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Background and purpose: Ulcerative colitis is a chronic inflammatory bowel disease that involves diffused inflammation of the large intestine. Omega-3 fatty acid (FA) has been known to regulate the inflammatory response associated with ulcerative colitis pathogenesis. Perilla frutescens is a valuable source of omega-3 FA and α-linolenic acid (ALA) contained in its seed oil. Therefore, the aim of this study was to evaluate the anti-inflammatory effect of Perilla seed oil (PSO) on colitis induced by dextran sulfate sodium (DSS) in a mouse model. Experimental approach: PSO was extracted using a cold-pressed extractor and FA composition of PSO was analyzed by GC-MS. Acute colitis in mice was induced with 3% DSS in drinking water for 7 days. Some mice were treated with PSO (20, 100, 200 mg/kg BW) for 3 weeks before the DSS administration. Sulfasalazine was used as a positive control. The clinical features, histopathologic, serum, and gene expression of proinflammatory cytokines in the colon were assessed. Finding/Results: PSO contained the highest proportion of ALA (61.51%). Furthermore, PSO pretreatment evidently reduced body weight loss, diminished diarrhea, gross bleeding, and DSS-induced colon shortening. PSO pretreatment attenuated histopathological changes in response to DSS-induced colitis. PSO pretreatment also markedly decreased inflammatory response in serum and the colon tissue of DSS-induced mice. Conclusion and implication: ALA in PSO is suggested to be mainly responsible for the reduction of DSS-induced colitis through suppressing inflammatory markers. PSO could be further developed as a functional health supplement, which would be beneficial for anti-inflammation in the colonic mucosa.

Industrially, after the removal of oil from perilla seeds (PS) by screw-type compression, the large quantities of residual perilla seed meal (PSM) becomes non-valuable waste. Therefore, to increase the health value and price of PS and PSM, we focused on the biological effects of perilla seed oil (PSO) and rosmarinic acid-rich fraction (RA-RF) extracted from PSM for their role in preventing oxidative stress and inflammation caused by TNF-α exposure in an A549 lung adenocarcinoma culture model. The A549 cells were pretreated with PSO or RA-RF and followed by TNF-α treatment. We found that PSO and RA-RF were not toxic to TNF-α-induced A549 cells. Both extracts significantly decreased the generation of reactive oxygen species (ROS) in this cell line. The mRNA expression levels of IL-1β, IL-6, IL-8, TNF-α, and COX-2 were significantly decreased by the treatment of PSO and RA-RF. The Western blot indicated that the expression of MnSOD, FOXO1, and NF-κB and phosphorylation of JNK were also significantly diminished by PSO and RA-RF treatment. The results demonstrated that PSO and RA-RF act as antioxidants to scavenge TNF-α induced ROS levels, resulting in decreased the expression of MnSOD, FOXO1, NF-κB and JNK signaling pathway in a human lung cell culture exposed to TNF-α.

Thai perilla (Perilla frutescens) extracts, which contain a substantial quantity of bioactive substances including phenolics and flavonoids, have shown marked anti-inflammatory activities in several investigated models. In the present study, the effect of perilla seed extract (PSE) and seed meal extract (PSME) on TNF-α-induced inflammatory response in human lung adenocarcinoma A549 cells was investigated. The total phenolic and flavonoid contents in PSME was lower than PSE. Markedly, rosmarinic acid was identified as the main constituent in both extracts. However, the DPPH and ABTS assays indicated that the antioxidant capacity of PSME was equal to PSE. Moreover, the iron-binding activity of PSE and PSME were exhibited by complex formation with Fe3+-NTA, indicating that the extracts may inhibit hydroxyl radical production via Fenton reaction. In vitro cytotoxicity analysis showed that both PSE or PSME co-treated with TNF-α, at 24 h exposure, were not toxic to the A549 cells. Interestingly, PSE and PSME dramatically exhibited an anti-inflammatory activity by inhibiting the mRNA expression of pro-inflammatory cytokines, IL-1β, IL-6, IL-8, and TNF-α, but did not influence iNOS and COX-2 mRNA expressions. Moreover, both extracts significantly reduced reactive oxygen species (ROS) production in TNF-α-induced A549 cells. The findings presented in this paper suggest that PSE and PSME could mitigate TNF-α-mediated inflammatory responses via limiting pro-inflammatory cytokine expressions and decreasing ROS production. Thus, perilla seed and seed meal, the by-product of a perilla seed oil cold-pressed extraction process, could be developed as food supplements or functional foods for the prevention of inflammation-induced lung carcinogenesis development. Keywords: Human lung adenocarcinoma cell line, Inflammation, Perilla seed, Perilla seed meal, Tumor necrosis factor-alpha (TNF-α)

Particulate matter from forest fires (PMFF) is an environmental pollutant causing oxidative stress, inflammation, and cancer cell metastasis due to the presence of polycyclic aromatic hydrocarbons (PAHs). Perilla seed meal contains high levels of polyphenols, including rosmarinic acid (RA). The aim of this study is to determine the anti-oxidative stress, anti-inflammation, and anti-metastasis actions of rosmarinic acid rich fraction (RA-RF) from perilla seed meal and its underlying molecular mechanisms in A549 cells exposed to PMFF. PMFF samples were collected via the air sampler at the University of Phayao, Thailand, and their PAH content were analyzed using GC-MS. Fifteen PAH compounds were detected in PMFF. The PMFF significantly induced intracellular reactive oxygen species (ROS) production, the mRNA expression of pro-inflammatory cytokines, MMP-9 activity, invasion, migration, the overexpression of c-Jun and p-65-NF-κB, and Akt phosphorylation. Additionally, the RA-RF significantly reduced ROS production, IL-6, IL-8, TNF-α, and COX-2. RA-RF could also suppress MMP-9 activity, migration, invasion, and the phosphorylation activity of c-Jun, p-65-NF-κB, and Akt. Our findings revealed that RA-RF has antioxidant, anti-inflammatory, and anti-metastasis properties via c-Jun, p-65-NF-κB, and Akt signaling pathways. RA-RF may be further developed as an inhalation agent for the prevention of lung inflammation and cancer metastasis induced by PM exposure.

