Formation of the thiocyanate conjugate of chlorogenic acid in coffee under acidic conditions in the presence of thiocyanate and nitrite: possible occurrence in the stomach

The objective of the present study was to elucidate how chlorogenic acid in coffee was transformed under acidic conditions simulating the mixture of saliva and gastric juice. When coffee was incubated in acidified saliva that contained nitrite and SCN-, in addition to nitric oxide (NO), four major components were detected. Two of the four components (components 3 and 4) were generated when chlorogenic acid was incubated in acidified saliva and when incubated in an acidic buffer solution in the presence of both nitrite and SCN-. By the incubation of chlorogenic acid in acidic nitrite in the absence of SCN-, components 3 and 4 were not formed but the quinone of chlorogenic acid and nitrated chlorogenic acid were formed. The result indicates that SCN- was indispensable for nitrous acid induced formation of components 3 and 4. Component 4 was isolated and its structure was determined to be (E)-5'-(3-(7-hydroxy-2-oxobenzo[d] [1,3]oxathiol-4-yl)acryloyloxy)quinic acid. Component 3, which was suggested to be 2-thiocyanatochlorogenic acid, seemed to be formed by the reaction between SCN- and the quinone of chlorogenic acid. As it has been reported that the quinone of chlorogenic acid can react with thiols and can decompose producing H2O2, the formation of component 4 can reduce the toxic effects of the quinone of chlorogenic acid.     

In vivo anti-influenza virus activity of an immunomodulatory acidic polysaccharide isolated from Cordyceps militaris grown on germinated soybeans

An acidic polysaccharide (APS) was isolated from the extract of Cordyceps militaris grown on germinated soybeans. Analyses of sugar composition indicated that APS consisted of d-galactose, L-arabinose, D-xylose, L-rhamnose, and D-galacturonic acid. On the basis of the result of methylation analysis, APS was considered to be mainly composed of Araf-(1-->, -->5)-Araf-(1-->, -->4)-Galp-(1--> and -->4)-GalAp-(1--> residues. When the polysaccharide was intranasally administered, it decreased virus titers in the bronchoalveolar lavage fluid and the lung of mice infected with influenza A virus and increased survival rate. Furthermore, APS increased TNF-alpha and IFN-gamma levels in mice when compared with those of untreated mice. APS enhanced nitric oxide (NO) production and induced iNOS mRNA and protein expressions in RAW 264.7 murine macrophage cells. The induction of mRNA expression of cytokines including IL-1beta, IL-6, IL-10, and TNF-alpha was also observed. These results demonstrated that APS might have beneficial therapeutic effects on influenza A virus infection at least in part by modulation of the immune function of macrophages.     

Restricted localization of claudin-16 at the tight junction in the thick ascending limb of Henle's loop together with claudins 3, 4, and 10 in bovine nephrons

Claudin-16 is one of the tight junction protein claudins and has been shown to contribute to reabsorption of divalent cations in the human kidney. In cattle, total deficiency of claudin-16 causes severe renal tubular dysplasia without aberrant metabolic changes of divalent cations, suggesting that bovine claudin-16 has some roles in renal tubule formation and paracellular transport that are somewhat different from those expected from the pathology of human disease. As the first step to clarify these roles, we examined the expression and distribution of claudin-16 and several other major claudin subtypes, claudins 1-4 and 10, in bovine renal tubular segments by immunofluorescence microscopy. Claudin-16 was exclusively distributed to the tight junction in the tubular segment positive for Tamm-Horsfall glycoprotein, the thick ascending limb (TAL) of Henle's loop, and was found colocalized with claudins 3, 4, and 10. This study also demonstrates that bovine kidneys possess segment-specific expression patterns for claudins 2-4 and 10 that are different from those reported for mice. Particularly, distribution of claudin-4 in the TAL and distal convoluted tubules was characteristic of bovine nephrons as were differences in the expression patterns of claudins 2 and 3. These findings demonstrate that the total lack of claudin-16 in the TAL segment is the sole cause of renal tubular dysplasia in cattle and suggest that the tight junctions in distinct tubular segments including the TAL have barrier functions in paracellular permeability that are different among animal species.     

Characteristics and epidemiologic genotyping of Staphylococcus aureus isolates from bovine mastitic milk in Hokkaido, Japan

Two hundred thirty one Staphylococcus aureus isolates from bovine mastitic milk were discriminated into 60 patterns and 16 lineages by pulsed-field gel electrophoresis (PFGE). The tested isolates were also investigated using coagulase and capsule serotyping and PCR for possession of genes that encode staphylococcal enterotoxins (sea to sei), enterotoxin-like toxins (selj to selr), and toxic shock syndrome toxin (tst). One hundred seventy three of the isolates (74.9%) possessed one or more toxin genes, while no egg-yolk factor was detected in most of them. The most common combinations of toxin genes possessed by the tested isolates were sec, seg, sei, sell, and tst, or seg and sei, or sec, seg, sei, sell, seln, and tst. Two hundred and ten of the isolates (91.0%) serotyped coagulase VI, and 207 of the isolates (89.6%) expressed serotype 5 or 8 capsules. These results suggested that isolates belonging to two major lineages have spread all over Hokkaido as bovine mastitic isolates. Additionally, no remarkable difference was recognized in the identification ratio of the isolates that belonged to the two major lineages between mastitis of subclinical origin and mastitis of clinical origin.     

