Transformation pathways of liposomes

Liposomes undergoing transformation were observed by dark-field light microscopy in order to study the role of lipid in morphogenesis of biological vesicular structures. Liposomes were found to transform sequentially in a well-defined manner through one of several transformation pathways. A circular biconcave form was an initial shape in all the pathways and it transformed into a stable thin flexible filament or small spheres via a variety of regularly shaped vesicles which possessed geometrical symmetry. The transformation was reversible up to a certain point in each pathway. Osmotic pressure was found to be the driving force for the transformations. Biological membrane vesicles such as trypsinized red cell ghosts also transformed by similar pathways.     

Mutations of Asp540 and the domain-connecting residues synergistically enhance Pyrococcus furiosus DNA ligase activity

The structure of Pyrococcus furiosus DNA ligase (PfuLig), which architecturally resembles human DNA ligase I (hLigI), revealed that the C-terminal helix stabilizes the closed conformation through several ionic interactions between two domains (adenylylation domain (AdD) and C-terminal OB-fold domain (OBD)). This helix is oriented differently in DNA-bound hLigI, suggesting that the disruption of its interactions with AdD facilitates DNA binding. Previously, we demonstrated that the replacement of Asp540 with arginine improves the ligation activity. Here we report that the combination of the Asp540-replacement and the elimination of ionic residues in the helix, forming interactions with AdD, effectively enhanced the activity.     

Global Identification of Genes Affecting Iron-Sulfur Cluster Biogenesis and Iron Homeostasis

Iron-sulfur (Fe-S) clusters are ubiquitous cofactors that are crucial for many physiological processes in all organisms. In Escherichia coli, assembly of Fe-S clusters depends on the activity of the iron-sulfur cluster (ISC) assembly and sulfur mobilization (SUF) apparatus. However, the underlying molecular mechanisms and the mechanisms that control Fe-S cluster biogenesis and iron homeostasis are still poorly defined. In this study, we performed a global screen to identify the factors affecting Fe-S cluster biogenesis and iron homeostasis using the Keio collection, which is a library of 3,815 single-gene E. coli knockout mutants. The approach was based on radiolabeling of the cells with [2-¹⁴C]dihydrouracil, which entirely depends on the activity of an Fe-S enzyme, dihydropyrimidine dehydrogenase. We identified 49 genes affecting Fe-S cluster biogenesis and/or iron homeostasis, including 23 genes important only under microaerobic/anaerobic conditions. This study defines key proteins associated with Fe-S cluster biogenesis and iron homeostasis, which will aid further understanding of the cellular mechanisms that coordinate the processes. In addition, we applied the [2-¹⁴C]dihydrouracil-labeling method to analyze the role of amino acid residues of an Fe-S cluster assembly scaffold (IscU) as a model of the Fe-S cluster assembly apparatus. The analysis showed that Cys37, Cys63, His105, and Cys106 are essential for the function of IscU in vivo, demonstrating the potential of the method to investigate in vivo function of proteins involved in Fe-S cluster assembly.     

A Simple Force-Motion Relation for Migrating Cells Revealed by Multipole Analysis of Traction Stress

For biophysical understanding of cell motility, the relationship between mechanical force and cell migration must be uncovered, but it remains elusive. Since cells migrate at small scale in dissipative circumstances, the inertia force is negligible and all forces should cancel out. This implies that one must quantify the spatial pattern of the force instead of just the summation to elucidate the force-motion relation. Here, we introduced multipole analysis to quantify the traction stress dynamics of migrating cells. We measured the traction stress of Dictyostelium discoideum cells and investigated the lowest two moments, the force dipole and quadrupole moments, which reflect rotational and front-rear asymmetries of the stress field. We derived a simple force-motion relation in which cells migrate along the force dipole axis with a direction determined by the force quadrupole. Furthermore, as a complementary approach, we also investigated fine structures in the stress field that show front-rear asymmetric kinetics consistent with the multipole analysis. The tight force-motion relation enables us to predict cell migration only from the traction stress patterns.     

