Investigation of Nostoc sp. HK-01, Cell Survival over Three Years during the Tanpopo Mission.

The survival of the terrestrial cyanobacterium Nostoc sp. HK-01 was tested as part of the Tanpopo mission experiment, which was conducted both outside and inside the International Space Station (ISS). The selection of Nostoc sp. HK-01 was based on the results of on-ground experiments that demonstrated that the cyanobacterium can survive simulated space environments. This study verified cell survival after exposure to the outside environment in low Earth orbit (LEO). We examined the cellular tolerance of Nostoc sp. HK-01 simultaneously outside and inside of the ISS over a 3-year period. After the experiments were conducted, we confirmed cell viability by fluorescein diacetate (FDA). Cell growth abilities for 3 years without sunlight in space-vacuum-exposed cells were not significantly different from those of cells kept in the dark of control cells in the ISS and on the ground. Though a few light-exposed cells in space vacuum survived outside the ISS after 3 years as judged by FDA staining assay, the survival could not be verified by testing the growth ability due to an insufficient number of cells. To the best of our knowledge, this is the first pure strain of Nostoc sp. HK-01 that survived in a space environment on the inside and outside of the ISS with and without sunlight for more than 3 years (1126 days).

Bioactive potential of the wild mushroom Astraeus hygrometricus in South-west India.

The wild mushroom Astraeus hygrometricus is a traditional delicacy during the monsoon season in South-western India. Bioactive principles and antioxidant potential of uncooked and pressure-cooked tender mushroom have been evaluated. Seven bioactive principles of mushroom (tannins, flavonoids, vitamin C, phytic acid, lycopene, ß-carotene and trypsin inhibition) were significantly higher, while total phenolics content was significantly lower in uncooked than in cooked samples. Mushroom was devoid of L-DOPA, whereas only uncooked samples showed haemagglutinin activity against A+ blood group. The principal component analysis of uncooked mushroom showed only two bioactive principles clustered with two antioxidant properties, while in cooked mushroom five bioactive principles clustered with three antioxidant properties depicting the nutraceutical potential of cooked mushroom. Future studies should focus on appropriate thermal treatment, which retain maximum bioactive and antioxidant potential to combat health- and lifestyle-related ailments. The A. hygrometricus is ectomycorrhizal, conservation of its host tree species is utmost importance in improvement and expansion of its yield to sustain food security and economic gains of local tribals.

Isolation and purification of an axenic diazotrophic drought-tolerant cyanobacterium, Nostoc commune, from natural cyanobacterial crusts and its utilization for field research on soils polluted with radioisotopes.

Nitrogen fixation and drought tolerance confer the ability to grow on dry land, and some terrestrial cyanobacteria exhibit these properties. These cyanobacteria were isolated in an axenic form from Nostoc commune clusters and other sources by modifying the method used to isolate the nitrogen-fixing and drought-tolerant cyanobacterium Nostoc sp. HK-01. Of these cyanobacteria, N. commune, which is difficult to isolate and purify, uses polysaccharides to maintain water, nitrogen fertilizers for nitrogen fixation, and can live in extreme environments because of desiccation tolerance. In this study, we examined the use of N. commune as biosoil for space agriculture and possible absorption of radioisotopes ((134)Cs, (137)Cs). This article is part of a Special Issue entitled: Photosynthesis Research for Sustainability: from Natural to Artificial.

Neuroprotective effects of flavonoids on hypoxia-, glutamate-, and oxidative stress-induced retinal ganglion cell death.

PURPOSE: This study was conducted to investigate the effect of flavonoids, a major family of antioxidants contained in foods, on retinal ganglion cell (RGC) death induced by hypoxia, excessive glutamate levels, and oxidative stress. Moreover, to assess the structure-activity relationships of flavonoids, three types of flavonoids with different numbers of hydroxyl groups and varieties of sugar chains were studied. METHODS: Three kinds of flavonoids-nicotiflorin, rutin, and quercitrin-were used. The death of neonatal rat purified RGCs was induced by hypoxic conditions (5% O(2), 5% CO(2), 37 °C) for 12 h, 25 µM glutamate over three days, or oxidative stress by depleting antioxidants from the medium for 24 h. RGC survival rates were calculated under each condition and compared with vehicle cultures. Modification of cell death signaling after stress-induced apoptosis and necrosis by flavonoids was assessed using caspase-3 and calpain immunoreactivity assays. RESULTS: Under hypoxic and glutamate stress, both nicotiflorin and rutin significantly increased the RGC survival rate at 1 nM or higher, while quercitrin increased it at 100 nM or higher. Under oxidative stress, nicotiflorin, rutin, and quercitrin also significantly increased the RGC survival rate at 1 nM, 0.1 nM, and 100 nM or higher, respectively. Rutin significantly inhibited the induction of caspase-3 under both hypoxia and excessive glutamate stress, as well as blocking the induction of calpain during oxidative stress. CONCLUSIONS: Nicotiflorin and rutin showed neuroprotective effects on hypoxia-, glutamate- or oxidative stress-induced RGC death at concentrations of 1 nM or higher. The presence of a specific sugar side chain (rutinoside) may enhance neuroprotective activity.

