Persistence of plant hormone levels in rice shoots grown under microgravity conditions in space: its relationship to maintenance of shoot growth

We investigated the effects of microgravity environment on growth and plant hormone levels in dark‐grown rice shoots cultivated in artificial 1 g and microgravity conditions on the International Space Station (ISS). Growth of microgravity‐grown shoots was comparable to that of 1 g‐grown shoots. Endogenous levels of indole‐3‐acetic acid (IAA) in shoots remained constant, while those of abscisic acid (ABA), jasmonic acid (JA), cytokinins (CKs) and gibberellins (GAs) decreased during the cultivation period under both conditions. The levels of auxin, ABA, JA, CKs and GAs in rice shoots grown under microgravity conditions were comparable to those under 1 g conditions. These results suggest microgravity environment in space had minimal impact on levels of these plant hormones in rice shoots, which may be the cause of the persistence of normal growth of shoots under microgravity conditions. Concerning ethylene, the expression level of a gene for 1‐aminocyclopropane‐1‐carboxylic acid (ACC) synthase, the key enzyme in ethylene biosynthesis, was reduced under microgravity conditions, suggesting that microgravity may affect the ethylene production. Therefore, ethylene production may be responsive to alterations of the gravitational force.     

A comparative evaluation of culture conditions for short-term maintenance (<24 hr) of human islets isolated using the Edmonton protocol

Once human islets are isolated, they are typically transplanted into type 1 diabetic recipients within 2 h of isolation. This time restriction makes it difficult for patients to travel from distant locations to receive an islet transplant and it also makes it difficult to complete pre-release quality control assessments (i.e., endotoxin and gram stain) before the expiration of the islet product. Therefore, there were two goals for this study. The first was to measure the stability of islets after a 24 h culture period using CMRL media 1066 (CMRL) supplemented with either fetal bovine serum (FBS); albumin or insulin transferrin and selenium (ITS). The second was to determine the impact of cell concentration and media depth on islet stability. The results of the study indicated that culture recoveries at 37 °C with CMRL + ITS (also known as Memphis media) were higher (64.1 ± 8.3%) than with CMRL supplemented with FBS (38.7 ± 9.7%) or albumin (47.6 ± 8.2%) and that post-culture islet viabilities, post-culture purities and stimulation indexes (SIs) were comparable. In the second series of experiments, the results showed that islets recoveries and SIs in cultures with low islet concentrations (300 IE/ml) were significantly better than cultures at high islet concentrations (1500 IE/ml). Additionally, at a shallow media depth (1.4 vs. 7 mm of media) the SI of the islets improved, and this effect was independent of the additive (i.e., FBS, albumin and ITS).     

Localization of Endogenous Biotin-Containing Proteins in Mouse Bergmann Glial Cells

A peroxidase-conjugated avidin–biotin complex was used to detect endogenous biotin-containing proteins in mouse cerebellum. By this method, Bergmann glial cells were found to be strongly labelled in the adult mouse cerebellum. Developmentally, cells in the granular layer, probably astrocytes, appeared to be labelled around postnatal 10-day (P10). Their labelling decreased after P20, although the positive-labelling remained in the Bergmann glial cells up to the adult stage. The findings were confirmed by using a Alexa Fluor 488-conjugated streptavidin technique. The labelling was not affected by routine hydrogen peroxide treatment, but it was eliminated by avidin–biotin blocking. By another transblot method, the reactive proteins in the mouse cerebellum were found to be 120?kDa (the strongest one) and 75?kDa. For electron microscopy, a gold-conjugated anti-biotin antibody was immunoreacted to the mitochondria of Bergmann glial cells. These results suggest that endogenous biotin-containing proteins are abundant in the Bergmann glial cells. Therefore, the avidin–biotin complex method is useful for detecting Bergmann glial cells, probably because of the difference of biotin metabolism in the cerebellar glial cells.     

