No abstract

Experimental hyperphenylalaninemia has been induced in 5-day-old chicks by dietary treatments with phenylalanine and alpha-methylphenylalanine. An increase of nearly 8-fold in plasma Phe/Tyr ratio was found after 4 days of supplementation the standard diet with 5% phenylalanine plus 0.4% alpha-methylphenylalanine. The increase in this ratio was about 13-fold after 9 days of the same treatment. Similar results were observed in brain and liver, although the increases were smaller than those found in plasma. Total body, brain and liver weight decreased after 9 days of treatment. Phenylalanine plus alpha-methylphenylalanine administration to 5-day-old chicks produced a significant decrease in the 3-hydroxy-3-methylglutaryl-CoA reductase and mevalonate-5-pyrophosphate decarboxylase specific activities from both brain and liver. These results demonstrated for the first time that experimental hyperphenylalaninemia inhibited different enzyme activities directly implicated in the regulation of cholesterogenesis. Therefore, a reduced cholesterol synthesis in brain may evidenciate the theory of an impaired myelination leading to mental retardation in phenylketonuria patients.

Phenylalanine, phenylpyruvate and phenylacetate produced a considerable inhibition of chick liver mevalonate 5-pyrophosphate decarboxylase while mevalonate kinase and mevalonate 5-phosphate kinase were not significantly affected. Phenolic derivatives of phenylalanine produced a similar inhibition of decarboxylase activity than that found in the presence of phenyl metabolites. The degree of inhibition was progressive with increasing concentrations of inhibitors (1.25-5.00 mM). Simultaneous supplementation of different metabolites in conditions similar to those in experimental phenylketonuria (0.25 mM each) produced a clear inhibition of liver decarboxylase and 3-hydroxy-3-methylglutaryl-CoA reductase. To our knowledge, this is the first report on the in vitro inhibition of both liver regulatory enzymes of cholesterogenesis in phenylketonuria-like conditions. Our results show a lower inhibition of decarboxylase than that of reductase but suggest an important regulatory role of decarboxylase in cholesterol synthesis.

There is growing evidence that overweight and obesity increase the risk of certain cancers. Studies in adults support the role of insulin-like growth factors (IGFs) and oestrogens in the pathogeneses of several cancers. We propose that hormone alterations described as risk factors for cancer in obese adults are present in prepubertal obese children. A group of obese children aged 6-9 years (n=40), and control group paired for age and sex, were used for the study. The obese children presented a significantly high level of IGF-I (P=0.0173) and insulin (P=0.0250), with a drop in sex hormone-binding globulin (SHBG) (P=0.0282). The 17 beta-oestradiol (E2)/SHBG ratio increase in obese subjects was marginally significant (P=0.0635). Grouping together all the children in quartiles for insulin and body mass index, the upper quartiles showed a rise in IGF-I and E2/SHBG. In a multivariant correlation analysis, only height (partial r=0.2464) and insulin (partial r=0.3002) were independent prediction variables for IGF-I concentration. The only variables statistically correlated with the E2/SHBG ratio were insulin (r=0.2879) and IGF-I (r=0.4140). The obese children in our study showed hormone changes described as risk factors for cancer in obese adults. These changes were significantly associated with the hyperinsulinaemia. We hypothesize that this potential risk should be taken into account given the long period of exposure involved in the presence of hormone alterations at such early ages.

Very small sample sizes frequently become the limiting factor in biochemical and biomembrane studies in which routine quantification of protein and bulk lipids are required. The procedure described here allows the simultaneous determination of protein and lipid without initial, multiple aliquots. The method is based on the quantitative precipitation of proteins from a defined hexane/isopropanol mixture. The liquid phase resulting after decanting and concentrating to dryness can then be used to assay the lipid content directly. Quantitative assay of protein can be achieved after resuspension of the pelleted material by addition of sodium dodecyl sulfate (0.1%) and deoxycholate (1%). The method is also applicable to other types of lipid- and protein-containing samples with a broad range of protein/lipid ratios and lipid compositions, as they occur, for example, in serum lipoproteins.