Exercise affects human immune functions through the hypothalamus-pituitary-adrenal (HPA) axis and reward systems in the brain. Most immune cells, including T cells, B cells and phagocytes, express functional hormone and neurotransmitter receptors and are regulated through the neuroendocrine immune network. Light-to-moderate exercise improves the defense activity against pathogens and prevents metabolic disorders. Many epidemiological and experimental studies have suggested there is an inverse relationship between moderate exercise training and illness risk. Furthermore, habitual exercise might improve immune regulation, reduce the activity of autoimmune disorders and delay age-related dysfunctions, such as sarcopenia. On the other hand, high-intensity exercise suppresses immune defense activity, especially against upper respiratory infections. Thus, high-intensity training and competitions possibly increase the risk of infection in athletes. Appropriate nutritional intake and hygiene control are also essential to protect both athletes and non-athletes.

Autoimmune disorders occur more frequently in women. This predominance is attributed to gonadal steroids related to immunoactivation, fetal microchimerism and/or X chromosome inactivation. Both in vivo and in vitro experiments have shown that estrogen up-regulates humoral immune responses, which are suppressed by androgen and progesterone. The histopathological similarities between Scleroderma (SSc) and Graft versus host disease (GVHD) suggest its pathophysiology as chronic GVHD between fetal cells and maternal tissues. This hypothesis was confirmed using highly sensitive PCR and in situ hybridization. However, other reports have suggested that chimeric fetal cells can restore damaged maternal tissues. Finally, recent advances in molecular biology have revealed skewed X chromosomal inactivation in patients with autoimmune disorders. Females exhibit chimeric X chromosome activation status between Xp (paternal) and Xm (maternal) cells. If thymic antigen presentation cells become skewed for Xp or Xm, central T cell selection lacks Xp or Xm directed regulatory T cells. Other explanations suggest reactivation of silenced X chromosome and subsequent CD40L transcription and/or lack of pseudoautosomal region relatedimmunoregulation in Xo monosomy lymphocytes. From the evolutionary antiquity of sex over the acquired immune system, we suggest that viviparity is the ultimate cause of inconsistencies in autoimmune recognition.

64 歳の女性がぶどう膜炎の原因精査目的で当院内科を受診した．症状として前腕の皮膚掻痒を訴えていた．血液検査では赤沈の亢進，胆道系酵素値の上昇，総コレステロール値の上昇，抗核抗体の上昇を認めた．胸部レントゲン写真では胸肋鎖関節部の異常骨化像を認めた．肝機能障害の精査中に抗ミトコンドリア M2 抗体の陽性が確認され，原発性胆汁性肝硬変症 (PBC) と診断した．ウルソデオキシコール酸の内服により皮膚掻痒は軽快し，肝機能異常も正常化した．超音波，腹部 CT など画像上，肝硬変症の所見は認められず，安定して経過している．胸肋鎖骨間異常骨化症 (ISCCO) は掌蹠膿疱症，乾癬などの皮膚疾患に合併することが多く，血清反応陰性脊椎関節症 (SpA) の一つと考えられている．PBCにはシェーグレン症候群や強皮症が時に合併し，関節症状を呈することがあるが，PBC と ISCCO との合併は文献上見当たらず稀である．

Rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) are the most common autoimmune disorders, although they each have very different pathophysiology. In general, RA is considered to be a Th1-mediated disease, while SLE is a Th2-mediated disease. Thus, their overlapping, in so called "rhupus", is a rare condition. In Rembrandt van Rijn's (1606-1669) portrait of the middle-aged Maria Bockenolle, we have what may be the earliest depiction of a case of rhupus syndrome: the coexistence of a butterfly rash and digital deformities. This suggests the possible historical importance of an RA epidemic which took place in the early 17th century.

High-speed counter-current chromatography has been successfully applied to the separation of the lac dye components. A 25-mg quantity of the sample was separated using a two-phase solvent system composed of tert.-butyl methyl ether-n-butanol-acetonitrile-water (2:2:1:5). The fractions were analyzed by high-performance liquid chromatography and electrospray tandem mass spectrometry. The separation yielded 2.6 mg of 97.2% pure laccaic acid C, 9.5 mg of 98.1% pure laccaic acid A, 3.6 mg of 98.2% pure laccaic acid B, and 0.5 mg of a 95.0% pure anthraquinonedicarboxylic acid with a molecular mass of 360.