Immunohistochemical Localization of Hemicelluloses and Pectins Varies During Tissue Development in the Bamboo Culm

The distribution of hemicelluloses and pectins in bamboo internodes was studied immunocytochemistrically at various stages of development. The ultra-structures of bamboo cell walls have been reported previously at various stages. The internodes were identically classified into three developmental phases: primary wall stage (phase I), unlignified secondary wall stage (phase II) and lignified wall stage (phase III), using the same bamboo culm. (1→,1→4)-β-Glucans were distributed in nearly all tissues in an actively elongating stage. Limited amounts of β-glucans were deposited in primary walls and the middle lamellae, but were limited to the phloem in secondary walls. This suggests that the function of β-glucans might be different in phloem vis-à-vis other tissues. Highly-substituted xylans were located in nearly all tissues of early phase I, but had disappeared in all tissues immediately prior to lignification. In contrast, low-branched xylan epitopes were present only in the protoxylem in phase I, but were present in all tissues immediately prior to lignification in phase II. In phase III, the epitopes were densely localized in lignified walls, suggesting that the substitution of xylans is closely related to maturation. Methyl-esterified (but not unesterified) pectins were present in all tissues of early phase I. Just before and after lignification, both types of pectins were concentrated in the phloem and protoxylem. Xyloglucans were largely distributed in the phloem and in lignified tissues, suggesting that they might be closely correlated with maturation. This represents the first account of the distribution of hemicelluloses and pectins at the tissue and ultrastructural level in bamboo internodes at various stages of development.     

Characteristics of gas exchange and morphology of a spring ephemeral, Erythronium japonicum, in comparison with a sun plant, Glycine max

Morphological characteristics and responses of gas exchanges to light intensity were examined in a typical vernal species, Erythronium japonicum Decne (E. japonicum), grown (i) on the floor of a deciduous broad-leaved Quercus mongolica forest (one of its native habitats, the Q. mongolica stand); (ii) bare land left undisturbed for 9 years after forest clearing (the bare stand); and (iii) in a sun crop, soybean, grown for 110 days in an experimental field and for 17 days in pots, in order to evaluate the adaptability of the photosynthetic process of this vernal species to its shady native habitats. The daytime solar radiation, ai and leaf temperatures and leaf–air vapor pressure difference (VPD) were significantly higher at the bare stand than at the Q. mongolica stand. When environmental factors observed at the Q. mongolica and bare stands were reproduced in an assimilation chamber, leaf temperatures of E. japonicum plants increased markedly with increased radiation, whereas those of soybean plants differed little from the respective air temperatures. The photosynthetic and transpiration rates and stomatal conductance in the former plants placed under conditions at the Q. mongolica stand increased with radiation and reached respective steady state values at maximum radiation at the site; whereas, under the conditions at the bare stand, they also increased and reached respective steady state values, but then continuously decreased to be lower than the respective value at the Q. mongolica stand. However, both rates and the conductance in the soybean plants under both conditions increased significantly with radiation and reached much higher respective values at the respective maximum radiations. Water use efficiency for E. japonicum plants was much higher under conditions at the Q. mongolica stand than at the bare stand and was practically equal to those for soybean plants under both conditions. Water potential in the leaves of E. japonicum at maximum radiation at the bare stand was one-third that of those at the Q. mongolica stand. The potential in soybean leaves differed little between both conditions and was roughly equal to the low value in E. japonicum leaves at the bare stand conditions. The stomatal densities on upper and lower leaf surfaces and the ratio of root weight to leaf area (R : L) differed little between E. japonicum plants grown at both stands as well as between young and adult soybean plants. However, the densities on the upper and lower surfaces of E. japonicum were 25% and 60% of the respective values of both soybean plants. The ratios of R : L of the E. japonicum plants were only one-quarter that of the young and adult soybean plants. The cooperation between these morphological and gas exchange characteristics in E. japonicum plants is discussed in relation to adaptation to the environment in native habitats.     

TT virus is distributed in various leukocyte subpopulations at distinct levels, with the highest viral load in granulocytes.

When TT virus (TTV) DNA was quantitated in whole blood and plasma aliquots from 27 viremic individuals by real-time detection PCR that can detect essentially all TTV genotypes, the TTV load was 6.9 +/- 3.5 (mean +/- standard deviation)-fold higher in the whole blood than in the plasma samples [P < 0.002 (paired t test)]. To clarify the reason for this difference, peripheral blood cells of various types including red blood cells, granulocytes (CD15+), B cells (CD19+), T cells (CD3+), monocytes (CD14+), and NK cells (CD3-/CD56+) were separated at a purity of 95.4-99.5% from each of three infected individuals with relatively high TTV viremia, and their TTV viral loads were determined. Red blood cells were uniformly negative, but the other cell types were positive for TTV DNA at various titers. In all three patients, the highest TTV load was found in granulocytes (4.2 x 10(4)-3.1 x 10(5) copies/10(6) cells), followed by monocytes (1.4-2.2 x 10(4) copies/10(6) cells) and NK cells (5.4-6.5 x 10(3) copies/10(6) cells); B and T cells were positive, with a low viral load (6.7 x 10(1)-2.7 x 10(3) copies/10(6) cells). These results indicate that TTV is distributed in various peripheral blood cell types at distinct levels, with the highest viral load in granulocytes, and that a significant proportion of the TTV DNA in peripheral blood is not identified by the standard plasma/serum DNA detection methods.     

