Translocation mechanism and the role of aerenchyma in nanoplastic translocation in Myriophyllum sp. "Roraima" and physiological responses.

Nanoplastics have become a growing concern due to their potential impact on freshwater vegetation. The uptake, translocation, and effects of 0.05-µm nanoplastics on Myriophyllum sp.

Cell-free translation system with artificial lipid-monolayer particles as a unique tool for characterizing lipid-monolayer binding proteins.

Methods for functional analysis of proteins specifically localizing to lipid monolayers such as rubber particles and lipid droplets are limited. We have succeeded in establishing a system in which artificially prepared lipid monolayer particles are added to a cell-free translation system to confirm the properties of proteins that specifically bind to lipid monolayers in a translation-coupled manner.

Acylated plastoquinone is a novel neutral lipid accumulated in cyanobacteria.

Although cyanobacteria do not possess bacterial triacylglycerol (TAG)-synthesizing enzymes, the accumulation of TAGs and/or lipid droplets has been repeatedly reported in a wide range of species. In most cases, the identification of TAG has been based on the detection of the spot showing the mobility similar to the TAG standard in thin-layer chromatography (TLC) of neutral lipids. In this study, we identified monoacyl plastoquinol (acyl PQH) as the predominant molecular species in the TAG-like spot from the unicellular sp.

Localization of Lipid Droplets in Embryonic Axis Radicle Cells of Soybean Seeds under Various Imbibition Regimes Indicates Their Role in Desiccation Tolerance.

Desiccation tolerance allows plant seeds to remain viable during desiccation and subsequent re-hydration. In this study, we tried to develop an experimental system to understand the difference between desiccation tolerant and desiccation sensitive radicle cells by examining excised embryonic axes after re-desiccation and subsequent imbibition under various regimes. Embryonic axes excised from soybean ( (L.

Using a novel CLEM system to localize lipid droplets and membranes in desiccated embryonic axis cells of soybean seeds.

Lipid droplets and membranes in radicle cells from desiccated embryonic axes of soybean (Glycine max) seeds were examined by a recently developed correlative light and electron microscopy system, which has been designed to facilitate the observation of identical locations using an upright reflected light microscope and compact SEM successively with minimum time lapse. Lipids are major components of membranes and are also stored in numerous lipid droplets lining plasma membranes in many seed cells. Fluorescently stained lipid droplets and membranes in the desiccated radicle cells were mainly located along the surface of shrunk protoplasm and around presumptive protein bodies, which will turn into vacuoles and increase their volume for radicle protrusion.

The NAD Kinase Slr0400 Functions as a Growth Repressor in Synechocystis sp. PCC 6803.

NADP+, the phosphorylated form of nicotinamide adenine dinucleotide (NAD), plays an essential role in many cellular processes. NAD kinase (NADK), which is conserved in all living organisms, catalyzes the phosphorylation of NAD+ to NADP+. However, the physiological role of phosphorylation of NAD+ to NADP+ in the cyanobacterium Synechocystis remains unclear.

TEM observation of compacted DNA of Synechococcus elongatus PCC 7942 using DRAQ5 labeling with DAB photooxidation and osmium black.

To visualize the fine structure of compacted DNA of Synechococcus elongatus PCC 7942, which appears at a specific time in the regular light/dark cycle prior to cell division, ChromEM with some modifications was applied. After staining DNA with DRAQ5, the cells were fixed and irradiated by red laser in the presence of 3,3'-diaminobenzidine and subsequently fixed with OsO4. A system with He-Ne laser (633 nm) was set up for efficient irradiation of the bacterial cells in aqueous solution.

Fine structure of Aldrovanda vesiculosa L: the peculiar lifestyle of an aquatic carnivorous plant elucidated by electron microscopy using cryo-techniques.

The aquatic carnivorous plant Aldrovanda vesiculosa L. is critically endangered worldwide; its peculiar lifestyle raises many questions and poses problems both intriguing on their own and relevant to conservation. While establishing a culture system for its propagation and restoring its natural habitat in Hozoji pond in Saitama, Japan, we conducted ultrastructural observations to examine the various aspects of Aldrovanda's way of life.

