Stimulation of Adrenal Chromaffin Cell Proliferation by Hypercalcemia Induced by Intravenous Infusion of Calcium Gluconate in Rats

Increased incidence of adrenal pheochromocytoma is frequently encountered in rat carcinogenicity studies. In some of the studies, the finding is judged to be due to a rat-specific mechanism of carcinogenesis caused by a disturbance of calcium homeostasis. However, direct evidence that the proliferation of chromaffin cells in the adrenal medulla is induced solely by hypercalcemia is not available. In this study, calcium gluconate was intravenously infused for 7 days to rat chromaffin cells by a tail cuff method, and cumulative labeling with bromodeoxyuridine (BrdU) was carried out to evaluate the proliferative activity. The serum calcium concentration was dose-dependently increased, and a high calcium concentration was stably sustained from day 2 to 7. In the adrenal medulla, BrdU-positive chromaffin cells increased in the calcium gluconate-treated animals, and the BrdU-labeling index increased in a dose-dependent manner. In addition, an increased BrdU-labeling index of chromaffin cells was shown to correlate with the serum calcium concentration. Our results demonstrate that hypercalcemia directly enhances the proliferative activity of chromaffin cells and that the proliferative activity is correlated with the serum calcium concentration.     

Roles of one-carbon metabolism in preimplantation period--effects on short-term development and long-term programming--

One-carbon metabolism (OCM) can be seen as integrated metabolic pathways centered on the metabolism of two nutritional substances, folate and methionine. Mammalian oocytes and preimplantation embryos express almost all enzymes that participate in OCM, suggesting that they can independently metabolize OCM nutrients. A deficiency or excess of OCM nutrients and their metabolites during in vitro culture affects preimplantation development of mammalian embryos. Recent in vivo studies have demonstrated that specific OCM dietary interventions during the periconceptional (mainly oocyte growth and preimplantation) period can cause epigenetic alterations in DNA of offspring and program the long-term consequences in their health in adulthood. The epigenetic processes are likely to be implicated in the effects of OCM nutrients; however, understanding their effects at the level of specific genes and their implications in assisted reproductive technology will require further investigations.     

Isolation and identification of C-19 fatty acids with anti-tumor activity from the spores of Ganoderma lucidum (reishi mushroom)

We previously showed that ethanolic extracts of spores of Ganoderma lucidum inhibit tumor cell proliferation and induce apoptosis of HL-60 cells. The active constituents appeared to be long-chain fatty acids, particularly carbon-19 (C-19) fatty acids which have not been reported in spores of Ganoderma lucidum. In the present study, two of these C-19 fatty acids which are key compounds in the activities, were identified as their 2-naphthyl ester derivatives after esterification of a mixture of fatty acids obtained from the spores. The active compounds were determines as nonadecanoic acid and cis-9-nonadecenoic acid. The location of the double bond of cis-9-nonadecenoic acid was demonstrated by GC-MS analysis, based on the fragmentation pattern of the adduct prepared from the fatty acid and dimethyl disulfide.     

Identification and kinetic study of tyrosinase inhibitors found in sake lees

The present study found that the n-hexane extract of freeze-dried sake lees inhibits tyrosinase activity and showed that the constituents isolated from the n-hexane extract are the mixture of triacylglycerols. The inhibitory effects of triolein and trilinolein found as the triacylglycerols were examined using tyrosinases from mushroom and Streptomyces castaneoglobisporus. The IC50 values of the triacylglycerol mixture for the oxidase activity on mushroom and Streptomyces tyrosinases were 20 and 0.14 microg/mL, respectively. The IC50 values of trilinolein for the oxidase activity on mushroom and Streptomyces tyrosinases were 8.4 and 0.1 microM, respectively. However, the inhibitory effect of triolein (IC50=30 microM) was lower than that of trilinolein, even when the Streptomyces tyrosinase was used for the assay. Kinetic analyses indicate that both trilinolein and triolein inhibit the tyrosinase activity noncompetitively. When transformed with a plasmid carrying the Streptomyces tyrosinase gene, the melanin-synthesizing ability of the transformed Escherichia coli host was dose-dependently interfered with by trilinolein.     