Discovery of the curcumin metabolic pathway involving a unique enzyme in an intestinal microorganism.

Polyphenol curcumin, a yellow pigment, derived from the rhizomes of a plant (Curcuma longa Linn) is a natural antioxidant exhibiting a variety of pharmacological activities and therapeutic properties. It has long been used as a traditional medicine and as a preservative and coloring agent in foods. Here, curcumin-converting microorganisms were isolated from human feces, the one exhibiting the highest activity being identified as Escherichia coli. We are thus unique in discovering that E. coli was able to act on curcumin. The curcumin-converting enzyme was purified from E. coli and characterized. The native enzyme had a molecular mass of about 82 kDa and consisted of two identical subunits. The enzyme has a narrow substrate spectrum, preferentially acting on curcumin. The microbial metabolism of curcumin by the purified enzyme was found to comprise a two-step reduction, curcumin being converted NADPH-dependently into an intermediate product, dihydrocurcumin, and then the end product, tetrahydrocurcumin. We named this enzyme "NADPH-dependent curcumin/dihydrocurcumin reductase" (CurA). The gene (curA) encoding this enzyme was also identified. A homology search with the BLAST program revealed that a unique enzyme involved in curcumin metabolism belongs to the medium-chain dehydrogenase/reductase superfamily.

Discovery of amide (peptide) bond synthetic activity in Acyl-CoA synthetase.

Acyl-CoA synthetase, which is one of the acid-thiol ligases (EC 6.2.1), plays key roles in metabolic and regulatory processes. This enzyme forms a carbon-sulfur bond in the presence of ATP and Mg(2+), yielding acyl-CoA thioesters from the corresponding free acids and CoA. This enzyme belongs to the superfamily of adenylate-forming enzymes, whose three-dimensional structures are analogous to one another. We here discovered a new reaction while studying the short-chain acyl-CoA synthetase that we recently reported (Hashimoto, Y., Hosaka, H., Oinuma, K., Goda, M., Higashibata, H., and Kobayashi, M. (2005) J. Biol. Chem. 280, 8660-8667). When l-cysteine was used as a substrate instead of CoA, N-acyl-l-cysteine was surprisingly detected as a reaction product. This finding demonstrated that the enzyme formed a carbon-nitrogen bond (EC 6.3.1 acid-ammonia (or amide) ligase (amide synthase); EC 6.3.2 acid-amino acid ligase (peptide synthase)) comprising the amino group of the cysteine and the carboxyl group of the acid. N-Acyl-d-cysteine, N-acyl-dl-homocysteine, and N-acyl-l-cysteine methyl ester were also synthesized from the corresponding cysteine analog substrates by the enzyme. Furthermore, this unexpected enzyme activity was also observed for acetyl-CoA synthetase and firefly luciferase, indicating the generality of the new reaction in the superfamily of adenylate-forming enzymes.

Total synthesis of (+)- and (-)-sundiversifolide via intramolecular acylation and determination of the absolute configuration.

Intramolecular acylation of an organolithium leads to an efficient stereocontrolled total synthesis of both enantiomers of sundiversifolide. The absolute configuration was determined by HPLC analysis and allelopathy assay. The gamma-lactone moiety resulted from a butenolide was obtained by the condensation of a bicyclic alpha-hydroxyhemiacetal with Ph3P=CMe(CO2R).

Flavonoids in the leaves of Oxalis corniculata and sequestration of the flavonoids in the wing scales of the pale grass blue butterfly, Pseudozizeeria maha.

Three C-glycosylflavones in the leaves of Oxalis corniculata, the host plant of the lycaenid butterfly pale grass blue (Pseudozizeeria maha), were identified as 6-C-glucosylluteolin (isoorientin), 6-C-glucosylapigenin (isovitexin) and isovitexin 7-methyl ether (swertisin). Comparative spectral and HPLC analyses between the leaf extract of the host plants and the wings of P. maha showed selective uptake of the host-plant flavonoid isovitexin to the wings of the butterfly.

An overview of challenges in modeling heat and mass transfer for living on Mars.