Perilla is a rich source of polyphenols, which exhibits antioxidant, anti-inflammatory activities, and a variety of biological effects. The effect of differential solvents on the polyphenols, flavonoids, rosmarinic acid (RA), antiinflammatory and antioxidant activities of perilla leaf require investigation. In this study, perilla leaf was extracted with 70% ethanol and sequentially fractionated according to the solvent’s polarity with hexane, dichloromethane, ethyl acetate, and water. Samples were subjected to the bioactive compound measurements. The antioxidant and antiinflammation nature of perilla was analyzed based on the scavenging effects on DPPH•, ABTS•+, O2•- and nitric oxide (NO), as well as FRAP assay, and determination of the inhibition effects on NO, inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2) production in the cell-based study. The results indicate that among all different solvents used for sequential fractionation, ethyl acetate (EtOAc) was most effective in the separation of anti-oxidative and antiinflammatory compounds in the perilla leaf extract. These properties can partly be due to the presence of polyphenols, flavonoids, and also RA. It can be demonstrated here that, the perilla leaf EtOAc fraction could be used as a natural active pharmaceutical ingredient for dietary supplements and nutraceuticals.

The aim of this study is to determine antioxidant and anti-inflammatory activities relating to the antiosteoporosis effects of various perilla seed meal (PSM) fractions. The remaining waste of perilla seed obtained from cold oil compression was extracted with 70% ethanol and sequentially fractionated according to solvent polarity with hexane, dichloromethane, ethyl acetate, and water. The results indicated that the seed-meal ethyl acetate fraction (SMEF) exhibited the highest antioxidant and anti-inflammatory activities, and rosmarinic acid (RA) content. The signaling pathways induced by the receptor activator of the nuclear factor kappa B (NF-κB) ligand (RANKL) that trigger reactive oxygen species (ROS) and several transcription factors, leading to the induction of osteoclastogenesis, were also investigated. The SMEF clearly showed attenuated RANKL-induced tartrate-resistant acid phosphatase (TRAP)-positive multinucleated osteoclasts and TRAP activity. A Western blot analysis showed that the SMEF significantly downregulated RANKL-induced NF-κB, AP-1 activation, and the nuclear factor of activated T-cell 1 (NFATc1) expression. SMEF also suppressed RANKL-induced osteoclast-specific marker gene-like MMP-9 using zymography. Furthermore, the SMEF showed inhibition of RANKL-induced ROS production in RAW 264.7 cells. The results suggest that the SMEF, which contained high quantities of RA, could be developed as a natural active pharmaceutical ingredient for osteoclastogenic protection and health promotion.

Gastric ulcers are a common problem in upper gastrointestinal tract (GI) disorders. Nonsteroidal anti-inflammatory drugs (NSAIDs) are one of the most aggressive factors leading to inducing gastric ulcers. Natural products with lower toxicity and safety are currently sought as a potential source to minimize the effect of the gastric ulcers.Perilla frutescensor Nga-mon (in Thai) leaves are rich in rosmarinic acid (RA), which has antioxidant, anti-inflammatory, and anticancer effects. This study investigates the protective effect of ethanolic extract (EE) and aqueous fraction (AF) fromPerilla frutescensleaves, which are rich in RA, on indomethacin- (IND-) induced gastric ulcer in a rat model. The EE at the doses of 50 and 500 mg/kg body weight, AF at the doses of 50, 250, and 500 mg/kg body weight, or famotidine (a standard drug) were administered for 14 days prior to ulcer induction. The ulceration was performed by intragastric administration of IND. Gross gastric ulcers and biological and histological parameters were examined. The pretreatment with AF had more significant effects than EE, including reduced ulcer index, decreased gastric secretion volume and decreased acidity, but it had an elevated gastric pH relative to the IND-induced gastric ulcer. In a histopathological study, the EE and AF decreased mucosal ulcer, inflammatory infiltration, and degenerative lining cells. The IND-induced expression of inflammatory mediators was significantly attenuated with EE and AF. The experiment also remarkably showed the preservation of mucus and apoptosis protection of EE and AF on a gastric mucosal ulcer. The findings demonstrated that the EE and AF of perilla leaves were capable of protecting the stomach against gastric ulcers induced by IND through anti-inflammatory and antiapoptotic mechanisms that should be further investigated. It is suggested thatPerilla frutescensleaf could be a potential alternative source of RA as a therapeutic agent and food supplement for NSAID-induced gastric injuries.

Perilla frutescens is commonly used as an n-3-fatty acid source for people living in Northern Thailand. However, cultivated areas are limited because it apparently requires specific growth conditions. Our results suggested that perilla seeds grown in higher altitude in Maehongson province contained more ALA contents than seeds collected from Chiangrai and Nan province. Furthermore, mass spectrometry (MS) proteomic and gene expression analysis revealed that the increase of KASII and FAD3 proteins associated with the mRNA expression and the ALA production. Predictive bioinformatics analysis demonstrated two important transcription factor binding sites, AP2 and B3, which responsible for KASII and FAD3 genes, respectively. The AP2 and B3 transcription factor families were known to be responsible for abiotic stress such as drought and temperature changes. In conclusion, high production of ALA by perilla apparently relies on the altitudes. Moreover, plant responses to abiotic stress environments contribute to the increase of KASII and FAD3 gene and protein expression. However, responsive transcription factors will be further studied for a proof of concept and additionally improving the growing methods in order to increase perilla productivity.

