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Adjuvants are essential substances for vaccines and immunotherapies that enhance antigen-specific immune responses. Single-stranded oligodeoxynucleotides containing an unmethylated CpG motif (CpG ODNs) are agonistic ligands for toll-like receptor 9 that initiate an innate immune response. They represent promising adjuvants for antiviral and antitumor immunotherapies; however, CpG ODNs have some limitations, such as poor nuclease resistance and low cell membrane permeability. Therefore, an effective formulation is needed to improve the nuclease resistance and immunostimulatory effects of CpG ODNs. Previously, we demonstrated the selective delivery of a small molecule toll-like receptor 7 ligand to immune cells through sugar-binding receptors using sugar-immobilized gold nanoparticles (SGNPs), which significantly enhanced the potency of the ligand. In this study, we examined SGNPs as carriers for partially phosphorothioated A-type CpG ODN (D35) and an entirely phosphorothioated B-type CpG ODN (K3) and evaluated the functionality of the sugar moiety on SGNPs immobilized with CpG ODN. SGNPs immobilized with D35 (D35-SGNPs) exhibited improved nuclease resistance and the in vitro and in vivo potency was significantly higher compared with that of unconjugated D35. Furthermore, the sugar structure on the GNPs was a significant factor in enhancing the cell internalization ability, and enhanced intracellular delivery of D35 resulted in improving the potencies of the A-type CpG ODN, D35. SGNPs immobilized with K3 (K3-SGNPs) exhibited significantly higher induction activities for both humoral and cellular immunity compared with unconjugated K3 and D35-SGNPs. On the other hand, sugar structure on K3-SGNPs did not affect the immunostimulatory effects. These results indicate that the sugar moiety on K3-SGNPs primarily functions as a hydrophilic dispersant for GNPs and the formulation of K3 to SGNPs contributes to improving the immunostimulatory activity of K3. Because our CpG ODN-SGNPs have superior induction activities for antigen-specific T-cell mediated immune responses, they may be effective adjuvants for vaccines and immunotherapies.

Polycyclic aromatic hydrocarbons (PAHs) are carcinogenic substances that are mainly generated during heating in food; therefore, the European Union (EU) has regulated the amount of benzo[a]pyrene and PAH4 in various types of food. In addition, the Scientific Committee on Food of the EU and the Joint Food and Agriculture Organization/World Health Organization Expert Committee on Food Additives have recommended that 16 PAHs should be monitored. Since coffee beverages and dark beer are roasted during manufacture, monitoring these 16 PAHs is of great importance. On the other hand, supercritical fluid chromatography (SFC) is a separation method that has garnered attention in recent years as a complement for liquid and gas chromatography. Therefore, we developed a rapid high-sensitivity analytical method for the above-mentioned 16 PAHs in coffee beverages and dark beer involving supercritical fluid chromatography/atmospheric pressure chemical ionization-mass spectrometry (SFC/APCI-MS) and simple sample preparation. In this study, we developed a novel analytical technique that increased the sensitivity of MS detection by varying the back-pressure in SFC depending on the elution of PAHs. In addition, analysis of commercially available coffee and dark beer samples in Japan showed that the risk of containing the 16 PAHs may be low., (Copyright © 2018 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.)

A method for simultaneous analysis of egg and milk allergens using LC-QTOF-MS was developed. The proteins measured were α-casein, β-lactoglobulin, and ovalbumin, which are the main protein allergens in milk and eggs. The proteins were digested using trypsin, and the digests were analyzed by LC-QTOF-MS. Sixteen peaks were detected that confirmed the amino acid sequences of the digests, and a MRM method with high resolution (MRM-HR) using product ions of these peaks was applied for quantification. Next, validation studies were performed using beverage products to which milk and egg standard protein solutions had been added. Good linearity was achieved over the concentration range of 1.25 to 20 μg/g of milk and egg protein, and acceptable reproducibility and accuracy were obtained at 10 μg/g. Moreover, good agreement was also observed between LC-QTOF-MS and ELISA. These findings suggest that this LC-QTOF-MS method may be useful for determining the milk and egg protein contents of beverages.