Para-crystalline membrane structures resembling prolamellar bodies in the invasion zones of indeterminate root nodules of Vicia faba L.

Novel para-crystalline structures resembling prolamellar bodies in etioplasts were found in the invasion zones of indeterminate root nodules of Vicia faba, which possess persistent meristems and exhibit sequential developmental stages. The para-crystalline structures existed in most cells in the area of the invasion zone and a hexagonal arrangement of tubular membranes was recognized. Extensive membranes, apparently procured from the structures, were often in contact with the bacteria in young infected cells.

One of the NAD kinases, sll1415, is required for the glucose metabolism of Synechocystis sp. PCC 6803.

Pyridine nucleotides (NAD(P)(H)) are electron carriers that are the driving forces in various metabolic pathways. Phosphorylation of NAD(H) to NADP(H) is performed by the enzyme NAD kinase (NADK). Synechocystis sp.

Backscattered electron imaging of high pressure frozen soybean root nodules visualizes formation of symbiosome membranes.

High-pressure frozen soybean root nodules were fractured and backscattered electron images were obtained from uncoated samples in a low vacuum scanning electron microscope equipped with a cryo-transfer system. Structures of infected cells were well preserved: numerous symbiosomes, as well as nuclei, plastids and mitochondria were observed without ice crystal damage. After appropriate sublimation of water, bacteria included in symbiosomes were visualized.

Effects of inactivation of the cyAbrB2 transcription factor together with glycogen synthesis on cellular metabolism and free fatty acid production in the cyanobacterium Synechocystis sp. PCC 6803.

Deletion of the cyAbrB2 (Sll0822) transcription factor in Synechocystis sp. PCC 6803 causes aberrant accumulation of glycogen. We previously tried to redirect the excess carbon stored as glycogen in the cyabrB2-disrupted (∆ cyabrB2) mutant by knockout of the glgC (slr1176) gene encoding glucose-1-phosphate adenylyltransferase.

Visualization of DNA Compaction in Cyanobacteria by High-voltage Cryo-electron Tomography.

This protocol describes how to visualize the transient DNA compaction in cyanobacteria. DNA compaction is a dramatic cytoplasmic event recently found to occur in some cyanobacteria before cell division. However, due to the large cell size and the transient character, it is difficult to investigate the structure in detail.

Manganese-mediated immobilization of arsenic by calcifying macro-algae, Chara braunii.

The restoration capability of charophyte Chara braunii was studied in arsenic-polluted water in the context of biogenic calcium and manganese depositions on the plant. In addition to calcite encrustation, formation of craterlike shape deposits of manganese oxides (MnO) with diameters of 5-10 μm was detected on the cell walls of the plants grown in Mn-rich media. Relative proportions of arsenic taken up by the plant biomass to those incorporated into the calcium and manganese biominerals were determined using a modified sequential chemical extraction method.

Backscattered electron imaging and elemental analysis of rapidly frozen plant cells using variable accelerating voltage.

Rapidly frozen rosemary leaves were observed at variable accelerating voltages in a low-vacuum scanning electron microscope equipped with a cryo transfer system. After water was sublimated from the fractured face of the leaf, distinct backscattered electron (BSE) images were obtained depending on the accelerating voltages applied. At 5 kV, surface cell wall structure was observed, whereas at 10 and 15 kV chloroplasts lining the inside of the cell wall and membrane were visualized.

Inositol Hexakis Phosphate is the Seasonal Phosphorus Reservoir in the Deciduous Woody Plant Populus alba L.

Seasonal recycling of nutrients is an important strategy for deciduous perennials. Deciduous perennials maintain and expand their nutrient pools by the autumn nutrient remobilization and the subsequent winter storage throughout their long life. Phosphorus (P), one of the most important elements in living organisms, is remobilized from senescing leaves during autumn in deciduous trees.

Buoyancy Limitation of Filamentous Cyanobacteria under Prolonged Pressure due to the Gas Vesicles Collapse.