FK506 enhanced osteoblastic differentiation in mesenchymal cells.

Bone morphogenetic protein (BMP) is a bone-derived growth factor capable of promoting the differentiation of mesenchymal cells into osteogenic lineage pathways. Recently, immunosuppressants were reported to cause a moderate increase in osteoblastic differentiation in a rat osteoblast-like osteosarcoma cell line. If immunosuppressants can induce osteoblastic differentiation, it will be useful for bone tissue transplantation. We assessed the effect of immunosuppressants with or without BMP-4 on inducing osteoblastic differentiation in osteoblast-like and other mesenchymal cells. FK506, an immunosuppressant often used clinically, induced a dose- and time-dependent increase in alkaline phosphatase (ALP) activity, one of the markers of osteoblast differentiation, in cells derived from mesenchyma. In the presence of BMP-4, ALP activity, mRNA levels of ALP and osteocalcin increased. FK506 was found to not only stimulate osteoblastic differentiation, but also to enhance BMP-4 induced osteoblastic differentiation. These results suggest that FK506 promotes differentiation of osteoblastic cells.     

Species separation in <Emphasis Type="Italic">Curvularia</Emphasis> “geniculata” group inferred from <Emphasis Type="Italic">Brn1</Emphasis> gene sequences

 Brn1, a reductase gene involved in the melanin biosynthetic pathway, was adopted for species delimitation among members in the “geniculata” group of Curvularia species and proved to be useful for this purpose. Phylogenetic trees of these fungal members were constructed from nucleotide sequences of this region. The so-called geniculata group of Curvularia was separated into several clusters. The conidial morphology of the members in each cluster is closely similar but clearly different among discrete clusters. The phylogenetic groups almost concurred with the morphological grouping. Thus, the synonymous treatment of Curvularia affinis, C. fallax, and C. senegalensis to C. geniculata in a previous study was supported. The isolates with warping hilum conidia were clearly different from C. geniculata and separated into two clusters. C. geniculata ATCC 6671 made an independent cluster situated near these clusters. The protuberant hilum species were located separately in the phylogenetic trees. For sound taxonomic treatment of these isolates, we should accumulate more information and retain our species determination for them. Received: September 26, 2002 / Accepted: March 12, 2003     

Marked reduction of Helicobacter pylori-induced gastritis by urease inhibitors, acetohydroxamic acid and flurofamide, in Mongolian gerbils.

Urease has been suggested to be essential for colonization and pathogenesis of Helicobacter pylori infection. In the present study, we evaluated the effects of urease inhibitors [acetohydroxamic acid (AHA) and flurofamide (FFA)] on H. pylori-induced gastritis in Mongolian gerbils. Animals were orally inoculated with H. pylori, and given urease inhibitors in their diet throughout the experimental period of six weeks or four weeks, starting from two weeks after H. pylori inoculation. With the administration of AHA at doses of 100, 500, and 2500 ppm throughout the experimental period, H. pylori-induced gastritis in animals was decreased in a dose-dependent manner, significantly so at 2500 ppm. Suppression of gastric lesions was also evident in animals administered 2500 ppm AHA after the H. pylori infection. Bacterial infection rates were reduced to 40-50% of the control value of 100%, by the highest dose of AHA. The potent urease inhibitor, FFA, also caused marked amelioration of H. pylori-associated gastritis on administration at 100 ppm throughout the six-week experimental period or for four weeks after H. pylori infection. Animals treated with FFA had few visible gastric lesions, and the proportion infected with H. pylori was reduced to less than 10%. Since antibiotic-resistant strains of H. pylori have become a serious problem, nonantibiotic urease inhibitors may be very useful to control H. pylori-associated gastroduodenal disease.