Microsatellite DNA in peach (Prunus persica L. Batsch) and its use in fingerprinting and testing the genetic origin of cultivars.

We isolated and sequenced 26 microsatellites from two genomic libraries of peach cultivar 'Redhaven', enriched for AC/GT and AG/CT repeats, respectively. For 17 of these microsatellites, it was possible to demonstrate Mendelian inheritance. Microsatellite polymorphism was assayed in 50 peach and nectarine cultivars. Of the 1300 PCRs carried out, all but two produced amplified products of the expected size. All microsatellites were polymorphic, showing 2-8 alleles per locus. Heterozygosity ranged from 0.04-0.74 (mean 0.47); the discrimination power (PD) ranged from 0.04-0.84 (mean 0.60). Cultivar heterozygosity varied greatly, with one cultivar ('Independence') being homozygous at all loci. The set of microsatellites discriminated all cultivars investigated, except several sport mutations, i.e., 'Dixitime' vs. 'Springcrest', 'Compact Redhaven' vs. 'Redhaven', and two pairs of cultivars, 'Venus' vs. 'Orion' and 'Elegant Lady' vs. 'Rome Star', whose pedigrees are controversial. We were able to analyze the paternity of several cultivars. In most cases, the parenthood was confirmed. The comparison of three long-living 'Redhaven' accessions supplied by different repositories did not provide any evidence of somatic instability of microsatellites. Hence, microsatellites, ranked according to their information content, are recommended as markers of choice for peach fingerprinting and suggestions are provided for interpreting band profiles and the correct sizing of alleles.     

Genetic variation in the Bribri and Cabecar Amerindians from Talamanca, Costa Rica.

A screening of more than 40 loci was performed in the Bribri and Cabecar Amerindian populations of Talamanca in Southeastern Costa Rica. Some differences were found in the distribution (presence or absence) of the variants (some of them private) of the GOT, PEPA, RH, TF and TPI loci when the results reported here were compared with those of the Bribris and Cabecars that inhabit in the Pacific side of the country. Admixture with non-Indians was detected at the ABO, ADA, and G6PD loci. The proportion of polymorphic loci (P) found were 17.1 in the Bribris and 19.4 in the Cabecars; their average heterozygosites (H) estimates were 0.039 and 0.049 respectively. Fst values (0.019 in the Bribris and 0.056 in the Cabecars) seem to indicate that random events have played important roles in the divergence of the Cabecars. Low effective population sizes of the emigrant groups to the Pacific side and probable gene flow in the case of the Bribris, could be the cause of these differences.     

L-thyroxine and L-triiodothyronine in the cerebral tissues of the adult rat. Effect of hypothyroidism]

The L-thyroxine and L-triiodothyronine concentrations in several brain areas (cerebral cortex, brain stem, hypothalamus, total brain and hypophysis) in normal and hypothyroid rats have been studied. Results show that L-thyroxine values at tissue level are inferior in the hypothyroid group, although non-significant with respect to the control group, whereas L-triiodothyronine presents values similar to the hypothyroid group and its control in all the brain regions studied with the exception of hypophysis. These results show that in hypothyroid situations exist a compensatory mechanism for maintaining the adequate L-triiodothyronine levels in several brain areas, although the serum levels are strongly decreased in hypothyroid animals.     

Effect of strontium and magnesium on blood calcium]

Twenty four male Wistar rats weighing 250 +/- 10 g, in three groups of 8 rats each, were used. Group A was used as control and the content of its drinking water was 6.5 mg/l Ca; 2.4 mg/l Mg. The drinking water of groups B and C was supplemented with 20 mM (SrCl2) and 20 mM (MgCl2), respectively. Once the 20 days of mineral supplementation had passed, arterial blood was extracted by puncture in the abdominal aorta. In the serum obtained after centrifugation, Ca, Mg, Sr and the total proteins (TP) were determined. Afterwards the serum was subjected to ultrafiltration. Concentrations of Ca, Mg and TP were measured in the obtained ultrafiltrates (u), with the above described techniques. The pH was measured before and after the ultrafiltration. The TP decreased significantly both in group B (supplemented with Sr), and in group C (supplement with Mg). Increases in Ca were found in group B and in Mg in group C. The Mg/Ca ratio increased 10% after the supplementation with Mg. At the ultrafiltrate a significant increase in Cau after supplementation with Sr and with Mg was observed. The Mgu/Cau ratio decreased 14% in the group supplemented with Sr and 38% after the supplementation with Mg. In conclusion, the supplementation with Sr (20 mM) in rats increases the Cau and could have the effect of reducing protein synthesis. These facts should be borne in mind when Sr is used for therapeutical purposes.     

Modelling of the disulphide-swapped isomer of human insulin-like growth factor-1: implications for receptor binding.

Insulin-like growth factor-1 (IGF-1) is a serum protein which unexpectedly folds to yield two stable tertiary structures with different disulphide connectivities; native IGF-1 [18-61,6-48,47-52] and IGF-1 swap [18-61,6-47, 48-52]. Here we demonstrate in detail the biological properties of recombinant human native IGF-1 and IGF-1 swap secreted from Saccharomyces cerevisiae. IGF-1 swap had a approximately 30 fold loss in affinity for the IGF-1 receptor overexpressed on BHK cells compared with native IGF-1.The parallel increase in dose required to induce negative cooperativity together with the parallel loss in mitogenicity in NIH 3T3 cells implies that disruption of the IGF-1 receptor binding interaction rather than restriction of a post-binding conformational change is responsible for the reduction in biological activity of IGF-1 swap. Interestingly, the affinity of IGF-1 swap for the insulin receptor was approximately 200 fold lower than that of native IGF-1 indicating that the binding surface complementary to the insulin receptor (or the ability to attain it) is disturbed to a greater extent than that to the IGF-1 receptor. A 1.0 ns high-temperature molecular dynamics study of the local energy landscape of IGF-1 swap resulted in uncoiling of the first A-region alpha-helix and a rearrangement in the relative orientation of the A- and B-regions. The model of IGF-1 swap is structurally homologous to the NMR structure of insulin swap and CD spectra consistent with the model are presented. However, in the model of IGF-1 swap the C-region has filled the space where the first A-region alpha-helix has uncoiled and this may be hindering interaction of Val44 with the second insulin receptor binding pocket.