Temporal and spatial transitions in collagen types during embryonic chick limb development : II. Comparison of the embryonic cartilage collagen molecule with that from adult cartilage

Analyses were performed to compare the (α1)₃ collagen molecule synthesized by the cartilaginous long bone anlage of 8-day chick embryos with that produced by adult cartilage, [α1(II)]₃. The embryonic molecule produced segment-long-spacing crystallites which had a banding pattern indistinguishable from that of authentic [α1(II)]₃. After carboxymethyl cellulose chromatography, the material in the α1 chain peak of the embryonic collagen was found to have the same amino acid composition as that of the α1(II) chain.     

A comparative study of the transport of pyruvate in liver mitochondria from normal and diabetic rats

A comparative study of the transport of pyruvate in liver mitochondria from normal and diabetic rats has been carried out. TheK _m for net pyruvate uptake in diabetic, ketotic mitochondria is practically equal to that measured in normal mitochondria, while theV _max is significantly lower. The lower activity of the pyruvate translocator in diabetic mitochondria compared to normal mitochondria is also shown by swelling experiments as well as by following the rate of pyruvate-supported respiration. Pre-exposure of mitochondria from normal rats to the ketone body acetoacetate and to 2-oxobutyrate results in a decrease of theK _m for pyruvate uptake. This effect is impaired in mitochondria from diabetic animals. The results indicate that the activity and the properties of the mitochondrial pyruvate translocator are modified in the diabetic, ketotic condition.Supported by a joint grant from Consiglio Nazionale delle Ricerche, Rome, Italy, and the Polish Academy of Sciences, Warsaw, Poland.     

Effects of copper and ceruloplasmin on iron transport in the Caco 2 cell intestinal model

Previous studies have implicated copper proteins, including ceruloplasmin, in intestinal iron transport. Polarized Caco2 cells with tight junctions were used to examine the possibilities that (a) ceruloplasmin promotes iron absorption by enhancing release at the basolateral cell surface and (b) copper deficiency reduces intestinal iron transport. Iron uptake and overall transport were followed for 90 min with 1 &mgr;M 59Fe(II) applied to the apical surface of Caco2 cell monolayers. Apotransferrin (38 &mgr;M) was in the basolateral chamber. Induction of iron deficiency with desferrioxamine (100 &mgr;M; 18 h) markedly increased uptake and overall transport of iron. Uptake increased from about 20% to about 65% of dose, and overall 59Fe transport from <1% to 60% of dose. On the basis of actual iron released into the basal chamber (measured with bathophenanthroline), transport increased 8-fold. Desferrioxamine pretreatment reduced cellular Fe by 55%. The addition of freshly isolated, enzymatically active human ceruloplasmin to the basolateral chamber during absorption had no effect on uptake or transport of iron by the cells. Unexpectedly, pretreatment with three different chelators of copper (18 h), which reduced cellular levels about 40%, more than doubled iron uptake and raised overall transport to 20%. This was so, whether or not cells were also made iron deficient with desferrioxamine. Acute addition of 1 &mgr;M Cu(II) to the apical chamber had no significant effect upon iron uptake, retention, or transport in iron deficient or normal cells, in the presence of absence of ascorbate. We conclude that intestinal absorption of Fe(II) is unlikely to depend upon plasma ceruloplasmin, and that cuproproteins involved in this form of iron transport must be binding copper tightly.     

