Divergence,demography and gene loss along the human lineage

Genomic DNA sequences are an irreplaceable source for reconstructing the vanished past of living organisms. Based on updated sequence data, this paper summarizes our studies on species divergence time, ancient population size and functional loss of genes in the primate lineage leading to modern humans (Homo sapiens sapiens). The inter- and intraspecific comparisons of DNA sequences suggest that the human lineage experienced a rather severe bottleneck in the Middle Pleistocene, throughout which period the subdivided African population played a predominant role in shaping the genetic architecture of modern humans. Also, published and newly identified human-specific pseudogenes (HSPs) are enumerated in order to infer their significance for human evolution. Of the 121 candidate genes obtained, authentic HSPs turn out to comprise only 25 olfactory receptor genes, four T cell receptor genes and nine other genes. The fixation of HSPs has been too rare over the past 6–7 Myr to account for species differences between humans and chimpanzees.     

Improvement of efficiency of the Ce(IV)-induced DNA scission--relationship between the kinetic parameters (k(cat) and Km) and the DNA structure.

The Michaelis constant (Km) for double-stranded DNA, single-stranded DNA, and dinucleotide hydrolysis by Ce(IV) ion are 4.4, 15, and more than 40 mM, respectively. The order of the k(cat), however, is dinucleotide > oligonucleotides. Not only the improvement of k(cat) but also that of Km is important for the design of an efficient artificial nuclease.     

Effects of rapid weight reduction diet on protein metabolism and physical performance

The purpose of this study was to provide information on the requirement of nutrition, especially of the protein during rapid weight reduction immediately before a competition of a weight-classification system sport, boxing. Weight reduction period was 9 days. Subjects were divided into three groups of free diet group (group A: n = 5), high protein diet group (group B: n = 5) and ordinary protein diet group (group C: n = 4). Group A had taken food ad libitum. Group B had taken 2.0 g/kg/day of protein in the first half and 1.5 g/kg/day of protein in the second half of weight reduction period, and Group C had taken 1.0 g/kg/day of protein throughout weight reduction period. Groups B and C had taken 2,000 kcal/day in the first half and 1,200 kcal/day in the second half of weight reduction period. Anthropometry and nutritional investigation were performed, and urine components were analyzed. The main results obtained were as follows: 1) Calorie and protein intake in Group A averaged 883 kcal/day and 0.9 g/kg/day in the second half of weight reduction period. 2) 3-methylhistidine and urea nitrogen in urine and 3-Me/Cr increased significantly at the end of weight reduction period in Group A, but decreased significantly in groups B and C. Nitrogen balance changed to a negative value only in group A. Differences in each of urine components were statistically significant between group A and the other two groups at the end of period. 3) Heart rate and oxygen intake at the same submaximal work load increased significantly in group A at the end of weight reduction period, but in groups B and C no noticeable changes were observed.(ABSTRACT TRUNCATED AT 250 WORDS)     

Changes in the subepithelial mesenchymal cell process meshwork in developing facial prominences in mouse embryos

We examined temporal and spatial changes in the subepithelial mesenchymal cell process meshwork (CPM) in normally developing medial (MNP) and lateral nasal prominences (LNP) in mouse embryos by light and scanning electron microscopy. Marked changes were found only in the MNP during the fusion of the MNP and LNP. The CPM density in the prospective fusion area of the MNP gradually increased as the epithelial surfaces approached each other, attained its maximum just before contact, and decreased after contact. The CPM density in the prospective fusion area of the LNP changed only slightly even when the epithelial surfaces approached each other. The increase in CPM density paralleled that in the density of mesenchymal cell bodies. The LNP grew more actively toward the line of fusion than did the MNP during the progressive fusion of the two prominences. A larger number of fusion-associated epithelial morphological changes--the appearance of superficial protruding cells and cell degeneration--occurred in the MNP than in the LNP. These findings suggest that the increased CPM density is closely related to the growth of the facial prominences and the fusion-associated epithelial morphology and that the CPM plays an important role in the epithelial-mesenchymal interaction during the formation of the upper lip and primary palate.     

Hydrolysis of DNA by Ce(IV)/EDTA complexes--the mechanism of DNA cleavage and design of artificial restriction enzymes.

Homogeneous Ce(IV) complex of EDTA promptly hydrolyzes oligonucleotides under physiological conditions. Moreover, the activity of Ce(IV)/EDTA for DNA hydrolysis is promoted by the addition of amines. When [Ce(IV)/EDTA] = 5 mumol dm3 and [ethylenediamine] = 100 mmol dm3, the catalytic activity is about 50 times as large as that of Ce(IV)/EDTA. The combination of Ce(IV)/EDTA and amines is eminent tools for the future molecular biology and biotechnology.     

The effect of transforming growth factor-beta on the alkaline phosphatase activity in rabbit renal cortical tubular cell cultures

The effect of various growth regulators including epidermal growth factor and transforming growth factor-beta on the alkaline phosphatase activity of rabbit renal cortical tubular cells has been investigated in a serum-free culture. As a result, it was found that transforming growth factor-beta, known to be a growth inhibitor of renal tubular cells, increased the alkaline phosphatase activity of the tubular cells dose-dependently and that cycloheximide blocked any increase in the activity of this factor. In contrast, epidermal growth factor decreased the alkaline phosphatase activity in the tubular cells.     

Transforming growth factor-beta-like activity is secreted by rabbit renal cortical tubular cells in primary culture.

The activity of an autocrine growth factor in a medium conditioned by cultured rabbit renal cortical tubular cells was investigated. Little stimulatory growth activity for tubular cells was observed in the conditioned medium, and inhibitory activity was seen only in acidified conditioned medium. This factor stimulated the colony formation of NRK 49F cells in soft agar only with epidermal growth factor and inhibited the DNA synthesis of primary cultured rat hepatocytes, and its molecular weight was about 25 kDa. The factor was neutralized by the specific antibody to transforming growth factor (TGF)-beta 1. These results indicate that renal tubular epithelial cells can produce latent TGF-beta in primary culture.