Cold Adaptation of a Mesophilic Subtilisin-like Protease by Laboratory Evolution

Enzymes isolated from organisms native to cold environments generally exhibit higher catalytic efficiency at low temperatures and greater thermosensitivity than their mesophilic counterparts. In an effort to understand the evolutionary process and the molecular basis of cold adaptation, we have used directed evolution to convert a mesophilic subtilisin-like protease from Bacillus sphaericus, SSII, into its psychrophilic counterpart. A single round of random mutagenesis followed by recombination of improved variants yielded a mutant, P3C9, with a catalytic rate constant (kcat) at 10 °C 6.6 times and a catalytic efficiency (kcat/KM) 9.6 times that of wild type. Its half-life at 70 °C is 3.3 times less than wild type. Although there is a trend toward decreasing stability during the progression from mesophile to psychrophile, there is not a strict correlation between decreasing stability and increasing low temperature activity. A first generation mutant with a >2-fold increase in kcat is actually more stable than wild type. This suggests that the ultimate decrease in stability may be due to random drift rather than a physical incompatibility between low temperature activity and high temperature stability. SSII shares 77.4% identity with the naturally psychrophilic protease subtilisin S41. Although SSII and S41 differ at 85 positions, four amino acid substitutions were sufficient to generate an SSII whose low temperature activity is greater than that of S41. That none of the four are found in S41 indicates that there are multiple routes to cold adaptation

Toward Mandatory Environmental Disclosure for Capital Markets: Discussion and Empirical Evidence from Japan

Increasing demand for environmental disclosure by capital markets hasamplified discussions calling for mandatory environmental disclosure. Towardmandating, capabilities and shortcomings of current voluntary environmentalreporting must be clarified from an investor’s perspective. The authors carriedout extensive empirical research in Japan, one of the most advanced countriesin the world in terms of environmental reporting. Global information vendorsare currently practicing a promising approach that enables investors to utiliseenvironmental data disclosed voluntarily. Relying on their approach, the authorsdeveloped an environmental database that covers 185 companies in the NikkeiIndex. This paper uses the information in this database to discuss the capabilitiesand shortcomings of voluntary environmental disclosure. The databaseenables investors to undertake simple analysis, but blanks and discrepanciesin boundaries damage the comparability and reliability of the data. Thesefundamental shortcomings stem largely from the adverse incentive: the morea company is engaged in environmental reporting, the worse it could appearin the database we developed. In contrast to the fact that an “invisible hand”could increase the number of sustainability reports published voluntarily, theadverse incentive faced by companies indicates that policy-making in the area ofenvironmental disclosure may become more vital in the future

Lyman Break Galaxies at z∼5z\sim5: Rest-Frame UV Spectra

We report initial results for spectroscopic observations of candidates of Lyman Break Galaxies (LBGs) at $z\sim5$ in a region centered on the Hubble Deep Field-North by using the Faint Object Camera and Spectrograph attached to the Subaru Telescope. Eight objects with $I_C\leq25.0$ mag, including one AGN, are confirmed to be at $4.5<z<5.2$. The rest-frame UV spectra of seven LBGs commonly show no or weak Lyalpha emission line (rest-frame equivalent width of 0-10\AA) and relatively strong low-ionization interstellar metal absorption lines of SiII $\lambda$1260, OI+SiII $\lambda$1303, and CII $\lambda$1334 (mean rest-frame equivalent widths of them are $-1.2 \sim -5.1$\AA). These properties are significantly different from those of the mean rest-frame UV spectrum of LBGs at $z\sim3$, but are quite similar to those of subgroups of LBGs at $z\sim3$ with no or weak Lyalpha emission. The weakness of Lyalpha emission and strong low-ionization interstellar metal absorption lines may indicate that these LBGs at $z\sim5$ are chemically evolved to some degree and have a dusty environment. Since the fraction of such LBGs at $z\sim5$ in our sample is larger than that at $z\sim3$, we may witness some sign of evolution of LBGs from $z\sim5$ to $z\sim3$, though the present sample size is very small. It is also possible, however, that the brighter LBGs tend to show no or weak Lyalpha emission, because our spectroscopic sample is bright (brighter than $L^{\ast}$) among LBGs at $z\sim5$. More observations are required to establish spectroscopic nature of LBGs at $z\sim5$.Comment: 16 pages, 3 figures, accepted by Ap