Tolbutamide alters glucose transport and metabolism in the embryonic mouse heart

BACKGROUND: Tolbutamide is a sulfonylurea oral hypoglycemic agent widely used for the treatment of non insulin-dependent diabetes mellitus. Tolbutamide produces dysmorphogenesis in rodent embryos and becomes concentrated in the embryonic heart after maternal oral dosing. Tolbutamide increases glucose metabolism in extra-pancreatic adult tissues, but this has not previously been examined in embryonic heart. METHODS: CD-1 mouse embryos were exposed on GD 9.5 to tolbutamide (0, 100, 250, or 500 microg/ml) for 6, 12, or 24 hr in whole-embryo culture. Isolated hearts were evaluated for (3)H-2DG uptake and conversion of (14)C-glucose to (14)C-lactate. Glut-1, HKI, and GRP78 protein levels were determined by Western analysis, and Glut-1 mRNA was measured by RT-PCR. RESULTS: Cardiac (3)H-2DG uptake increased after exposure to 500 microg/ml tolbutamide for 6 hr, and 100, 250, or 500 microg/ml tolbutamide for 24 hr, compared to controls. Glycolysis increased after exposure to 500 microg/ml tolbutamide for 6 or 24 hr compared to controls. Glut-1 protein levels increased in hearts exposed to 500 microg/ml tolbutamide for 12 or 24 hr, and Glut-1 mRNA increased in hearts exposed to 500 microg/ml tolbutamide for 24 hr compared to controls. HKI protein levels increased in hearts exposed to 500 microg/ml tolbutamide for 6 hr, but not 12 or 24 hr. There was no effect on GRP78 protein levels in hearts exposed to tolbutamide for 6, 12, or 24 hr. CONCLUSIONS: Tolbutamide stimulates glucose uptake and metabolism in the embryonic heart, as occurs in adult extra-pancreatic tissues. Glut-1 and HKI, but not GRP78, are likely involved in tolbutamide-induced cardiac dysmorphogenesis.     

A comparative study of the effects of neonatal androgenization and estrogenization on prolactin secretion in adult female rats

Effects of neonatal androgenization (NA) and estrogenization (NE) were compared especially in terms of the prolactin (PRL) secretion in female rats. Twenty-four h after birth, a total of seven groups of newborn female rats were treated as follows. Three NA groups received a single s.c. injection of 10, 100 or 1000 micrograms of testosterone, respectively. Similarly, three NE groups received 1, 10 or 100 micrograms of estradiol-17 beta, respectively. The remaining one group was injected with oil vehicle only, and served as controls. At 8 weeks of age, animals were killed by rapid decapitation. PRL, estradiol and progesterone were measured in the plasma. Anterior pituitary (AP) was weighed, and AP PRL content was measured. NA and NE, at the highest doses, resulted in a similar degree of hyperprolactinemia and hyperestrogenemia showing an effect ratio of about 1:10. This ratio was, however, not true with the lower doses. Furthermore, there was no dose-dependency in the effect of NE on the plasma PRL and estradiol levels. In turn, plasma progesterone levels were dose-dependently decreased by both NA and NE. AP PRL content, expressed per AP, was significantly higher than control values in only NA (1000 micrograms) and NE (100 micrograms) groups. AP weight was increased by NA (1000 micrograms) but not by any NE treatment. These results indicate that NA and NE do not always exert similar effects on the PRL secretion or on several other related parameters. Therefore, aromatization of testosterone to estradiol does not appear to be the sole mechanism mediating the neuroendocrine consequences of NA.     

A comparative analysis of adult body size and its correlates in acanthocephalan parasites

Adult acanthocephalan body sizes vary interspecifically over more than two orders of magnitude; yet, despite its importance for our understanding of the coevolutionary links between hosts and parasites, this variation remains unexplained. Here, we used a comparative analysis to investigate how final adult sizes and relative increments in size following establishment in the definitive host are influenced by three potential determinants of acanthocephalan sizes: initial (cystacanth) size at infection, host body mass, and the thermal regime experienced during growth, i.e. whether the definitive host is an ectotherm or an endotherm. Relative growth from the cystacanth stage to the adult stage ranged from twofold to more than 10,000-fold across acanthocephalan species, averaging just over 100-fold. However, this relative increment in size did not correlate with host mass, and did not differ between acanthocephalan species using ectothermic hosts and those growing in endothermic hosts. In contrast, final acanthocephalan adult sizes correlated positively with host mass, and after correction for host mass, final adult sizes were higher in species parasitising endotherms than in those found in ectotherms. The relationship between host mass and acanthocephalan adult size practically disappears, however, once phylogenetic influences are taken into account. Positive relationships between adult acanthocephalan size, cystacanth size and egg size indicate that a given relative size is a feature of an acanthocephalan species at all stages of its life cycle. These relationships also suggest that adult size is to some extent determined by cystacanth size, and that the characteristics of the definitive host are not the sole determinants of parasite life history traits.     

