Trehalose synthesis in Saccharomycopsis fibuligera does not respond to stress treatments.

Synthesis of trehalose in Saccharomycopsis fibuligera sdu under various stress conditions was investigated. Neither the activation of trehalose-6-phosphate synthase (SfTPS1) nor the change in trehalose content was observed under stress exposure of S. fibuligera sdu cells. The results of reverse transcription polymerase chain reaction, which was performed with the specific primers designed to target the SfTPS1 gene fragment cloned from this strain, also showed that all stress treatments did not increase the expression of SfTPS1 gene. These results demonstrated that synthesis of trehalose in response to stress conditions in S. fibuligera sdu clearly differs from that of Saccharomyces cerevisiae and most other fungi. The phylogenetic analysis of the amino acid sequence deduced from the SfTPS1 gene fragment showed that the SfTPS1 sequence formed a separate family that was far related to S. cerevisiae TPS1. The yeast strain, which can accumulate a large amount of trehalose under normal growth conditions, has many applications and TPS1 gene in such strain may have unique use in transgenic organisms.

Trehalose accumulation in a high-trehalose-accumulating mutant of Saccharomycopsis fibuligera sdu does not respond to stress treatments.

The isolation of high-trehalose-accumulating mutant A11 from Saccharomycopsis fibuligera sdu has been previously described. In this paper, accumulation of trehalose under various stress conditions in S. fibuligera A11 was investigated. Neither activation of trehalose-6-phosphate synthase (SfTps1) nor change in trehalose content was observed under stress exposure of S. fibuligera A11 cells. A fragment of the Sftps1 gene in this strain was also cloned by degenerate PCR using the CoDeHOP strategy and multiply-aligned Tps1 sequences. This sequence allowed us to investigate the expression of the Sftps1 gene, which was also kept constant under the various stress conditions. Altogether, these results indicate that trehalose metabolism in S. fibuligera A11 in response to stress conditions clearly differs from that of Saccharomyces cerevisiae and most other fungi. The expression of the Sftps1 gene was not responsive to different stress treatments.

Bone turnover response to changes in calcium intake is altered in girls and adult women in families with histories of osteoporosis.

Heredity and environmental factors contribute to the development of osteoporosis. Because calcium is the major mineral in bone and adolescence is a key period in bone acquisition, we hypothesized that bone turnover would be less responsive to alterations in dietary calcium intake in both girls and adult women from families with histories of osteoporosis. To address this issue, we studied calcium kinetics in the maternal grandmother (age range 56-81 years), mother (age range 32-47 years), and granddaughter (age range 8-15 years) in 10 multigenerational families. In five families, the mother and/or grandmother had osteoporosis (bone mineral density > or = 2 SD below the age-specific mean). To examine both active and passive calcium absorption, families consumed low- (279 +/- 64 mg/day) and high- (1580 +/- 385) calcium diets for 10 days prior to administration of oral (46Ca) and intravenous (42Ca) stable isotopes. Using repeated measures analysis of variance, fractional calcium absorption, true calcium absorption, bone calcium deposition, and the balance in bone calcium turnover were all significantly affected by diet (p < 0.01). Females from nonosteoporotic families had decreased bone calcium resorption with little change in bone calcium deposition during the high-calcium study. In contrast, girls and adult women from osteoporotic families had increased both bone calcium deposition and resorption during the high-calcium period, leading to a less positive balance in bone calcium turnover. A significant interaction between bone status and diet was found for bone calcium resorption (p < 0.05) and approached significance for bone calcium deposition (p < 0.07), effects which were independent of generation. We conclude that girls and women from osteoporotic families have a significantly altered bone turnover response to acute changes in calcium intake.

Variables related to urinary calcium excretion in young girls.

The relations among dietary and calcium kinetic factors and 24-h urinary mineral excretion were evaluated in a group of 89 healthy girls (51 white and 38 black) aged 4.9-16.7 years. Nutrient intakes were calculated for each participant using a weighed intake of all food and beverage on the day of the 24-h urine collection study and two subsequent 24-h food records. A significant relation was noted between urinary calcium and sodium excretion (r = 0.55; p < 0.0001). No significant relations were found between urinary calcium and (a) calcium intake (r = 0.08), (b) protein intake (r = 0.14), or (c) phosphorus intake (r = 0.11). Urinary calcium was not significantly related to fractional calcium absorption (r = 0.03) or net calcium absorption (r = 0.11), but was significantly associated with the bone calcium deposition rate (r = 0.24; p < 0.03). Using a multiple regression model, both urinary sodium and the bone calcium deposition rate were independent predictors of urinary calcium excretion in this population (r = 0.57; p = 0.0001). A substantial number of the children in this population had urinary calcium excretion > 4 mg/kg/day (12%). The incidence of hypercalciuria differed between the racial groups and was markedly higher in the white than in the black children (17.6 vs. 5.3%). Over a range of usual calcium intakes, during the rapid-bone-growth period in childhood and early adolescence, urinary calcium appears relatively unaffected by calcium intake and is most strongly associated with urinary sodium levels.

Increased efficiency of calcium absorption during short periods of inadequate calcium intake in girls.