Mitochondrial inhibitor sodium azide inhibits the reorganization of mitochondria‐rich cytoplasm and the establishment of the anteroposterior axis in ascidian embryo

In ascidian eggs, cytoplasmic and cortical reorganization, previously called ooplasmic segregation, occurs in two phases during the first cell cycle. In the second phase of reorganization, the mitochondria‐rich cytoplasm (myoplasm) moves to the future posterior side, concurrent with sperm aster migration along the egg cortex. Although this reorganization is the critical step for establishing the anteroposterior axis, its molecular mechanism is not fully understood. In this study, we showed that low concentrations of the mitochondrial inhibitor sodium azide (NaN₃), which showed the low toxicity in sperm, inhibited the second phase of reorganization without the microtubule depolymerization. In the NaN₃‐treated embryo, the sperm aster was not attracted to the cortex and altered its migration pathway; therefore, the myoplasm remained at the vegetal pole. Consequently, the anteroposterior axis was not established. Another mitochondrial inhibitor, oligomycin, did not affect these processes. These results suggest that NaN₃ inhibits unknown molecules that are important for the second phase of reorganization. Identifying the target molecule of NaN₃ will lead to a molecular understanding of cytoplasmic and cortical reorganization.     

Phenology of annual kelp Eckloniopsis (Phaeophyceae,Laminariales) forest on a Diadema barren in Uchiura Bay,Central Pacific Coast of Honshu,Japan

On the southern coast of Uchiura Bay, central Pacific, Japan, Diadema barrens have expanded since the 1980s but Eckloniopsis radicosa (annual kelp endemic to Japan) has remained in deeper waters (>10 m in depth). Phenology of the kelp was studied on isolated boulders from December 2011 for a year. Young sporophytes appeared in December and rapid growth from April brought the maximum blade length (83.3?±?13.9 cm) and width (56.8?±?12.7 cm) and standing crop (7.4 kg m^?2) in May and June, respectively. Sorus formation began in June and spore release occurred from July to September. Blade length decreased from August and disappeared in November though holdfast remained. The unique holdfast-like spiny ball was found to provide habitats for mobile animals; its forests have an important role to maintain the biodiversity on barrens. During the period, water temperature was between 14.6 and 27.8 °C, salinity was stable around 34–35?‰, and nutrients were never depleted. Tolerance to large and frequent fluctuation of water temperature (7 °C in a day), rapid growth in winter to spring, and occurrence on limited boulders in soft substrata may be the reasons for the success in the maintenance of its forest in Diadema barrens.     

Design of recycling system for poly(methyl methacrylate) (PMMA). Part 2: process hazards and material flow analysis

Introduction

In this series of papers, we present a design of poly(methyl methacrylate) (PMMA) recycling system considering environmental impacts, chemical hazards, and resource availability. We applied life cycle assessment (LCA), environment, health, and safety (EHS) assessment as well as material flow analysis to the evaluation of the recycling system.

Purpose

Recycling systems for highly functional plastics such as PMMA have not been studied sufficiently. Along with the popularization of PMMA-containing products such as liquid crystal displays (LCDs), the use of PMMA is steadily increasing, which will result in more waste of PMMA in the next decades. In this study, pyrolysis process for recycling waste PMMA into methyl methacrylate (MMA) monomer was examined, considering not only general environmental impacts quantified by life cycle assessment but also local environment, health, and safety hazards, and raw material availability.

Methods

Process EHS hazards assessment was applied to quantify the local effects of the PMMA monomer recycling process. Process hazards are strongly connected with the hazardous properties of chemical substances and stream conditions within the process. Two alternative cooling methods exist, and their difference was analyzed by LCA and EHS assessment. Besides the process hazard, the availability of waste PMMA must be an important point for the feasibility of implementing the PMMA monomer recycling process. The available amount can be quantified by analyzing the material flow of PMMA-containing products. PMMA contained in LCDs as light guide panels was selected as a feasible source of waste PMMA, and the quantity of PMMA flows in the society was evaluated.

Results and discussion

In the case of PMMA, monomer recycling has less process hazard than the production of fresh MMA from crude oil. The implementation of circulated cooling water could significantly decrease the process hazard in PMMA pyrolysis attributable to chemical hazards. Material flow analysis revealed that the availability of waste PMMA shows a fluctuating trend in the next 20 years because of the sharp peak demand for LCD television sets. The fluctuation is strongly dependent on the lifetime of LCD television sets.

Conclusions

PMMA monomer recycling has a potential to reduce environmental impacts with a less process hazards than fresh MMA production from crude oil. The availability of waste PMMA has a strong relationship with the lifetime of LCD television sets. The multiple and comprehensive assessments can reveal various aspects of a process technology.