High-oil-load encapsulation of medium-chain triglycerides and D-limonene mixture in modified starch by spray drying

Oil mixtures of medium-chain triglycerides (MCT) and D-limonene in mixing ratios from 10 to 100 wt% were encapsulated in modified starch (wall material) by spray drying to produce oil-rich powders. The oil load (mass ratio of oil mixture to wall material) of the infeed emulsion markedly influenced the properties of the infeed liquid and the characteristics of the resulting powder. The viscosity of the infeed liquid and the particle size of the powder exponentially decreased with increasing oil load, while the emulsion droplet size in the infeed liquid increased. In addition, retention of D-limonene during spray drying also decreased markedly with increasing oil load. Irrespective of the different oil loads and concentrations of the wall material, D-limonene retention was well correlated with the emulsion droplet diameter of the infeed liquid. The encapsulation efficiency of the oil mixture exhibited a maximum value (almost 100%) at an oil load between 0.5 and 1.0, before decreasing at higher oil loads. At an oil load of 2.0, the encapsulation efficiency of D-limonene was reduced to almost zero, while around 40% of the initial MCT was encapsulated in the powder. The increase in oil load also led to increased amounts of surface oil of MCT and D-limonene in the resulting powder due to the increasing emulsion droplet diameter of the infeed liquids. PRACTICAL APPLICATION: This study proposes the microencapsulation of medium-chain triglycerides under high-oil-load conditions by spray drying. The powders prepared by this process provide significant benefits in terms of rapid energy conversion after consumption without accumulation in the body. Important quality factors of the powder products such as the encapsulation efficiency and the amount of surface oil were examined to understand the optimum process conditions for spray drying.     

Application of electrospray ionization ion trap/time-of-flight mass spectrometry for chemically-synthesized small RNAs

In this study, we have demonstrated an accurate and rapid small RNA analytical method with both sequence determination and detailed modification analysis by electrospray ionization-ion trap/time-of-flight mass spectrometry (ESI-IT/TOFMS). To develop this ideal method, we have examined the performance of ESI-IT/TOFMS using various chemically-synthesized model sequences of modified or unmodified microRNAs (miRNAs). The deconvoluted mass of a 22-nucleotide (nt) miRNA was obtained from a multiply charged precursor ion (MS(1)). The ion exhibited high mass accuracy (< 7 ppm) and high mass resolution (a value of m/Δm=10,000) and was therefore very useful in RNA composition assignment. The optimized MS(2) method using ion trap collision-induced dissociation, as well as automatic annotation analysis of product ions based on the accurate mass information, enabled the precise sequencing determination of intact miRNAs. Further, the detailed structural analysis of 3'-terminal modified nucleic acid in intact methylated miRNA was carried out using the MS(3) capability of the hybrid IT/TOFMS. The direct infusion method also provided a high throughput and good sensitivity because the analytical time and sample concentration needed in a series of experiments with reliable data were only 3 min and 100 nM, respectively. This study provides a novel approach for characterizing the intact chemically-synthesized small RNA without chemical and enzymatic digestions and would be widely applicable for the structural analysis of complicated modified small RNAs.     

Chain‐length Distribution Profiles of Amylopectin Isolated from Endosperm Starch of Waxy and Low‐amylose Bread Wheat (Triticum aestivum L.) Lines with Common Genetic Background

Large A‐type endosperm starch granules were isolated from near‐isogenic waxy and non‐waxy lines and low‐amylose mutant lines of bread wheat with a common genetic background. The amylose contents of A‐type starch ranged from 2.6% to 23.6%. Amylopectin was isolated by concanavalin A (Con A) precipitation from the isolated starch. The λ_max (range: 532‐538 nm) and blue values at 680 nm (range: 0.026‐0.037) of the iodine‐amylopectin complex were not significantly different among the isolated amylopectins, indicating that amylopectins from non‐waxy and low‐amylose lines did not contain such long chains as amylose or extra‐long chains of amylopectin affecting iodine complex properties. Chain‐length distribution profiles measured by both high‐performance size‐exclusion chromatography (HPSEC) and high‐performance anion‐exchange chromatography (HPAEC) showed that the amylopectin structures of these lines were indistinguishable from each other. Extra‐long chains were not detected in the amylopectins by HPSEC measurement. The side‐chains measured by HPAEC were classified into four groups according to their degree of polymerization (DP), and the proportion of each group were in the following ranges: DP 6‐12, 26.5‐27.5%; DP 13‐24, 43.6‐44.1%; DP 25‐36, 13.6‐14.2%, and DP 37‐60, 11.0‐11.7%. The alleles on the Wx‐D1 locus, i.e., Wx‐D1a, Wx‐D1d, Wx‐D1f, and Wx‐D1g, responsible for granule‐bound starch synthase (GBSS I) biosynthesis had no influence on the properties of iodine‐amylopectin complex and the chain‐length distribution profiles of amylopectin.     