A simple, rapid and reproducible analytical method for thiabendazole (TBZ) and imazalil (IMA) in citrus fruit and banana has been developed. The method involves the use of an ion-exchange cartridge for sample clean-up followed by ion-pair high-performance liquid chromatography with ultraviolet detection. The recoveries of TBZ and IMA from citrus fruits spiked at levels of 10 mu g/g and 5 mu g/g were in the range of 94-98% and 93-98% with coefficients of variation of 0.5-2.2% and 1.6-2.7%, respectively. The recoveries of TBZ and IMA from banana spiked at levels of 3 mu g/g and 2 mu g/g were 94% and 94% with coefficients of variation 1.1% and 4.9%, respectively. The detection limits for TBZ and IMA were 0.1 mu g/g in citrus fruit and 0.05 mu g/g in banana. (C) 1998 Elsevier Science B.V. All rights reserved.

Tissues were collected to survey the incidence of tetracycline antibiotics (TCs) residues in slaughtered animals that did not pass inspection at slaughterhouses in Aichi Prefecture, Japan, April, 1985-March, 1996, because of disease symptoms. Tissues were analyzed by liquid chromatography. Among 218 animals (85 cattle and 133 hogs), 79 (30.9%) were positive for TCs, including 43 (19.7%) for oxytetracycline (OTC), 3 (1.4%) for tetracycline, 28 (12.8%) for chlortetracycline, and 1 (0.5%) for doxycycline (DC). One sample (cattle kidney) was positive for both OTC and DC. Cattle and hog samples collected in 1991-1995 showed significantly higher incidence of TC and CTC residues than those in 1985-1986 (X2-test), respectively. Kidney samples showed higher incidence of TCs residues and 3-4 times higher residual concentrations than liver samples.

The purity test of the Japanese Pharmacopoeia Absorbent Gauze for fluorescent whitening agents is regulated to perform by a method of irradiating the gauze with ultraviolet rays in a dark place. This method has been reported to have a difficulty in detecting the existence of fluorescent whitening agents in several cases. We, therefore, tried to establish a method to examine the existence of fluorescent whitening agents using physical and chemical methodologies. After cutting the Japanese Pharmacopoeia Absorbent Gauze samples in small pieces, fluorescent whitening agents were extracted by 30% pyridine solution at 100°C for 30 min. We analysed the fluorescent agents using reversed phase TLC, spectrofluorophotometer and HPLC with a fluorescence detector using 8 standard fluorescent whitening agents as controls. Conditions of reversed phase TLC : plate, RP-18W (Merck) ; solvent system, A) acetonitrile-water (1 : 4), B) acetonitrile-methanol-water (3 : 3 : 10), C) methyl ethyl ketone-methanol-water (1 : 1 : 1). Conditions of HPLC with fluorescence detection : column, Wakosil 5C18 (i.d. 4.6×150 mm) ; mobile phase, 0.01M ammonium acetate-acetonitrile (7 : 3) ; flow rate, 1.0 ml/min ; injection volume, 20 μl ; detections, fluorescence detector (excitation 345 nm and emission 435 nm).

A reliable identification method for laccaic acids which are the main components in lac dye has been established using ESI LC/MS/MS with a volatile mobile phase containing acetylacetone. Addition of acetylacetone to the mobile phase enables us to separate laccaic acids without reduction of peak resolution. The volatile mobile phase, acetonitrile-0.05% aqueous TFA solution (1-4) containing 0.005 M acetylacetone, is applicable to directly interfaced ESI LC/MS/MS without clogging problems. In all ES tandem mass spectra of laccaic acids, [M-H-CO2]- and [M-H-2CO2]- appeared in considerable abundance, together with the weak ions of [M-H]-, [M-H-H2O]-, [M-H-CO2-H2O]-, and [M-H-2CO2-H2O]- when [M-H]- was selected as the precursor ion. Laccaic acids can be easily identified by these product ions. The method has been successfully applied to identification of laccaic acids in a jelly and in "Shi-kou," which is a stick lac imported 1,200 years ago.