Engineering a life-support system for living on Mars requires the modeling of heat and mass transfer. This report describes the analysis of heat and mass transfer phenomena in a greenhouse dome, which is being designed as a pressurized life-support system for agricultural production on Mars. In this Martian greenhouse, solar energy will be converted into chemical energy in plant biomass. Agricultural products will be harvested for food and plant cultivation, and waste materials will be processed in a composting microbial ecosystem. Transpired water from plants will be condensed and recycled. In our thermal design and analysis for the Martian greenhouse, we addressed the question of whether temperature and pressure would be maintained in the appropriate range for humans as well as plants. Energy flow and material circulation should be controlled to provide an artificial ecological system on Mars. In our analysis, we assumed that the greenhouse would be maintained at a subatmospheric pressure under 1/3-G gravitational force with 1/2 solar light intensity on Earth. Convection of atmospheric gases will be induced inside the greenhouse, primarily by heating from sunlight. Microclimate (thermal and gas species structure) could be generated locally around plant bodies, which would affect gas transport. Potential effects of those environmental factors are discussed on the phenomena including plant growth and plant physiology and focusing on transport processes. Fire safety is a crucial issue and we evaluate its impact on the total gas pressure in the greenhouse dome.

[Natural history of flowers and gravity].

Many flowers have coevolved with their pollinator animals. Gravity has been one of selection pressure for the evolution of flowers. Gravity rules morphology and other features of flowers in many aspects. Pair matching between the flower and its specific pollinator is one of factors that determine the fitness of both sides. Evolution of flower morphology and its molecular basis are reviewed briefly. Anemophilous flowers are also under the influence of gravity. Shape and other features of entomophilous flowers have been highly diversed. Gravitropic response and its mechanism are summarized. Recent findings on gravitropism and phototropism of pistils and stamens are presented in this article.

Fate of allelopathic substances in space--allelopathy of velvet bean plant and gravity.

Interactions between organisms and species have been long known. It has not been known that the interactive phenomena (allelopathy) may (or may not) differ in space from those on earth. We have studied the gravitational effects on allelopathy by exposing a plant-plant system to pseudo-microgravity, which was generated by a 3D-clinostat. We hypothesized that allelopathy would be modified under altered gravity. L-DOPA (L-3,4-dihydroxyphenylalanine) is known to be the major substance in the allelopathy of velvet bean, released from its root. It has been found that there have been some differences of the allelopathic action of velvet bean plant under pseudo-microgravity. Biosynthesis, release, transport and sensing mechanism associated with allelopathy might be affected by gravity.

Up and down asymmetrical world for flower blooming.

Form of plant body shows vertical polarity. Photosynthetic organs deploy upwards to seek light. Root system extends downward for water. In addition to this major polarity, flowers has similar features, because they bloom in the world where up and down asymmetry dominates. Many flowering plants co-evolved with pollinator animals. Success of their reproduction is linked to the shape of flower organs in many ways. Orientation of inflorescence and each individual flowers, arrangement and shape of flower organs exhibit various up and down asymmetry. Some flowers mimic female of its pollinator animal. In such case, posture of the target animal is copied to the flower. Since animal posture and shape have vertical polarity, flower happens to equip same kind of polarity. Tropic response of pistils is another up and down feature that improves fitness of flowering plants. Certain lily flowers show phototropism to bend pistil upward. Azalea flower depends on gravity as the major environmental cue, and light as the secondary signal for up. Molecular machinery for those tropic responses seems to be shared with other tropism expressed in shoot and root. However, certain differences are found in distribution of sediment amyloplast, or spatial allocation of photo-sensing and bending site. Tropic responses result in adaptation of those flowers against behavior of pollinator animals and terrestrial environment, where gravity affects living organisms and interaction among them.

Lily pistils bend toward light.

Flower shape shows up and down features in many cases. Pistils of lily flower, Lilium cv. 'Casablanca', bend upward by sensing exclusively light. There found no sediment amyloplast in pistil corresponding to the absence of gravitropism. Fine granules of amyloplast disappear from pistil and tepal after opening of flower, except stomatal guard cells distributed on their epidermis. Bending of pistil caused by phototropism occurs when rate of its local elongation passes maximum and decreases. Elongation of pistil starts from its basal part close to ovary, and propagate toward distal tip afterwards. Bending of pistil follows this propagation. Because of this feature, curvature of pistil tends to be rather steep at the basal part. From spot irradiation experiment, site of sensing light was found not spatially separated by twice of diameter from site of bending.

Distribution of L-DOPA in the root of velvet bean plant (Mucuna pruriens L.) and gravity.

