Comparative immunohistological study on using capsaicin,piperine, and okadaic acid for the transepithelial passage of the inactivated viral and bacterial vaccines in fish

The practical difficulty of parenteral application of fish vaccines against devastating fish diseases diverted the interest toward oral vaccination. Search for effective methods to enhance the oral uptake of viral and bacterial vaccines is continuing. The current research focus on a new role of mucosal fish vaccine adjuvants inducing the antigen uptake by enhancing vascularity or increasing intestinal permeability. Some inflammatory substances cause reversible pathology to the intestinal epithelium, which could be employed for the transepithelial passage of vaccine particles. The natural inflammatory substances used were capsaicin, piperine, and okadaic acid as 1 mg, 2 mg, and 1 μg/fish, respectively. Two inactivated vaccines were used as antigens to test the effect of these inflammatory substances in two different fish hosts. Tested vaccines were inactivated redspotted grouper nervous necrosis virus vaccine in sevenband grouper (Epinephelus septemfasciatus) and inactivated Edwardsiella tarda vaccine in red sea bream (Pagrus major) fish models. The inflammatory substances and each vaccine were anally intubated to fish. Capsaicin proved to be effectively aiding the transepithelial passage of vaccine particles more than piperine, while okadaic acid had no detectable effect.     

Development of a robot assisted carotid blood flow measurement system

The ultrasound diagnosis of the carotid artery is one of the most common non-invasive methods to detect early stage heart and cerebrovascular diseases. However, the precision and repetitiveness of the probe positioning depend exclusively on the operator's skills and dexterity. For this purpose, we propose the development of a robot assisted system to enhance the accuracy and repetitiveness of the probe positioning to measure the wave intensity (WI) index. In this paper, the Waseda-Tokyo Women's Medical-Aloka Blood Flow Measurement Robot System No. 2 Refined (WTA-2R) is presented. The WTA-2R consists of a conventional ultrasound diagnosis system, a 6-DOFs parallel link slave manipulator, a 6-DOFs serial link passive arm, and a master device. Experiments were carried out to verify its effectiveness in terms of accuracy and required time to perform the task. From the experimental results, the positioning accuracy and reduction of required time were confirmed.     

Investigation of localized deformation in partially saturated sand under triaxial compression using microfocus X-ray CT with digital image correlation

In this paper, localized deformation in partially saturated sand was investigated quantitatively using microfocus X-ray computed tomography (CT) and an image analysis of the CT images. Triaxial compression tests on a partially saturated dense Toyoura sand specimen were carried out under a low confining pressure and under drained conditions for both air and water. The development of localized deformation was observed macroscopically using microfocus X-ray CT, and the displacement field over the entire specimen was quantified by an image analysis of the CT images with the digital image correlation (DIC) technique. The progressive development of shear bands is discussed with reference to these images. In addition, the region of localization was observed microscopically by partial CT scanning on a micron scale with high spatial resolution. Changes in the particulate structures are also discussed herein. The DIC image analysis of the partial CT images provided a microscopic displacement field and indicated that very fine localized shear deformation developed before the shear bands had become visible in the macroscopic investigation.     

Performance decay of proton-exchange membrane fuel cells under open circuit conditions induced by membrane decomposition

The degradation in performance of proton-exchange membrane fuel cells (PEMFCs) under open circuit conditions was investigated. The oxygen reduction reaction (ORR) kinetic current density at 0.9 V was found to decrease from 36 to 4 mA cm⁻² (geometric) without significant crossover increase or loss in the electrochemically active surface area. Cyclic voltammograms for the electrodes show characteristic changes, e.g. appearance of peaks at ∼0.2 V and shift of the onset of platinum oxide formation to higher potentials. It was identified that the large ORR kinetic decay has its origins in the reduction of available Pt sites due to adsorption of anions, which are postulated to be membrane decomposition products such as sulfate ions. Procedures carried out to condense water in the fuel cell led to the expulsion of anions out of the membrane electrode assembly (MEA) resulting in the partial recovery of ORR kinetic current density to 15 mA cm⁻². In order to attain complete performance recovery of the catalyst, a more effective and practical method to flush out the anions is desirable.     

A spatial evaluation of forest biomass usage using GIS

We conducted a spatial evaluation of forest biomass usage using a geographic information system (GIS) for the Japanese town of Yusuhara. In Japan, over 60% of the land is covered with forest, of which at least 40% is artificial forest. However, because of high labor costs, the profitability of forestry is decreasing, so timber cultivation is not done to the extent that it could be, and thinning has to be subsidized. Under these circumstances, much of the forest is deteriorating. Most of the thinning is accounted for by throwaway thinning, in which the resulting wood is not used. However, with the steep rise in oil prices and the intensification of global warming concerns, expectations are rising for the use of biomass energy from thinned timber that has previously been discarded. If thinned timber, logging residues, and offcuts are utilized for biomass energy and their economic value becomes apparent, profitability will improve for both final cutting and thinning. And in addition to forestry activities being invigorated, it will be possible for some of the deteriorating forests (which have associated dangers such as landslides) to recover. However, using thinned timber and logging residues is problematic in that profitability is affected by harvesting costs. Harvesting costs are largely determined by geographic factors and are higher for more distant stands. Accordingly, in this article, operational costs for different stands are calculated using GIS and matched with total demand in the subject region. In addition, stands with lower operational costs are identified and an investigation of a highly feasible use of forest biomass is carried out.     