We herein present a method for the analysis of 10 dithiocarbamate fungicides (DTCs) in beer, fruit juice, and malt samples, where the DTCs were converted into water-soluble sodium salts in the presence of NaHCO 3 prior to methylation using dimethyl sulfate. Extraction of the methylated compounds from matrices was performed using a "quick, easy, cheap, effective, rugged, and safe" (QuEChERS) method. Following a dispersive solid-phase extraction as a clean-up step, the methylated compounds were detected by liquid chromatography-tandem mass spectrometry. Performance evaluation was carried out on beer, fruit juice, and malt samples using representative compounds. Accuracies of the spiked compounds from the various matrices ranged from 92.2 to 112.6%, and the limits of quantification of propineb, mancozeb, and thiuram were <0.52, <0.55, and <6.97 µg/kg, respectively. The developed method was then applied in the determination of dithiocarbamate fungicide contents in commercial beer and fruit juice samples.

A high-resolution liquid chromatography-Orbitrap mass spectrometry (LC-Orbitrap MS) method was developed for simultaneous determination of 20 Fusarium toxins (nivalenol, fusarenon-X, deoxynivalenol, 3-acetyl deoxynivalenol, 15-acetyl deoxynivalenol, HT-2 toxin, T-2 toxin, neosolaniol, diacetoxyscirpenol, fumonisin B1, fumonisin B2, fumonisin B3, fumonisin A1, fumonisin A2, fumonisin A3, zearalenone, α-zearalenol, β-zearalenol, α-zearalanol, and β-zearalanol) in cereals. The separation of 20 Fusarium toxins with good peak shapes was achieved using a pentafluorophenyl column, and Orbitrap MS was able to detect accurately from cereal matrix components within ±0.77 ppm. The samples were prepared using a QuEChERS kit for extraction and a multifunctional cartridge for purification. The linearity, repeatability, and recovery of the method were >0.9964, 0.8%-14.7%, and 71%-106%, respectively. Using this method, an analysis of 34 commercially available cereals detected the presence of deoxynivalenol, 15-acetyl deoxynivalenol, fumonisin B1, fumonisin B2, fumonisin B3, fumonisn A1, fumonisin A2, fumonisin A3, and zearalenone in corn samples with high concentration and frequency. Trichothecenes was detected from wheat samples with high frequency; in particular, the concentration of deoxynivalenol was high. Conversely, α-zearalenol, β-zearalenol, α-zearalanol, and β-zearalanol were not detected in any of the samples.

Three compounds, hypothesized as fumonisin A1 (FA1), fumonisin A2 (FA2), and fumonisin A3 (FA3), were detected in a corn sample contaminated with mycotoxins by high-resolution liquid chromatography-Orbitrap mass spectrometry (LC-Orbitrap MS). One of them has been identified as FA1 synthesized by the acetylation of fumonisin B1 (FB1), and established a method for its quantification. Herein, we identified the two remaining compounds as FA2 and FA3, which were acetylated fumonisin B2 (FB2) and fumonisin B3 (FB3), respectively. Moreover, we examined a method for the simultaneous analysis of FA1, FA2, FA3, FB1, FB2, and FB3. The corn samples were prepared by extraction using a QuEChERS kit and purification using a multifunctional cartridge. The linearity, recovery, repeatability, limit of detection, and limit of quantification of the method were >0.99, 82.9%-104.6%, 3.7%-9.5%, 0.02-0.60 μg/kg, and 0.05-1.98 μg/kg, respectively. The simultaneous analysis of the six fumonisins revealed that FA1, FA2, and FA3 were present in all corn samples contaminated with FB1, FB2, and FB3. The results suggested that corn marketed for consumption can be considered as being contaminated with both the fumonisin B-series and with fumonisin A-series. This report presents the first identification and quantification of FA1, FA2, and FA3 in corn samples.

