Lactase phlorhizin hydrolase turnover in vivo in water-fed and colostrum-fed newborn pigs.

We have estimated the synthesis rates in vivo of precursor and brush-border (BB) polypeptides of lactase phlorhizin hydrolase (LPH) in newborn pigs fed with water or colostrum for 24h post partum. At the end of the feeding period, piglets were anaesthetized and infused intravenously for 3h with L-[4-3H]- phenylalanine. Blood and jejunal samples were collected at timed intervals. The precursor and BB forms of LPH were isolated from jejunal mucosa by immunoprecipitation followed by SDS/PAGE, and their specific radioactivity in Phe determined. The kinetics of precursor and BB LPH labelling were analysed by using a linear compartmental model. Immunoisolated LPH protein consisted of five polypeptides [high-mannose LPH precursor (proLPHh), complex glycosylated LPH precursor (proLPHe), intermediate complex glycosylated LPH precursor (proLPH1i) and two forms of BB LPH]. The fractional synthesis rate (Ks) of proLPHh and proLPHc (approx. 5%/min) were the same in the two groups but the absolute synthesis rate (in arbitrary units, min-1) of proLPHh in the colostrum-fed animals was twice that of the water-fed animals. The Ks values of proLPHi polypeptides were significantly different (water-fed, 3.89%/min; colostrum-fed, 1.6%/min), but the absolute synthesis rates did not differ. The Ks of BB LPH was not different between experimental treatment groups (on average 0.037%/min). However, the proportion of newly synthesized proLPHh processed to BB LPH was 48% lower in colostrum-fed than in water-fed animals. We conclude that in neonatal pigs, the ingestion of colostrum stimulates the synthesis of proLPHh but, at least temporarily, disrupts the processing of proLPH polypeptides to the BB enzyme.

Lenten cell: ultrastructure, absorptive properties, and enzyme expression of a novel type of cell in the newborn and suckling pig intestinal epithelium.

BACKGROUND: The small intestinal epithelium is made up of columnar absorptive enterocytes and a smaller number of specialized non-absorptive cells, including goblet cells, enteroendocrine cells, M cells, cup cells, and tuft cells. During a study on milk protein absorption in newborn pigs, we identified an enterocyte that showed no uptake of milk proteins and that could be found only in the jejunum and ileum of pigs during the first 2 weeks of life. We call this previously undescribed enterocyte the lenten cell. METHODS: We used light microscopy and transmission electron microscopy in conjunction with immunolabelling and cytochemical techniques to determine the occurrence, ultrastructure, absorptive properties, and brush border hydrolase expression of lenten cells. RESULTS: Lenten cells constituted approximately 1-2% of the villous epithelium. They were seen in newborn and suckling pigs 1-9 days of age, but were not found in weaned pigs. Morphologically, lenten cells were spindle- or wineglass-shaped, with a ventrally sited nucleus and an electron-dense cytoplasm with numerous cytokeratin filaments. Lenten cells had a normal brush border with microvilli that were slightly thicker than those of absorptive enterocytes, but they did not express the brush border hydrolases lactase, aminopeptidase N, and alkaline phosphatase. Lenten cells did not endocytose milk proteins or horseradish peroxidase, but contained some endocytic or secretory vacuoles and a few dense granules. CONCLUSIONS: No role for lenten cells has been identified in this study, but presence of these cells during the neonatal period, when growth and differentiation of the gastrointestinal tract is at a peak, clearly suggests that lenten cells may play a role in this process.

Pathogenesis of small-intestinal mucosal lesions in chronic diarrhea of infancy: II. An electron microscopic study.

