[Detection of allergenic substances (shrimp, crab) in processed seafood].

We have carried out a study (2009-2012) on processed seafood products in order to determine the level of contamination with shrimp and crab. In 2010-2012, after the Allergy Labeling Regulation went into effect, the detection rate of crustacean protein in processed seafood products including small fish, such as niboshi, tukudani and so on (both boiled and dried), was 63%. Detection rates for processed seafood products in which crustacean protein levels were below 1 µg/g were 36% with and 58% without advisory labels, allowing us to conclude that 60% of labels were adequate. On the other hand, the detection rate for processed seafood products with crustacean protein levels higher than the baseline of 10 µg/g was 9%, of which 60% carried no advisory labels. The rate of shrimp DNA detection using the Akiami primer in processed foods containing shrimp and crab was high (73%). This suggests that it is necessary to test these products using the Akiami primer for supplemental analyses of shrimp DNA. The PCR analysis for crab DNA detection failed due to combined detection of mantis shrimp DNA, which accounted for 8% of the total detected.

Changes in immune functions during a peaking period in male university soccer players.

The concept of peaking ensures that athletes have trained to attain their absolute peak performance levels prior to a competition. This study investigates the effects of peaking on the functions of neutrophils and lymphocytes in university soccer players during a five-day soccer training camp followed by two weeks of tapering. The study subjects were 22 soccer players who were members of a university soccer club. We carried out our investigation during a five-day training camp and two weeks after the training camp (i.e. the tapering period). We measured body composition, immune-related parameters (leukocyte count, neutrophil count, lymphocyte count, immunoglobulins and complements), myogenic enzymes, superoxide dismutase activity (SOD) and neutrophil functions [reactive oxygen species (ROS) production, phagocytic activity, serum opsonic activity and lymphocyte subtypes]. Leukocyte and neutrophil counts tend to increase after the training camp compared with values before the training camp, and recovered during the conditioning period, although the final values were still lower than those before the training camp. The amount of ROS production per neutrophil and level of SOD decreased significantly during the conditioning period compared with before the training camp (p < 0.05, p < 0.01, respectively). Levels of Th1 cells decreased significantly during the conditioning period compared with the training camp (p < 0.05). In conclusion, the period of two weeks for the tapering period was considered insufficient to allow the athletes' immune function to recover completely after an intensive training camp.

[In vivo screening of illegal drug].

The neuro-behavioral observation scorebook that improved the previous observation methods of Irwin was followed, the test material was administered to 5 mice per each group, and the mean value of the obtained score was determined. The behavior of a normal animal was assumed to be point 0, animals showing suppressive behavior were scored in the minus region, and animals that showed excitement behavior were scored in the plus region. Each score was divided into three stages, according to the level of strength of the biological effect. The score of each observation item was totaled, and the level of the strength of the biological effect in the item was judged according to its mean value. These test methods of neuro-behavioral observations we proposed were able to detect the biological effects of a drug simply and promptly, and contributed sufficient data to support an administrative measure aimed at anticipating and improving the prevention of health damage in humans by non-regulated drugs from a scientific perspective. Recently, we developed a method of serial measurement of the quantity of monoamine in the mouse central nervous system by microdialysis, and performed it. The Kanagawa Prefectural Institute of Public Health conducted a study of the biological effect of non-regulated drugs. A characteristic here is what they examined about drug-dependency other than observing the behavior of the animal.

Facilitative effects of Eucommia ulmoides on fatty acid oxidation in hypertriglyceridaemic rats.

