IL-12 and IL-18 Induction and Subsequent NKT Activation Effects of the Japanese Botanical Medicine Juzentaihoto

In this study, we first measured some cytokine concentrations in the serum of patients treated with Juzentaihoto (JTT). Of the cytokines measured interleukin (IL) -18 was the most prominently up-regulated cytokine in the serum of patients under long term JTT administration. We next evaluated the effects of JTT in mice, focusing especially on natural killer T (NKT) cell induction. Mice fed JTT were compared to control group ones. After sacrifice, the liver was fixed, embedded and stained. Transmission electron microscope (TEM) observations were performed. Although the mice receiving the herbal medicine had same appearance, their livers were infiltrated with massive mononuclear cells, some of which were aggregated to form clusters. Immunohistochemical staining revealed that there was abundant cytokine expression of IL-12 and IL-18 in the liver of JTT treated mice. To clarify what the key molecules that induce immunological restoration with JTT might be, we next examined in vitro lymphocyte cultures. Mononuclear cells isolated and prepared from healthy volunteers were cultured with and without JTT. Within 24 hours, JTT induced the IL-12 and IL-18 production and later (72 hours) induction of interferon (IFN)-gamma. Oral administration of JTT may induce the expression of IL-12 in the early stage, and IL-18 in the chronic stage, followed by NKT induction. Their activation, following immunological restoration could contribute to anti-tumor effects

Spectrophotometric determination of hydrogen peroxide by FIA with Bindschedler's Green leuco base as color reagent

ビンドシェドラーズグリーンロイコ塩基(LBG)を色原体とし,Fe(II)を触媒としたFIA系による過酸化水素の定量法について検討した.弱酸性溶液中,Fe(II)が存在するとLBGは過酸化水素によって725nmに吸収極大を持つ緑色のビンドシェドラーズグリーン(BG(+))に酸化され,この波長での吸光度の増加を利用して過酸化水素を定量した.pH及び共存させる塩酸ヒドロキシルアミンの濃度を調節することにより,過酸化水素が0~1ppmの範囲で検量線は直線性を示す.過酸化水素0.58ppmでの10回繰り返し測定の相対標準偏差は0.4%で,検出限界は5ppb(S/N=2)であった.1時間当たりの分析速度は30検体であった.市販の各種水あめ中のグルコース(3~96%)を,グルコースオキシターゼで反応させた後,FIAで測定した.A FIA method for the determination of hydrogen peroxide with a Bindschedler's Green leuco base (LBG) as color reagent and iron(II) as catalyst is described. LBG was oxidized by hydrogen peroxide to Bindschedler's Green (BG(+)) which showed a maximum absorbance at 725 nm. The increase in the absorbance at 725 nm was measured for the determination of hydrogen peroxide. The reagent solution (1) consisted of 10 mM hydrochloric acid containing 1 mM LBG, and the reagent solution (2) consisted of 0.1 M acetate buffer (pH 4.6) containing 50 mM ammonium iron(II) sulfate and 0.65 M hydroxylamine hydrochloride. Reagent solutions (1) and (2), and a carrier solution (deionized water) were propelled by two double plunger pumps (flow rate: 0.8 ml/min), and a sample solution (50 μl) was injected into the carrier stream. After mixing the reagent solution (1) and (2) in the preheating coil, the mixture stream was mixed with the carrier stream in the reaction coil (37℃). The linear relationship between the peak height and the concentration of hydrogen peroxide was obtained in the range of 0 to 1 ppm. The relative standard deviation of 0.58 ppm hydrogen peroxide was 0.4% (n=10). The detection limit was 5 ppb (S/N=2). The sampling rate was 30 samples per hour. The glucose in the range of 0 to 10 ppm was determined by the FIA method after the reaction with glucose oxidase