Induction of immune response in macaque monkeys infected with simian–human immunodeficiency virus having the TNF-α gene at an early stage of infection

AbstractTNF-α has been implicated in the pathogenesis of, and the immune response against, HIV-1 infection. To clarify the roles of TNF-α against HIV-1-related virus infection in an SHIV-macaque model, we genetically engineered an SHIV to express the TNF-α gene (SHIV-TNF) and characterized the virus's properties in vivo. After the acute viremic stage, the plasma viral loads declined earlier in the SHIV-TNF-inoculated monkeys than in the parental SHIV (SHIV-NI)-inoculated monkeys. SHIV-TNF induced cell death in the lymph nodes without depletion of circulating CD4+ T cells. SHIV-TNF provided some immunity in monkeys by increasing the production of the chemokine RANTES and by inducing an antigen-specific proliferation of lymphocytes. The monkeys immunized with SHIV-TNF were partly protected against a pathogenic SHIV (SHIV-C2/1) challenge. These findings suggest that TNF-α contributes to the induction of an effective immune response against HIV-1 rather than to the progression of disease at the early stage of infection

末梢神経内血流量の測定に関する研究-水素クリアランス法を用いて-

金沢大学医学部整形外科研究課題/領域番号:61921029, 研究期間(年度):1986出典：研究課題「末梢神経内血流量の測定に関する研究-水素クリアランス法を用いて-」課題番号61921029（KAKEN：科学研究費助成事業データベース（国立情報学研究所）） （https://kaken.nii.ac.jp/ja/grant/KAKENHI-PROJECT-61921029/）を加工して作

Theoretische Grundlage der okologischen Buchhaltung : Rechnung der Umweltbelastungen durch die doppelte Okobilanzierung(Study Note 2,Japan Environmental Policy Priorities Index)

The studies of EcoBalance in the field of social science are not necessarily very successful up to today. The authors think that one of the main grounds for it would be the large gab, lying between the calculation of EcoBalance and that of business economics. The calculation of EcoBalance is so far based on the substantial flows, on the other hand that of business economics is executed with the concepts of revenue and expense, deviating from the substantial flows. There could be two approaches to overcome this situation. The first approach is to adjust accounting in business economics to the calculation of EcoBalance. The practice of EcoBalance with monetary impact assessment is a typical example of this approach. The second approach is to adjust the calculation of EcoBalance to accounting in business economics, namely to introduce the principle of double-entry bookkeeping in EcoBalance, which constitutes the theoretical basis of accounting in business economics. This approach is not yet explored in academic or in practical field. In this thesis, therefore, the second approach will be chosen and a calculation system of double-entry EcoBalance will be developed and summarised. The whole picture of the calculation system will be shown, basing on the example of a car manufacturing by using JEPIX, which enables to make different types of environmental impacts comparable. In the framework of double-entry EcoBalance the five different accounts shown below will play the fundamental roll. (1) "Environmental Impacts (UB)" represents the products or service, to which environmental impacts are attached, such as finished products or works in process. This account would correspond to the asset in accounting of business economics. (2) "Unrealised Responsibility of the Environmental Impacts (UVdU)" represents the responsibility related to the environmental impacts. The responsibility is recognised in accounting procedure but substantially not yet realised. This account would correspond to the liability, namely allowance in accounting of business economics. (3) "Responsibility for the Environmental Impacts (VfU)" represents the responsibility for the environmental impact which the company has produced. This account would correspond to the capital in accounting of business economics. (4) "Establishment of Responsibility for the Environmental Impacts (EdVfU)" represents the cause for the increase of the VfU. This account would correspond to the revenue in accounting of business economics. (5) "Release of Responsibility for the Environmental Impacts" represents the cause for the decrease of the VfU. This account would correspond to the expense in accounting of business economics. The important key to the development and execution of this system is the equivalent coefficients which enable the calculation of different types of environmental impacts in a single system and the authors believe that JEPIX will be playing the important roll for it. The basic ideas of this paper came from the problem consciousness and guestions of Kentaro Azuma about periopdical belonging and matching of environmental impacts, and they have fundamentally been developed thought the continuous discussions between Nobuyuki Miyazaki and Azuma. The theory in this paper was designed and wrote by miyazaki, and translated into German by Azuma. This paper was reprinted from the Journal of Social Science NO.51. March 2004