Freshwater cyanobacterium Pseudanabaena galeata were cultured in chambers under artificially generated pressures, which correspond to the hydrostatic pressures at deep water. Variations occurred in gas vesicles volume, and buoyancy state of cells under those conditions were analyzed at different time intervals (5 min, 1 day, and 5 days). Variations in gas vesicles morphology of cells were observed by transmission electron microscopy images.

Effects of tiotropium on lung function in current smokers and never smokers with bronchial asthma.

The effects of tiotropium, an inhaled long-acting muscarinic antagonist, on lung function were investigated in current smokers and nonsmokers with asthma treated with inhaled corticosteroids (ICSs) and other asthma controllers: inhaled long-acting β agonists, leukotriene receptor antagonists, and/or theophylline. We conducted a double-blind, placebo-controlled study of an inhaled single dose of tiotropium in 9 asthmatics currently smoking and 9 asthmatics who have never smoked in a crossover manner. Lung function was measured before and 1, 3, and 24 h after inhalation of 18 μg of tiotropium or a placebo.

Ultrastructure of compacted DNA in cyanobacteria by high-voltage cryo-electron tomography.

Some cyanobacteria exhibit compaction of DNA in synchrony with their circadian rhythms accompanying cell division. Since the structure is transient, it has not yet been described in detail. Here, we successfully visualize the ultrastructure of compacted DNA in the cyanobacterium Synechococcus elongatus PCC 7942 under rigorous synchronized cultivation by means of high-voltage cryo-electron tomography.

Prevalence of Airflow Limitation Defined by Pre- and Post-Bronchodilator Spirometry in a Community-Based Health Checkup: The Hisayama Study.

Spirometry in health checkup may contribute to early diagnosis of chronic obstructive pulmonary disease (COPD) and asthma. Although post-bronchodilator airflow limitation is essential for definite diagnosis of COPD and post-bronchodilator normalization of airflow is suggestive of asthma, this test has not been prevailed in health checkup. The objective of this study was to estimate the prevalence of airflow limitation defined by pre- and post-bronchodilator spirometry in health checkup.

Free fatty acid production in the cyanobacterium Synechocystis sp. PCC 6803 is enhanced by deletion of the cyAbrB2 transcriptional regulator.

The cyAbrB2 (Sll0822) transcriptional regulator in Synechocystis sp. PCC 6803 is involved in coordination of carbon and nitrogen metabolism and its deletion causes distinct phenotypes such as decreased expression levels of nitrogen-regulated genes and high accumulation of glycogen granules. From the viewpoint of metabolic engineering, the highly accumulated glycogen granules in the ΔcyabrB2 mutant could be a valuable source for the production of biofuels.

The Effects of Dark Incubation on Cellular Metabolism of the Wild Type Cyanobacterium Synechocystis sp. PCC 6803 and a Mutant Lacking the Transcriptional Regulator cyAbrB2.

The cyAbrB2 transcriptional regulator is essential for active sugar catabolism in Synechocystis sp. PCC 6803 grown under light conditions. In the light-grown cyabrB2-disrupted mutant, glycogen granules and sugar phosphates corresponding to early steps in the glycolytic pathway accumulated to higher levels than those in the wild-type (WT) strain, whereas the amounts of 3-phosphoglycerate, phosphoenolpyruvate and ribulose 1,5-bisphosphate were significantly lower.

Abscisic acid-induced rearrangement of intracellular structures associated with freezing and desiccation stress tolerance in the liverwort Marchantia polymorpha.

The plant growth regulator abscisic acid (ABA) is known to be involved in triggering responses to various environmental stresses such as freezing and desiccation in angiosperms, but little is known about its role in basal land plants, especially in liverworts, representing the earliest land plant lineage. We show here that survival rate after freezing and desiccation of Marchantia polymorpha gemmalings was increased by pretreatment with ABA in the presence of increasing concentrations of sucrose. ABA treatment increased accumulation of soluble sugars in gemmalings, and sugar accumulation was further increased by addition of sucrose to the culture medium.

Photoassimilation, assimilate translocation and plasmodesmal biogenesis in the source leaves of Arabidopsis thaliana grown under an increased atmospheric CO2 concentration.

Using 18-day-old Arabidopsis thaliana seedlings grown under increased (780 p.p.m.