Our electron microscopic study of biopsies taken from 10 infants with protracted diarrhea was conducted in an effort to determine the pathogenesis of the disorder. In this article, the ultrastructure of the jejunal mucosa of the infants is described in relation to overlying or adherent bacteria of unidentified type. In addition to the known changes on the enterocyte surface caused by adherent bacteria (cupping and effacement), other cytopathic changes, not previously reported, are documented. Included are widespread loss of enterocytes, including intraepithelial lymphocytes, into the bowel lumen; cytopathological changes within the enterocytes; and marked thickening of the basal laminae of the enterocytes and the endothelium of lamina propria blood vessels. In addition, we noted deposition of collagen fibrils in the lamina propria below the basal laminae, active phagolysis within macrophages, and lack of cisternal material (immunoglobulin) in the plasma-cell cytoplasm. Although these changes are nonspecific, they may be related in part to the presence of the nonadhering and adhering bacteria, and their identification may further our understanding of the "sick mucosa" that occurs in chronic diarrhea of infancy.

Human lactoferrin stimulates thymidine incorporation into DNA of rat crypt cells.

In a search for dietary factors that might stimulate enterocyte proliferation, we developed an assay for thymidine incorporation into DNA using harvested crypt cells from mature rat small intestine. Human colostrum stimulated a significant increase in thymidine incorporation into rat crypt cell DNA during a 60-min period of incubation. When the protein with biological activity was purified to a single peak by sequential ion exchange and gel filtration chromatography, it was found to have the characteristics of lactoferrin. The protein was identical to lactoferrin standards by sodium dodecyl sulfate polyacrylamide gel electrophoresis, isoelectric focusing, and double-diffusion immunologic precipitation. All available human lactoferrins stimulated thymidine uptake and all reacted with a lactoferrin polyclonal antibody. Human lactoferrin appears to be a potent activator of thymidine incorporation into DNA in incubated rat crypt cells, a nutritional function not previously reported.

Randomized study of sunshine exposure and serum 25-OHD in breast-fed infants in Beijing, China.

PIP: The hypothesis that breastfed infants in Beijing, China, have low vitamin D status and that sunshine exposure increases serum 25-hydroxyvitamin D (25-OHD) concentrations was tested in a randomized prospective study involving 42 healthy infants 1-8 months of age. 24 controls were exposed to the usual amount of sunshine, while the 18 cases were provided with 2 hours of sunshine/day. All infants were being breastfed without vitamin D supplementation. Serum 25-OHD concentrations at entry into the study ranged from 3-61 ng/ml and increased with age. 3 infants in each group were diagnosed as having rickets. Serum 25-OHD concentrations did not change significantly during the 2-month study period among controls, while serum 25-OHD values increased significantly in the experimental group. Final infant serum 25-OHD concentrations correlated with ultraviolet exposure scores (p.001). The estimate of ultraviolet score needed to maintain a serum 25-OHD of 11 ng/ml was 2.4 (24 minutes/day with only the face uncovered). Rickets has been reported in as many as 18% of Chinese infants and seems to especially affect those 2-4 months of age. The fact that not all infants with rickets in this study had low 25-OHD concentrations suggests 2 possibilities: 1) not all rickets is necessarily related to a vitamin D deficiency, or 2) serum 25-OHD concentrations are not the best indicator of vitamin D status. The low serum 25-OHD concentrations in many Chinese infants may be related to low maternal milk vitamin D content resulting from the lack of vitamin D food fortification. Overall, these results suggest that the diet of breastfed Chinese infants should be supplemented with orally administered vitamin D.^ieng

Fortified mothers' milk for very low birth weight infants: results of growth and nutrient balance studies.

Mothers' milk, fortified daily with skim and cream components derived from mature donor human milk, was fed fresh during the first 2 postnatal months to 18 (group FMM) healthy, very low birth weight (VLBW) infants. Following the same feeding protocol, 16 additional VLBW infants were fed commercial formula that had a similar range of energy and nitrogen. Infants were evaluated for growth, metabolizable energy, balances of nitrogen and fat, and biochemical markers of nutritional status. Growth measurements from birth to 1800 gm were similar in the two groups. Nutritional balance was performed at approximately 2.5 (first study period) and 6 (second study period) weeks. Metabolizable energy (average 109 kcal/kg/day) was similar in both groups and met theoretical needs of the growing VLBW infant. Apparent nitrogen retention was similar and equivalent to estimates of intrauterine nitrogen accretion (approximately 325 mg/kg/day). Fat absorption increased from the first (70% of intake) to the second study (80% of intake) and was similar in both groups. Serum values for albumin, total protein, prealbumin, hemoglobin, and creatinine were also similar, and declined with increases in postnatal age. Significant differences were observed between feeding groups for serum phosphorus and calcium. These results indicate that healthy VLBW infants maintain adequate growth and macronutrient balance for the first 2 months postnatally when fed mothers' milk fortified with additional skim and cream components.

