Alpha-tocopherol content and alpha-tocopherol transfer protein expression in leukocytes of children with acute leukemia

-Tocopherol (a form of vitamin E) is a fat-soluble vitamin that can prevent lipid peroxidation of cell membranes. This antioxidant activity of -tocopherol can help to prevent cardiovascular disease, atherosclerosis and cancer. We investigated the -tocopherol level and the expression of -tocopherol transfer protein (-TTP) in the leukocytes of children with leukemia. The plasma and erythrocyte -tocopherol levels did not differ between children with leukemia and the control group. However, lymphocytes from children with leukemia had significantly lower -tocopherol levels than lymphocytes from the controls (58.4±39.0 ng/mg protein versus 188.9±133.6, respectively; p0.05), despite the higher plasma -tocopherol/cholesterol ratio in the leukemia group (5.83±1.64 μmol/mmol versus 4.34±0.96, respectively; p0.05). No significant differences in the plasma and leukocyte levels of isoprostanes (the oxidative metabolites of arachidonic acid) were seen between the leukemia patients and controls. The plasma level of acrolein, a marker of oxidative stress, was also similar in the two groups. Investigation of -TTP expression by leukocytes using real-time PCR showed no difference between the two groups. These findings suggest that there may be comparable levels of lipid peroxidation in children with untreated leukemia and controls, despite the reduced -tocopherol level in leukemic leukocytes.     

Nitrate reductase from the magnetotactic bacterium Magnetospirillum magnetotacticum MS-1: purification and sequence analyses

We purified the nitrate reductase from the soluble fraction of Magnetospirillum magnetotacticum MS-1. The enzyme was composed of 86- and 17-kDa subunits and contained molybdenum, non-heme iron, and heme c. These properties are very similar to those of the periplasmic nitrate reductase found in Paracoccus pantotrophus. The M. magnetotacticum nap locus was clustered in seven open reading frames, napFDAGHBC. The phylogenetic analyses of NapA, NapB, and NapC suggested a close relationship between M. magnetotacticum nap genes and Escherichia coli nap genes, which is not consistent with the 16S rDNA data. This is the first finding that the alpha subclass of Proteobacteria possesses a napFDAGHBC-type nap gene cluster. The nap gene cluster had putative fumarate and nitrate reduction regulatory protein (Fnr) and NarL protein binding sites. Furthermore, we investigated the effect of molybdate deficiency in medium on the total iron content of the magnetosome fraction and discussed the physiological function of nitrate reductase in relation to the magnetite synthesis in M. magnetotacticum.     

Mutation rate and novel tt mutants of Arabidopsis thaliana induced by carbon ions

Irradiation of Arabidopsis thaliana by carbon ions was carried out to investigate the mutational effect of ion particles in higher plants. Frequencies of embryonic lethals and chlorophyll-deficient mutants were found to be significantly higher after carbon-ion irradiation than after electron irradiation (11-fold and 7.8-fold per unit dose, respectively). To estimate the mutation rate of carbon ions, mutants with no pigments on leaves and stems (tt) and no trichomes on leaves (gl) were isolated at the M2 generation and subjected to analysis. Averaged segregation rate of the backcrossed mutants was 0.25, which suggested that large deletions reducing the viability of the gametophytes were not transmitted, if generated, in most cases. During the isolation of mutants, two new classes of flavonoid mutants (tt18, tt19) were isolated from carbon-ion-mutagenized M2 plants. From PCR and sequence analysis, two of the three tt18 mutant alleles were found to have a small deletion within the LDOX gene and the other was revealed to contain a rearrangement. Using the segregation rates, the mutation rate of carbon ions was estimated to be 17-fold higher than that of electrons. The isolation of novel mutants and the high mutation rate suggest that ion particles can be used as a valuable mutagen for plant genetics.     