High-speed countercurrent chromatography was successfully applied to the isolation of colistin-A and colistin-B from a commercial colistin preparation. As the first step, TLC and HPLC analysis conditions for the colistin components were established. Using these techniques, a two-phase solvent system composed of n-butanol-0.04M aqueous trifluoroacetic acid (TFA) (1:1) was selected for high-speed CCC, where the concentration of TFA was the major factor in controlling the partition coefficients of the colistin components. Yields of colistin-A and colistin-B were 9 mg each from 20 mg of the commercial sample, and the purity of each component was over 90 %. Fast atom bombardment (FAB) mass spectrometry was utilized to confirm the nature of the isolated components.

Structural characterization of the colistin (CL) components were carried out using Frit-fast atom bombardment liquid chromatography/mass spectrometry (Frit-FAB LC/MS), tandem mass spectrometry (MS/MS) and the amino acid analysis proposed by MARFEY, and the total structures of 4 minor components including the absolute configuration of the constituent amino acids were proposed. The structures of the minor components were the same as those of the main component colistin A or B except that L-leucine is replaced by L-valine or L-isoleucine.

To reliably identify the residual tetracycline antibiotics (TCs), oxytetracycline (OTC), tetracycline, chlortetracycline (CTC) and doxycycline (DC), in bovine tissues, we have established a confirmation method using electrospray ionization liquid chromatography-tandem mass spectrometry (ESI LC-MS-MS) with daughter ion scan. All TCs gave [M+H-NH3]+ and [M+H-NH3-H2O]+ as the product ions, except for DC when [M+H]+ was selected as the precursor ion. The combination of C18 cartridge clean-up and the present ESI LC-MS-MS method can reliably identify TCs fortified at a concentration of 0.1 ppm in bovine tissues, including liver, kidney and muscle, and has been successfully applied to the identification of residual OTC in bovine liver and residual CTC in bovine muscle samples previously found at concentrations of 0.58 ppm and 0.38 ppm by LC, respectively.

In order to facilitate the application of the foam counter-current chromatography (CCC) technique to various natural products, we have introduced two parameters, i.e., “foaming power” and “foam stability”, which reliably indicate applicability of the sample to foam CCC. In each test, the sample was delivered into a graduated cylinder with a ground stopper followed by vigorous shaking. The foaming power was expressed by the volume ratio of the resulting foam to the remaining solution, and the foam stability by the duration of the foam. A set of data obtained from various samples including bacitracin, gardenia yellow, phloxine B, rose bengale and senega methanol extract revealed a significant correlation between these parameters and the degree of foam enrichment by foam CCC. The overall results indicated that a sample with the following properties may be effectively enriched by foam CCC: the foaming power is greater than 1.0 and the foam stability is over 250 min. We applied these experimental results to the enrichment of microcystins in a cyanobacteria bloom sample extract using foam CCC.

A simple and rapid method for the determination of heme iron in foods was investigated. A sample was weighed in a centrifuge tube and the fat in the sample was removed with diethyl ether. The residue was mixed well with 1-2 ml water, and added 10 ml of methyl isobutyl ketone (MIBK) and 5 ml of concentrated hydrochloric acid. The tube was placed in an ultrasonic water bath for 10 min. Five ml of water was added to separate the MIBK layer from the aqueous layer. After shaking for 5 min, the mixture was centrifuged for 5 min at 3000 rpm. Heme iron in the MIBK layer was determined by spectrophotometer at a wavelength of 640 nm. The calibration curve was linear from 2.5 to 15 mu g Fe/ml. The recoveries of heme iron added to foods were within the range from 80.5% to 90.0%. An interlaboratory study of this method for the determination of heme iron in cookie, liver and meat was conducted in 3 laboratories. The coefficient of variation for each sample was within the range of 6.29-7.15%. This method is simple and useful for the determination of heme iron in foods.

High-speed counter-current chromatography (HSCCC) has been successfully applied to the separation of the ivermectin components. A 25-mg quantity of the sample was separated using a two-phase solvent system of n -hexane-ethyl acetate-methanol-water (19:1:10:10, v/v). The fractions were analyzed by high-performance liquid chromatography, nuclear magnetic resonance, and fast-atom bombardment mass spectrometry. The separation yielded 18.7 mg of 99.9% pure ivermectin B1a, 1.0 mg of 96.0% pure ivermectin B1b, and 0.3 mg of 98.0% pure avermectin Bla (precursor of ivermectin).