BACKGROUND: Tea made from Eucommia ulmoides leaves is widely consumed as a health food, since recent studies have revealed various pharmacological effects of the tea, e.g. a hypotriglyceridaemic effect. This study was conducted to elucidate the mechanisms underlying the plasma triglyceride-lowering effect of E. ulmoides leaves. RESULTS: Rats were divided into four groups: a normal group, a group fed a high-fat/high-fructose diet (untreated group) and two groups fed a high-fat/high-fructose diet and E. ulmoides tea (4 or 20 g L(-1) extract, treated groups). Plasma triglyceride concentrations were reduced in treated groups in a dose-dependent manner compared with the untreated group. DNA microarray analysis revealed that genes involved in hepatic α-, ß- and ω-oxidation, mainly related to the peroxisome proliferator-activated receptor α and δ signalling pathway, were up-regulated in the treated group. High-performance liquid chromatography analysis showed that E. ulmoides leaves contain three phytochemicals that make up 60 mg g(-1) of the material and are likely to be the active components. CONCLUSION: This study indicates that the promotion of fatty acid oxidation, probably by the action of phytochemicals, participates in the ameliorative effect of E. ulmoides leaves on hypertriglyceridaemia. These findings provide the scientific evidence for the functionality of E. ulmoides.

Ameliorative effects of mulberry (Morus alba L.) leaves on hyperlipidemia in rats fed a high-fat diet: induction of fatty acid oxidation, inhibition of lipogenesis, and suppression of oxidative stress.

To determine the effects of mulberry (Morus alba L.) leaves on hyperlipidemia, we performed gene expression profiling of the liver. Rats were fed a high-fat diet and administered mulberry leaves for 7 weeks. Plasma triglyceride and non-esterified fatty acid levels were significantly lower in the rats treated with mulberry leaves as compared with the untreated rats. DNA microarray analysis revealed that mulberry leaves upregulated expression of the genes involved in α-, ß- and ω-oxidation of fatty acids, mainly related to the peroxisome proliferator-activated receptor signaling pathway, and downregulated the genes involved in lipogenesis. Furthermore, treatment with mulberry leaves upregulated expression of the genes involved in the response to oxidative stress. These results indicate that consumption of fatty acids and inhibition of lipogenesis are responsible for the reduction in plasma lipids caused by mulberry administration. In addition, mulberry treatment maintains the body's oxidative state at a low level despite enhancing fatty acid oxidation.

Effects of 2.5-hour sumo training on serum opsonic activity.

Sumo is a traditional Japanese sport, but the effect of actual daily training on neutrophil function is unknown. We evaluated the effect of sumo training on serum opsonic activity (SOA), which is one of the main neutrophil-related functions. Seventeen male university sumo wrestlers participated in the study. Changes in anthropometric parameters, concentrations of serum immunoglobulins (IgG, IgA and IgM), complements (C3 and C4), myogenic enzymes (lactate dehydrogenase, asparate aminotransferase, alanine aminotransferase and creatine kinase), white blood cell/neutrophil counts and SOA were measured immediately before and after actual daily training for 2.5 h. Compared with the pre-values, immunoglobulins and complements, myogenic enzymes and white blood cell/neutrophil counts significantly increased (p < 0.01 for all). As for SOA, the values of the peak height and the area under the curve significantly increased after the training when assessed using lucigenin as a chemiluminigenic probe (p < 0.01 for all), but showed no significant change when luminol was used as the chemiluminigenic probe. In conclusion, daily actual sumo training for 2.5 h increases SOA, thus possibly activating the reactive oxygen species production of neutrophils.

[Study on processed senna found in health teas].

Sennoside A and B were detected in 21 commercial health tea products surveyed in 2000-2007, but there were 8 products in which the leaves could not be identified as senna because the leaves had become discolored. The results of assay of sennoside levels and TLC chromatograms suggested that processed senna had been used in these products. Next, with reference to tea and health tea manufacturing methods, pharmaceutical senna was roasted or wet-processed experimentally. The results indicated that the discolored leaves contained in commercial health tea were most likely derived from senna leaves. Moreover, sennosides in medicinal doses were detected in some processed senna samples, and were determined to have a cathartic action in mice. Based on morphological confirmation and the results of component analysis, including sennoside, the discolored leaves found in commercial health teas were therefore determined to be senna leaves. There may be possible health risks, including diarrhea.