Velvet bean (Mucuna pruriens L.) has been found that the degree of suppression on the lettuce root growth by velvet bean was less on the 3D-clinorotation. The number and growth of adventitious root in velvet bean differed among the clinostated and control group. L-DOPA (L-3,4-dihydroxyphenylalanine) is known to be the major substance in the allelopathy of velvet bean plant, released from its root. Since L-DOPA is a precursor of melanin pigment, and is easily converted to melanin by oxidation, locality of L-DOPA production in the plant body can be seen through pigmentation. The amount of L-DOPA was analyzed by HPLC and LC-ESI/MS. The distribution of L-DOPA in the root was different among the ground control condition and pseudo-microgravity.

Pistil and stamen of lily flowers bend upward by light.

Pistil and stamen of lily flowers bend upward. Such tropic response at sensing external stimuli increases the success of pollination in these flowers and improve their adaptability. They do not bend under the dark, but toward light irradiated. From these observation, lily flowers is concluded that they equip phototropic mechanism to sense direction of incident light and control differential elongation of pistil and stamen. Action spectrum of light for this tropism was found similar to those for the phototropism shown in coleoptiles of monocotyledonous plants. Image analysis of pistil and stamen indicates that elongation and bending does not proceed uniformly over those organs. In the case of pistil, elongation starts at its basal part and propagates towards its top after opening of perianth. Steep bending occurs at the basal zone of pistil as long as differential incidence of light is given at its part.

Volatile allelopathy in velvet bean (Mucuna pruriens [correction of pruiens] L.) and gravity.

Allelopathy caused by volatile compounds was studied with velvet bean (Mucuna pruriens [correction of prueiens] L.). Preparatory experiment was conducted to identify the relationship between action of volatiles and gravity. The experimental protocol was defined and tested to study on volatile allelopathy of velvet bean. Root growth of lettuce (Lactuca sativa L.) seedlings was inhibited by volatile gas from velvet bean seedlings planted in Agripot. In this study, it was shown that ten-day velvet bean seedling could exhibit the volatile allelopathic ability. Growth chambers for volatile experiment were designed. Velvet bean seed can germinate in a 3-ml small vial.

Growth of endophyte (Neotyphodium) during seed germination of tall fescue (Festuca arundinacea).

Growth of a filamentous fungus endophyte, Neotyphodium, was examined during the seed germination process of its host plant, tall fescue, Festuca arundinacea, by the ELISA method. Distribution of endophyte differed from that of substances induced by endophyte.

Reduced allelopathic inhibition of lettuce (Lactuca sativa) growth caused by velvet bean (Mucuna pruriens) under 3D-clinorotation.

Allelopathy between Mucuna pruriens (velvet bean) and Lactuca sativa (lettuce) was studied under 3D-clinorotation. Growth of both roots and shoots of lettuce seedlings was suppressed by the presence of velvet bean. The degree of suppression was less on the clinostat compared to the normal static earth gravity. L-DOPA (L-3, 4-dihydroxyphenylalanine) is known to be a major substance in allelopathy of velvet bean. Amount of L-DOPA diffused out from a sintered filter paper into agar medium was compared between clinorotation and control group, and found no significant difference. It was concluded that some factors related to release, transport, and sensing phenomena of allelopathic substances may be responsible to the new findings in this study.

Growth of endophyte, Neotyphodium, and its host plant, tall fescue (Festuca arundinacea), under 3D-clinorotation.

Growth of a filamentous fungus endophyte, Neotyphodium, and its host plant, tall fescue, Festuca arundinacea, was examined during the seed germination process under pseudo-microgravity [correction of micrgravity] generated by three dimensional (3D-) clinorotation. The shoot growth of tall fescue infected with the endophyte was remarkably suppressed on a 3D-clinostat compared with that of the ground control. Without being infected, shoot growth of tall fescue was not strongly affected by the 3D-clinorotation. Many aggregated hyphae were observed in the plant seed incubated for 1-day on the 3D-clinostat [correction of clinost] than in those kept on the ground. These results indicate that the clinorotation induces responses in the endophyte and its host plant different from those under normal gravity.

Plant growth-promoting oligosaccharides produced from tomato waste.

Tomato juice waste was hydrolyzed with acid. Tomato juice waste (500 g; wet weight) was heated with 0.5 N HCl (2.5 l) at 70 degrees C for 4 h. After neutralization, the growth-promoting extracts (300 g; dry weight) in the plants were produced from the tomato waste. The acid extract significantly promoted the growth of cockscomb (Celosia argentea L.) and tomato (Lycopersicon esculentum L.) seedlings. We have recognized potent plant growth-promoting substances in the acid extract from tomato waste. The most effective components in the active fraction were almost all oligogalacturonic acids (DP 6-12). This paper is the first report that plant growth-promoting oligosaccharides can be directly produced from tomato juice waste. It is possible that the substances from the tomato waste can become useful plant growth regulators in the agriculture field in the future.