Microbial Reduction of 1,2-Diketones to Optically Active α-Hydroxyketones

We have attempted to develop an intraoral method which can measure the textural changes in foodstuffs during chewing by using electromyography (EMG). Forty-three foodstuffs with variable textural attributes were used.

Total chewing energy for these foodstuffs during chewing varied from 3 to 108 for the masseter muscle and 13 to 154 for the digastric muscle, respectively. Large differences in total chewing energy could be observed by EMG among the foodstuffs. The chewing energy for many foodstuffs revealed distinct differences throughout the chewing process. Foodstuffs could be categorized into six groups according to the changing patterns of chewing energy. EMG data and the number of strokes were influenced by masticatory index and salivary flow rate.     

Gelation and Its Related Changes in Amylose Solution

The phase change for an amylose solution in the binary solvent system of dimethylsulfoxide (DMSO) with water was investigated under various conditions from sol to gel. The phase change was determined with measurements of the fluorescent depolarization and other methods by varying the solvent constitution at 25°C, and then varying the temperature at 10% of DMSO concentration.

The phase diagrams obtained with both variables were substantially similar and were also similar to those for an aqueous agarose solution. This similarity in phase diagram suggests a similar gel formation mechanism of amylose to agarose.

It was found that the phase separation point for the amylose solution agreed with the gel formation point and also with the starting point of retrogradation.     

Characterization of Lactobacillus brevis L62 strain, highly tolerant to copper ions

Lactic acid bacteria (LAB) as starter culture in food industry must be suitable for large-scale industrial production and possess the ability to survive in unfavorable processes and storage conditions. Approaches taken to address these problems include the selection of stress-resistant strains. In food industry, LAB are often exposed to metal ions induced stress. The interactions between LAB and metal ions are very poorly investigated. Because of that, the influence of non-toxic, toxic and antioxidant metal ions (Zn, Cu, and Mn) on growth, acid production, metal ions binding capacity of wild and adapted species of Leuconostoc mesenteroides L3, Lactobacillus brevis L62 and Lactobacillus plantarum L73 were investigated. The proteomic approach was applied to clarify how the LAB cells, especially the adapted ones, protect themselves and tolerate high concentrations of toxic metal ions. Results have shown that Zn and Mn addition into MRS medium in the investigated concentrations did not have effect on the bacterial growth and acid production, while copper ions were highly toxic, especially in static conditions. Leuc. mesenteroides L3 was the most efficient in Zn binding processes among the chosen LAB species, while L. plantarum L73 accumulated the highest concentration of Mn. L. brevis L62 was the most copper resistant species. Adaptation had a positive effect on growth and acid production of all species in the presence of copper. However, the adapted species incorporated less metal ions than the wild species. The exception was adapted L. brevis L62 that accumulated high concentration of copper ions in static conditions. The obtained results showed that L. brevis L62 is highly tolerant to copper ions, which allows its use as starter culture in fermentative processes in media with high concentration of copper ions.     

N-glucosides as human sodium-dependent glucose cotransporter 2 (hSGLT2) inhibitors

Inhibition of renal sodium-dependent glucose cotransporter 2 (SGLT2) increases urinary glucose excretion (UGE), and thus reduces blood glucose levels in hyperglycemia. A series of N-glucosides was synthesized for biological evaluation as human SGLT2 (hSGLT2) inhibitors. Among these compounds, N-glucoside 9d possessing an indole core structure showed good in vitro activity (IC₅₀ = 7.1 nM against hSGLT2). Furthermore, 9d exhibited favorable in vivo potency with regard to UGE in rats based on good pharmacokinetic profiles.     