The ethanolic extracts of leaves of Psidium guajava (PG), Pandanous odorus (PO),Rhinacanthus nasutus (RN) contained polyphenolic compounds in the order of PG > RN > PO. All the extracts exhibited 2,2-diphenyl-1-picryhydrazyl (DPPH) and 2,2’-azino-bis 3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radicals scavenging activities. PG showed a marked antioxidant effect whereas PO and RN were comparable. Interestingly, they were able to bind with ferric (Fe3+) and ferrous (Fe2+) ion and completed within 5 min of incubation times. In accordance with previous results, PG extract could effectively protect erythrocytes from iron induced hemolysis with the activity higher than PO and RN. Based on all results, tested leaf extracts of three Thai medicinal plants might be used for alleviating or preventing harmfulness in many oxidative stress disorders. Key words: Psidium guajava, Pandanous odorus, Rhinacanthus nasutus, antioxidant, iron binding capacity, anti-hemolysis activity.

There has been increasing interest in finding natural antioxidants to prevent free radical damage and retard the progress of chronic inflammatory diseases. Our previous data demonstrated the strong antioxidant properties of polyphenolics in Mamao seed (MS) and Mamao marc (MM) extracts. In this study we further investigated the effect of MS and MM polyphenolics on hydrogen peroxide (H 2 O 2 )-induced apoptosis and tumour promoter 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced inflammation, using human breast epithelial (MCF10A) cells. MS and MM extracts conferred dose-dependent protection against H 2 O 2 -induced apoptosis by inhibiting PARP/caspase-3 cleavage, inducing anti-apoptotic Bcl-2 expression, and down-regulating pro-apoptotic Bax. Moreover, MS and MM polyphenolics inhibited TPA-induced COX-2 and NF-κB activation by blocking the degradation of cytoplasmic IκBα, as well as subsequent nuclear translocation of p65 and attenuation of the activation of ERK, but not JNK and p38. These data establish the molecular mechanism for the anti-apoptotic and anti-inflammatory effects of MS and MM polyphenolics.

Purslane weed has long been used for animal feed in Thailand but has been underutilized, and no published data exist on the chemical and nutritional qualities of Thai wild purslane. In this study, we aimed to determine the microchemical components of purslane stem, leaf, and flower, including the phenolic acid, flavonoid, ascorbic acid, β-carotene, and fatty acid content. The water extract of the flower fraction contained the highest total phenolic acid content and had the highest O2-scavenging activities, whereas leaf contained the highest amount of total flavonoids and ascorbic acid. The β-carotene content was not significantly different in leaf and flower fractions but was significantly greater than that in stem (P < 0.05). The predominant phenolic acid was chlorogenic acid for all fractions. Rutin was the major flavonoid found in leaf, and myricetin was highest in flower and stem. Alpha linolenic acid (18:3n-3) content ranged from 16% (149 mg per 100-g sample) of total fatty acid in stem to 50% (523 mg per 100-g sample) in leaf. We suggest that Thai wild purslane could be considered a nutritional source for animal feed or an excellent vegetable in the human diet.

The objective of this study was to investigate the in vitro bioactivities associated with the content of phytochemicals in the extracts from perilla seed meal extract (PSME) compared with dietary seed (PSE). PSE had higher total phenolics and flavonoids content than PSME. However, hydrophilic phytochemical contents in PSME were quantitatively equivalent to PSE. Rosmarinic acid was predominantly found in both extracts. Cell viability and anti‐mutagenicity testing demonstrated that PSE and PSME were biosecured and non‐genotoxic. Both extracts strongly scavenged free radicals and significantly reduced reactive oxygen species (ROS) production. The extracts drastically diminished nitric oxide (NO) production of LPS‐treated RAW 264.7 cells via iNOS mRNA expression. The expression of IL‐6 and COX‐2 were evidently inhibited by these extracts. It could be concluded that PSE and PSME clearly showed in vitro anti‐mutagenicity, antioxidant and anti‐inflammatory capacities. In particular, the by‐product perilla seed meal could be considered as a high nutritive functional food. PRACTICAL APPLICATIONS: This study suggests that the seed meals, a by‐product from seed oil industry, can be utilized as a valuable dietary source for humans and animals. The high content of polyphenols and their bioactivities can be developed as functional foods, and excipients and fillers in pharmaceuticals and nutraceuticals production. Moreover, recycling of the by‐product seed meals should also reduce environmental and sanitary pollution.