The ultrasonic cleaning of artificially soiled fabrics with and without shake was carried out in an aqueous anionic surfactant solution. The polyester, cotton and polyester/cotton (65/35) fabrics were soiled with oleic acid or carbon black as a model soil, and cleaned together with their original fabrics with applying ultrasound for 5min. The detergency and the soil redeposition were determined from the change in the Kubelka-Munk function of the soiled and original fabric surfaces due to the cleaning. For any fabric, the removal of oleic acid and carbon black from the soiled fabric and their redeposition onto the original fabric increased with increasing electric power consumption of ultrasound. When ultrasound and shake were applied at the same time, the detergency further increased for any electric power consumption. The maximum detergency obtained with combination of ultrasound 340W and shake 160spm was compared with detergency obtained with Wascator, a horizontal axis drum type washer. It was found that the ultrasound/shake combination cleaning enabled efficient removal of both soils from any fabric and the detergency of the polyester fabrics was comparable to that with Wascator. The mechanical action during the washing was evaluated by two mechanical action test pieces commercially available, which indicated that the ultrasound/shake combination cleaning provided gentle mechanical action to the fabric in comparison with the drum type washer. The SEM observations showed the damage of the fabric and fiber surfaces was negligibly small after the ultrasound/shake combination washing., (Copyright © 2014 Elsevier B.V. All rights reserved.)

The ultrasound was applied to textile washing as a mechanical action for soil removal. The polyester fabric was soiled with carbon black or oleic acid as a model contaminant, and washed with the original fabric in aqueous solutions without and with alkali or surfactant by applying ultrasound, shaking or stirring action. The detergency and soil redeposition were evaluated from the change in the surface reflectance of artificially soiled fabrics and the original fabric due to washing. In comparison with shaking and stirring actions, ultrasound was found to remove the particulate and oily soils efficiently in a short time and at low bath ratio. With increasing ultrasound power, the detergency of both soils increased and exceeded that obtained with Wascator, a horizontal axis drum type washer. Using three standard fabrics for determining mechanical action during washing, it was shown that ultrasound washing caused little mechanical damage to the fabric. However, the soil redeposition was frequently observed for ultrasonic washing, especially at low bath ratio., (Copyright © 2012 Elsevier B.V. All rights reserved.)

A simple method for analysis of trichothecenes [Type A: diacetoxyscirpenol, neosolaniol, HT-2 toxin, and T-2 toxin, Type B: deoxynivalenol, nivalenol, fusarenon-X, 3-acetyldeoxynivalenol, 15-acetyldeoxynivarenol] in barley tea and beer using liquid chromatography tandem mass spectrometry (LC/MS/MS) was developed. Trichothecenes were extracted with ethyl acetate-methanol (19:1). The solvent was evaporated to dryness and the residue was dissolved in water-methanol (3:1) for injection into the LC/MS/MS. The LC separation was performed with an octadecylated silica column at a flow-rate of 0.2 mL/min, using a mobile phase consisting of water, methanol and acetonitrile. MS/MS was used in multiple reaction monitoring, employing electrospray ionization (ESI-MRM). The recoveries of trichothecenes from drinks at 1 microg/L (Type A) and 10 microg/L (Type B) were 52.5-115.2% (barley tea) and 68.1-127.5% (beer). Five barley tea and ten beer samples were analyzed by this method. Trichothecenes were not detected in them. This method may have applications in quality assurance.

The effects of supplementing 8% casein or 10% soy protein isolate (SPI) diets with graded levels of oligo-L-methionine (a mixture of hexa- and heptapeptides, OM) or L-methionine (Met) were studied in rats to determine the reason for the difference in nutritional quality between proteins and corresponding amino acid mixtures. As the OM concentration of the casein-based diet was increased from 0.02% to 0.6%, maximum weight gain was attained at 0.2%, and the growth-promoting activity of OM was comparable to Met at all the corresponding levels tested. Liver fat began to accumulate when supplemental Met reached a level of 0.08% of the casein diet, but OM addition did not produce a fatty liver at dietary levels of less than 0.3%. When SPI was used as the dietary protein source, the effect of supplemental OM was significantly less than that of Met. Digestibility of OM (assessed by incremental portal plasma Met concentration) was measured 30 min after feeding the casein or SPI diet supplemented with 3% OM using rats fasted for 24 h. Plasma Met concentration was greatly increased in rats fed the casein plus OM diet compared with that of rats fed the SPI + OM diet. Similarly, the 30-min portal Met concentration significantly increased in response to the casein + OM diet compared with the SPI + OM diet regardless of the prefed proteins (25% casein and 25% SPI for 2 wk).(ABSTRACT TRUNCATED AT 250 WORDS)

