Genetic Analysis, Expression in Eschericia coli of Aconitase from Chemolithotrophic Acidithiobacillus thiooxidans

An aconitase from Acidithiobacillus thiooxidans was purified and characterized, and its gene was cloned. The cloned aconitase gene (acn) was expressed in Escherichia coli JM 109; aconitase activity was found in the cell extarct. The acn gene encodes a 646-amino acid polypeptide and is located upstream of the isocitrate dehydrogenase gene (icd). A. thiooxidans aconitase showes high sequence similar to pig heart aconitase and E.coli aconitase B. Twenty-five of twenty-seven active site residues assigned in pig heart aconitase are conserved in A. thiooxidans aconitase. The enzyme was purified by DEAE-Toyopearl 650M column chromatogrophy. The purified enzyme had an optimum pH of 7.5 and an optimum temperature of 60 C. Thermal inactivation studies of the purified enzyme revealed the enzyme activity to be uninfluenced after one hour incubation at 40 c. Enzyme activity was retained 100% after incubation of the enzyme at pH 6.0-9.0 for 60min. The A. thiooxidans aconitase was composed of a single polypeptide chain with a molecular mass of 66 kDa

Physical performance disorder（Locomotive syndrome）closely relates to both hypertension and diabetes mellitus which are components of metabolic syndrome

Locomotive syndrome is physical performance disorder in the elderly person. The physical performance disorder can become cause of metabolic syndrome. This study focused on the locomotive syndrome’s relationship to the hypertension, diabetes mellitus, and hyperlipidemia which are components of the metabolic syndrome. To evaluate the locomotive syndrome, questionnaire score system : brand-new loco-check-score and established locomo-５-score were adopted. In the questionnaire, current medications were also listed especially about hypertension, diabetes mellitus, and hyperlipidemia. A correlation between loco-check-score and locomo-５-score was statistically evaluated by Spearman rank correlation coefficient. Moreover correlations between body mass index（BMI）and loco-check-score, BMI and locomo-５-score were statistically evaluated by Spearman rank correlation coefficient. In a health consultation event, １４６ persons checked the questionnaire （mean age ７２．５yo）. Differences of the loco-check-score and the locomo-５-score in the hypertension-group, diabetes mellitus group, and the hyperlipidemia group were statistically evaluated by the Mann-Whitney U test respectively. Age matched persons with no medications comprised control groups. Significant differences were observed in the hypertension group and diabetes mellitus group both in the loco-check-score and locomo-５-score systems. These two groups showed significantly high loco-check-score and locomo-５-score than the control groups. The loco-check-score closely related to the locomo-５-score. On the other hand, no relations were observed between BMI and the loco-check-score, and between BMI and the locomo-５-score

Physical performance disorder（Locomotive syndrome）closely relates to both hypertension and diabetes mellitus which are components of metabolic syndrome

Locomotive syndrome is physical performance disorder in the elderly person. The physical performance disorder can become cause of metabolic syndrome. This study focused on the locomotive syndrome’s relationship to the hypertension, diabetes mellitus, and hyperlipidemia which are components of the metabolic syndrome. To evaluate the locomotive syndrome, questionnaire score system : brand-new loco-check-score and established locomo-５-score were adopted. In the questionnaire, current medications were also listed especially about hypertension, diabetes mellitus, and hyperlipidemia. A correlation between loco-check-score and locomo-５-score was statistically evaluated by Spearman rank correlation coefficient. Moreover correlations between body mass index（BMI）and loco-check-score, BMI and locomo-５-score were statistically evaluated by Spearman rank correlation coefficient. In a health consultation event, １４６ persons checked the questionnaire （mean age ７２．５yo）. Differences of the loco-check-score and the locomo-５-score in the hypertension-group, diabetes mellitus group, and the hyperlipidemia group were statistically evaluated by the Mann-Whitney U test respectively. Age matched persons with no medications comprised control groups. Significant differences were observed in the hypertension group and diabetes mellitus group both in the loco-check-score and locomo-５-score systems. These two groups showed significantly high loco-check-score and locomo-５-score than the control groups. The loco-check-score closely related to the locomo-５-score. On the other hand, no relations were observed between BMI and the loco-check-score, and between BMI and the locomo-５-score

Drug retention rates and relevant risk factors for drug discontinuation due to adverse events in rheumatoid arthritis patients receiving anticytokine therapy with different target molecules