Freeze-fracture replica of the rat cornea

The fine structures of the rat cornea, with special reference to their intercellular junctions, were studied using the freeze-fracture technique. At the corneal epithelium, gap junctions could be observed between the adjacent cells. At the stroma, crater-shaped depressions (between 300 and 500 A diameter) with pipe-like appearing structures connecting the lamellae were found. Intercellular junctions existing between the endothelial cells at the area near the anterior chamber are postulated to be the fascia (macula) occludens.     

Kinetic resolution of an indan derivative using Bacillus sp. SUI-12: synthesis of a key intermediate of the melatonin receptor agonist TAK-375

The chiral indan derivative (S)-2 (2-[(8S)-1,6,7,8-tetrahydro-2H-indeno[5,4-b]furan-8-yl]ethyl-amine) was synthesized by enzyme-catalyzed asymmetric hydrolysis of the racemic acetamide 1 (N-[2-(1,6,7,8-tetrahydro-2H-indeno[5,4-b]furan-8-yl)ethyl]acetamide). The reaction was carried out using Bacillus sp. SUI-12 screened for the ability to hydrolyze 1 to give (S)-2 with high enantioselectivity. In a scaled-up experiment, a low reaction rate was observed. However, by changing the culture medium and the reaction conditions, it became possible to run the reaction to 40% conversion on a 10-g or more scale, obtaining (S)-2 at >;99% enantiomeric excess (ee). The (S)-2 obtained was available for the synthesis of the melatonin receptor agonist TAK-375 (N-[2-[(8S)-1,6,7,8-tetrahydro-2H-indeno[5,4-b]furan-8-yl]ethyl]propanamide).     

Spatial distributions and profiles of atmospheric polycyclic aromatic hydrocarbons in two industrial cities in Japan

The spatial distribution and concentration profiles of 39 vapor and particulate polycyclic aromatic hydrocarbons (PAHs) have been investigated in two Japanese industrial cities (Fuji and Shimizu; a summer and winter season in each). The concentrations of particulate PAHs in winter tended to be higher than those in the summer, but for vapor PAHs, this was not the case. Significant correlations (p < 0.01) were found between most of the PAH concentrations monitored in winter, suggesting the presence of common emission sources. To identify PAH spatial distributions and emission sources in the area, we created contour maps for PAHs monitored; this indicates that the distinctive local distributions correspond to the emission sources. PAH profiles based on benzo[e]pyrene (BeP) concentration, especially for certain relatively heavy molecular weight PAHs, showed differential behaviors among divided areas related to potential regional emission sources such as paper-making plants, power plants, and traffic. We conclude that the origins of atmospheric PAHs in the surveyed areas were dominated by not only traffic but also by stationary emission sources such as paper-making plants and power plants and that local distributions were dependent on the local wind direction.     

Prognostic predictor with multiple fuzzy neural models using expression profiles from DNA microarray for metastases of breast cancer

Gene expression profiling data from DNA microarray were analyzed using the fuzzy neural network (FNN) modeling method for predicting the distant metastases of breast cancer. The best model consisting of five genes was able to predict metastases of breast cancer with 94% accuracy. Furthermore, 100% accuracy was achieved by majoritarian decision using only 25 genes from five noninferior models which were constructed independently. From the constructed model, gene expression rules, which may cause distant metastases, were explicitly extracted and 60% of the metastases cases could be explained by this rule. The FNN modeling method described in this paper enables precise extraction of significant biological markers affecting prognosis without prior knowledge.