Cryptosporidium infection in juvenile pet rabbits

Cryptosporidium infection was confirmed by fecal examination for the first time in pet rabbits in a wholesale store located in Kanagawa Prefecture, Japan. Fecal samples were obtained postmortem from juvenile rabbits (n=66), which had died after developing diarrhea. Feces from healthy rabbits (n=30) were also collected and examined as controls. Two types of Cryptosporidium oocysts distinctive in size and shape were found (Type A and B). Types A and B oocysts were detected from 16.7% and 13.6% of the diarrheic, and 3.3% and 0% of the normal feces, respectively. Since Cryptosporidium oocysts were detected at a higher rate in the diarrheic rabbits than in the healthy rabbits, special caution should be taken when handling a pet rabbit presenting with diarrhea.     

Cardiovascular effects of continuous propofol infusion in horses

We examined the influence of propofol infusion on cardiovascular system at the rate of 0.14, 0.20 and 0.30 mg/kg/min in six adult Thoroughbred horses. The cardiovascular parameters were heart rate (HR), mean arterial pressure (MAP), mean right atrial pressure (MRAP), stroke volume (SV), cardiac output (CO), systemic vascular resistance (SVR), pre-ejection period (PEP) and ejection time (ET). In order to keep the ventilation conditions constantly, intermittent positive pressure ventilation was performed, and the partial arterial CO(2) pressure was maintained at 45 to 55 mmHg during maintenance anesthesia. SV showed a significant dose-dependent decrease however, CO did not show significant change. SVR decreased significantly at higher dose. PEP was prolonged and PEP/ET increased significantly at the highest dose. From these results, it became clear that SV decreases dose-dependently due to decrease of cardiac contractility during anesthesia with continuous propofol infusion in horses. On the other hand, since MAP and CO did not show significant changes, total intravenous anesthesia with propofol was suggested to be suitable for long-term anesthesia in horses.     

Controlling the electronic structure of bilayer graphene

We describe the synthesis of bilayer graphene thin films deposited on insulating silicon carbide and report the characterization of their electronic band structure using angle-resolved photoemission. By selectively adjusting the carrier concentration in each layer, changes in the Coulomb potential led to control of the gap between valence and conduction bands. This control over the band structure suggests the potential application of bilayer graphene to switching functions in atomic-scale electronic devices.     

Hyperproduction and application of alpha-agarase to enzymatic enhancement of antioxidant activity of porphyran

The nucleotide sequence of the gene for the alpha-agarase, AgaA33, from Thalassomonas sp. strain JAMB-A33 was determined. The open reading frame for AgaA33 was revealed to encode 1463 amino acid residues. We succeeded in extracellular production of recombinant -agarase (AgaA33) efficiently using Bacillus subtilis as a host. This is the first report of recombinant production of -agarase. Furthermore, we demonstrated that hydrolysis of alpha-1,3 linkages in porphyran, a sulfated polysaccharide from marine red algae, by alpha-agarase is an important step for improvement of its antioxidant activity with regard to free-radical-scavenging capacity and superoxide radical anion scavenging activity, whereas the hydrolysis of beta-1,4 linkages in porphyran by beta-agarase did not increase on the antioxidant activity markedly.     

Anisotropy and corotation of galactic cosmic rays

The intensity of Galactic cosmic rays is nearly isotropic because of the influence of magnetic fields in the Milky Way. Here, we present two-dimensional high-precision anisotropy measurement for energies from a few to several hundred teraelectronvolts (TeV), using the large data sample of the Tibet Air Shower Arrays. Besides revealing finer details of the known anisotropies, a new component of Galactic cosmic ray anisotropy in sidereal time is uncovered around the Cygnus region direction. For cosmic-ray energies up to a few hundred TeV, all components of anisotropies fade away, showing a corotation of Galactic cosmic rays with the local Galactic magnetic environment. These results have broad implications for a comprehensive understanding of cosmic rays, supernovae, magnetic fields, and heliospheric and Galactic dynamic environments.     

Eye fatigue-reduction effect of wood rim cover on liquid crystal display

The purpose of this study is to confirm objectively by an instrumental measurement if there is less eye fatigue in VDT work when using a wood rim covered display than when using a plastic housing display. The subjects were 26 graduate and undergraduate university students. Each subject did the same 60-min VDT activity with both a plastic housing and wood-covered display. Before and after VDT activity, each subject’s near-point accommodation distance was measured by a near-point ruler. As a result, it was confirmed that variation rate of the near-point accommodation distance by the VDT work with the wood-covered display was smaller than with the plastic housing display. From sensory tests, progression of subjective symptoms such as eye pain and headache was less with the wood-covered display than with the plastic housing display.