Hypoxia down-regulates sFlt-1 (sVEGFR-1) expression in human microvascular endothelial cells by a mechanism involving mRNA alternative processing

sFlt-1 (soluble Flt-1) potently inhibits angiogenesis by binding extracellularly to VEGF (vascular endothelial growth factor). In the present paper, we report that hypoxia down-regulates sFlt-1 expression in HMVECs (human microvascular endothelial cells), a constituent of microvessels where angiogenesis occurs. Hypoxia (5-1% O?) increased VEGF expression in HMVECs. In contrast, the levels of sFlt-1 mRNA and protein in HMVECs decreased significantly as the O? concentration fell, whereas mFlt-1 (membrane-bound Flt-1) mRNA and protein remained unchanged. This suggested that hypoxia selectively regulates alternative 3'-end processing of sFlt-1 pre-mRNA. We have also demonstrated that sFlt-1 overexpression in lentiviral-construct-infected HMVECs counteracted VEGF-induced endothelial cell growth. We next identified cis-elements involved in sFlt-1 mRNA processing in HMVECs using a human Flt-1 minigene and found that two non-contiguous AUUAAA sequences function as the poly(A) signal. Furthermore, we identified a cis-element in intron 13 that regulates sFlt-1 mRNA processing. Mutagenesis of the U-rich region in intron 13 caused a significant decrease in the soluble-form/membrane-form RNA ratio in the minigene-transfected HMVECs. These results suggest that decreased sFlt-1 expression due to hypoxia contributes to hypoxia-induced angiogenesis and reveals a novel mechanism regulating angiogenesis by alternative mRNA 3'-end processing.     

Characterization of lipovitellin 2 as a tyrosine-phosphorylated protein in oocytes, eggs and early embryos of Xenopus laevis

A tyrosine-phosphorylated protein of 33 kDa is shown to be present in the solubilized yolk fraction of Xenopus laevis oocytes, eggs, and early embryos. The phosphoprotein is lipovitellin 2 as demonstrated by immunoprecipitation and mass spectrometry studies, and is termed pp33/LV2. Sub-cellular fractionation and immunoblotting studies demonstrate that pp33/LV2 is stably present in the Triton X-100-resistant and SDS-soluble yolk fractions during oogenesis, oocyte maturation, and early embryogenesis. From after the swimming tadpole stages (stage 39～), however, it becomes partly soluble to Triton X-100-containing buffer and all disappear thereafter (stage 48～49). In vitro enzyme assays with the use of the tyrosine phosphatase LAR and the tyrosine kinase Src demonstrate the reversible nature of the tyrosine phosphorylation of pp33/LV2. Microinjection studies demonstrate that the solubilized yolk fractions, but not those immunodepleted of pp33/LV2 or those pretreated with LAR, inhibit progesterone- or insulin-induced oocyte maturation. A pp33/LV2-like protein seems to present in two Xenopus subspecies, one other frog species, and two fish species, but not in other amphibian species, such as newt and salamander. These results suggest that LV2, in its tyrosine-phosphorylated form, serves in a cellular function in a species-specific manner, but the mechanism is still unknown.     

Fluctuation and variation in stream-fish assemblages after a catastrophic flood in the Miyagawa River,Japan

In the autumn of 2004, a typhoon caused a catastrophic flood of the Miyagawa River in Japan. Based upon snorkeling surveys conducted every autumn from 2005 to 2009, we monitored the post flood fluctuation of the local fish assemblages at nine sites of both the main stream and subsidiary streams of the river. Results revealed that species richness significantly increased from 2005 to 2009. In addition, the fish densities of eight species significantly increased over the same period, whereas the density of one species decreased, and that of eight others remained unchanged. Categorization based on Euclidean distance revealed five main clusters from the nine sites. Among these sites, fish assemblages within subsidiary streams were stable as they remained within the same clusters while those in the main stream were dynamically variable through time as they changed cluster membership. In addition, the Euclidean distance between two arbitrary fish assemblages was positively correlated with environmental distance (the Euclidean distance calculated based on river width, depth, velocity and pebble size), time distance, and spatial distance along the river. In conclusion, the fish assemblages were dynamically and regularly altered and varied in the five years after the flood, except for those in the subsidiary streams, and such variation was related to environmental, temporal and spatial variation.