Molecular characterization of <Emphasis Type="Italic">Potato spindle tuber viroid</Emphasis> in dahlia

A new variant of Potato spindle tuber viroid (PSTVd) was detected for the first time from dahlia grown in Japan. The dahlia isolate of PSTVd formed a quasi-species and a major sequence variant consisting of 361 nucleotides in length, including five substitutions, three insertions, and one deletion, when compared to the intermediate strain from potato. In bioassays with the new isolate, Rutgers tomato developed mild stunting and leaf curling.     

Biomass allocation and nitrogen limitation in a Cryptomeria japonica plantation chronosequence

We investigated soil net nitrogen mineralization rate, above- and belowground biomass allocation, and nitrogen use in a Cryptomeria japonica plantation chronosequence. Total biomass accumulation showed an asymptotic accretion pattern, and the peak total biomass accumulation rate occurred approximately 30 years after afforestation. Soil net nitrogen mineralization rate was lowest 30 years after afforestation. Between years 30 and 88, net nitrogen mineralization increased again. These results indicate that an imbalance in soil nitrogen supply and plant nitrogen demand occurred approximately 30 years after afforestation. Furthermore, leaf nitrogen concentration, which was used as an index of plant nitrogen status, was lower in mature forest than in young forest, suggesting that mature stands did not take up nitrogen as successfully. If soil resources such as nitrogen limit plant growth, plants may increase biomass allocation to fine root structure; however, fine root biomass was not higher in 30- and 88-year-old stands than in younger stands, suggesting that changes in biomass allocation may not be effective against nitrogen deficiency in a C. japonica plantation chronosequence.     

Polar cardenolide monoglycosides from stems and twigs of Nerium oleander and their biological activities

Twelve polar cardenolide monoglycosides, 1, 2, 4?C13, and oleagenin (3) were isolated from the methanol extract of stems and twigs of Nerium oleander. Among these, oleagenin (3) and cardenolide monoglycosides named cardenolide B-1 (1) and cardenolide B-2 (2) were isolated from natural sources for the first time. The in vitro antiinflammatory activity of compounds 1?C13 was examined on the basis of inhibitory activity against the induction of the intercellular adhesion molecule-1 (ICAM-1). Compounds 4?C7 were active at an IC₅₀ value of less than 0.4 ??M. The cytotoxic activity of compounds 1?C13 was evaluated against three human cell lines: normal human fibroblast cells (WI-38), malignant tumor cells derived from WI-38 (VA-13), and human liver tumor cells (HepG2). Compounds 4, 6, and 7 were active toward these three cell lines at IC₅₀ values of less than 0.7 ??M, and compounds 5 and 8 were active toward the cell lines at IC₅₀ values of less than 1.5 ??M. The multidrug resistance (MDR) cancer-reversal activity of compounds 1?C13 was evaluated on the basis of the amount of calcein accumulated in MDR human ovarian cancer 2780AD cells in the presence of each compound. Compound 1 and 12 showed significant effects on calcein accumulation.     

Sodium iodoacetate-induced experimental osteoarthritis and associated pain model in rats

Degenerative lesions were induced in the knee joint of Wistar rats by intraarticular injection of chondrocyte metabolism inhibitor mono-iodoacetate (MIA) at doses of 0, 0.3 or 3 mg/joint. Histopathological examination and the measurement of hind paw weight ratio as an index of joint pain by incapacitance tester were performed. Histological findings that are similar to those observed in human osteoarthritis (OA), such as disorganization of chondrocytes, erosion and fibrillation of cartilage surface, and subchondral bone exposure etc., were observed in a MIA-dose-dependent manner. Saflanin-O fast green staining revealed that marked diffuse reduction of proteoglycan in cartilage tissue of rats treated with MIA. The clinical scores of the joint pain were closely correlated to the grade of histological findings. We conclude that the present experimental model in combination with a novel dual channel weight averager would be very useful for the study of human OA, and could be applied for estimation of therapeutic effect of new anti-OA drugs.     

Nucleotide Sequence of Citrus Tristeza Virus Seedling Yellows Isolate

The complete nucleotide sequence of a seedling-yellows-inducing isolate NUagA of Citrus tristeza virus (CTV) was determined. It consisted of 19302 nucleotides and contained 12 open reading frames (ORF) organized identically to those of previously sequenced isolates. This genome is the largest among the CTV genome sequenced so far ; it is 6 nucleotides (nt), 76 nt, 43 nt, and 53 nt longer than that of T36 (quick decline, Florida), VT (seedling yellows, Israel), T385 (mild, Spain), and SY568 (stem pitting, California), respectively. Sequence comparison of NUagA and the other isolates revealed approximately 90% identities throughout the 3′ half of the genome. The 5′ half of the genome was only about 70% identical to that of T36 but still high at about 90% to those of VT, SY568, and T385. Comparison of amino acid sequences on ORF1a encoding polyproteins, the most variable region, reflects the CTV isolate relationship ; NUagA is closely related to VT, SY568, and T385, but distantly related to T36. Received 29 May 2000/ Accepted in revised form 16 November 2000     

Chemical Characteristics of Fog Water at Mt. Tateyama, Near the Coast of the Japan Sea in Central Japan

