摘要:

Patients with cystic fibrosis are severely affected by an infection withPseudomonas aeruginosa,a microbe known to synthesize phospholipase C. This study was designed to determine whether that enzyme would affect the function of pulmonary surfactant phospholipids. Mucoid and nonmucoid strains ofP. aeruginosa, freshly obtained from patients with cystic fibrosis, were cultured for 12 h on agar plates. The bacteria were suspended in saline solution and then pelleted by centrifugation. The supernatant was used to dilute the surfactant preparation, calf lung surfactant extract, from 35 to 2 mg/mL. Surfactant function, before and after incubation, was examined with a capillary surfactometer, an instrument specifically developed for an evaluation of the ability of surfactant to maintain patency of a narrow glass tube, simulating a terminal conducting airway. Phospholipid hydrolysis was also evaluated biochemically by determining the total content of phospholipids in surfactant before and after incubation. In five experiments, the lipids were separated with thin-layer chromatography, and the phosphorus content was determined in the diacylphosphatidylcholine band before and after incubation for 6, 24, and 48 h. Capillary openness and phospholipid concentration decreased as enzyme concentration and time of incubation increased (p< 0.0001). Linear regression showed a significant correlation between time of capillary openness and phospholipid concentration (r= 0.957;p< 0.0001). Calf lung surfactant extract hydrolysis was catalyzed by extracts of the bacteria, particularly the nonmucoid, analogous to the catalysis observed with phospholipase C. Surfactant hydrolysis catalyzed by enzymes fromP. aeruginosamight severely affect surfactant function provided enzyme concentration is high and time of incubation is long.

ISSN:0031-3998    

出版商:OVID    

年代:2000

数据来源: OVID

摘要:

The present study was designed to evaluate the effect of gestational age and intrauterine growth on the long chain polyunsaturated fatty acid (LCP) synthesis from dietary precursors in neonates as reflected by plasma pools. These have been considered conditionally essential nutrients for normal growth, sensory maturation, and neurodevelopment.In vivoelongation/desaturation of deuterated d5-linoleic acid (d5-LA) to form arachidonic acid (AA), and d5-&agr;-linolenic acid (d5-LNA) to form docosahexaenoic acid (DHA), was studied in 19 preterm appropriate-for-gestational-age (AGA) infants, 11 AGA term, and 11 intrauterine growth-retarded (IUGR) infants. They received a dose of 50 mg/kg body weight of d5-LA and d5-LNA enterally during the first days of life; d5-labeled derivatized fatty acids were determined in blood samples obtained at 24, 48, and 96 h after dosing. Lipids were extracted and fatty acids analyzed by gas chromatography and negative ion mass spectrometry. Mean concentrations, &mgr;g/mL, and d5/d0 for n-3 and n-6 precursor and products were computed at various times and were also integrated over the complete study period. Significantly higher time-integrated concentration of d5-AA and d5-DHA were observed in preterm infants relative to the other two groups. Time-integrated enrichment of DHA relative to LNA was 100-fold lower in preterms, 410-fold lower in term, and 27-fold lower in IUGR infants. Similar significant declines in product to precursor enrichments were noted for the n-6 series. A significant negative correlation of AA and DHA formation based on time-integrated d5/d0 ratios with gestational age was noted; product/precursor enrichmentversusgas chromatography for the n-6 series had anrof −0.5,p= 0.001, and for the n-3 series had anrof −0.6,p= 0.0001. Birth weight or weight adequacy did not add further strength to the relationship. We conclude that LCP formation from deuterated precursors occurs as early as 26 wk gestation, and in fact is more active at earlier gestational ages; growth retardation appears to slow down or diminish LCP formation. No quantitative estimates of LCP synthesis or nutritional sufficiency can be derived from these data.

ISSN:0031-3998    

出版商:OVID    

年代:2000

数据来源: OVID

摘要:

To explore the roles of lactogens in islet function, we generated a stable line of rat insulinoma (INS-1) cells that express rat placental lactogen II (rPLII) constitutively in culture. We used this cell line (Ins-rPLII) to examine the effects of endogenous rPLII on &bgr;-cell growth, islet formation, and the expression of glucose transporter 2 (glut-2) and insulin mRNA. Growth and maturation of Ins-rPLII cells were compared with that of cells transfected stably with an empty expression plasmid (control) and of INS-1 cells treated with exogenous prolactin. The Ins-rPLII cells proliferated more rapidly than control cells in serum-free medium and showed distinct morphologic characteristics in culture. Whereas the control cells flattened readily on plastic and formed a branching monolayer, the Ins-rPLII cells remained more rounded, sent out fewer projections, and formed more numerous (p< 0.01) and larger (p< 0.01) &bgr;-cell clusters. Larger clusters assumed a spherical form with well-delineated smooth borders and detached more readily from the culture plates. Maturational progression of the Ins-rPLII cells was associated with a 40% increase in preproinsulin mRNA (p< 0.05) and a 2–3-fold increase in glut-2 mRNA (p< 0.01). Induction of glut-2 mRNA was accompanied by a 1.4–2.4-fold increase (p< 0.01) in the uptake of radiolabeled 2-deoxyglucose. Similar effects were observed in INS-1 cells exposed for 48 h to exogenous prolactin. These findings suggest novel roles for the lactogenic hormones in the maturation and growth of pancreatic islets. Lactogen induction of &bgr;-cell aggregation coupled with localized &bgr;-cell growth may contribute to the expansion of islet mass that occurs in pregnancy and during the perinatal period. The induction of insulin and glut-2 mRNA provides a mechanism by which the lactogens may increase fetal and maternal insulin production and enhance the sensitivity of the pancreas to glucose.