Ethanolic extracts of 30 Thai medicinal plants, traditionally used as alternative treatments in diabetes, were evaluated for antioxidative activity by the 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) method. They were evaluated in vitro for oxidative stress by thiobarbituric acid-reactive substance (TBARS) assay in pooled plasma of diabetic patients compared to without treatment of the extracts (control). The extracts were also assayed for protein glycation. The results showed that five plants had strong antioxidant activity: Phyllanthus emblica Linn. (PE), Terminalia chebula Retz. (TC), Morinda citrifolia Linn. (MC), Kaempferia parviflora Wall. (KP) and Houttuynia cordata Thunb.(HC), respectively. Thirty plant extracts were good correlation between total antioxidant activity and antiradical activity by TBARS as well as by glycation (r = 0.856, p

Ethanolic extracts from various parts of 26 Thai indigenous plants were examined for phenolic constituents and free radical scavenging capacity, to determine their potential as a source of natural antioxidants. Total phenolic content and total flavonoid content were evaluated according to the Folin-Ciocalteu procedure, and a colorimetric method, respectively. The results showed that total phenolic compounds and flavonoid content were higher in seed extracts of berries used in wine production, while the levels in extracts obtained from herbs and vegetables were lower. Chewing plants which have an astringent taste gave a significantly higher total phenolic content and flavonoid content. Antiradical activity determined from 1/EC50 by the DPPH radical-scavenging method was highest in wine production seeds and chewing plants. The correlation coefficient from regression analysis showed a positive relationship between total phenolic and total flavonoid content (r = 0.9). The results suggest that ethanolic extracts of some Thai indigenous plants exhibit a potential for use as natural antioxidants.

Effects of extracts of a plant, which has been used as a traditional medicine for treating diabetes on glucose transport activity was evaluated in cultured L8 muscle cells. The aqueous extract of Canna indica root (CI) at doses of 0.1-0.5 mg/ml, which contains total phenolic compounds equivalent to 6-30 microg of catechin caused a dose- and time-dependent induction of 2-deoxy-[3H]glucose (2-DG) uptake activity. The induced 2-DG uptake was significantly increased within 8 h and reached a maximum by 16 h. The CI extract increased the amount of glucose transporter isoforms 1 (GLUT1) and 4 (GLUT4) at the cell surface and enhanced expression of GLUT1 protein. Cycloheximide treatment almost completely reversed CI-induced 2-DG uptake to the basal level. Exposure of muscle cells to wortmannin and SB203580 diminished CI-mediated glucose uptake by 38 and 14%, respectively. The effect of CI and insulin was partially additive. Phytochemical analysis detected the presence of flavonoids and catechol in the CI. Taken together, these data provide evidence for differential effects of CI on regulated-glucose transport in muscle cells. Our findings suggest that GLUT1 protein synthesis and the activation of phosphatidylinositol 3-kinase (PI3K) are critical for the increase in glucose transporter activity at the plasma membrane and essential for the maximal induction of glucose transport by CI in L8 muscle cells.

The recent case-control studies in Thailand indicate that a high incidence of liver cancer in Thailand has not been associated with common risk factors such as hepatitis B infection, aflatoxin intake and alcohol consumption. While the infestation by the liver fluke Opisthorchis viverrini (OV) accounted for the high risk in north-east Thailand, there was no such exposure in the other regions of the country where the incidence of liver cancer is also high. Case-control studies suggest that exposure to exogenous and possibly endogenous nitrosamines in food or tobacco in betel nut and cigarettes may play a role in the development of hepatocellular carcinoma (HCC), while OV infestation and chemical interaction of nitrosamines may also be aetiological factors in the development of cholangiocarcinoma (CCA). Over 1800 samples of fresh and preserved food were systematically collected and tested between 1988 and 1996. All the food items identified by anthropological studies to be consumed frequently in four major regions of Thailand were analysed for volatile nitrosamines using gas chromatography combined with a thermal energy analyser. Relatively high levels of N -nitrosodimethylamine (NDMA), N -nitrosopiperidine (NPIP) and N -nitrosopyrrolidine (NPYR) were detected in fermented fish (“Pla-salid”). NDMA was also detected at levels ranging from trace amounts to 66.5 μ g/kg in several salted and dried fish (“Larb-pla” and “Pla-siu”). NDMA and NPYR were frequently detected in several vegetables, particularly fermented beans (“Tau-chiau”) at levels ranging between 1 and 95.1 μ g/kg and 0–146 μ g/kg, respectively. The possible role of nitrosamines in Thai food in the aetiology of liver cancer (HCC, CCA) is discussed.

This study investigated the antioxidant and anti-inflammatory activities of g-oryzanol rich extracts from Thai glutinous purple rice bran. Five purple rice bran samples, PAH E-KAW,GAM PEUAK, NIAW DAM, GAM THOR and GAM BOUNG were obtained from local glutinous cultivars in Northern Thailand. The g-oryzanol rich extracts were prepared by soxhlet extraction and semi-purified by column chromatography. The extracts contained g-oryzanol in the range of 1.23 to 9.14% w/w. The extracts containing more than 5% w/w of g-oryzanol (PAH E-KAW, GAM THOR and GAM BOUNG) were selected for investigation of antioxidant and anti-inflammatory activities; and first standardized by adding the equivalent amount of each extract to give 10% w/v g-oryzanol. All of extracts exerted moderate ABTS·+ and superoxide anion scavenging activity. GAM BOUNG exhibited the highest ABTS·+ scavenging activity with VCEAC value of 201.3 ± 1.84 mg L-ascorbic acid/ gram extract, TEAC value of 267.2 ±2.96 mg Trolox/ gram extract and QEAC value of 137.4 ± 3.41 mg Quercetin/ gram extract followed by PAH E-KAW and GAM THOR, respectively. GAM BOUNG also showed the highest activity on superoxide anion scavenging at IC50 = 12.56 ± 0.13 mg/ml followed by GAM THOR and PAH E-KAW. Furthermore, GAM BOUNG exhibited a strong inhibition effect on linoleic acid peroxidation (IC50 = 36.79 ± 2.97 mg/ml). Additionally, all of the extracts showed a strong anti-inflammatory activity through inhibitory effect on nitric oxide (NO) production in combined LPS-IFN-g-activated RAW 264.7 murine macrophage cells. GAM BOUNG exhibited the highest inhibitory effects on NO production (IC50 = 29.32 ± 2.21 mg/ml), without exerting cytotoxicity followed by GAM THOR and PAH E-KAW. These results indicate that g-oryzanol rich extracts from Thai glutinous purple rice bran are acting as a lipophilic radical scavenger better than a hydrophilic radical scavenger. Moreover, these extracts exert a potent anti-inflammatory activity. Key words: Purple rice bran, g-oryzanol, antioxidation, anti-inflammatory.

