Changes in the temporal and spatial expression of H beta 58 during formation and maturation of the chorioallantoic placenta in the Rat.
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Cloning and sequencing of a cDNA amplified by RNA fingerprinting at the implantation site of pregnant rats revealed 80% similarity with H beta 58, previously shown to be essential for formation of the chorioallantoic placenta in the mouse. H beta 58 mRNA was detected in the endometrium of hormonally sensitized rats stimulated to undergo decidualization and in the contralateral uterine horns lacking a decidual stimulus, indicating that uterine expression of H beta 58 mRNA did not require decidualization or the presence of a blastocyst. Immunodetection in the early postimplantation uterus (Days 6-8 of pregnancy) showed H beta 58 localized in the luminal and glandular epithelia and some stromal cells. Decidual cells at Day 6 of pregnancy expressed H beta 58, and by Day 9 of pregnancy, the protein localized throughout the maternal decidua. The temporal and spatial distribution of H beta 58 in the developing chorioallantoic placenta was assessed at Days 10, 12, and 14 of pregnancy. Immunoreactive H beta 58 localized to erythroid cells within the developing fetal vasculature of the chorioallantoic primordia at Day 10 of pregnancy. By Day 12, the fetal vasculature extended into the placental labyrinth, and the erythroid stem cells continued to strongly express H beta 58. At Day 14 of pregnancy, immunoreactivity became evident in the trophoblast giant cells and syncytiotrophoblast of the fetal placenta. As the chorioallantoic placenta matured (Day 18), H beta 58 mRNA was 3.6-fold higher in the labyrinth compared with the junctional region. Stable cell lines (HRP/LRP) isolated from the rat labyrinthine placenta expressed H beta 58 mRNA and protein. The expression pattern of H beta maternal and fetal placental tissues and its early expression in fetal erythroid stem cells during formation and maturation of the chorioallantoic placenta suggest that H beta 58 plays key roles in the regulatory networks that control hematopoietic development and placentation. (+info)
Leaf ureide degradation and N(2) fixation tolerance to water deficit in soybean.
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Accumulation of ureides in leaves is associated with the sensitivity of N(2) fixation in soybean to soil water deficit. Consequently, ureide degradation in leaves may be a key to increasing soybean tolerance to dry soils. Previous research indicated that allantoic acid degradation is catalysed by different enzymes in cultivars Maple Arrow and Williams. The enzyme found in Williams requires manganese as a cofactor. The first objective of this study was to determine if the two degradation pathways were associated with differences in N(2) sensitivity to soil water deficits. N(2) fixation of Williams grown on low-Mn soil was sensitive to stress, but it was relatively tolerant when grown on soil amended with Mn. N(2) fixation in Maple Arrow was relatively tolerant of soil drying regardless of the Mn treatment. The second objective of this study was to expand the study of the degradation pathway to nine additional genotypes. Based on ureide degradation in the presence and absence of Mn, these genotypes also segregated for the two degradation pathways. Those genotypes with the Mn-dependent pathway tended to have drought-sensitive N(2) fixation, but there was one exception. The genotypes not requiring Mn for ureide degradation were drought-tolerant except for one genotype. These results demonstrated the possibility for increasing N(2) fixation tolerance to soil water deficits in soybean by selection of lines with high ureide degradation rates, which were commonly associated with the Mn-independent pathway. (+info)
Quantitative design of drug compatibility by weighted modification method.
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AIM: To set up a new method for designing and quantitatively analyzing drug compatibility. METHODS: Drugs for compatibility were divided into 6 dose levels which were evenly distributed to 6 compound groups according to a fixed design. A new mathematical model was set up to fit the dose-effect data of 6 groups. The coefficients, obtained from the model, reflected the dose-effect relationship and the important degree of every drug in combination. According to the coefficients, the drugs in compatibility could be distinguished into principal drug, synergist, inferior, antagonist, and assistant. Because compatibility in the maximal effect group was nearly (or was) an optimal one in 6 groups, the doses in the group were taken as a base for further modification which considered interaction among drugs. The results of the modification were demonstrated by further experiment. This method was applied to design and to quantitatively analyze the compatibility of allantoin, metronidazole, and dexamethasone sodium phosphate by 2 effect indices in mice. RESULTS: This new method was able to effectively determine important degree of drugs in combination, and to optimize their doses for designing compatibility. CONCLUSION: This weighted modification method is a highly efficient, accurate, and practical means for designing and quantitatively analyzing drug compatibility. (+info)
The influence of the extracellular fluid volume on the tubular reabsorption of uric acid.
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Changes is tubular reabsorption of uric acid in response to alterations in the extracellular fluid volume (ECFV) were examined in rats by clearance studies and by direct intratubular microinjections. Contraction of the ECFV led to a rise in the serum uric acid concentration and a 47% decrease in the clearance of uric acid. The ratio of uric acid to inulin clearance also fell, indicating an increase in the net tubular reabsorption of urate. Volume expansion resulted in an increase in the urate clearance and a 37% decrease in the net tubular reabsorption of uric acid. To localize the site in the nephron where these changes occur, microinjections of [2-14C]urate were performed. The lack of conversion of radioactive urate to allantoin after microinjections was demonstrated by thin-layer chromatography. After contraction of the ECFV, urinary recoveries of uric acid were significantly decreased after microinjections into proximal tubular sites. In contrast, recoveries were increased from these proximal sites after volume expansion. No evidence for distral reabsorption was obtained in any group of animals. These studies demonstrate that net urate reabsorption is influenced by the state of hydration of the ECFV and that these alterations are mediated by changes in the rates of reabsorption in the proximal tubule. (+info)
Metabolism of xanthine and hypoxanthine in the tea plant (Thea sinensis L.).