Effects at early stage of life of elevated milk replacer feeding on growth rate,plasma IGF‐I concentration and intestinal nutrient transporter expression in Holstein bull calves

In order to evaluate the effects of an elevated amount of modified milk replacer on body weight, daily gain, starter intake, plasma endocrine parameters and expression of nutrient transporters in small intestinal epithelia, Holstein bull calves (n = 24) were fed for 60 days either with the usual amount of 24% crude protein (CP) and 20% fat milk (CF) replacer (C group), or with a double amount of a modified milk replacer of 28% CP and 16% CF (E group). Body weight from D20 to D60 and daily gain before D40 was greater or tended to be greater for the E group than the C group. Plasma concentrations of insulin‐like growth factor‐1 (IGF‐I) and insulin were greater for the E group than the C group on D28 but not on D56, without changing plasma growth hormone levels. Gene expression for sodium‐dependent glucose transporter 1 and fatty acid translocase (CD36) was altered in day‐ and intestine‐dependent manners. From these findings, we conclude that an elevated intake of milk replacer given up to 40 days old is sufficient to enhance body weight, which may be associated with increased plasma IGF‐I concentrations, in Holstein bulls.     

Relationships between immunoglobulin and fat‐soluble vitamins in colostrum of Japanese Black multiparous cows

Data from 19 Japanese Black multiparous cows were collected to clarify the relationships among immunoglobulin (Ig) G, IgA, β‐carotene, vitamin A and α‐tocopherol contents in colostrum of cows in order to evaluate the role of fat‐soluble vitamins on colostral IgG and IgA production. Mean colostral IgG was 141 mg/mL, ranging from 65 to 208 mg/mL, whereas mean colostral IgA was 8.7 mg/mL, ranging from 1.0 to 34.6 mg/mL. Colostral IgG increased with aging in multiparous cows. There were positive correlations between colostral IgG and colostral vitamin A or colostral α‐tocopherol in cows, and the higher adjusted R² was obtained in the prediction model of colostral IgG from age and colostral vitamin A. Colostral vitamin A was positively correlated with colostral β‐carotene or colostral α‐tocopherol in cows, but there were no relationships between colostral IgA and colostral IgG or colostral fat‐soluble vitamins. These results indicate that fat‐soluble vitamin contents in colostrum of cows may change in similar patterns and high colostral vitamin A is related with high colostral IgG.     

Complex patterning by vertical interchange atom manipulation using atomic force microscopy

The ability to incorporate individual atoms in a surface following predetermined arrangements may bring future atom-based technological enterprises closer to reality. Here, we report the assembling of complex atomic patterns at room temperature by the vertical interchange of atoms between the tip apex of an atomic force microscope and a semiconductor surface. At variance with previous methods, these manipulations were produced by exploring the repulsive part of the short-range chemical interaction between the closest tip-surface atoms. By using first-principles calculations, we clarified the basic mechanisms behind the vertical interchange of atoms, characterizing the key atomistic processes involved and estimating the magnitude of the energy barriers between the relevant atomic configurations that leads to these manipulations.     

Methylene Blue Stainability of Root-Tip Protoplasts as an Indicator of Aluminum Tolerance in a Wide Range of Plant Species, Cultivars and Lines

We previously developed a new simple technique of methylene blue (MB) staining for the discrimination of aluminum (Al)-tolerant protoplasts from 4 plant species (rice, oats, maize and pea). The objectives of the present study were to confirm the applicability of this technique to a wider range of plant species, cultivars and lines, and to identify a common strategy for the early stage of Al tolerance. A total of 10 plant species, two Brachiaria spp., two Oryza spp., buckwheat, maize, pea, triticale, wheat and barley, corresponding to 18 different plant samples (species, cultivars, and lines), were used. Al tolerance (relative net root elongation of the longest root), which was screened at 20 μM AICI₃ in 0.2 mM CaCl₂ (pH 4.9) for 24 h, ranged widely from 10 to 88. Among cultivars and lines within the same species, Al accumulation in the root tip portion was higher in Al-sensitive plants, corresponding to more severe permeabilization of the plasma membranes (PM). Protoplasts isolated mainly from the epidermis, and outer and central parts of the cortex were stained to different degrees by MB, and the blue color was observed both on the surface and inside the protoplasts. Color pictures obtained after staining for 3 min with 0.1% MB were analyzed by Image Hyper II. The ratio of the heavily stained area at threshold 95 to the entire area stained with MB at threshold 125 was defined as MB stainability. MB stainability was negatively correlated with Al tolerance ( y = 48.6e^{−0.02 x} , R ²= 0.676**) suggesting the common importance of permeation characteristics of PM, in addition to PM negativity for Al tolerance in a wide range of plant species, cultivars and lines. Analysis of the PM lipid composition was proposed as an important topic for future studies on the negativity and permeation of PM.