Adequate calcium intake is essential for skeletal integrity, particularly during the period of peak bone mass acquisition from 9 to 17 y of age. Currently, the calcium intake of many adolescent girls is below the recommended dietary allowance. The purpose of this study was to evaluate the ability of girls to respond to acute periods of inadequate dietary calcium intake. Calcium absorption was evaluated in 11 girls aged 11.6 +/- 2.4 y after 10 d on both a low-calcium (7.05 +/- 2.03 mmol/d) and a high-calcium (35.30 +/- 2.28 mmol/d) diet. Fractional calcium absorption was determined by using oral (46Ca) and intravenous (42Ca) stable isotopes of calcium. During a low calcium intake, fractional calcium absorption was significantly greater (0.582 +/- 0.087 compared with 0.260 +/- 0.068, P < 0.0001) and urinary calcium excretion was significantly lower (1.30 +/- 0.83 compared with 3.08 +/- 1.98 mmol/d, P < 0.004) than values obtained during a high calcium intake. Concentrations of 1,25-dihydroxyvitamin D (combination of cholecalciferol and ergocalciferol) were greater during the low calcium intake, although the difference was not significant (108.7 +/- 30.6 compared with 90.0 +/- 25.1 pmol/L, P < 0.1; n = 9). Excretion of N-telopeptide was significantly greater during the low calcium intake (761 +/- 508 compared with 413 +/- 341 nmol bone collagen equivalent (BCM)/mmol creatinine, P < 0.02; n = 9), indicating that bone resorption was increased. These results suggest that during short periods of inadequate calcium intake, girls are able to significantly increase the efficiency of calcium absorption and decrease urinary calcium losses to conserve calcium required for bone mineral acquisition.

Absorption by 1-year-old children of an iron supplement given with cow's milk or juice.

A paucity of data are available on toddlers for the evaluation of optimal strategies of Fe supplementation. In this study, we used a two-tracer stable isotope technique to determine Fe absorption from a 5-mg dose of stable isotopically enriched (57Fe or 58Fe) ferrous sulfate given with cow's milk (CM) compared with the same dose given with apple juice. Ten children (age 13 +/- 1 mo, weight 10.8 +/- 1.1 kg) who had recently discontinued formula feeding and begun on CM were studied. Red blood cell (RBC) iron incorporation of the isotope was determined 14 d after dosing with 57Fe and 58Fe. Fe absorption was calculated based on the assumption that 90% of absorbed Fe is incorporated into RBC. Absorption of Fe was significantly greater (13.7 +/- 6.4%) when given with juice than with milk (5.7 +/- 4.0%), p < 0.01 by paired t tests. Fe absorption from the dose given with juice was significantly negatively correlated with serum ferritin (n = 9, r = -0.70, p < 0.05). These results indicate that 1) a small supplement of Fe is better absorbed when given with juice than with CM, and 2) a large variability in Fe absorption exists in healthy 1-y-old infants, which is related to their existing Fe stores.

Differences in calcium absorption and kinetics between black and white girls aged 5-16 years.

To assess racial differences in calcium (Ca) metabolism, we measured Ca absorption and kinetics in 89 girls (38 black and 51 white) aged 4.9-16.7 years using a dual-tracer stable isotope technique. We found significantly greater rates of fractional (0.44 +/- 0.13 vs. 0.25 +/- 0.08, p < 0.0001) and total calcium absorption (406 +/- 142 vs. 234 +/- 82 mg/day, p < 0.0003) in black than in white postmenarcheal girls. Fractional absorption of Ca was also greater in black than in white premenarcheal girls (0.39 +/- 0.14 vs. 0.30 +/- 0.10, p = 0.01). Fractional absorption of Ca was more closely correlated to Ca intake in white (r = -0.42, p = 0.002) than in black girls (r = -0.25, p = 0.14). Urinary Ca excretion was significantly lower in pre- but not postmenarcheal black girls than in white girls. Calcium kinetic values associated with bone calcium deposition were greater in black girls, indicating a greater rate of bone Ca deposition in both pre- and postmenarcheal black girls. These results suggest that the greater bone mass accumulated during childhood and adolescence in black than in white females is due, in part, to greater rates of Ca absorption in black girls.

Application of magnetic sector thermal ionization mass spectrometry to studies of erythrocyte iron incorporation in small children.

The optimal evaluation of iron metabolism requires the administration of two isotopes of iron. However, high-precision measurement of isotopic ratios from blood samples obtained after administration of two stable isotopes of iron to human subjects has not previously been reported. Using a cation-exchange system to isolate iron from blood samples, we found that high-precision (< 0.2%) measurements of 58Fe/56Fe and 57Fe/56Fe could be performed using magnetic sector thermal ionization mass spectrometry. Clinical studies in four 1-year-old infants showed that this technique could be used to demonstrate a lower rate of iron absorption in small children given an iron supplement (57Fe) with milk compared to those given iron (58Fe and ferrous sulfate) with ascorbic acid. This technique will enable the evaluation of iron metabolism in populations in whom the use of radioactive iron tracers is not appropriate.