Impacts of moso bamboo (Phyllostachys pubescens) invasion on dry matter and carbon and nitrogen stocks in a broad‐leaved secondary forest located in Kyoto,western Japan

In western and central Japan, the expansion of exotic moso bamboo (Phyllostachys pubescens Mazel ex J. Houz.) populations into neighboring vegetation has become a serious problem. Although the effects of bamboo invasion on biodiversity have been well studied, shifts in nutrient stocks and cycling, which are fundamental for ecosystem functioning, are not fully understood. To explore the effects of P. pubescens invasion on ecosystem functions we examined above‐ and below‐ground dry matter and carbon (C) and nitrogen (N) stocks in a pure broad‐leaved tree stand, a pure bamboo stand, and two tree–bamboo mixed stands with different vegetation mix ratios in the secondary forest of Kyoto, western Japan. In the process of invasion, bamboo shoots offset broad‐leaved tree deaths; thus, no clear trend was apparent in total above‐ or below‐ground biomass or in plant C and N stocks during invasion. However, the ratio of above‐ground to below‐ground biomass (T/R ratio at the stand level) decreased with increasing bamboo dominance, especially in the early stages of invasion. This shift indicates that rapid bamboo rhizomatous growth is a main driver of substantial changes in stand structure. We also detected rises in the C/N ratio of forest‐floor organic matter during bamboo invasion. Thus major impacts of P. pubescens invasion into broad‐leaved forests include not only early shifts in biomass allocation, but also changes in the distribution pattern of C and N stored in plants and soil.     

Structural analysis of telechelic polymer solution using dissipative particle dynamics simulations

A telechelic polymer is an amphiphilic polymer that can form micellar structures when dissolved in water. A telechelic polymer solution shows viscoelastic behaviour owing to the formation of characteristic networks, i.e. loops, bridges and dangling chains. For industrial purposes, telechelic polymers have many applications as thickening agents, such as in paints and cosmetics. Thus, it is desirable to predict and control the rheological properties of telechelic polymers. However, detailed studies at the molecular level have not yet been performed. In this study, I use the dissipative particle dynamics (DPD) method to investigate the relationship between the characteristic structural properties and the molecular structure in telechelic polymer solutions. I show that the morphology of telechelic polymer solutions depends on the concentration and chain length, the distribution of the end-to-end distance, the mean square end-to-end distance, the mean square radius of gyration and the time-averaged mean square displacement. Although an effect of entanglement is important for properties of polymer melts, the polymer chain composed of DPD particles cannot reproduce it. Therefore, I compare telechelic polymer solutions with and without the segmental repulsive potential (SRP), which can simulate the effect of entanglement in DPD simulations. The results indicate that it is necessary to include the SRP in DPD simulations to correctly analyse the behaviour of telechelic polymer solutions.     

Generation of insulin-producing β-like cells from human iPS cells in a defined and completely xeno-free culture system

Human induced pluripotent stem （hiPS） cells are considered a potential source for the generation of insulin-producing pancreatic β-ceUs because of their differentiation capacity. In this study, we have developed a five-step xeno-free culture system to efficiently dif- ferentiate hiPS cells into insulin-producing cells in vitro. We found that a high NOGGIN concentration is crucial for specifically inducing the differentiation first into pancreatic and duodenal homeobox-1 （PDX1）-positive pancreatic progenitors and then into neurogenin 3 （NGN3）-expressing pancreatic endocrine progenitors, while suppressing the differentiation into hepatic or intestinal cells. We also found that a combination of 3-isobutyl-l-methylxanthine （IBMX）, exendin-4, and nicotinamide was important for the differentiation into insulin single-positive cells that expressed various pancreatic β-cell markers. Most notably, the differentiated cells contained en- dogenous C-peptide pools that were released in response to various insulin secretagogues and high levels of glucose. Therefore, our results demonstrate the feasibility of generating hiPS-derived pancreatic β-ceUs under xeno-free conditions and highlight their poten- tial to treat patients with type I diabetes.     

Flexible and rigid structures in HIV-1 p17 matrix protein monitored by relaxation and amide proton exchange with NMR

The HIV-1 p17 matrix protein is a multifunctional protein that interacts with other molecules including proteins and membranes. The dynamic structure between its folded and partially unfolded states can be critical for the recognition of interacting molecules. One of the most important roles of the p17 matrix protein is its localization to the plasma membrane with the Gag polyprotein. The myristyl group attached to the N-terminus on the p17 matrix protein functions as an anchor for binding to the plasma membrane. Biochemical studies revealed that two regions are important for its function: D14–L31 and V84–V88. Here, the dynamic structures of the p17 matrix protein were studied using NMR for relaxation and amide proton exchange experiments at the physiological pH of 7.0. The results revealed that the α12-loop, which includes the 14–31 region, was relatively flexible, and that helix 4, including the 84–88 region, was the most protected helix in this protein. However, the residues in the α34-loop near helix 4 had a low order parameter and high exchange rate of amide protons, indicating high flexibility. This region is probably flexible because this loop functions as a hinge for optimizing the interactions between helices 3 and 4. The C-terminal long region of K113-Y132 adopted a disordered structure. Furthermore, the C-terminal helix 5 appeared to be slightly destabilized due to the flexible C-terminal tail based on the order parameters. Thus, the dynamic structure of the p17 matrix protein may be related to its multiple functions.     

