Use of the modified relative dose response (MRDR) assay in rats and its application to humans for the measurement of vitamin A status

After an appropriate dose of 3,4-didehydroretinol (vitamin A2, DR) is given orally in corn oil to retinol (R)-depleted rats, the ratio of plasma DR/R values at 3.5 h is inversely related to the liver concentration of vitamin A (Tanumihardjo & Olson, 1988). In the present study, a similar procedure, termed the modified relative dose response (MRDR) assay, was employed with rats containing a much broader range of liver reserves; ie, less than 2 to 107 nmol of vitamin A per g wet weight of liver (less than 1 to 30 micrograms/g). The DR/R ratio for 15 of 16 rats with liver reserves less than 17 nmol/g (less than 5 micrograms/g) was greater than 0.15, whereas the ratio was less than 0.15 for 7 of 8 rats with liver reserves greater than 17 nmol/g. This distribution provides sensitivity, specificity and positive predictive values of 94, 88 and 94 per cent, respectively. The DR/R ratios reached a maximal plateau in two other groups of rats between 3.5 and 8 h. At all times up to 24 h, mean DR/R ratios for vitamin A-depleted rats (45 +/- 6 nmoles of vitamin A/g liver) were approximately twice those for vitamin A-sufficient rats (230 +/- 40 nmoles of vitamin A/g liver). In three well-nourished adults, presumably with liver reserves of greater than 300 nmol/g wet weight liver (greater than 80 micrograms retinol equivalents/g), and in two young (1- and 3-years old) well-nourished children, maximal DR/R ratios were less than 0.023. In these cases, peak DR/R ratios were observed, with one possible exception, between 8 and 12 h.(ABSTRACT TRUNCATED AT 250 WORDS)     

Androgen receptor YAC transgenic mice carrying CAG 45 alleles show trinucleotide repeat instability

X-linked spinal and bulbar muscular atrophy (SBMA) is caused by a CAG repeat expansion in the first exon of the androgen receptor (AR) gene. Disease-associated alleles (37-66 CAGs) change in length when transmitted from parents to offspring, with a significantly greater tendency to shift size when inherited paternally. As transgenic mice carrying human AR cDNAs with 45 and 66 CAG repeats do not display repeat instability, we attempted to model trinucleotide repeat instability by generating transgenic mice with yeast artificial chromosomes (YACs) carrying AR CAG repeat expansions in their genomic context. Studies of independent lines of AR YAC transgenic mice with CAG 45 alleles reveal intergenerational instability at an overall rate of approximately 10%. We also find that the 45 CAG repeat tracts are significantly more unstable with maternal transmission and as the transmitting mother ages. Of all the CAG/CTG repeat transgenic mice produced to date the AR YAC CAG 45 mice are unstable with the smallest trinucleotide repeat mutations, suggesting that the length threshold for repeat instability in the mouse may be lowered by including the appropriate flanking human DNA sequences. By sequence-tagged site content analysis and long range mapping we determined that one unstable transgenic line has integrated an approximately 70 kb segment of the AR locus due to fragmentation of the AR YAC. Identification of the cis - acting elements that permit CAG tract instability and the trans -acting factors that modulate repeat instability in the AR YAC CAG 45 mice may provide insights into the molecular basis of trinucleotide repeat instability in humans.      

Cryopreservation of human oocytes: a review of current problems and perspectives

It is well recognized that the ability to cryopreserve unfertilizedhuman oocytes would make a significant contribution to infertilitytreatment. However, despite considerable interest, very fewsuccessful pregnancies have arisen from cryopreserved oocytesafter thawing, insemination and transfer of the subsequent embryo.The reasons for this lack of progress may well result from adearth of information on how the various biophysical changesduring a cryopreservation regimen affect human oocyte function.Recently, fundamental studies on the effects of cooling, membranepermeability, cryoprotectant addition and ice formation havebeen performed on human oocytes by a number of groups, and theseform the basis of the current review. It is likely that successfulhuman oocyte cryopreservation will only follow once these factorsare fully understood, but the existing base of knowledge shouldprovide a platform for further improvements in the techniquescurrently employed.