Growth and differentiation potentials in confluent state of culture of human skeletal muscle myoblasts

The transitional behaviors of myoblasts toward differentiation were investigated in the cultures at the low and high seeding densities (respectively, X₀ = 1.0 × 10³ and 2.0 × 10⁵ cells/cm²). In the culture at the low seeding density, an increase in confluence degree accompanied a decrease in growth potential (R_p), being R_p = 0.85 and 0.11 at t = 48 and 672 h, respectively. Myoblasts seeded at the high density resulted in the immediate cessation of growth with keeping the low range of R_p = 0.02–0.09 throughout the culture. The reduction of R_p led to the generation of three subpopulations of cells in proliferative, quiescent and differentiated states. Close cell contacts in the confluent state of high seeding culture induced cell quiescence to a higher extent with suppressing differentiation.     

Purification, characterization, and gene cloning of glycerol dehydrogenase from Hansenula ofunaensis, and its expression for production of optically active diol

Optically active alcohol is an important building block as a versatile chiral synthon for the asymmetric synthesis of pharmaceuticals and agrochemicals. We purified and characterized glycerol dehydrogenase from Hansenula ofunaensis and prepared optically active 1,2-octanediol using a recombinant Escherichia coli harboring the glycerol dehydrogenase gene. The deduced amino acid sequence was investigated for identities with those of other alcohol dehydrogenases in the NCBI databank. The identification of the unknown product of a resting-cell reaction was performed by GC-MS. In the deduced amino acid sequence composed of 376 residues, the NAD(H) binding pattern and cysteine residues that correspond to the cysteine ligands at the zinc atom were conserved as they are in alcohol dehydrogenases from other origins. Glycerol dehydrogenase from Hansenula polymorpha DL-1 (Pichia angusta, DDBJ/EMBL/GenBank accession no. BAD32688) had the highest identity to our enzyme, showing 73% identity. Our glycerol dehydrogenase catalyzed the NAD(+)-dependent oxidation of long-chain secondary alcohols such as 1,2-pentanediol, 1,2-hexanediol, 1,2-heptanediol, and 1,2-octanediol. Activities toward 2,4-pentanediol and 2,5-hexanediol were hardly detected. From these results, it was confirmed that our enzyme requires two hydroxyl groups on adjacent carbon atoms for oxidation. 2,3-Pentanedione, 2,3-hexanedione, and 3,4-hexanedione were significantly reduced. The transformants oxidized only (R)-1,2-octanediol in 50 mM racemate (R:S=52:48), and produced (S)-1,2-octanediol (24 mM, <99.9% e.e.) after 24 h of incubation. The reaction product was suggested to be 1-hydroxy-2-octanone by GC-MS, which showed secondary hydroxyl groups oxidized. Glycerol dehydrogenase from H. ofunaensis could be useful for the production of long-chain optically active secondary alcohols.     

Simultaneous estimation of the thermophysical properties of three kinds of fruit juices based on the measured result by a transient heat flow probe method

The thermophysical properties of three kinds of fruit juices (grape juice, orange juice, and pineapple juice) were measured at various temperatures (10–50 °C) and concentrations (10–50%). A new method for the simultaneous determination of thermophysical properties using a modified version of current probe theory method was proposed. The temperature changes of the probe upon insertion in the sample were fitted to an approximate solution of the heat conduction equation, and the values of two parameters in that solution were determined. Using the values of these parameters, the thermal conductivity and thermal diffusivity of each sample were determined. The specific heat of each sample was estimated from the definition of thermal diffusivity. These thermophysical properties were expressed as a function of concentration and temperature.     

Application of microalgae hydrolysate as a fermentation medium for microbial production of 2-pyrone 4,6-dicarboxylic acid

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Noncompetitive inhibition by L-cysteine and activation by L-glutamate of the iron-oxidizing activity of a mixotrophic iron-oxidizing bacterium strain OKM-9

A mesophilic, mixotrophic iron-oxidizing bacterium strain OKM-9 uses ferrous iron as a sole source of energy and L-glutamate as a sole source of cellular carbon. Uptake of L-glutamate into OKM-9 cells is absolutely dependent on ferrous iron oxidation. Thus, the Fe(2+)-dependent L-glutamate uptake system of strain OKM-9 is crucial for the bacterium to grow mixotrophically in iron medium with L-glutamate. The relationship between iron oxidation and L-glutamate transport activities was studied. Iron oxidase containing cytochrome a was purified 9-fold from the plasma membrane of OKM-9. A purified iron oxidase showed one rust-colored band following disc gel electrophoresis after incubation with Fe(2+). The Fe(2+)-dependent L-glutamate transport system was also purified 14.5-fold from the plasma membrane using the same purification steps as for iron oxidase. Fe(2+)-dependent L-glutamate and L-cysteine uptake activities of OKM-9 were 0.36 and 0.24 nmol/mg/min, respectively, when a concentration of 18 mM of these amino acids was used as a substrate. Both uptake activities were completely inhibited by potassium cyanide (KCN), suggesting that cytochrome a in the iron oxidase is involved in the transport process. The iron-oxidizing activity of strain OKM-9 was activated 1.7-fold by 80 mM L-glutamate. In contrast, the activity was noncompetitively inhibited by L-cysteine. The Michaelis constant of iron oxidase for Fe(2+) was 12.6 mM and the inhibition constant for L-cysteine was 41.6 mM. A marked inhibition of iron oxidase by 50 mM L-cysteine was completely reversed by the addition of 60 mM L-glutamate. The results suggest the possibility that iron oxidase has a binding site for L-cysteine and the cysteine first bound to the iron oxidase was replaced by the added L-glutamate.