Planting geometry as a pre‐screening technique for identifying CO2 responsive rice genotypes: a case study of panicle number

Identifying CO₂ responsive genotypes is a major target for enhancing crop productivity under future global elevated atmospheric CO₂ concentration ([CO₂]). However, [CO₂]‐fumigation facilities are extremely expensive and are not easily accessible, and are limited in space for large‐scale screening. Hence, reliable donors for initiating [CO₂]‐responsive breeding programs are not in place for crops, including rice. We propose a simple and novel phenotyping method for identifying [CO₂]‐responsive genotypes, and quantify the responsiveness to low planting density over 4‐year trials across both temperate and tropical conditions. Panicle number per plant is the key determinant of grain yield and hence was the focus trait across all our trials. In temperate climate, a 3‐season field screening using 127 diverse rice genotypes and employing two planting densities (normal and low density) was conducted. Two japonica genotypes were selected based on their higher responsiveness to low planting density as candidates for validating the proposed phenotyping protocol as a pre‐screen for [CO₂]‐responsiveness. The approach using the two selected candidates and three standard genotypes was confirmed using a free‐air CO₂ enrichment facility and temperature gradient chambers under elevated [CO₂]. In tropical climate, we grew three rice cultivars, previously identified for their [CO₂]‐responsiveness, at two planting densities. The experiments provided confirmation that responsiveness to low planting density was correlated with that of [CO₂]‐responsiveness across both the temperate and tropical conditions. The planting density would be useful pre‐screening method for testing large panels of diverse germplasm at low cost complemented by available CO₂‐control facilities for final validation of candidates from the pre‐screens.     

Importance of specific hydrogen bonds of archaeal rhodopsins for the binding to the transducer protein

Four rhodopsins, bacteriorhodopsin (bR), halorhodopsin (hR), sensory rhodopsin (sR) and phoborhodopsin (pR) exist in archaeal membranes. bR and hR work as a light-driven ion pump. sR and pR work as a photo-sensor of phototaxis, and form signaling complexes in membranes with their respective cognate transducer proteins HtrI (with sR) and HtrII (with pR), through which light signals are transmitted to the cytoplasm. What is the determining factor(s) of the specific binding to form the complex? Binding of the wild-type or mutated rhodopsins with HtrII was measured by isothermal titration calorimetric analysis (ITC). bR and hR could not bind with HtrII. On the other hand, sR could bind to HtrII, although the dissociation constant (K(D)) was about 100 times larger than that of pR. An X-ray crystallographic structure of the pR/HtrII complex revealed formation of two specific hydrogen bonds whose pairs are Tyr199(pR)/Asn74(HtrII) and Thr189(pR)/Glu43(HtrII)/Ser62(HtrII). To investigate the importance of these hydrogen bonds, the K(D) value for the binding of various mutants of bR, hR, sR and pR with HtrII was estimated by ITC. The K(D) value of T189V(pR)/Y199F(pR), double mutant/HtrII complex, was about 100-fold larger than that of the wild-type pR, whose K(D) value was 0.16 microM. On the other hand, bR and hR double mutants, P200T(bR)/V210Y(bR) and P240T(hR)/F250Y(hR), were able to bind with HtrII. The K(D) value of these complexes was estimated to be 60.1(+/-10.7) microM for bR and to be 29.1(+/-6.1) microM for hR, while the wild-type bR and hR did not bind with HtrII. We concluded that these two specific hydrogen bonds play important roles in the binding between the rhodopsins and transducer protein.     

Phylogeography of Potentilla fruticosa,an alpine shrub on the Qinghai-Tibetan Plateau

Aims Our objectives were (i) to elucidate the phylogeography of chloroplast DNA (cpDNA) in Potentilla fruticosa in relation to Quaternary climate change and postglacial colonization, (ii) to infer historical population range expansion using mismatch distribution analyses and (iii) to locate the refugia of this alpine species on the Qinghai-Tibetan plateau during glacial–interglacial periods.Methods Potentilla fruticosa is a widespread species distributed on the Qinghai-Tibetan Plateau. We sampled leaves of P. fruticosa from 10 locations along a route of ~1?300 km from the northeastern plateau (Haibei, Qinghai) to the southern plateau (Dangxiong, Tibet). We examined the cpDNA of 15 haplotypes for 87 individuals from the 10 populations based on the sequence data from ~1?000 base pairs of the trnS–trnG and rpl20–rps12. Phylogenetic relationship of haplotypes was analyzed using the Phylip software package and the program TCS. The diversity of populations indices was obtained using the program ARLEQUIN.Important findings With the limited samples, we found that (i) higher nucleotide diversity often occurs in high-altitude populations, (ii) the ancestral haplotypes distribute in the populations with higher nucleotide diversity than recent haplotypes, (iii) the expansion time of population in the high altitudes was estimated to be approximately at 52–25 ka BP (1000 years Before Present, where “Present” is AD 1950) and that in the low altitudes to be ~5.1–2.5 ka BP and (iv) the source location of P. fruticosa is at the high altitudes, which might provide refugia for the species during the interglacial warm periods. The species expanded from the high-elevated locations on the Tanggula Mountains during the Holocene.     