A comparative study of elemental composition of human breast milk and infant milk substitutes

Instrumental neutron activation analysis (INAA) and protoninduced X-ray emission (PIXE) analysis have been employed to determine the concentration of 13 elements in human breast milk, various infant formulas, and locally produced cereals from Nigeria, as well as from various infant formulas and natural cow and goat milk available in the UK. The study shows that if the locally produced cereal is to be used on a regular basis for babies in Nigeria, then their diet must be supplemented with essential trace elements. Furthermore, parents should be discouraged from giving their infants cow and goat milk because of the high concentration of major elements compared to human breast milk.     

Biliary excretion of copper in fischer rats treated with copper salt and in long-evans cinnamon (LEC) rats with an inherently abnormal copper metabolism

Increased biliary Cu excretion was found in Fischer rats injected with Cu. The biliary Cu was located at the void (large-molecule region) and total (small-molecule region) volume of a Sephadex G-75 column. The most Cu was found in the total volume. The two Cu peaks comigrated with absorbance at 280 nm. Although the bile from Cu-untreated Fischer rats did not show Cu absorbance in the total volume, absorbance at 280 nm was also found in this region. Even though Long-Evans Cinnamon (LEC) rats deposited a gross amount of Cu (194.0±27.8 μg/g liver) in the liver, they conversely showed reduced Cu excretion into the bile. LEC bile did not show Cu absorbance but rather absorbance at 280 nm in the total volume. Therefore, it seems unlikely that the small molecules found in the Sephadex G-75 regulate biliary Cu excretion in Cu-loaded rats, although the molecules bind to Cu. When the bile from Cu-untreated fischer and LEC rats was incubated with CuCl₂ solution, the most Cu was recovered in the total volume of this column. Our results suggest that reduced biliary Cu excretion in LEC rats is not related to the small molecules, and that Cu cannot be excreted in the form of macromolecules in rats to decrease Cu from the Cu-loaded liver.     

Lack of ceruloplasmin expression alters aspects of copper transport to the fetus and newborn, as determined in mice

Copper transport and accumulation were studied in virgin and lactating C57BL/6 mice, with and without expression of ceruloplasmin (Cp), to assess the importance of Cp to these processes. One hour after i.p. injection of tracer ⁶⁴Cu, liver and kidney accounted for 80% of the radioactivity, and mammary gland 1%, while in lactating Cp+/+ mice 2–4 days post partum, uptake by mammary gland was 9-fold higher and that of liver and other organs was decreased, with ⁶⁴Cu rapidly appearing in milk. Parallel studies in Cp−/− mice (siblings from same colony) gave virtually identical results. However, their milk contained less ⁶⁴Cu, and actual copper contents determined by furnace atomic absorption were less than half those for milk from normal dams. Liver copper concentrations of pups born to Cp−/− dams also were half those of pups from wild type dams. Copper in pup brains was unaffected; but iron concentrations were reduced. We conclude that absence of Cp, while not affecting entry of exchangeable copper from the blood into the mammary gland, does have a significant effect on the availability of this metal to the newborn through the milk and in the form of stores accumulating in gestation.     

A comparative study of vitamin D binding globulins in milk.

1. The occurrence of 25-hydroxy vitamin D binding protein in human, bovine, monkey and porcine milk was investigated. 2. Sucrose gradient ultracentrifugation revealed the presence of 4.2 S and 5.7 S binding globulins in the whey of human, monkey and porcine milk. 3. Although bovine plasma also contains a 4.2 S globulin only a 5.7 S protein was found in bovine milk. 4. The 4.2 S and 5.7 S globulins in milk could not be resolved by polyacrylamide gel electrophoresis or by isoelectric focusing. 5. Plasma and whey binding proteins of any one species had the same isoelectric point but there were small differences among species (4.5-4.8). 6. Competitive displacement studies showed that the binding proteins in milk have high affinity for 25-hydroxy-cholecalciferol and 24,25-dihydroxycholecalciferol.