The purpose of this study was to determine the effect of polyphenols extracted from the tamarind seed coat (PETSC) on glutathione peroxidase (GPx) activity, red blood cell parameters and bilirubin in heat-stressed broilers. One hundred forty-seven broilers, 18-days old were divided into two groups. In group 1, broilers were maintained at an environmental temperature of 26 ± 2 °C throughout the experimental period. In group 2, the broilers were maintained at 38 ± 2 °C (cyclic temperature: 26 ± 2 °C; −38 ± 2 °C; and −26 ± 2 °C, and broilers were maintained at 38 ± 2 °C for 6 h/ day) and received PETSC at a concentration of 0, 100, 200, 300, 400 or 500 mg/kg in their diet ad libitum. Parameters were investigated on days 1, 7, 14 and 21 of the experimental period. Results showed that GPx activity of heat-stressed broilers that received 100 mg/kg of PETSC in their diet was lower (P

Ethnopharmacological relevance Many Thai traditional herbs have been used for hundreds of years for hair treatment and nourishment, including hair loss. However, scientific evidence about their mechanisms of action has not yet been elucidated. Aims of the study The purpose of this research is to define the possible mechanisms involved in hair loss treatment of the selected plants by determining the 5α-reductase enzyme inhibition and hair growth promoting activities, and the relationship between these two activities. Materials and methods Seventeen Thai plants traditionally used for hair treatment were selected. The plants were dried, ground and extracted by maceration with ethyl alcohol. These extracts were further tested for 5α-reductase inhibition using enzymes from rat livers. Hair growth promoting activity was tested in C57BL/6 mice. Results Carthamus tinctorius L. was the most potent 5α-reductase inhibitor, with a finasteride equivalent 5α-reductase inhibitory activity (FEA) value of 24.30 ± 1.64 mg finasteride equivalent per 1 g crude extract. Phyllanthus emblica L. was the second most potent inhibitor, with FEA of 18.99 ± 0.40. Rhinacanthus nasutus (L.) Kurz. was the least potent 5α-reductase inhibitor (FEA 10.69 ± 0.96). Carthamus tinctorius also was the most potent hair growth promoter in C57BL/6 mice. There were strong relationships between 5α-reductase inhibitory activity and hair growth promoting activity ( r = 0.719), and between 5α-reductase inhibitory activity and hair follicle count ( r = 0.766). Conclusions Ethanolic extract of Carthamus tinctorius was the most potent 5α-reductase inhibitor and hair growth promoter. This discovery may lead to the development of new alternative medicines for hair loss prevention and treatment.

We investigated the effects of antioxidant activity of fermentation product (FP) of five Thai indigenous products on oxidative stress in Wistar rats with streptozotocin (STZ)-induced diabetes type II. The rats were fed with placebo and with the FP (2 and 6 mL/kg body weight/day) for 6 weeks. Rutin, pyrogallol and gallic acid were main compounds found in the FP. Plasma glucose levels in diabetic rats receiving the higher dose of the FP increased less when compared to the diabetic control group as well as the group receiving the lower FP dose (13.1%, 29%, and 21.1%), respectively. A significant dose-dependent decrease in plasma levels of thiobarbituric acid reactive substance (P<.05) was observed. In addition, the doses of 2 and 6 mL FP/kg/day decreased the levels of erythrocyte ROS in diabetic rats during the experiment, but no difference was observed when compared to the untreated diabetic rat group. Results imply that FP decreased the diabetes-associated oxidative stress to a large extent through the inhibition of lipid peroxidation. The FP also improved the abnormal glucose metabolism slightly but the difference was not statistically significant. Thus, FP may be a potential therapeutic agent by reducing injury caused by oxidative stress associated with diabetes.

The aim of this experiment was to determine the effect of polyphenols extracted from tamarind (Tamarindus indica L.) seed coat on physiological changes, oxidative stress and production of male broilers maintained at high environmental temperatures. The results found that body temperature and respiratory rate of broilers maintained at 38 +/- 2 degrees C was higher than broilers maintained at 26 +/- 2 degrees C (P < 0.05). On day 1, the heterophil/ lymphocyte ratio of broilers maintained at 38 +/- 2 degrees C and received polyphenols at 300 and 400 mg/kg in diets was lower than broilers that received polyphenols at 0 and 200 mg/kg in diets (P < 0.05). At week 1, the malondialdehyde of the broilers maintained at 38 +/- 2 degrees C who received polyphenols at 400 mg/ kg in their diet was lower than broilers that received polyphenols at 100 and 200 mg/kg in diets (P < 0.05). At week 1, the body weights of broilers that were maintained at 38 +/- 2 degrees C who received polyphenols at 100-500 mg/ kg in diets, and broilers maintained at 26 +/- 2 degrees C were higher than that of the control group which had not been treated with a polyphenol diet (P < 0.05). This study indicated that polyphenols could reduce heat stress, oxidative stress and improve the growth rate of heat-stressed broilers.