Microhabitat distribution and behaviour of Branchiobdellidan Holtodrilus truncatus found on the freshwater shrimp Neocaridina spp. from the Sugo River, Japan

A study was performed on the microhabitat distribution and some aspects of behaviour of the ectosymbiotic branchiobdellidan Holtodrilus truncatus (Annelida, Clitellata) found on the freshwater shrimp that inhabit the Sugo River, Hyogo Prefecture, western Japan. Observations on shrimp that were collected from the Sugo River (2003 to 2011) confirmed that the host shrimp is Neocaridina spp. (Atyidae). The attachment location on the host shrimp was predominately between the 1^st pleopod and the 5th pereopod (55.3%). The reproductive method of H. truncatus is hemaphroditism. The cocoon was found only inside the carapace of the host shrimp. The cocoon was transparent and contained a maximum of 14 juvenile worms (developing embryos). When hatching approached, H. truncatus’s worms became elongated and slender, and only one worm hatched out at a time. When Holtodrilus truncatus was removed from its host and was maintained in river water without any food, it survived for a maximum of 46 days. In a host exchange experiment, where we provided several other freshwater shrimp species, Palaemonidae fed on H. truncatus. Moreover, Palaemon paucidens and Macrobrachium nipponense from Lake Biwa also preyed upon H. truncatus. The possible symbiotic relationship between H. truncatus and Neocaridina spp. (family Atyidae) is further discussed.     

Preliminary observations of spawning behavior of white-streaked grouper (Epinephelus ongus) in an Okinawan coral reef

Epinephelus ongus forms spawning aggregations at specific reef sites at Okinawa. The aim of the present study was to describe the spawning behavior of E. ongus based upon underwater video imagery. Epinephelus ongus spawning is usually paired with courting males and females rushing upwards towards the surface during the spawning ascent, with a subsequent release of eggs at the apex of the ascent. In two of four spawning events we observed, however, the ascending pair was joined by additional males. Fish eggs collected at the site were identified as those of E. ongus and confirmed that this species spawns mainly at night during a slack tide. This study is the first documentation of E. ongus spawning behavior in the field.     

Oxysterol-binding protein (OSBP) is required for the perinuclear localization of intra-Golgi v-SNAREs

Oxysterol-binding protein (OSBP) and OSBP-related proteins (ORPs) have been implicated in the distribution of sterols among intracellular organelles. OSBP regulates the Golgi cholesterol level, but how it relates to Golgi function is elusive. Here we report that OSBP is essential for the localization of intra-Golgi soluble vesicle N-ethylmaleimide-sensitive fusion attachment protein receptors (v-SNAREs). Depletion of OSBP by small interfering RNA causes mislocalization of intra-Golgi v-SNAREs GS28 and GS15 throughout the cytoplasm without affecting the perinuclear localization of Golgi target-SNARE syntaxin5 and reduces the abundance of a Golgi enzyme, mannosidase II (Man II). GS28 mislocalization and Man II reduction are also induced by cellular cholesterol depletion. Three domains of OSBP—an endoplasmic reticulum–targeting domain, a Golgi-targeting domain, and a sterol-binding domain—are all required for Golgi localization of GS28. Finally, GS28 mislocalization and Man II reduction in OSBP-depleted cells are largely restored by depletion of ArfGAP1, a regulator of the budding of coat protein complex (COP)-I vesicles. From these results, we postulate that Golgi cholesterol level, which is controlled by OSBP, is essential for Golgi localization of intra-Golgi v-SNAREs by ensuring proper COP-I vesicle transport.     

Amphidinol 3 preferentially binds to cholesterol in disordered domains and disrupts membrane phase separation

Amphidinol 3 (AM3), a polyhydroxy-polyene metabolite from the dinoflagellate Amphidinium klebsii, possesses potent antifungal activity. AM3 is known to interact directly with membrane sterols and permeabilize membranes by forming pores. Because AM3 binds to sterols such as cholesterol and ergosterol, it can be assumed that AM3 has some impact on lipid rafts, which are membrane domains rich in sphingolipids and cholesterol. Hence, we first examined the effect of AM3 on phase-separated liposomes, in which raft-like ordered and non-raft-like disordered domains are segregated. Consequently, AM3 disrupted the phase separation at 22 μM, as in the case of methyl-β-cyclodextrin, a well-known raft-disrupter that extracts sterol from membranes. The surface plasmon resonance measurements and dye leakage assays show that AM3 preferentially recognizes cholesterol in the disordered membrane, which may reflect a weaker lipid-cholesterol interaction in disordered membrane than in ordered membrane. Finally, to gain insight into the AM3-induced coalescence of membrane phases, we measured membrane fluidity using fluorescence correlation spectroscopy, demonstrating that AM3 significantly increases the order of disordered phase. Together, AM3 preferentially binds to the disordered phase rather than the ordered phase, and enhances the order of the disordered phase, consequently blending the separated phases.