Two hundred twenty undamaged human fetuses without external malformations were supplied by the human embryo and fetus collection at the Department of Anatomy, Kyoto University. The diameters of the cornea and eyeball were measured by slide calipers under a stereomicroscope. The various diameters of the eyeball showed a parallel, linear increase from the 12th to the 28th week of menstrual age. Corneal diameters appeared to increase in parallel with the eyeball as a whole until the 16th week of menstrual age; later on their rate of growth declined. The corneal limbus could be recognized macroscopically at the 17th week of menstrual age and was distinctly seen from the 20th week of menstrual age. The corneal curve at the limbus began to be a little convex at the 21st week of menstrual age and became distinctly convex from the 23rd week. The cornea was transparent from the 12th to the 28th week of menstrual age. The sclera lost its transparency from the 21st week. It is significant that during fetal life the sagittal diameter of the eyeball is shorter than the vertical and transverse diameters.

Patients treated successfully with retinal detachment surgery 11-39 years earlier were called back and examined. The average age was 35 years at the time of surgery and the average interval between the operation and this follow-up examination was 16 years. The visual acuity had decreased in many patients. The most frequent causes of this fall in visual acuity after surgery were cataract and macular degeneration. Chorioretinal atrophy with blackish pigment spots developed in the reattached retina a long time after surgery and caused defects in the visual fields. This atrophy seems to be related to the long duration of retinal detachment before and after operation, which may cause malnutrition of the detached retina and result in incomplete recovery of visual cells, leading to chorioretinal atrophy. Macular degeneration was caused or accelerated by detachment of the macula. There was no difference in postoperative ocular findings between only diathermy and combination of diathermy with segmental scleral-buckling procedures. However, Arruga's ring suture method induced cataract more frequently than did other surgical procedures.

The relationship between the 'diabetic state' and massive periretinal proliferation (MPP) in idiopathic retinal detachment was examined. In 18 (3.5%) of 508 cases of idiopathic retinal detachment, MPP occurred before or after retinal detachment surgery. In 24 cases of retinal detachment, a diabetic or suspected diabetic state was found. 11 of the 24 were receiving treatment for diabetes and never developed MPP, while 5 of the 13 patients who were not being treated had MPP. This suggests that MPP in retinal detachment may easily be induced by the diabetic state and prevented by treatment for diabetes. Additional causes of MPP in retinal detachment were giant tears, massive vitreous hemorrhages at operation and frequent recurrence.

In 600 patients with successfully operated unilateral retinal detachment, the visual acuity prognosis of the operated eye was studied in relation to that of the normal eye. Within 2 years after operation, most of these patients were in the following three groups: (1) visual acuity worse than 0.5 in the non-detached retina and worse than 0.1 in the reattached one; (2) visual acuity better than 1.0 in the non-detached retina and worse than 0.3 in the reattached one; and (3) visual acuity better than 1.0 in the non-detached retina and better than 0.8 in the reattached one. Some of the patients in group 2 showed greater improvement of the visual acuity in the reattached retina, which occasionally became superior to that in the non-detached one. In spite of possible improvement until 2 years after operation, the visual acuity in the reattached retina was apt to decrease after more than 5 years following operation. The presence of macular detachment was ill for the recovery of visual acuity. About 30% of the patients with a visual acuity of about 1.0 in the non-detached retina showed no improvement of the visual acuity in the reattached retina. About 50% of the patients with successful surgery for retinal detachment had a difference of more than 0.6 in visual acuity between the non-detached and the detached retinas.

Two hundred fifty-two undamaged human fetuses without external malformations were supplied by the human embryo and fetus collection of the Department of Anatomy, Kyoto University. The sagittal, vertical and horizontal diameters of the eyeball were measured by micrometer, and the transverse, vertical and horizontal circumferences of the eyeball were measured by mapmeter from photographs of the side views of the eye. All the diameters of the eyeball showed a parallel linear increase from the 12th to the 40th week of menstrual age. The increase of circumferences of the eyeball paralleled each other throughout the 12th to the 40th week. It is significant that the sagittal diameter of the eyeball is shorter than the vertical and transverse diameters during fetal life. The vertical meridian circumference of the eyeball is also shorter than the transverse and horizontal meridian circumferences during fetal life. These findings suggest that many infants are apt to have hyperopia at birth.