Objective: To compare reasons for discontinuation and drug retention rates per reason among anticytokine therapies, infliximab, etanercept and tocilizumab, and the risk of discontinuation of biological agents due to adverse events (AE) in patients with rheumatoid arthritis (RA). Method: This prospective cohort study included Japanese RA patients who started infliximab (n=412, 636.0 patientyears (PY)), etanercept (n=442, 765.3 PY), or tocilizumab (n=168, 206.5 PY) as the first biological therapy after their enrolment in the Registry of Japanese Rheumatoid Arthritis Patients for Long-term Safety (REAL) database. Drug retention rates were calculated using the Kaplan-Meier method. To compare risks of drug discontinuation due to AE for patients treated with these biological agents, the Cox proportional hazard model was applied. Results: The authors found significant differences among the three therapeutic groups in demography, clinical status, comorbidities and usage of concomitant drugs. Development of AE was the most frequent reason for discontinuation of biological agents in the etanercept and tocilizumab groups, and the second most frequent reason in the infliximab group. Discontinuation due to good control was observed most frequently in the infliximab group. Compared with etanercept, the use of infliximab (HR 1.69; 95% CI 1.14 to 2.51) and tocilizumab (HR 1.98; 95% CI 1.04 to 3.76) was significantly associated with a higher risk of discontinuation of biological agents due to AE. Conclusions: Reasons for discontinuation are significantly different among biological agents. The use of infliximab and tocilizumab was significantly associated with treatment discontinuation due to AE compared with etanercept

EV Range Extender in a Two-Battery HEECS Chopper-Based Powertrain

This paper first presents a new powertrain based on a two-battery High-Efficiency Energy Conversion System (HEECS) chopper that is suitable for electric vehicles (EVs). The HEECS chopper is based on the principle of a partial power conversion circuit, and the overall efficiency is over 99% in a wide load range. The efficiency of this powertrain was measured in the steady state by two types of powertrains, a non-chopper powertrain and an HEECS chopper-based powertrain, using a motor test bench. On the basis of these data, several driving tests, such as the Worldwide-harmonized Light vehicles Test Cycle (WLTC), were simulated, and four driving cycle patterns were included. A 6.4% reduction in energy consumption was observed in WLTC low mode compared with the energy consumed by the non-chopper powertrain in the experiments. Thus, the HEECS chopper-based powertrain is more suitable for low-speed driving ranges than high-speed ranges

A specific form of prefibrillar aggregates that functions as a precursor of amyloid nucleation

Non-fibrillar protein aggregates that appear in the earlier stages of amyloid fibril formation are sometimes considered to play a key role in amyloid nucleation; however, the structural features of these aggregates currently remain unclear. We herein identified a characteristic pathway of fibril formation by human insulin B chain, in which two major species of prefibrillar aggregates were identified. Based on the time-resolved tracking of this pathway with far-UV circular dichroism (CD) spectroscopy, dynamic light scattering (DLS), and 1H-NMR spectroscopy, the first prefibrillar aggregate with a hydrodynamic diameter of approximately 70 nm accumulated concomitantly with the formation of a β-sheet structure, and the size further evolved to 130 nm with an additional structural development. These prefibrillar aggregates were metastable and survived at least 24 hours as long as they were maintained under quiescent conditions. The energy barrier for nucleation was overcome by shaking or even by applying a single short ultrasonic pulse. Furthermore, an investigation where nucleation efficiency was monitored by fibrillation rates with varying the timing of the ultrasonic-pulse treatment revealed that the second prefibrillar aggregate specifically produced amyloid nuclei. These results suggest that the second form of the prefibrillar aggregates acts as a direct precursor for the amyloid nucleation

De Novo Design of Foldable Proteins with Smooth Folding Funnel Automated Negative Design and Experimental Verification

AbstractDe novo sequence design of foldable proteins provides a way of investigating principles of protein architecture. We performed fully automated sequence design for a target structure having a three-helix bundle topology and synthesized the designed sequences. Our design principle is different from the conventional approach, in that instead of optimizing interactions within the target structure, we design the global shape of the protein folding funnel. This includes automated implementation of negative design by explicitly requiring higher free energy of the denatured state. The designed sequences do not have significant similarity to those of any natural proteins. The NMR and CD spectroscopic data indicated that one designed sequence has a well-defined three-dimensional structure as well as α-helical content consistent with the target

Development of Shooting Method for Impact Systems

A shooting method is a very powerful numerical method to obtain periodic solutions of nonlinear systems. However, as a variational equation of motion is needed in the shooting method and it is very difficult to obtain it in the impact systems, the shooting method for impact systems has not been developed. In this report, a shooting method for impact systems is presented by solving this problem of variational equation. Namely, the variational equation with the delta function and its differentiation is derived. It is shown that the calculation speed of this method is very fast and complicated periodic solutions are easily obtainable in high accuracy. The stabilities of periodic solutions obtained in the shooting method are in good accordance with those obtained by the analytical method. The discontinuities in the stability of the periodic solutions are shown using characteristic multiplier. Lyapunov exponents are also calculated by applying the integral technique of variational equation