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1. The metabolism of xanthine and hypoxanthine in excised shoot tips of tea was studied with micromolar amounts of [2(-14)C]xanthine or [8(-14)C]hypoxanthine. Almost all of the radioactive compounds supplied were utilized by tea shoot tips by 30 h after their uptake. 2. The main products of [2(-14)C]xanthine and [8(-14)C]hypoxanthine metabolism in tea shoots were urea, allantoin and allantoic acid. There was also incorporation of the label into theobromine, caffeine and RNA purine nucleotides. 3. The results indicate that tea plants can catabolize purine bases by the same pathways as animals. It is also suggested that tea plants have the ability to snythesize purine nucleotides from glycine by the pathways of purine biosynthesis de novo and from hypoxanthine and xanthine by the pathway of purine salvage. 4. The results of incorporation of more radioactivity from [8(-14)C]hypoxanthine than from [2(-14)C]xanthine into RNA purine nucleotides and caffeine suggest that hypoxanthine is a more effective precursor of caffeine biosynthesis than xanthine. The formation of caffeine from hypoxanthine is a result of nucleotide synthesis via the pathway of purine salvage. (+info)
Targeted mutagenesis of Smad1 reveals an essential role in chorioallantoic fusion.
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The Smad family of intracellular signaling intermediates transduce signals downstream from the transforming growth factor beta (TGF-beta) family of receptor serine threonine kinases. The original member of this family, Smad1, has been shown to mediate signals from receptors for the bone morphogenetic proteins (BMPs), a large group of ligands in the TGF-beta superfamily that mediate important developmental events. We have targeted the Smad1 gene in mice and created mutants null at this locus. Smad1 mutant mice die at approximately 9.5 days postcoitum due to defects in allantois formation. In Smad1 mutant mice, the allantois fails to fuse to the chorion, resulting in a lack of placenta and failure to establish a definitive embryonic circulation. Although vasculogenesis is initiated in the mutant allantois, the vessels formed are disorganized, and VCAM-1 protein, a marker for distal allantois development, is not expressed. Smad1 null fibroblasts are still able to respond to BMP2, however, suggesting that the defect observed in the developing extraembryonic tissue is caused by a very specific loss of transcriptional activity regulated by Smad1. Our data further demonstrate that although highly similar structurally, Smad proteins are not functionally homologous. (+info)
Microbial protein production determined by urinary allantoin and renal urea sparing in normal and low protein fed Corriedale sheep.
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The aim of the present study was to compare the amount of microbial N entering the duodenum and the efficiency of N utilisation for microbial protein synthesis in normal (NP, 17.4 g N/d) and low protein (LP, 7.5 g N/d) fed Corriedale sheep. Renal functional tests for urea handling studies, and determination of urinary allantoin as an indirect method to estimate the microbial protein production were used. Although the N intake was 57% lower in LP sheep, the microbial N production was not very different between both diets (NP: 3.99 +/- 0.01 vs. LP: 3.79 +/- 0.02 g/d, P < 0.05). The efficiency of the microbial protein synthesis in the rumen, expressed as grams of microbial N per kg of digestible organic matter apparently digested in the rumen, was not statistically different for both diets. The urinary elimination of urea was reduced by 84% in LP sheep, essentially due to an important decrease in both renal plasma flow (-63%) and glomerular filtration rate (-71%). These haemodynamic changes would also reduce the filtered load and the urinary elimination of allantoin, thereby leading to an underestimation of the amount of microbial protein entering in the duodenum. Since the renal urea spared by the kidneys remains in the blood, it limits the drop ofthe available urea for ruminal recycling consecutive to a low nitrogen diet. (+info)
Induction of angiogenesis by a fragment of human tyrosyl-tRNA synthetase.
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The first step of protein synthesis is catalyzed by aminoacyl-tRNA synthetases. In addition, certain mammalian tRNA synthetases link protein synthesis to cytokine signaling pathways. In particular, human tyrosyl-tRNA synthetase (TyrRS) can be split by proteolysis into two fragments having distinct cytokine activities. One of the TyrRS fragments (mini TyrRS) contains features identical to those in CXC chemokines (like interleukin-8) that also act as angiogenic factors. Here mini TyrRS (but not full-length TyrRS) is shown to stimulate chemotaxis of endothelial cells in vitro and stimulate angiogenesis in each of two in vivo animal models. The angiogenic activity of mini TyrRS can be opposed by anti-angiogenic chemokines like IP-10. Thus, a biological fragment of human tyrosyl-tRNA synthetase links protein synthesis to regulation of angiogenesis. (+info)