A Saccharomyces cerevisiae genome-wide mutant screen for altered sensitivity to K1 killer toxin

Using the set of Saccharomyces cerevisiae mutants individually deleted for 5718 yeast genes, we screened for altered sensitivity to the antifungal protein, K1 killer toxin, that binds to a cell wall beta-glucan receptor and subsequently forms lethal pores in the plasma membrane. Mutations in 268 genes, including 42 in genes of unknown function, had a phenotype, often mild, with 186 showing resistance and 82 hypersensitivity compared to wild type. Only 15 of these genes were previously known to cause a toxin phenotype when mutated. Mutants for 144 genes were analyzed for alkali-soluble beta-glucan levels; 63 showed alterations. Further, mutants for 118 genes with altered toxin sensitivity were screened for SDS, hygromycin B, and calcofluor white sensitivity as indicators of cell surface defects; 88 showed some additional defect. There is a markedly nonrandom functional distribution of the mutants. Many genes affect specific areas of cellular activity, including cell wall glucan and mannoprotein synthesis, secretory pathway trafficking, lipid and sterol biosynthesis, and cell surface signal transduction, and offer new insights into these processes and their integration.     

Different accumulation of elements in the rami of the coronary arteries of Thai

To analyze the accumulation of elements in the rami of the coronary arteries, the authors determined element contents in both the coronary arteries and their rami of the Thai by inductively coupled plasma-atomic emission spectrometry. The subjects consisted of 13 men and 6 women, ranging in age from 34 to 83 yr. It was found that a higher accumulation of calcium and phosphorus occurred in the proximal sites of coronary arteries such as the anterior descending, right coronary, left coronary, and circumflex arteries than the distal sites such as the diagonal, lateral, conus, acute marginal, and posterior descending arteries. The accumulation of calcium and phosphorus in the anterior descending and right coronary arteries of the Thai increased remarkably in the fifties, thereafter decreased, and increased again in the eighties. It should be noted that a very high accumulation of calcium and phosphorus in the coronary arteries occurred in the fifties. In regard to relationships among the average contents of elements in the coronary arteries and their rami, it was found that there were very significant direct correlations between calcium and phosphorus contents, between calcium and magnesium contents, and between phosphorus and magnesium contents. These results suggested that as calcium and phosphorus increased in the coronary arteries and their rami, magnesium increased simultaneously in all of the coronary arteries and their rami.     

Accumulation of elements in the arteries and cardiac valves of Thai with aging

To elucidate whether the extent of element accumulation in the arteries and cardiac valves with aging was different between different races, the authors investigated the accumulation of elements in the arteries and cardiac valves of the Thai with aging and the relationships among elements in the cardiac valves. After ordinary dissection at Chiang Mai University was finished, 16 arteries and 4 cardiac valves were resected and element contents were determined by inductively coupled plasma-atomic emission spectrometry. In the 16 arteries, the average content of calcium was the highest in the site of the abdominal aorta ramifying into the common iliac arteries, and it decreased in the order internal iliac, coronary, abdominal aorta, common iliac, external iliac, superior mesenteric, inferior mesenteric, thoracic aorta, brachial, radial, common carotid, subclavian, ulnar, axillary, renal, and internal thoracic arteries. The average contents of phosphorus and magnesium in respective arteries were parallel with the average contents of calcium, except for the coronary artery. In comparison with the arteries of the Japanese, the trend of calcium accumulation in the arteries of the Thai was almost similar to that in the arteries of the Japanese, except for the coronary artery and thoracic aorta. The calcium accumulation in the coronary artery was much higher in the Thai than in the Japanese, whereas that in the thoracic aorta was lower in the Thai than in the Japanese. Regarding elements in the cardiac valves, the calcium content increased remarkably in the seventies in the aortic valve and in the nineties in the pulmonary valve, but it hardly increased in both the mitral and tricuspid valves with aging. The average content of calcium was the highest in the aortic valve, and it decreased in the order pulmonary, tricuspid, and mitral valves. Regarding the relationship among elements in the aortic valves, it was found that there were extremely significant direct correlations among the contents of calcium, phosphorus, and magnesium, whereas there were significant direct correlations between zinc and either calcium or phosphorus contents. Although significant correlations were found between sulfur and the other element contents in the aortic valves of the Japanese, no significant correlations were found between them in the aortic valves of the Thai. In the mitral valves, extremely or very significant direct correlations were found among the contents of calcium, phosphorus, magnesium, and sulfur, with some exceptions that there were no significant correlations between phosphorus and either magnesium or sulfur contents. In addition, no significant correlation was found in the calcium content between the aortic valve and coronary artery in the same individuals.     