Measurements of the chemical composition of fog water at Murododaira (altitude, 2,450 m), on the western slope of Mt. Tateyama near the coast of the Japan Sea, were performed each autumn from 2004 through 2007. Strong acidic fogs (pH?<?4) containing high concentrations of nssSO ₄ ^2? were frequently observed in the autumn of 2005, when the air mass at Mt. Tateyama originated mainly from the polluted regions of Asia. The ratio of NO ₃ ^? /nssSO ₄ ^2? in fog water was relatively high in 2004 and 2007. High concentrations of nssCa²⁺ derived from dust particles were detected in 2006. Background Kosa particles might have been predominant in the free troposphere and could have neutralized acidic fogs in the autumn of 2006. High concentrations of sea-salt components were also observed in October 2005. The sea-salt particles might have been transported from the Pacific Ocean by a strong typhoon, and significant Mg²⁺ loss was observed. Peroxides higher than 100 μM, which are seriously harmful to vegetation were sometimes detected.     

The biological activities of cardenolide triglycosides from stems, twigs, and leaves of Nerium oleander

Sixteen cardenolide triglycosides (1?C16) were isolated from stems, twigs, and leaves of Nerium oleander. Among them, 3??-O-(4-O-gentiobiosyl-d-diginosyl)-7??,8-epoxy-14-hydroxy-5??,14??-card-20(22)-enolide, named cardenolide B-3 (16), was isolated from natural sources for the first time. The in vitro anti-inflammatory activities of compounds 1?C16 were examined on the basis of inhibitory activity against the induction of intercellular adhesion molecule-1 (ICAM-1). Compounds 1?C5 were active at an IC50 value of less than 7 ??M. The cytotoxic activity of isolated compounds was evaluated against three human cell lines: normal human fibroblast cells (W-38), malignant tumor cells induced from WI-38 (VA-13), and human liver tumor cells (HepG2). Compounds 1?C5 were active toward WI-38 cells; compounds 1, 3, and 5 were active toward VA-13 cells; and compounds 1?C5 were active toward HepG2 cells at IC₅₀ values of less than 10 ??M. The multidrug-resistant (MDR) cancer-reversal activity of compounds 1?C16 was evaluated on the basis of the amount of calcein accumulated in MDR human ovarian cancer 2780AD cells in the presence of each compound. Compounds 13 and 14 showed significant effects on calcein accumulation.     

Stability to light, heat, and hydrogen peroxide at different pH values and DPPH radical scavenging activity of acylated anthocyanins from red radish extract

The stability of red radish extract to light, heat, and hydrogen peroxide at different pH values (3, 5, and 7) was examined, in which major anthocyanins were pelargonidin glycosides acylated with a combination of p-coumaric, ferulic, or caffeic acids. The light irradiation (fluorescence light, 5000 lx; at 25 degrees C) indicated that the red radish extract was more stable at lower pH than at higher pH. The HPLC analyses revealed that diacylated anthocyanins in the extract were more stable to light at pH 3 than monoacylated anthocyanins. No significant difference in degradation rates of acylated anthocyanins at pH 5 was observed, whereas anthocyanins acylated with p-coumaric or ferulic acids were more stable at pH 7 than ones with caffeic acids. The stability to heat (at 90-95 degrees C) showed a tendency similar to that for light. The number of intramolecular acyl units contributes to stability to light and heat at lower pH, whereas the characteristics of intramolecular acyl units influence the stability at higher pH. The degradation behavior of red radish extract to H2O2 were almost the same to those of light and heat, depending on the pH. However, HPLC analyses revealed that the stability of individual acylated anthocyanins were independent of the pH. These data suggest that the characteristics, the number, and the binding site of intramolecular acyl units affect the stability of anthocyanin to H2O2. DPPH radical scavenging activity of all acylated anthocyanins was higher than those of pelargonidin and perlargonidin-3-glucoside. The activity of acylated anthocyanins mostly depended on the activity of intramolecular acyl units (caffeic acid > ferulic acid > p-coumaric acid). However, the activity was highly affected by the binding site of intramolecular acyl units even if anthocyanins have common acyl units.     

Medium without ammonium accumulation supports the developmental competence of human embryos

L-Glutamine has been shown to play an important role during in vitro culture of mammalian embryos. However, it is easily decomposed into ammonium, which is believed to have deleterious effects on preimplantation embryos. In this study, we assessed prospectively the developmental competence of human embryos cultured in medium containing L-glutamine or a novel stable glutamine derivative and vitamins. The subjects of this study were 41 women who underwent IVF/ET treatment from September to November 2006 and from whom 6 or more oocytes were retrieved. Sibling oocytes were randomly divided into EA/BA (EmbryoAssyst and BlastAssyst containing a novel stable glutamine derivative and vitamins), and BAS groups (BlastAssyst system containing L-glutamine). There was no difference in pronuclear formation rate between EA/BA and BAS (74 vs. 69%). The blastulation rates of embryos based on the number of zygotes cultured in EA/BA on Days 5 (Day 0=insemination, 54%) and 6 (63%) were significantly higher (P<0.05) than those cultured in BAS (Day 5: 33% and Day 6: 45%, respectively). The present data indicate that a medium containing a novel stable glutamine derivative and vitamins supports the developmental competence of human embryos.