of monobromobimane derivatives separated by SDS-PAGE. The mechanisms responsible for the observed effects and the possible implications for the adverse effects of hyperoxic therapies are not known and need to be investigated.

Total absence of the small bowel in a premature neonate

We report here an interesting unique case of total loss of small bowel in the absence of associated malrotation or gastroschisis. Total loss of small bowel, acquired in utero in the absence of associated anomalies such as malrotation or gastroschisis, has not been previously reported. Several reports of congenital short bowel exist. However, this uncommon finding is typically associated with malrotation. Babies with this condition often present with functional intestinal obstruction. Several cases in which infarction of gastroschisis and autolysis of the bowel, followed by in utero resolution of the abdominal wall defect, have been reported as a cause of congenital absence of the small bowel. We present here the first report, to our knowledge, of an infant with total absence of the small bowel without gastroschisis or malrotation.