Mass ratios of magnesium to calcium and phosphorus in the arteries of Japanese and Thai

To elucidate quantitative changes of Ca, P, and Mg in the arteries with aging, the authors investigated changes of the mass ratios of Mg to Ca and P in the arteries of Japanese and Thai by inductively coupled plasma-atomic emission spectrometry. The arteries of Japanese that were used were the thoracic and abdominal aortas, coronary, common iliac, internal iliac, external iliac, and femoral arteries, in which very high accumulations of Ca and P occurred in old age. The arteries of Thai that were used were the abdominal aorta, ramifying site of the abdominal aorta, coronary, common iliac, internal iliac, and external iliac arteries. It was found that there were extremely significant correlations both between Ca and Mg contents and between P and Mg contents in all of the arteries of the Japanese and the Thai. With regard to the mass ratio, the mass ratios of Mg to Ca ranged from 1.5% to 2.1% in the six arteries of the Japanese, except for the thoracic aorta at 3.1%, at an advanced stage of atherosclerosis, being similar to each other. In the arteries of the Thai, the mass ratios of Mg to Ca ranged from 1.9% to 3.0%, except for the coronary artery at 0.5%, at an advanced stage of atherosclerosis. The mass ratios of Mg to P ranged from 2.5% to 2.7% in the six arteries of the Japanese, except for the coronary artery at 1.8%, at an advanced stage of atherosclerosis. With regard to the arteries of the Thai, the mass ratios of Mg to P ranged from 1.9% to 3.3%, except for the coronary artery at 0.7%, at an advanced stage of atherosclerosis. These results revealed that both the mass ratios of Mg to Ca and Mg to P were almost similar among the arteries of Japanese and Thai, except for the coronary arteries. Therefore, these results suggested that the inorganic deposits in the coronary arteries of Japanese and Thai were similar to those in the intimal tunica of the thoracic aorta, whereas in the other arteries, they were similar to those in the middle tunica of the thoracic aorta.     

A special comment on trace metals in the viscera of Turbo marmoratus, a toxic snail, on Ryukyu Island

Trace metals in the viscera of Turbo marmoratus, a huge conical spire, in the coral reef of Ryukyu Island were analyzed. The highest amount of Zn and Fe was detectable in the midgut gland known to be polluted by saxitoxin. This intestine maintained a considerable amount of Si (higher than Ca) which fluctuated in parallel with the taurine amount. These data suggest that the viscera of the alimentary tract accumulate some metals, especially Zn and Fe.     

Isoflavonoids from Dalbergia olivari

Two isoflavonoids, named olibergin A (1) and B (2) were isolated from the stem bark of Dalbergia oliveri (Leguminosae). Along with three previously known compounds, they are inhibitors of Epstein-Barr virus early antigen activation induced by 12-O-tetradecanoylphorbol-13-acetate in Raji cells. Their structures were elucidated on the basis of spectroscopic analyses.     

Crystal structure of a conger eel galectin (congerin II) at 1.45A resolution: implication for the accelerated evolution of a new ligand-binding site following gene duplication

The crystal structure of congerin II, a galectin family lectin from conger eel, was determined at 1.45A resolution. The previously determined structure of its isoform, congerin I, had revealed a fold evolution via strand swap; however, the structure of congerin II described here resembles other prototype galectins. A comparison of the two congerin genes with that of several other galectins suggests acceralated evolution of both congerin genes following gene duplication. The presence of a Mes (2-[N-morpholino]ethanesulfonic acid) molecule near the carbohydrate-binding site in the crystal structure points to the possibility of an additional binding site in congerin II. The binding site consists of a group of residues that had been replaced following gene duplication suggesting that the binding site was built under selective pressure. Congerin II may be a protein specialized for biological defense with an affinity for target carbohydrates on parasites' cell surface.