[Analysis of the chemical drugs among structural isomer].

Drugs that have a pharmacological effect similar to legal drugs such as narcotics and stimulants are available in the market and widely used. 5-methoxy-N,N-di-iso-propyl-tryptamine (5MeO-DIPT) and alpha-methyl-tryptamine (AMT) were categorized as narcotics and were specified as legal drugs in April 2005, and also 2,5-dimethoxy-4-n-propylthiophenethylamine (4C-T-7) and N-methyl-alpha-ethyl-3,4-methylenedioxy-phenethylamine (MBDB) were categorized as narcotics and were specified as legal drugs in April 2006, in Japan. We are analyzing these chemical drugs by investigating the market research. It is recognized that during the analysis of chemical drugs, drugs that resemble a structural isomer of a target substance, such as 5MeO-DIPT and 5-methoxy-N,N-di-n-propyl-tryptamine (5MeO-DPT) or 4C-T-7 and 2,5-dimethoxy-4-iso-propylthiophenethylamine (4C-T-4), should be distinguished. The results of TLC, IR, GC-MS and HPLC analyses were compared. 5MeO-DIPT and 5MeO-DPT could be distinguished by TLC and HPLC analyses, but not by IR and GC-MS analysis. The drugs 4-hydroxy-N-methyl-N-iso-propyl-tryptamine (4HO-MIPT) and 4-hydroxy-N,N-di-ethyl-tryptamine (4HO-DET) or could not be distinguished. Moreover, the isomers of 4-hydroxy-N-methyl-N-n-propyl-tryptamine (4HO-MPT) was not found to be present. Thus, we have demonstrated that the chemical drug could be distinguished from each other, and we have also shown that NMR data is essential for the analysis.

Inhibitory effects of mulberry leaf extract on postprandial hyperglycemia in normal rats.

We examined the inhibitory effects of aqueous ethanol extract from mulberry leaves (ME) on postprandial hyperglycemia in normal Wistar rats. ME dose-dependently suppressed the postprandial rise of blood glucose in rats, when ME (0.02-0.5 g/kg) was given 0.5 h before the administration of carbohydrates such as sucrose, maltose and starch. The ME dose showing 50% inhibition of the increment of blood glucose (ED50) was 0.11 g/kg for sucrose, 0.44 g/kg for maltose, and 0.38 g/kg for starch. ME and its basic fraction (MB) containing 1-deoxynojirimycin were assayed for their inhibitory effects (IC50) on disaccharidase derived from the small intestine of rats. The IC50 value of ME was 3.2 microg/mL for sucrase, 10 microg/mL for isomaltase, and 51 microg/mL for maltase. The IC50 value of MB was 0.36 microg/mL for sucrase, 1.1 microg/mL for isomaltase, and 6.2 microg/mL for maltase. The IC50 value of 1-deoxynojirimycin as the principle component in ME was 0.015 microg/mL for sucrase and 0.21 microg/mL for maltase, and this value was comparable to the IC50 of voglibose. The inhibitory activity of ME in a-amylase was weak. These results suggest that ME strongly suppresses postprandial hyperglycemia after carbohydrate loading by inhibiting the activity of disaccharidases in the small intestine of rats.

[Ninety-day dietary toxicity study of mulberry leaf extract in rats].

Mulberry leaf extract was studied toxicologically in male and female SD rats. The extract was administered orally at concentrations of 0% (control group), 0.1%, 0.4% and 1% in basal diet for 90 days. No remarkable change in test animals of both sexes was observed in terms of body weight gain or at necropsy. Hematology and blood chemistry revealed no abnormalities. Pathological examination revealed no toxic change in any organ observed. These findings indicate that dietary intake of 1% mulberry leaf extract for 90 days (884.5 mg/kg/day for males and 995.7 mg/kg/day for females as mean daily intake) causes no toxicological change in rats.