Studies on the Polymorphism of L-Glutamic Acid

The growth rate of the α-crystal of L-glutamic acid was measured under various degrees of supersaturation and temperatures. The rate constants and the activation energies for (0 0 1) and (1 1 1) faces were measured and the latter values were 6.7 and 11.5 kcal/mol, respectively. The controlling process of the α-crystal growth was investigated by comparison of Sherwood numbers of dissolution and crystallization, and the crystallization process was found to be controlled by the surface reaction.     

Synthesis, structure, and hypochromism of (2,9)(6,9)purinophane

The title compound 1 was synthesized via 8 steps from phthalimide derivative 2. The molecular structure of 1 was determined by X-ray analysis and the relationship between the hypochromism and the stacking mode of two purine rings was discussed.     

Characterization of two isozymic forms of heart fructose 6-phosphate, 2-kinase:fructose 2,6-bisphosphatase.

Occurrence of two isozymic forms of fructose 6-P, 2-kinase: fructose 2,6-bisphosphatase in bovine heart was investigated by transcribing mRNAs and amplifying the cDNAs with polymerase chain reactions. Analysis of the PCR products revealed 1.7 Kb and 1.5 Kb DNAs, and the determination of their nucleotide sequences showed that these DNAs are identical except for the lack of 180 base pairs near the 3' of the bovine heart enzyme DNA previously reported (6). This missing nucleotide sequence encodes Asn451-Gln510 and contains the phosphorylation sites for cAMP dependent protein kinase and protein kinase C.     

Presence of a novel exon 2E encoding a putative transmembrane protein in human IL-33 gene

Interleukin-33 (IL-33) is a dual-function molecule that regulates gene expression in nuclei and, as a cytokine, conveys proinflammatory signals from outside of cells via its specific receptor ST2L. There are still a lot of questions about localization and processing of IL-33 gene products. In the course of re-evaluating human IL-33 gene, we found distinct promoter usage depending on the cell type, similar to the case in the ST2 gene. Furthermore, we found a novel exon 2E in the conventional intron 2 whose open reading frame corresponded to a transmembrane protein of 131 amino acids. Dependence of exon 2E expression on differentiation of HUVEC cells is of great interest in relation to human IL-33 function.     

Developmental changes in proliferative activity of cells of the murine Rathke's pouch

Summary To clarify proliferative activity in the cells of the Rathke's pouch of the rat, we studied the labeling index using bromodeoxyuridine-immunohistochemistry. Rat fetuses were removed 1 h after transplacental injection of bromodeoxyuridine on day 11.5–21.5 of gestation, and were subsequently used to examine cellular proliferation. Although the labeling index within the Rathke's pouch was 30% on day 12.5, it decreased with development and by day 18.5 of gestation had a value of about 5%. The labeling index within Rathke's pouch was not homogeneous throughout the entire pouch, but tended to be higher in regions where cells were more densely packed. This heterogeneous pattern of distribution of labeling index values continued until day 15. On that day, immunoreactive ACTH cells first appeared in the region where the labeling index was low. From day 17 of gestation, the uneven distribution of the labeling index became vague and, simultaneously, the distribution of ACTH cells became homogeneous throughout the pouch. It was concluded that proliferation of the epithelium of Rathke's pouch is heavily involved in the growth of the pouch until at least the appearance of ACTH cells.     

Synergistic stimulatory effect of glucocorticoid, EGF and insulin on the synthesis of ribosomal RNA and phosphorylation of nucleolin in primary cultured rat hepatocytes.

Effects of dexamethasone, EGF and insulin on the synthesis of rRNA and phosphorylation of nucleolin in primary cultures of adult rat hepatocytes were studied. Hepatocytes were incubated for 8 h with EGF (20 ng/ml) plus insulin (0.1 microM) and/or for 20 h with dexamethasone (1 microM) before the end of incubation. The incorporation of [3H]uridine into acid-insoluble materials and the nuclear activity of RNA polymerase I were stimulated approx. 2-fold with EGF plus insulin and these were further enhanced 2-3-times by dexamethasone, although dexamethasone alone exerted no stimulation. When hepatocytes were incubated with [32P]orthophosphate, similar enhancement by these hormones was also observed in the phosphorylation of a nucleolar protein, nucleolin, which was detected by immunoprecipitation with anti-nucleolin antibodies. The amount of nucleolin was slightly increased by EGF plus insulin in the presence of dexamethasone, but scarcely changed by treatment with EGF plus insulin or dexamethasone alone. Cycloheximide inhibited RNA synthesis to a greater or lesser degree in the case of all hepatocytes which were cultured with or without these hormonal treatments. These results indicate that the in vivo effect of glucocorticoid on rRNA synthesis and nucleolin phosphorylation in liver is primarily a direct action on parenchymal cells and requires other growth factors such as EGF and insulin.     

Human endogenous retrovirus (HERVK9) structural polymorphism with haplotypic HLA-A allelic associations

The frequency and HLA-A allelic associations of a HERVK9 DNA structural polymorphism located in close proximity to the highly polymorphic HLA-A gene within the major histocompatibility complex (MHC) genomic region were determined in Japanese, African Americans, and Australian Caucasians to better understand its human population evolutionary history. The HERVK9 insertion or deletion was detected as a 3' LTR or a solo LTR, respectively, by separate PCR assays. The average insertion frequency of the HERVK9.HG was significantly different (P < 1.083e(-6)) between the Japanese (0.59) and the African Americans (0.34) or Australian Caucasians (0.37). LD analysis predicted a highly significant (P < 1.0e(-5)) linkage between the HLA-A and HERVK9 alleles, probably as a result of hitchhiking (linkage). Evolutionary time estimates of the solo, 5' and 3' LTR nucleotide sequence divergences suggest that the HERVK9 was inserted 17.3 MYA with the first structural deletion occurring 15.1 MYA. The LTR/HLA-A haplotypes appear to have been formed mostly during the past 3.9 MY. The HERVK9 insertion and deletion, detected by a simple and economical PCR method, is an informative genetic and evolutionary marker for the study of HLA-A haplotype variations, human migration, the origins of contemporary populations, and the possibility of disease associations.