It is our working hypothesis that the high rate of the liver and gastric cancers in North and Northeast Thailand is associated with increased daily dietary intake of nitrate, nitrite, and nitrosodimethylamine (NDMA). Samples of fresh and preserved Thai foods were systematically collected and analyzed from 1988 to 1996 and from 1998 to 2005. Consumption frequencies of various food items were determined on the basis of a dietary questionnaire given to 467 adults (212 males and 255 females) from 1998 to 2005. Food consumption data for the preceding and current year were collected and intakes (day, week, and month) of nitrate, nitrite, and NDMA were calculated. The trends in liver and stomach cancer age-standardized incidence rates (ASR) in four regions of Thailand were compared with the dietary intake of nitrate, nitrite, and NDMA in those same geographic regions. Mean daily intakes of nitrate of 155.7 mg/kg, of nitrite of 7.1 mg/kg, and of NDMA of 1.08 microg/kg per day were found. Significant differences in dietary nitrate, nitrite, and NDMA intakes were seen between various Thai regions (P < 0.0001), and these corresponded to the variations in liver and stomach cancer ASR values between the regions. Dietary factors are likely to play key roles in different stages of liver and stomach carcinogenesis in Thailand.

To overcome the problem of aflatoxins (AF) in Thai foods, a sensitive in-house direct cELISA using monoclonal antibody (mAb) was established and compared with a commercial ELISA kit and thin-layer chromatography for the determination of AFB(1) levels in corn and peanuts. Among eight in-house mAbs (AF1-8), AF5 was used in the direct cELISA owing to its excellent specificity and sensitivity with the detection limit of 4 microg kg(-1). The recovery of AFB(1) spiked at 5, 10, 20, 40 and 80 microg kg(-1) ranged from 88.1 to 99.5%. Correlation coefficients of the ELISA with the commercial ELISA kit and thin-layer chromatography were 0.912 and 0.802 for corn, and 0.941 and 0.832 for peanuts, respectively (p

As part of a survey of anti-tumorpromotive chemicals, garcinone E (1) and nine known xanthones, 8-deoxygartanin, gartanin, xanthone I, β-mangostin, garcinone B, 6-deoxy-γ-mangostin, 1, 5, 8-trihydroxy-3-methoxy-2-(3-methyl-2-butenyl)xanthone, γ-mangostin and α-mangostin, together with egonol, were isolated from the fruit hulls of Garcinia mangostana L. collected in Thailand. The structure of garcinone E was elucidated as 1, 3, 6, 7-tetrahydroxy-2, 5, 8-tri-(3-methyl-2-buteny)xanthone by spectroscopic methods, including the application of a number of two dimensional (2D)NMR techniques (1H-1H, 13C-1H correlation spectroscopy (COSY), nuclear Overhauser effect spectroscopy (NOESY) and correlation via long-range coupling (COLOC).

In Thailand, smoking of commercial cigarettes and of handmade cigarettes has drastically increased in recent decades. Cancer of the lung and of the upper aero-digestive tract have also increased in Thailand as they have in many other countries. It is our working hypothesis that the increase of primary cancer of the liver, especially of cholangiocarcinoma in the north-eastern provinces of Thailand is associated with the use of tobacco in men infested with the liver fluke Opisthorchis viverrini (OV). Bioassays have shown that volatile nitrosamines and tobacco-specific nitrosamines induce cholangiocarcinoma in laboratory animals and that the hepatocarcinogenic action of nitrosodimethylamine in hamsters is significantly increased by infestation with the liver fluke OV. The endogenous formation of nitrosamines is significantly increased by OV infestation. This report presents analytical data on the concentration of volatile nitrosamines and tobacco-specific nitrosamines in mainstream smoke of nine leading brands of commercially produced Thai cigarettes which represent approximately 85% of the market share in Thailand. Observed ranges (ng/cigarette) were 8.5-31.9 for nitrosodimethylamine, 8.8-49.6 for nitrosopyrrolidine and 4.2-18.9 for nitrosodi-n-butylamine. These values are exceptionally high compared with the smoke of light and blended cigarettes from North America and Western Europe. Among the tobacco-specific nitrosamines, the range was 28-730 for nitrosonornicotine and 16-370 for 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone. There was a correlation between volatile and tobacco-specific nitrosamines, and tar and nicotine deliveries in the mainstream smoke. The analytical data are in line with the rate for lung cancer and support our working hypothesis that nitrosamines, and especially the tobacco-specific nitrosamines, are associated with the increased risk for primary liver cancer among those Thai people who smoke cigarettes and also carry OV infestation.

By using a highly sensitive enzyme-linked immunosorbent assay (ELISA) based on a monoclonal antibody, microcystin (MC) concentration was analyzed in environmental water samples (total, 134), collected in 1993-1995 from ponds, lakes, reservoirs, and rivers in Japan, Thailand, Germany, and Portugal. MCs detected in the water samples filtered over a glass filter were designated as free MCs, and those samples that were freeze-thawed twice before the filtration were designated as total MCs. MCs (> 50 pg/ml) were detected in 14 of 24 samples collected from the lakes that were used as recreation and water supply in Japan in different regions. In the MC-positive samples, the concentration of free MCs was only a few percentages of the total MCs, indicating that the most part of MCs found in the water samples was present in algal cells. An additional trial on 33 samples collected continuously from Lake Inbanuma, Japan, during June-September 1994-1995 revealed that the total MCs were in a range of 52-52,000 pg/ml. In Chiang Mai, Thailand, 6 of 10 samples were positive, with the mean and highest of 161 and 354 pg/ml, respectively. In the Frankfurt area. Germany, 4 of 10 and 7 of 8 samples collected in the same lakes for recreation in July 1993 and November-December 1994 showed the presence of MCs, with their mean and highest values of 257 and 407 pg/ml, respectively. Another survey of MCs in dense bloomed samples collected with plankton net revealed a contamination of MCs up to 36,000 pg/ml. In Portugal, 28 of 29 samples from 4 lakes, 20 rivers, and 5 reservoirs were positive for MCs, with the respective means of 13,664, 11,048, and 2,278 pg/ml. These data indicated that MCs contaminate environmental water in ponds, rivers, lakes, and reservoirs worldwide. The present ELISA is considered to be a reliable tool for the mass monitoring and risk assessment of MCs in water supplies.