When the content of methanol in certain types of waterproofing aerosol products was determined according to the official method for household products containing harmful substances, the existence of an interfering substance in the products was found. The interfering substance was identified to be tert-butanol by GC-MS,In order to avoid this interference, we developed a simple headspace (HS)-gas chromatographic (GC) method. For this method, one gram of the sample sprayed into a 200 mi flask under ice cooling was weighed into a centrifugal tube. Each 10 mi of water and chloroform was added to the centrifugal tube. After shaking vigorously and:centrifuging (3500 rpm, 5 min), 0.5 g of the aqueous layer was weighed into a 24 mi bottle. The bottle was incubated for 30 minutes at 25 degrees C and then 50 mu l of the HS gas was injected into a GC with a fused silica capillary column coated with methyl phenyl cyanopropyl silicone.The proposed method was successfully applied to the screening analysis of methanol in 70 kinds of commercial household aerosol products.

Analysis of gardenia yellow in food products has been carried out by the detection of crocin and/or geniposide as indicator compounds. However, neither indicator compound was present in a few food products. As a new analytical strategy for gardenia yellow, we adopted crocetin, which is produced from colored components of gardenia yellow by alkaline hydrolysis, as an indicator compound. Crude extracts from food products were hydrolyzed in 0.1N NaOH for 30min at 50°C and the resultant hydrolyzates were analyzed by HPLC. Recoveries from jelly, candy and rice crackers through the whole analytical procedure were 85.6, 86.0 and 80.5%, respectively. When 20 food products labeled as containing gardenia yellow were analyzed, 11 of them had different chromatographic patterns from an authentic preparation and crocin was not detected at all in 5 samples. However, crocetin was found in every food product after alkaline hydrolysis, in the range of 0.1-13.4μg/g (as crocetin). Geniposide was found in only 3 food products.

Orange II (O-II), which is a non-permitted dye for use in food in Japan, was found in a pickled vegetable. It was difficult to identify O-II by usual analytical methods for food dyes such as paper and thin layer chromatographies, because there are three isomers in total, Orange I (O-I), Orange RN (O-RN), and O-II. In order to reliably identify the dye in a pickled vegetable, thin layer chromatography (TLC), thin layer chromatography/fast-atom bombardment mass spectrometry (TLC/FABMS) and high performance liquid chromatography with a photodiode-array detector (PDA-HPLC) were employed. The Rf value and TLC/FAB mass spectrum of the dye suggested that the dye was O-II or O-RN. The retention time and ultraviolet/visible spectrum of the dye coincided with those of standard O-II, but were different from those of O-RN. Therefore, we concluded that the dye in the pickled vegetable was O-II.

To detect trace amounts of arsenic, lead, cadmium and tin in soft drinks, a simple and rapid determination without sample pretreatment was developed. A sample of a soft drink was diluted with distilled water and examined by electrothermal atomic absorption spectrometry with addition of nickel nitrate as a matrix modifier. Detection limits of arsenic, lead, cadmium and tin in the samples were 0.025 ppm (as As2O3), 0.025 ppm, 0.005 ppm and 0.3 ppm, respectively.The developed method was applied to the determination of arsenic, lead, cadmium and tin in 13 kinds of soft drinks (total 150 samples). The amounts of arsenic and lead in the soft drinks were less than 0.025 ppm in all samples: cadmium and tin were detected in only 2 (0.006 similar to 0.019 ppm) and 5 (6.9 similar to 86 ppm) of 150 samples, respectively. The proposed method is simple and rapid for determination of arsenic, lead, cadmium and tin in soft drinks.

A technique for the analysis of coal tar dyes using TLC is described. The 12 coal tar dyes permitted for use in food in Japan were separated on an amino-modified silica gel plate using ethanol-acetonitrile-15% aqueous ammonia-10% aqueous sodium chloride solution (3:3:1:1) as the solvent system. To investigate the applicability of the present amino TLC for the analysis of coal tar dyes in food, 70 foods were analyzed and the chromatographic behavior of the dyes was investigated. The separation was not affected by coexisting substances from the food and the spots showed no tailing and always gave the same Rf Values as the standards with good reproducibility. These results indicated that the present amino TLC makes it easier to identify the dyes in food, and this should therefore be a useful technique for the analysis of coal tar dyes in food.