Galectin-3 but not galectin-1 induces mast cell death by oxidative stress and mitochondrial permeability transition

Galectin-1 and galectin-3 are the most ubiquitously expressed members of the galectin family and more importantly, these two molecules are shown to have opposite effects on pro-inflammatory responses and/or apoptosis depending on the cell type. Herein, we demonstrate for the first time that galectin-3 induces mast cell apoptosis. Mast cells expressed substantial levels of galectin-3 and galectin-1 and to a lesser extent the receptor for advanced glycation end products (RAGE) on their surfaces. Treatment of cells with galectin-3 at concentrations of > or =100 nM for 18-44 h resulted in cell death by apoptosis. Galectin-3-induced apoptosis was completely prevented by lactose, neutralizing antibody to RAGE, and the caspase-3 inhibitor z-DEVD-fmk. Galectin-3-induced apoptosis was also completely abolished by dithiothreitol and superoxide dismutase, but not inhibited by catalase. Moreover, galectin-3 but not galectin-1 induced the release of superoxide, which was blocked by lactose, anti-RAGE, and dithiothreitol. Finally, galectin-3-induced apoptosis was blocked by bongkrekic acid, an antagonist of the mitochondrial permeability transition pore (PTP), while atractyloside, an agonist of the PTP, greatly facilitated galectin-1-induced apoptosis. These data suggest that galectin-3 induces oxidative stress, PTP opening, and the caspase-dependent death pathway by binding to putative surface receptors including RAGE via the carbohydrate recognition domain.     

Involvement of Aquaporin-5 Water Channel in Osmoregulation in Parotid Secretory Granules

Aquaporins (AQPs) are a family of channel proteins that allow water or very small solutes to pass, functioning in tissues where the rapid and regulated transport of fluid is necessary, such as the kidney, lung, and salivary glands. Aquaporin-5 (AQP5) has been demonstrated to localize on the luminal surface of the acinar cells of the salivary glands. In this paper, we investigated the expression and function of AQP5 in the secretory granules of the rat parotid gland. AQP5 was detected in the secretory granule membranes by immunoblot analysis. The immunoelectron microscopy experiments confirmed that AQP5 was to be found in the secretory granule membrane. Anti-AQP5 antibody evoked lysis of the secretory granules but anti-aquaporin-1 antibody did not and AQP1 was not detected in the secretory granule membranes by immunoblot analysis. When chloride ions were removed from the solution prepared for suspending secretory granules, the granule lysis induced by anti-AQP5 antibody was inhibited. Furthermore, 4,4′-diisothiocyanatostilbene-2,2′-disulfonic acid, an anion channel blocker, blocked the anti-AQP5 antibody-induced secretory granule lysis. These results suggest that AQP5 is, expressed in the parotid gland secretory granule membrane and is involved in osmoregulation in the secretory granules.     

Oxygen tension regulates reactive oxygen generation and mutation of Helicobacter pylori

Although both bacillary and coccoid forms of Helicobacter pylori reside in human stomach, the pathophysiological significance of the two forms remains obscure. The present work describes the effect of oxygen tension on the transformation and reactive oxygen species (ROS) metabolism of this pathogen. Most H. pylori cultured under an optimum O2 concentration (7%) were the bacillary form, whereas about 80% of cells cultured under aerobic or anaerobic conditions were the coccoid form. The colony-forming unit of H. pylori decreased significantly under both aerobic and anaerobic culture conditions. The bacillary form of H. pylori generated predominantly superoxide radical, whereas the coccoid form generated preferentially hydroxyl radical. Specific activities of cellular respiration, urease, and superoxide dismatase decreased markedly after transformation of the bacillary form to the coccoid form, with concomitant generation of protein carbonyls and 8-hydroxyguanine. The frequency of mutation of cells increased significantly during culture under nonoptimum O2 conditions. These results indicate that ROS generated by H. pylori catalyze the oxidative modification of cellular DNA, thereby enhancing the transformation from the bacillary to the coccoid form. The enhanced generation of mutagenic hydroxyl radicals in the coccoid form might accelerate mutation and increase the genetic diversity of H. pylori.     

