The p53 isoforms are differentially modified by Mdm2

The discovery that the single p53 gene encodes several different p53 protein isoforms has initiated a flurry of research into the function and regulation of these novel p53 proteins. Full-length p53 protein level is primarily regulated by the E3-ligase Mdm2, which promotes p53 ubiquitination and degradation. Here, we report that all of the novel p53 isoforms are ubiquitinated and degraded to varying degrees in an Mdm2-dependent and -independent manner, and that high-risk human papillomavirus can degrade some but not all of the novel isoforms, demonstrating that full-length p53 and the p53 isoforms are differentially regulated. In addition, we provide the first evidence that Mdm2 promotes the NEDDylation of p53β. Altogether, our data indicates that Mdm2 can distinguish between the p53 isoforms and modify them differently.     

Autoantibody to vasoactive intestinal peptide in human circulation

In a radioassay for Vasoactive Intestinal Peptide (VIP)-binding, eight out of 33 plasma samples from healthy human subjects exhibited specific binding ranging from 2.6% to 46.7% of total [125 I]VIP. This binding was competitively displaced by unlabeled VIP. The structurally homologous peptides, Peptide Histidine Isoleucine (PHI) and secretin, were, respectively, 72-fold and 413-fold less potent than VIP in displacing bound [125 I]VIP, whereas the unrelated peptides, neurotensin, eledoisin, bombesin and metenkephalin, were without effect on the binding. The antibody nature of the VIP-binding factor was suggested by its precipitation with ammonium sulfate, attenuation after absorption with Staphylococcus aureus preparations, precipitation with antisera against human IgG and IgM, and coelution with standard IgG and IgM on anion-exchange and high-performance gel-filtration columns. Pepsin treatment of purified IgG fraction yielded a VIP-binding species with apparent molecular weight of 108 +/- 13 kDa that was precipitated by antiserum against the F(ab)2 fragment of the IgG molecule. These results demonstrate the existence in some human plasmas of an autoantibody that binds VIP.     

Some characteristics of pollen wall cytochemistry and ultrastructure in Japanese larch (Larix leptolepis Gord.)

Summary The mature pollen of Larix leptolepis Gord. (Conifer) contains five different cell types, and the plasma membrane of the vegetative cell is continuous and organized. The pollen wall is composed of two morphologically and cytochemically distinct domains: the exine and the intine. In the multilayered exine, the ektexine appears granular and the endexine, lamellar. The intine is thick and bilayered with a microfibrillar structure occupying its inner portion. Cytochemical reactions of the exine and the intine are similar to those found in angiosperms. Pollen wall involvement in the male female recognition system is discussed with respecl to the angiosperms.     

Effect of nutrition and endoparasitic treatment on growth, onset of puberty and reproductive activity in Menz ewe lambs

Ewe lambs of the Ethiopian Menz breed were assigned at weaning (90+/-3 days) to four levels of nutrition (poor, low, medium and high) to achieve different premating growth rates with or without treatment for endoparasites. A concentrate mixture providing 2.5 Mcal/kg dry matter (DM) metabolizable energy and 15.2 g/kg DM digestible protein was used. Improved nutrition increased lamb postweaning average daily weight gain up to puberty by 6 to 26 g/day and the conception rate to first estrus by 9 to 16% while it reduced the mortality rate by 24 to 31% and age at first lambing by 2 to 5 months. Lambs reached puberty (age at first estrus) at 16.9+/-0.1 kg (+/-SEM) or 60% of mature body weight and 350+/-12 days of age. The onset of puberty was advanced by weaning weight (P<0.05), itself being well correlated with birth weight (r = 0.51, P<0.001), and by level of nutrition (high=299+/-19, medium=301+/-18, low=383+/-23 and poor=454+/-31 days, P<0.001) through enhanced growth rate (r = -0.82, P<0.001). No independent effect of drenching for endoparasites on pubertal development was observed (P>0.05), but its interaction with season-of-birth improved the growth of lambs born during the period of short rains (P<0.05). Overall mean litter size at first lambing was 1.07; the twinning rate was 6.5% and the birth weight was 1.9+/-0.1 kg. Up to 13.4% of newborn lambs, averaging 1.3+/-0.6 kg, died on the day of parturition. The results indicate that improved growth rate and body weight, resulting from better postweaning nutrition, affects the attainment of puberty in Menz ewe lambs. Mitigation of nutrition stress and endoparasitic infection depending on season-of-birth would thus increase the annual reproductive rate of breeding ewes and flock productivity.     