The effects of stevioside and steviol (a product of enzy-matic hydrolysis of stevioside) on intestinal glucose absorption were examined in the hamster jejunum in vitro. By using the jejunal rings technique, we found that stevioside at a high dose of 5 mM had no inhibitory effect on glucose absorption. In contrast, glucose absorption was inhibited 43% by 1 mM steviol. The inhibition of glucose absorption by steviol was related to steviol concentration and incubation time. The inhibitory effect of steviol compared to phlorizin and ouabain was also investigated. Steviol, which caused a decrease in glucose accumulation in the intestinal ring tissues, possibly acts on the brush border membrane as does phlorizin. Furthermore, it was also found that steviol altered the morphology of the intestinal absorptive cells. These results suggest that the possible site of inhibitory action of steviol might be on the mucosal side and/or at the intracellular organelles of intestinal absorptive cells.

The high content (about 39%) of polymeric tannins in tamarind ( Tamarindus indica L.) seed husk (TSH) was demonstrated, and an extract (crude TSE) with a high content (about 94%) of polymeric tannins was prepared from TSH with a one pot extraction using ethanol/water (3:2, v/v). The crude TSE was further purified with Sephadex LH20 to give one fraction (metTSE) eluted with methanol/water (3:2, v/v) and another (acTSE) eluted with acetone/water (3:2, v/v). The tannins of acTSE were established as polymeric proanthocyanidins (PA) by 13C NMR spectroscopy; this was further confirmed by IR and UV spectroscopy, n-BuOH/HCl and vanillin assays, and from HPLC pattern. The ratio of procyanidins to prodelphinidins was 2:3, and the average degree of polymerization of acTSE was 7. Galloylated flavan-3-ols were not detected in acTSE. The main ingredients of metTSE were confirmed to be polymeric PA by 13C NMR spectroscopy. The antioxidant activities using DPPH and ABTS assays were investigated. The IC50 values of acTSE were 4.2±0.2 (DPPH assay) and 6.2±0.3 μg/mL (ABTS assay).

We studied the effect of soybean milk protein (SMP) in a two-stage carcinogenesis experiment on mouse skin. Mice were given soybean protein isolate (SPI) diet or SPI diet supplemented with SMP. After 4 weeks on the diets, the mice were shaved and a tumor initiator was applied. A tumor promotor was then applied twice a week on the same area of the skin throughout the experiment. After 20 weeks on the treatment, the percentage of tumor-bearing mice and the volume of tumor tended to be lower in the mice on the SMP diet than those on the SPI diet. The number of tumor was also significantly lower in the former group as compared to the latter group. There was no difference in growth between mice of the SPI and SMP groups. The results indicate the safety and the anti-carcinogenic effect of SMP in mice.

Leaves of Stevia rebaudiana Bertoni have been popularly used as a sweetener in foods and beverages for diabetics and obese people due to their potent sweetener stevioside. In this report, stevioside and steviol were tested for mutagenicity in Salmonella typhimurium strains TA98 and TA100 and for chromosomal effects on cultured human lymphocytes. Stevioside was not mutagenic at concentrations up to 25 mg/plate, but showed direct mutagenicity to only TA98 at 50 mg/plate. However, steviol did not exhibit mutagenicity in either TA98 or TA100, with or without metabolic activation. No significant chromosomal effect of stevioside and steviol was observed in cultured blood lymphocytes from healthy donors (n = 5). This study indicates that stevioside and steviol are neither mutagenic nor clastogenic in vitro at the limited doses; however, in vivo genotoxic tests and long-term effects of stevioside and steviol are yet to be investigated.

The tumor promoter okadaic acid binds specifically to a particulate as well as a cytosolic fraction of various mouse tissues, e.g., skin, brain, lung and colon. The KD value was 21.7 nM for receptors in the particulate fraction and 1.0 nM for those in the cytosolic fraction of mouse skin. The specific binding of [3H]okadaic acid to the particulate fraction of mouse skin was inhibited dose-dependently by okadaic acid, but not okaidaic acid tetramethyl ether, an inactive compound, or by other tumor promoters, such as 12-O-tetradecanoylphorbol-13-acetate and teleocidin. The results suggest a new pathway of tumor promotion mediated through the okadaic acid receptor(s).

The action of N-acetylneuraminic acid aldolase on 7- and 8-carbon analogues of N-acetylneuraminic acid has been studied. The 8-carbon analogue was slowly and reversibly cleaved by the aldolase to yield 2-acetamido-2-deoxy-d-lyxose and pyruvic acid. The 7-carbon analogue was not cleaved by the enzyme. The 8-carbon analogue competed with N-acetylneuraminic acid for the active site of the enzyme, but the 7-carbon analogue exhibited no inhibitory effect even when present at a 10-fold greater concentration than N-acetylneuraminic acid.