Longitudinal changes in milk composition of mothers delivering preterm and term infants.

The concentrations of protein nitrogen (PN), non-protein nitrogen (NPN), energy, fat, sodium (Na), calcium (Ca), phosphorus (P), magnesium (Mg), and zinc (Zn) were determined in human milk from mothers giving birth to full-term (n = 13) and preterm infants (n = 8). Milk samples were collected under controlled conditions at two-week intervals for 12 weeks postpartum. Statistically significant differences in PN, Ca, and P concentrations were detected between the milk from mothers of preterm and term infants. The mean PN concentration in the preterm milk was statistically higher than that of term milk (198 vs. 164 mg N/dl), in contrast to the lower mean Ca (220 vs. 261 mg/l) and P (125 vs. 153 mg/l) concentrations detected in the preterm milk. No other differences in mean nutrient concentration were observed between the two groups. Concentrations of PN, NPN, Na, P, and Zn decreased over time. The concentration of Mg increased slightly. The content of fat, energy, and Ca did not change.

Studies of human milk relevant to milk banking.

Issues regarding the efficacy of feeding human milk to premature infants include the development of optimal protocols for collecting, storing, and processing human milk. Studies of the nutritional and immunologic composition of milk produced by women who delivered term or premature infants and who weaned their infants gradually from human milk have been studied to identify optimal donors. The effects of specific collection, storage, and processing conditions on the composition of mature human milk also have been evaluated. Collection, storage, and processing conditions have distinct effects on specific functional components. The caloric content of milk, the content of nutrients carried in the lipid portion of milk, and selected enzymatic activities depend on the completeness with which the breast is emptied. Storing milk in polyethylene, polypropylene, and pyrex containers influences key immunologic components in human milk as do storage temperatures. None of the nutrient compositions of milks studied matched current estimates of the nutritional needs of premature infants. Importantly, both the concentrations and the pattern of change in nutrient and immunologic contents are distinct in milks of women delivering infants at term or prematurely. Further changes are seen during the process of gradual weaning.

Immunologic factors in human milk during the first year of lactation.

The effects of the duration of lactation upon lactoferrin, lysozyme, total IgA, SIgA, SIgA antibodies to Escherichia coli somatic antigens and leukocytes in human milk were investigated. Longitudinal and cross-sectional studies were performed with milk collected from women 20 to 35 years of age during te first year of lactation. Collection and storage conditions and immunologic analyses were controlled to minimize confounding variables. The concentrations of lactoferrin, total IgA, and leukocytes and the uptake of 3H-thymidine by phytohemagglutinin-stimulated lymphocytes fell during the first several weeks of lactation; afterward, the levels of lactoferrin and IgA stabilized. Approximately 90% of total IgA in human milk during the year was SIgA. Secretory IgA antibody titers to E. coli increased in some individuals studied longitudinally suggesting that the enteromammary gland pathway of SIgA antibody production was active after several weeks of lactation. Moreover, the concentrations of lysozyme, after falling to a nadir of 20 to 30 micrograms/ml at 2 to 4 weeks, rose to 200 to 300 micrograms/ml by six months and remained elevated. The immunologic system in human milk undergoes remarkable changes which may represent adaptations for the recipient infant.

Intracellular concentration of elements in normal and dystrophic skeletal muscles of the chicken.