Identification of Glycolytic Enzyme Polypeptides on the Two-Dimensional Protein Map of Saccharomyces cerevisiae and Application to the Study of Some Wine Yeasts

Using a modification of the basic two-dimensional polyacrylamide gel electrophoresis technique, the polypeptides of the protein map of Saccharomyces cerevisiae involved in glycolysis were investigated. This study resulted in a reassignment of two of the seven glycolytic enzyme polypeptides previously identified (Ludwig et al., Mol. Cell. Biol. 2:117-126, 1982), those corresponding to phosphoglycerate kinase and to alcohol dehydrogenase. It also resulted in the identification of two additional glycolytic polypeptides, the enolase B monomer and the glyceraldehyde phosphate dehydrogenase B monomer. The glycolytic enzymes polypeptides so identified were investigated in 5 laboratory strains (all S. cerevisiae) and in 11 commerical strains used for wine making (S. cerevisiae and Saccharomyces bayanus). It appeared highly significant that a particular electrophoretic variant of the glyceraldehyde phosphate dehydrogenase B monomer was found only in the wine yeasts. Furthermore, it was strongly suggested that S. cerevisiae and S. bayanus strains are distinguishible on the basis of a different electrophoretic migration of the enolase B monomer.     

Further studies on prevalence of hydatidosis in slaughtered animals from North Jordan

Examination of 471 sheep, 118 goats, 157 cattle and 56 camels slaughtered in abattoirs in North Jordan was carried out during March-May 1984. Drought conditions that prevailed during the preceding winter led to slaughtering old female sheep (greater than or equal to 4 years) due to scarcity of food, which allowed us to analyse the prevalence of hydatidosis in various age groups of sheep. An overall infection rate of 27.8, 1.7, 5.8 and 10.7 percent was found in sheep, goats, cattle and camels, respectively. The infection rate was as low as 1.5 percent in male and 1.9 percent in female sheep under 2 years of age. However, the rate of hydatid infection increased with age and reached as high as 63.7 percent in ewes 4 years of age and older. The percentage of animals with fertile cysts was also highest in sheep (68.7 percent of infected animals) and increased with age reaching 100 percent in ewes which were 10 years of age or older. Analysis of all cysts recovered from the livers and lungs of infected ewes from various age groups revealed a sharp increase in the mean total number of cysts in age groups over 8 years of age. The fertility rate of the cysts in the liver was significantly greater in ewes 6 years old or more (64.8--78.6 percent) than in younger age groups (8.7-46.2 percent). In the lung, the fertility rate increased progressively with age reaching as high as 97.9 percent in ewes 10 years old or more. These findings of high infection and fertility rates of hydatid disease in sheep, particularly of older age groups, prompt plans for further epidemiological studies and control programmes.     

Biological and physicochemical characterization of recombinant human erythropoietins fractionated by Mono Q column chromatography and their modification with sialytransferase

Ten erythropoietin (EPO) fractions differing in sialic acid content, ranging from 9.5 to 13.8 mol mol^–1 of EPO, were obtained from baby hamster kidney cell-derived recombinant human EPO by Mono Q column chromatography. The mean pI values of the EPO fractions determined by IEF-gel electrophoresis systematically shifted from 4.11 to 3.31, coinciding with the sialic acid content, without a change in the constitution of asialo N-linked oligosaccharides of each fraction. Although a linear relationship between thein vivo bioactivity and the sialic acid content of the fractionated, samples was observed until 12.1 mol mol^–1 of EPO, there was no further increase in their activity over 12.4 mol mol^–1 of EPO. On the other hand, an inverse relationship between thein vitro bioactivity and sialic acid content of EPO was observed. Also, we showed that thein vivo bioactivity of some fractions with low sialic acid contents was increased after treatment with 2,6-sialyltransferase, but thein vivo bioactivity of the other fractions with high sialic acid contents was either decreased or not affected.Abbreviations EPO erythropoietin - rHuEPO recombinant human erythropoietin - hCG human chorionic gonadotropin - BHK baby hamster kidney - CHO Chinese hamster ovary - NeuAc N-acetyl neuraminic acid - Gal galactose - HRCs hemolyser-resistant cells - WST-1 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium Na - IEF isoelectric focusing - pI isoelectric point