Glycoprotein substrates for neuraminidase and glycoprotein inhibitors of the agglutination of erythrocytes by influenza virus were subjected to oxidation by very low concentrations of periodate under conditions which selectively oxidize N-acetylneuraminic acid (NANA), to the 8carbon and 7-carbon aldehydes. Molar ratios of periodate to NANA were varied from 0.2 to 4.5. Reduction of the oxidized glycoproteins with sodium borohydride yielded glycoproteins containing the 8- and 7-carbon analogues of NANA as well as the parent 9-carbon compound in amounts depending on the quantity of periodate employed. These were quantitated in the mixture by means of gas-liquid chromatography. Under the conditions used, periodate oxidation of other sugars in the glycoproteins could not be detected. The capacity of neuraminidases from Vibrio cholerae, Clostridium perfringens, and PR-8 influenza virus to cleave N-acetylneuraminic acid or its 8-carbon (NANA-8) or 7-carbon (NANA-7) analogues from native and modified orosomucoid and from NANA-, NANA-8-, and NANA-7-N-acetylgalactosaminitol were studied. NANA-8 was hydrolyzed from the glycoproteins and the reduced disaccharide at an initial rate about 30% of that for NANA. The 7-carbon analogue of NANA was not cleaved from the reduced disaccharide, but was slowly cleaved from modified orosomucoid at an initial rate of 5 to 10% of that for NANA. It was concluded that the optimum activity of neuraminidase requires the intact structure of the polyhydroxy side chain of NANA. When the molar ratio of periodate to NANA exceeded 1.5, all of the NANA was converted to the 8- and 7-carbon analogues, and the capacity of the glycoproteins to inhibit agglutination of human erythrocytes by PR-8 influenza virus was reduced by 95 to 100%. It is concluded that carbon atoms 7, 8, and 9 in the polyhydroxy side chain of NANA acid in glycoproteins are involved in their binding to influenza virus.

Ovine and human follicle-stimulating hormone (FSH) were modified by periodate oxidation followed by borohydride reduction, yielding FSH-containing analogues of N-acetylneuraminic acid (NANA) shortened by 1 or 2 carbon atoms. The FSH activity of the modified preparations was assayed by the human chorionic gonadotropin augmentation assay. The treatment decreased the biological activity of FSH somewhat, but, even when no unmodified NANA remained, the potency was still about half that of untreated FSH. Treatment of either native or modified FSH with neuraminidase resulted in the release of all NANA and NANA-8 and some NANA-7, and the loss of hormonal activity. It appears, therefore, that an intact 3-carbon polyhydroxy side chain in NANA is not required for full FSH activity.

The 8-carbon and the 7-carbon analogues of N-acetyl-neuraminic acid were necessary in order to study some of the structural requirements of N-acetylneuraminic acid in its role as substrate for certain enzymes. The 8-carbon analogue of N-acetylneuraminic acid was synthesized by the alkaline condensation of 2-acetamido-2-deoxy-d-lyxose and di-tert-butyloxaloacetate. This same 8-carbon compound and the 7-carbon analogue of N-acetylneuraminic acid were isolated after acid hydrolysis of Collocalia mucoid which had been modified by periodate oxidation followed by borohydride reduction. The two analogues could also be detected when periodate oxidation and borohydride reduction were applied to crystalline β-methoxy-N-acetylneuraminic acid methyl ester.

The widely used thiobarbituric acid technique for the quantitation of N -acetylneuraminic acid (NANA) was improved to eliminate the interference of the ubiquitous 2-deoxyribose. The 2-deoxyribose chromogen was completely removed by cyclohexanone extraction at pH 5.6–6.0. After readjusting the pH to 1.7–1.9, the chromogen representative of NANA was quantitatively extracted with cyclohexanone. All other aspects of the original technique (L. Warren 1959 J. Biol. Chem. 534 , 1971–1975) remained unchanged. The technique has been applied to determine total as well as neuraminidase-susceptible NANA in the preparation of the immunogen (neuraminidase-treated myeloblasts) utilized to stimulate specific immunity in patients with myeloblastic leukemia and certain solid tumors; NANA levels significantly affect immunogenicity. Data obtained from a variety of tumors using pH-dependent extraction as compared to the thiobarbituric acid method, isoamyl alcohol extraction, and ion-exchange purification showed that 2-deoxyribose interference may cause as much a two- to threefold error in the quantitation of NANA.

Purpose In 1988, we first reported the complete chemical structure of a new type of phorbol ester, abbreviated to DHPB, found in seed oil of Jatropha curcas L. (Saboodam in Thai) and its tumor-promoting activity on mouse skin. Although this seed oil contains toxic phorbol ester, it was planned to use it as a feasible renewable oil and the extracted seed cake as fertilizer. This utilization value opened a new science of Jatropha curcas. Methods The main experimental results are cited from our publications, and the relevant literature screened from journals and PubMed. Results and discussion This paper begins with our original work on the structural elucidation of a new phorbol ester, 12-deoxy-16-hydroxyphorbol (DHPB): its tumor-promoting activity was compared with that of TPA. We think that it is timely to review the following research advances with Jatropha curcas, so numerous topics are classified as follows: (1) historical development of phorbol esters in seed oil; (2) toxicity of phorbol ester based on various bioassays; (3) degradation of phorbol ester; (4) a new pharmaceutical compound in seed; and (5) tumor promotion and progression with endogeneous tumor promoters in human carcinogenesis. The discovery of phorbol ester in seed oil raised awareness of the danger of public use of seed oil and seed cake in Thailand, and also indicated the necessity of discussing the concept of primary and tertiary cancer preventions. Conclusion It is worthwhile to study the future benefits and cancer risks of globally distributed Jatropha curcas L.