Selective reaction of hydroxylamine with chromophore during the photocycle of pharaonis phoborhodopsin

Crystals of alpha-momorcharin (MMC) were cross-linked and soaked in an 80% acetonitrile--water mixture and X-ray data were collected to 2.2 A resolution. MMC is a ribosome-inactivating protein with a sugar chain on Asn-227. In previous studies, the whole conformation of the sugar chain could not be obtained in the aqueous system. Here the structure of MMC in a low water system is shown to be similar to the native one, but the sugar chain on Asn-227 is defined by the electron density map. Several oxygen atoms of the oligosaccharide formed intramolecular hydrogen bonds to the protein moiety. The conformation of the residues in the active center is similar to that in the aqueous system. Our results show conformational alteration of the tetrasaccharide of MMC in organic media. They indicate that the oligosaccharides are more rigid in organic solvents. X-ray determination in organic media may be used to solve some structures of oligosaccharides linked to glycoproteins.     

The role of shrub (Potentilla fruticosa) on ecosystem CO2 fluxes in an alpine shrub meadow

Aims Recent studies have shown that alpine meadows on the Qinghai-Tibetan plateau act as significant CO₂ sinks. On the plateau, alpine shrub meadow is one of typical grassland ecosystems. The major alpine shrub on the plateau is Potentilla fruticosa L. (Rosaceae), which is distributed widely from 3 200 to 4 000 m. Shrub species play an important role on carbon sequestration in grassland ecosystems. In addition, alpine shrubs are sensitive to climate change such as global warming. Considering global warming, the biomass and productivity of P. fruticosa will increase on Qinghai-Tibetan Plateau. Thus, understanding the carbon dynamics in alpine shrub meadow and the role of shrubs around the upper distribution limit at present is essential to predict the change in carbon sequestration on the plateau. However, the role of shrubs on the carbon dynamics in alpine shrub meadow remains unclear. The objectives of the present study were to evaluate the magnitude of CO₂ exchange of P. fruticosa shrub patches around the upper distribution limit and to elucidate the role of P. fruticosa on ecosystem CO₂ fluxes in an alpine meadow.Methods We used the static acrylic chamber technique to measure and estimate the net ecosystem productivity (NEP), ecosystem respiration (R e), and gross primary productivity (GPP) of P. fruticosa shrub patches at three elevations around the species' upper distribution limit. Ecosystem CO₂ fluxes and environmental factors were measured from 17 to 20 July 2008 at 3 400, 3 600, and 3 800 m a.s.l. We examined the maximum GPP at infinite light (GPP max) and maximum R e (R emax) during the experimental time at each elevation in relation to aboveground biomass and environmental factors, including air and soil temperature, and soil water content.Important findings Patches of P. fruticosa around the species' upper distribution limit absorbed CO₂, at least during the daytime. Maximum NEP at infinite light (NEP max) and GPP max of shrub patches in the alpine meadow varied among the three elevations, with the highest values at 3 400 m and the lowest at 3 800 m. GPP max was positively correlated with the green biomass of P. fruticosa more strongly than with total green biomass, suggesting that P. fruticosa is the major contributor to CO₂ uptake in the alpine shrub meadow. Air temperature influenced the potential GPP at the shrub-patch scale. R emax was correlated with aboveground biomass and R emax normalized by aboveground biomass was influenced by soil water content. Potentilla fruticosa height (biomass) and frequency increased clearly as elevation decreased, which promotes the large-scale spatial variation of carbon uptake and the strength of the carbon sink at lower elevations.     

Effects of chemically- and environmentally-induced hypothermia on micronucleus induction in rats

We investigated micronucleus induction in rats treated with chlorpromazine and reserpine, drugs that induce hypothermia. We administered chlorpromazine (31.3--250mg/kg) or reserpine (500--2000 mg/kg) intraperitoneally and measured temperature rectally. Chlorpromazine at 62.5-250mg/kg and reserpine at all doses significantly decreased rectal temperature, although the hypothermic response was weaker than previously reported in mice. Only chlorpromazine at 250mg/kg decreased rectal temperature transiently to <33 degrees C for 20h and induced a statistically significant increase in micronucleated polychromatic erythrocyte frequency. When rats treated with reserpine at 500mg/kg were exposed to an environmental temperature of 16 degrees C for 6, 12, or 24h to keep their body temperature under 33 degrees C, only the 24h treatment group significantly induced micronuclei. In addition, relatively large micronuclei (diameter of micronucleus> or = 1/4 diameter of cytoplasm) accounted for 33.0% of the induced micronuclei, suggesting that hypothermia affected the mitotic apparatus. The hypothermic response to chlorpromazine and reserpine was weaker in rats than in mice, and it was correspondingly more difficult to induce micronuclei in rats with those drugs.     

Successful treatment of persistent bronchorrhea by gefitinib in a case with Recurrent Bronchioloalveolar Carcinoma: a case report

Background  

Bronchorrhea is one of late complaints in patients with bronchioloalveolar carcinoma (BAC) and hampers their quality of life. Although an effective treatment for bronchorrhea in these patients has not been established, recently we have treated effectively one case of persistent bronchorrhea associated with clinical recurrent BAC with gefitinib (ZD1839, 'Iressa™'; AstraZeneca Japan; Osaka, Japan).