ISSN:0031-3998    

出版商:OVID    

年代:2000

数据来源: OVID

摘要:

There is increasing evidence that the intrauterine milieu and corticosteroid exposure play a role in the etiology of hypertension. We examined adrenocortical gene expression and circulating corticosteroids in the d 21 fetal spontaneously hypertensive rat (SHR) and its normotensive genetic control, the Wistar-Kyoto (WKY) rat. By RNase protection assays, we found no differences in the relative abundances of mRNAs for P450scc and P450c11&bgr;, and barely detectable P450c11AS mRNA in the adrenals of fetal SHR and WKY rats. P450c11B3 RNA was undetectable by reverse transcription polymerase chain reaction in both SHR and WKY fetuses. The zonal expression of P450c11 mRNA was comparable in SHR and WKY fetuses byin situhybridization histochemistry. There were no significant differences in peripheral levels of aldosterone and corticosterone by radioimmunoassay in fetal SHR and WKY rats. Based upon the absence of distinct differences in the aspects of adrenocortical activity examined, it is unlikely that they are integral in the programming of hypertension in this model.

ISSN:0031-3998    

出版商:OVID    

年代:2000

数据来源: OVID

摘要:

Premature infants are at risk of developing metabolic bone disease mainly because of low calcium and phosphorus intake. We have examined the effect of different mineral supplements on bone mineral content at term in 127 premature infants with gestational age <32 wk in a double-blinded randomized trial. We used either phosphate supplementation of human milk as recommended by the European Society of Pediatric Gastroenterology and Nutrition or fortified supplementation with protein, calcium, and phosphorus or preterm formula as recommended by the American Academy of Pediatrics. The intervention period was from 1 week old until 36 wk of gestational age, and the infants were fed approximately 200 mL·kg−1·d−1. Bone mineral content was measured at term by dual-energy x-ray absorptiometry scan. Surprisingly, neither phosphate, fortifier, nor preterm formula supplementation had any significant effect on bone mineral content at term compared with infants fed their own mother’s milk only. There was a tendency to higher total bone mineral content in infants fed preterm formula compared with infants fed their own mother’s milk only (p= 0.05), but when the bone mineral content was corrected for the size of the infant, there was no difference (p= 0.68). Infants fed preterm formula had a significantly higher weight at term compared with infants fed their own mother’s milk only (p= 0.02), but did not differ significantly in length or head circumference. In a regression analysis, the amount of supplemented phosphorus was significantly associated with weight at term (p= 0.008). We conclude that when feeding 200 mL·kg−1·d−1, mineral supplementation of human milk or use of preterm formula does not significantly improve bone mineralization outcome at term.

ISSN:0031-3998    

出版商:OVID    

年代:2000

数据来源: OVID

摘要:

The allergy-preventing effect of breast-feeding remains controversial, possibly because of individual variations in the composition of the breast milk. The aim of this study was to investigate the concentrations of cytokines involved in allergic reactions and IgA antibody production in breast milk from allergic and nonallergic mothers. The cytokine concentrations were determined in colostrum and 1-mo milk samples from 24 mothers with, and 25 mothers without, atopic symptoms, using commercial ELISA kits. The immunosuppressive cytokine transforming growth factor-&bgr; was predominant and was detectable in all milk samples. IL-6 was detected in the majority of colostral and mature milk samples, whereas the other cytokines were less commonly detected. The concentrations of IL-6, IL-10, and transforming growth factor-&bgr;, which are all involved in IgA synthesis, correlated with each other and with total IgA concentrations in colostrum. The concentrations of IL-4 were higher in colostrum from allergic than nonallergic mothers, and similar trends were seen for IL-5 and IL-13. In conclusion, transforming growth factor-&bgr; and IL-6 were the predominant cytokines in human milk. The correlation between the concentrations of cytokines involved in IgA synthesis,i.e.IL-10, IL-6, and transforming growth factor-&bgr;, may explain the stimulatory effect on IgA production in breast-fed babies. Varying concentrations of IL-4, IL-5, and IL-13 may explain some of the controversy regarding the possible allergy-preventive effect of breast-feeding.

ISSN:0031-3998    

出版商:OVID    

年代:2000

数据来源: OVID

摘要:

Whole-body degradation rates of transfer, ribosomal, and messenger RNA were determined noninvasively in 3-, 6-, 10-, 14-, and 18-y-old female and male subjects (n= 14 per age group per sex) under normal living conditions. The method for determining the RNA degradation rates is based on measuring the renal excretion rates of special RNA catabolites (modified ribonucleosides and nucleobases) by HPLC. Resting metabolic rates were calculated for the same subjects by their body weights using formulas taken from literature. We found high correlations between the degradation rates of the different RNA classes (micromoles per day per kilogram body weight) and the resting metabolic rate (kilojoules per day per kilogram body weight): in females (n= 70),r= 0.75–0.82 and in males (n= 70),r= 0.68–0.79 (p< 0.0001). We conclude that a causal relationship exists between the whole-body degradation rates of the different RNA classes and the resting metabolic rate. Therefore, in healthy subjects noninvasive determinations of RNA degradation rates could be very useful to assess the resting metabolic rate.

ISSN:0031-3998    

出版商:OVID    

年代:2000

数据来源: OVID