Determination of ammoniacal nitrogen in water by FIA with a gas-diffusion unit

水中のアンモニア態窒素のガス拡散/吸光光度FIAについて検討した.キャリヤー液として水酸化ナトリウム溶液を用いることにより,試料中のアンモニウムイオンをガス状のアンモニアとし,多孔質ポリテトラフルオロエチレンチューブを使用したガス拡散装置で分離した.アンモニアガスはチモールブルー溶液に吸収させ,pHの変化を596nmで測定する吸光度の変化に変えて定量した.ガス拡散装置はフランジ型の接続方法を採用し,液漏れしにくいものを製作した.本法によれば5ppm以下のアンモニア態窒素を1時間当たり25試料の速さで定量できた.又,検出限界は2ppbであり,20及び200ppbのアンモニア態窒素の9回繰り返し定量の相対標準偏差はそれぞれ3.8及び1.6%であった.環境水をFIA装置に直接注入してアンモニア態窒素を定量した結果とミクロケルダール蒸留法で不純物を取り除いた後本法で定量した結果は良く一致した.In this paper, the authors present an assembly of a gas-diffusion unit with a tubular microporous PTFE (polytetrafluoroethylene) membrane, constructed without any glue, and also present the determination of ammoniacal nitrogen in water. In this method, the sample was injected into an alkaline stream (carrier stream) in which the ammonium ions injected were converted to ammonia molecules, then the carrier stream flowed into the gas-diffusion unit. Reagent solution, which contained Thymol Blue and was adjusted to pH 8.4, was also fed to the gas-diffusion unit, and flowed through the microporous PTFE membrane tubing. Ammonia passed through the PTFE membrane and reacted with the acidic form of Thymol Blue to change it to the basic form. The color change to the basic form was measured at 596 nm in a flow-through cell. The detection limit was about 2 ppb as ammoniacal nitrogen, the response was linear up to 5 ppm, the sampling rate was 25 h(-1), and the relative standard deviations obtained by nine injections of 20 ppb and 200 ppb solutions were 3.8% and 1.6%, respectively. The results obtained by directly injecting water samples were in good agreement with that obtained by injecting the solutions prepared according to the Micro-Kjeldahl's distillation method

Determination of total carbonate-carbon in water by FIA with gas-diffusion unit

水中の総炭酸(二酸化炭素,炭酸塩及び炭酸水素塩)のガス拡散/吸光光度FIA法について検討した.キャリヤー液として硫酸を用い,試料中の総炭酸をガス状の二酸化炭素にしてガス拡散装置で分離しガス状二酸化炭素のみ試薬溶液に吸収させる.試薬溶液として酸-塩基指示薬であるクレゾールレッドを含む水溶液を使用し,pHの変化による吸光度の変化から定量する.ガス拡散装置はフランジ型の接続方法を採用しており,接着剤を用いる必要がなく接続部から液漏れしにくい.本法は1.2×10(-3)M以下の総炭酸が定量でき1時間当たり15試料の分析が可能であった.又,検出限界は5×10(-6)Mであり,水道水(4.7×10(-4)M)の10回繰り返し定量の相対標準偏差は,0.8%と良好であった.環境水を本法で定量した結果と間接吸光検出イオンクロマトグラフィーで定量した結果はよく一致した.In FIA, a gas-diffusion technique is highly selective, because there are only a few species which are generated as gases at room temperature. In this paper, the authors determined the total carbonate-carbon content in water by FIA with gas-diffusion unit assembly with a tubular microporous polytetrafluoroethylene (PTFE) membrane, which was reported previously. In this method, the sample was injected into a carrier stream of sulfuric acid in which carbonate and hydrogencarbonate ions injected were converted to gaseous carbon dioxide. The reagent stream which contained Cresol Red and was adjusted to pH 9.0, and the carrier stream, were fed to the gas-diffusion unit. Carbon dioxide passed through the PTFE membrane: the color change of Cresol Red to the acidic form was measured at 410 nm in a flow-through cell. The detection limit was 5×10(-6) M (S/N=3), the response was linear up to 1.2×10(-3) M, the sampling rate was 15 h(-1), and the relative standard deviation obtained by ten injections of tap water (4.7×10(-4) M) was 0.8%. The results obtained by the proposed method were in good agreement with those obtained by indirect photometric ion chromatography

Macular choroidal thickness and volume in eyes with angioid streaks measured by swept source optical coherence tomography.

[Purpose]: To study the mean choroidal thickness and volume of the macula in eyes with angioid streaks using swept source optical coherence tomography (OCT) in the 1050-nm wavelength range. [Design]: Prospective case series. [Methods]: The macular area of 39 eyes of 23 patients with angioid streaks and of 20 normal eyes of 20 matched controls (Group 1) was studied with a swept source OCT prototype system. Eyes with angioid streaks were classified into 1 of 4 groups: those without choroidal neovascularization (CNV) (Group 2); those with CNV that had no history of treatment (Group 3); those with CNV that had previously received only anti–vascular endothelial growth factor treatments (Group 4); and those with CNV that had previously received photodynamic therapy (Group 5). Using a raster scan protocol with 512 × 128 A-scans, we produced a macular choroidal thickness map (6 × 6 mm2). [Results]: There were no significant differences in age, axial length, or refractive error among the 5 groups. Mean choroidal thickness of the macula in Group 2 (218.9 ± 46.8 μm) was as great as that in Group 1 (218.8 ± 69.2 μm). However, the macular choroidal thickness in Group 3 (119.7 ± 49.2 μm), Group 4 (140.1 ± 64.9 μm), and Group 5 (144.0 ± 52.6 μm) was significantly less than that of Group 1 (P < .05). There were no statistical differences between Groups 3 through 5. In each group, the choroid of the nasal quadrant was significantly thinner compared to that in other quadrants (P < .05). [Conclusions]: The choroid in eyes with angioid streaks without CNV was as thick as that in normal controls, but was significantly thinner in eyes with angioid streaks that had developed CNV