In this study, the intracellular concentrations of six elements (mmole/kg dry weight) were directly measured in the muscle fibers of pectoralis major muscles of eight week old, genetically dystrophic and normal chickens by the X-ray microanalysis technique. The extent of muscle degeneration was evaluated by morphometric measurements of muscle fiber diameter and other histological changes. A significant increase in the concentration of intracellular sodium and chlorine was evident in dystrophic muscles. The concentration of intracellular sodium was 127.0 +/- 35.0 in the muscle fibers of dystrophic chicks compared to 65.7 +/- 16.5 in normal controls. The concentration of chlorine was 90.5 +/- 27.5 and 54.1 +/- 5.5 in the muscle fibers of dystrophic and normal chicks respectively. The intracellular concentrations of potassium, magnesium, phosphorous, and sulfur remained unchanged in the dystrophic condition. Morphometric studies revealed that the dystrophic pectoralis muscles contain fewer but thicker fibers per unit area compared to normal pectoralis muscles. The importance of these findings are discussed in relation to the results of earlier investigations.

Fiber, intestinal sterols, and colon cancer.

It has been postulated that dietary fiber's protective effect against the development of colon cancer, diverticular disease, and atherosclerosis may be due to the adsorption and/or dilution of intestinal sterols such as bile acids and neural sterols and their bacterial metabolites by component(s) of fiber. Dietary fiber is made up of four major components-cellulose, hemicellulose, lignin, and pectin. There is evidence that hemicellulose and pectin may induce an increase in fecal bile acid excretion in man which may be accompanied by a decrease in serum cholesterol. Natural fibers, such as rolled oats, alfalfa, guar gum, and Bengal gram have been shown to have hypocholesterolemic properties of alfalfa, wheat straw, and some other fibers found considerable amounts of bile acids in vitro. On the other hand, wheat bran, oat hulls, and all the synthetic fibers tested bound only negligible amounts of bile acids under the same conditions. Vegetarians in the United States have lower plasma lipids and different plasma lipoprotein patterns than those of comparable control populations on regular mixed diet. They also have smaller daily fractional turnover rates of cholic acid and deoxycholic acid pool size. In addition, populations on a mixed Western diet, where the rate of large bowel cancer is high (North American, English, Scottish, etc.) degraded and excreted cholesterol and bile acid metabolites to a greater degree than populations where the rate of colon cancer is comparatively low (Ugandan, Japanese, etc). It cannot be denied that the fiber theory linking fiber deficiency with the development of colon cancer and other diseases, is simple, attractive and appears to be firmly based in common sense. When subjected to research studies, however, the situation appears much more complex than expected. Although some progress is being made, the data are often contradictory and confusing, probably due to lack of adequate documentation of fiber intake (e.g., use of dietary fiber instead of crude fiber) and/or the absence of detailed information on the chemistry of the fiber itself.

Magnesium supplementation in protein-calorie malnutrition.

The widespread observation of magnesium depletion in edematous malnutrition has been confirmed in Guatemalan children. The magnesium requirement during initial stages of therapy has been estimated as 2.7 mEq/kg per day. This may be achieved by adding 0.5% MgSO4.7H2O to a solution containing 15% dextromaltase and 1.5% KCl which is used to dilute whole milk; two parts milk and one part dilution mixture. The replacement of magnesium deficits was not essential for recovery from edematous malnutrition, however, the present evidence suggested that the rate of recovery was accelerated by approximately 2 weeks in those children who received the supplement.

Electrolyte metabolism of muscle in the salt-losing form of congenital adrenal hyperplasia.

The study of metabolism of muscle electrolyte in children with the salt-losing form of congenital adrenal hyperplasia reveals two types of alterations. After admission and during initial therapy with salt and desoxycorticosterone, the changes are typical of those seen in experimental animals with adrenalectomy and excessive replacement therapy. Discontinuation of the sodium supplement after three months of therapy resulted in a return of muscle electrolyte values to normal. During the period of poor growth common to these patients a different pattern was observed. Sodium and water accumulated without alteration in tissue potassium. The mechanism of this alteration is not clear; however, it is consistent with the known effects of excess cortisone on muscle composition. These observations permit the conclusion that at least two fractions of sodium are present in muscle fibers, that which exchanges potassium and that which is independent of potassium metabolism.