Synthesis and expression of genes encoding tuna, pigeon, and horse cytochromes c in the yeast Saccharomyces cerevisiae.

Genes encoding tuna, pigeon, and horse cytochromes c were constructed with synthetic oligodeoxyribonucleotides having preferred codons and portions of the iso-1-cytochrome c-encoding gene from the yeast Saccharomyces cerevisiae. The genes were ligated into an expression vector, which contains the normal 5'- and 3'-untranslated regions of the yeast iso-1-cytochrome c gene, and were integrated in single copy into the chromosome. Yeast strains were also constructed with multiple integrated copies of the pigeon gene. The heterologous and normal mRNA levels of the single-copy strains were equivalent. Although the N-terminal methionines were completely cleaved in the heterospecific proteins, the levels of trimethylation of Lys72 and acetylation of N-terminal glycines ranged from 39-78% and 10-70%, respectively. Horse cytochrome c was produced at a nearly normal level, whereas the pigeon and tuna cytochromes c were produced at approx. 40% of the normal levels. The levels of the cytochromes c and growth of the mutant yeast strains indicated that the heterospecific cytochromes c had approx. 50% specific activity in vivo.     

Lactate dehydrogenase isoenzymes are present in matrix vesicles

Matrix vesicles were isolated from epiphyseal growth plates of young rabbits. Lactate dehydrogenase activity was detected in the isolated matrix vesicles only in the presence of detergents, suggesting that NADH, the cofactor for the assay, does not penetrate the membrane of matrix vesicles. In contrast, the activity of alkaline phosphatase, a marker enzyme of the outer surface of matrix vesicles, was detected in the matrix vesicles using p-nitrophenyl phosphate as the substrate both in the presence and absence of detergents. Lactate dehydrogenase activity was detected only in the cytosol of chondrocytes of the epiphyseal growth plates but not in other subcellular fractions, showing that lactate dehydrogenase is not from the plasma membrane and membranes of intracellular organelles of chondrocytes. The isolated matrix vesicles contained all five lactate dehydrogenase isoenzymes but did not possess other cytosolic enzymes. These results show that lactate dehydrogenase is located in the matrix vesicles and suggest the presence of a mechanism for the specific uptake of cytosolic lactate dehydrogenase and the possibility of enzymatic quantification of the matrix vesicles at various calcification sites.     

Electrochemical characterization of immobilized NAD+

Nicotinamide adenine dinucleotide (NAD⁺) has been covalently attached to alginic acid using carbodiimide coupling, thereby producing a macromolecular adduct of NAD, which can be rendered either soluble or insoluble by adjustment of pH. It was found that this NAD⁺ · alginic acid complex was enzymatically active, and also that the oxidized form could be electrochemically reduced without loss in enzymatic activity. This NAD⁺ adduct has now also been polarographically characterized as to its two-step reduction waves, which are slightly shifted toward more cathodic potential as compared to free NAD⁺. When controlled electrolysis was conducted to reduce the bound NAD⁺ at the cathode, the NADH so formed by electrochemical action was found to be again oxidizable either enzymatically or electrochemically without loss in co-enzymic function. The NADH adduct produced by electrochemical reduction of the NAD⁺ adduct has also been characterized by voltammetry.     

Age-related changes in blood pressure in the senescence-accelerated mouse (SAM): aged SAMP1 mice manifest hypertensive vascular disease

Age-related changes in systolic blood pressure were assessed, using the senescence-accelerated mouse (SAM) model for aging research with strains SAMR1, SAMP1, and SAMP8. Each of the strains manifested a characteristic change in blood pressure with age. The SAMR1 strain, with normal aging, did not have chronologic changes from 2 to 27 months of age. The SAMP1 strain, with accelerated senescence, had a significant increase in blood pressure with age, and some (8 of 39) mice manifested hypertensive vascular disease characterized by high blood pressure, cardiac hypertrophy, and arteriolar fibrinoid necrosis at 11 to 14 months of age. The gradual increase in blood pressure after 8 to 10 months was considered to be preceded by progressive renal changes, from glomerulonephritis to contraction of the kidney, suggesting that the high blood pressure in the SAMP1 strain was of renal origin. Blood pressure in the SAMP8 strain, with age-related deficits in learning and memory, gradually decreased after 5 to 7 months of age, and was suggested to be due to the astrogliotic changes in response to spongiform degeneration in the medulla oblongata at 11 to 14 and 15 to 18 months of age.     

Immediate-Early Expression of the Gene Causing Superinfection Breakdown in Bacteriophage T4B

Superinfection breakdown appears to belong to the "immediate-early" functions induced by T4 phage.     

On the synchorological and floristic trends and discontinuities in regard to the Japan-Liukiu-Formosa area

Conclusion and summary The author discussed the floristic and vegetational differences (a) between Formosa and the Philippines, especially between Botel Tobago and Formosa, (b) between Formosa and Continental China, (c) between Japan proper and Continental China together with Corea, Mandschuria and the eastern part of Siberia, (d) between Formosa and Japan proper, and then he discussed (e) the flora of the Liukius, especially the differences between the Yaeyama Islands and Formosa as well as between Yaku Shima and the Amami Islands. In addition he briefly outlined the vegetation of Formosa and Japan, especially the altitudinal distribution of various forest-communities.The conclusions confirmed the floristic discontinuity between Formosa and the Philippines already set forth by E. D. Merrill and several other botanists. Concerning the phytogeographical position of Botel Tobago, the writer confirms the opinion of T. Kano and R. Kanehira that the flora of Botel Tobago is an extension of that of the Batan-Babuyan Islands, the northern border of the Philippines.The floristic close-relation between Japan proper and Continental China is evident from the phytogeographical distribution of the genera of higher plants found in Japan.Data show that the Formosan flora, especially of the highland, is more closely related to the south-western part of China, and also to the Himalayan region, than to the other surrounding regions.The writer considers that the relationship of the flora of Japan proper to that of Formosa is rather weak. He thinks, however, that their floras and vegetations are related to each other through their mutual relation to the lowland and lower montane flora of Continental China situated besides both of them.It is concluded that the flora of the Liukius is an extension of the Formosan flora, judging by the distribution of the genera and species. Looking over the Japan-Liukiu-Formosa area, a conspicuous floristic and synchorological discontinuity is recognized between Yaku Shima and the Amami Islands, and it is conceivable that the Yaku Shima flora is a south-western extension of the flora of Kyushu or Japan proper, as well as the vegetation of Yaku Shima is closely related essentially to that of Japan proper.The writer and his cooperators adopted a new name of Ardisieto-Shiietum Sieboldi to the Shiia forest of Yaku Shima.The writer is unable to make any synchorological or plant-sociological precise comparison between Japan and Continental China along with the eastern part of Siberia and moreover between Formosa and Continental China, because of few or no plant-sociological detailed data of the continental area in the Far Eastern Asia.The author delivered a special lecture on this subject before the botany class in the National Taiwan University, Taipei, Formosa (Taiwan), on December 4^th 1953. — Contributions from the Department of Biology, Faculty of Science, Kyushu University, No. 38.     

Genetic typing of the Senescence-Accelerated Mouse (SAM) strains with microsatellite markers

The Senescence-Accelerated Mouse (SAM) strains constitute a murine model of accelerated senescence originating from the ancestral AKR/J strains and consist of nine senescence-prone (SAMP) strains and four senescence-resistant (SAMR) strains. The chromosomes (Chrs) of the SAM strains were typed with 581 microsatellite markers amplified by PCR, and the fundamental genetic information of the SAM strains was obtained. One-third of the examined markers displayed polymorphism among the strains, and only two alleles were detected in almost all loci among the SAM and AKR/J strains. However, in 12 loci (5.6% of total 215 polymorphic markers), the third allele was detected among the SAM strains. The genetic typing and developmental history suggested that the SAM strains were related inbred strains developed by the accidental crossing between the AKR/J strain and other unknown strain(s). Comparison of the distribution of the loci in the SAMP and the SAMR series revealed notable differences in the four regions on Chrs 4, 14, 16, and 17. This indicated that some of these chromosomal sites might contain the genes responsible for accelerated senescence in the SAMP series. Received: 17 July 1998 / Accepted: 17 November 1998