White and gray matter integrity evaluated by MRI-DTI can serve as noninvasive and reliable indicators of structural and functional alterations in chronic neurotrauma.

We aimed to evaluate whether white and gray matter microstructure changes observed with magnetic resonance imaging (MRI)-based diffusion tensor imaging (DTI) can be used to reflect the progression of chronic brain trauma. The MRI-DTI parameters, neuropathologic changes, and behavioral performance of adult male Wistar rats that underwent moderate (2.1 atm on day "0") or repeated mild (1.5 atm on days "0" and "2") traumatic brain injury (TBI or rmTBI) or sham operation were evaluated at 7 days, 14 days, and 1-9 months after surgery. Neurobehavioral tests showed that TBI causes long-term motor, cognitive and neurological deficits, whereas rmTBI results in more significant deficits in these paradigms. Both histology and MRI show that rmTBI causes more significant changes in brain lesion volumes than TBI. In vivo DTI further reveals that TBI and rmTBI cause persistent microstructural changes in white matter tracts (such as the body of the corpus callosum, splenium of corpus callus, internal capsule and/or angular bundle) of both two hemispheres. Luxol fast blue measurements reveal similar myelin loss (as well as reduction in white matter thickness) in ipsilateral and contralateral hemispheres as observed by DTI analysis in injured rats. These data indicate that the disintegration of microstructural changes in white and gray matter parameters analyzed by MRI-DTI can serve as noninvasive and reliable markers of structural and functional level alterations in chronic TBI.

Mesenchymal stem cells reduce long-term cognitive deficits and attenuate myelin disintegration and microglia activation following repetitive traumatic brain injury.

The underlying mechanisms for the beneficial effects exerted by bone marrow-mesenchymal stem cells (BM-MSCs) in treating repetitive traumatic brain injury (rTBI)-induced long-term sensorimotor/cognitive impairments are not fully elucidated. Herein, we aimed to explore whether BM-MSCs therapy protects against rTBI-induced long-term neurobehavioral disorders in rats via normalizing white matter integrity and gray matter microglial response. Rats were subjected to repeated mild lateral fluid percussion on day 0 and day 3. On the fourth day post-surgery, MSCs groups received MSCs (4 × 106 cells/ml/kg, intravenously) and were assessed by the radial maze, Y maze, passive avoidance tests, and modified neurological severity scores. Hematoxylin & eosin, and Luxol fast blue stainings were used to examine the histopathology and white matter thickness. At the same time, immunofluorescence staining was used to investigate the numbers of tumor necrosis factor-alpha (TNF-α)-containing microglia in gray matter. Three to nine months after neurotrauma, rats displayed sensorimotor and cognitive impairments, reduced thickness in white matter, and over-accumulation of TNF-α-containing microglia and cellular damage in gray matter. Therapy with BM-MSCs significantly attenuated the rTBI-induced sensorimotor and cognitive impairments and all their complications. Mesenchymal stem cell therapy might accelerate the recovery of sensorimotor and cognitive impairments in rats with rTBI via normalizing myelin integrity and microglia response.

Analysis of Chlorogenic Acid in Sweet Potato Leaf Extracts.

Sweet potato (Ipomoea batatas L.) is one of the most important food crops worldwide, with leaves of different varieties showing purple, green and yellow, and these leaves provide a dietary source of nutrients and various bioactive compounds. The objective of this study was to identify the active constituents of chlorogenic acids (CGAs) in different methanolic extract of leaves of three varieties of sweet potato (purple CYY 98-59, green Taoyuan 2, and yellow CN 1927-16) using liquid chromatography-tandem mass spectrometry. Genotype-specific metabolite variations were observed; CGAs and three isomeric peaks were detected in sweet potato leaf extracts (SPLEs). Among them, the yellow SPLE contained the highest contents of 3,5-dicaffeoylquinic acid (3,5-di-CQA) and 3,4-dicaffeoylquinic acid (3,4-di-CQA), followed by the green SPLE, whereas the purple SPLE retained lower 3,5-di-CQA content compared to yellow and green SPLEs. All three SPLEs contained lower 4,5-dicaffeoylquinic acid (4,5-di-CQA) and CGA contents compared to 3,5-di-CQA and 3,4-di-CQA, although CGA constituents were not significantly different in genotypes, whereas purple SPLE contained higher 4,5-di-CQA content compared to yellow and green SPLEs. This study indicates that SPLs marketed in Taiwan vary widely in their biological potentials and may impart different health benefits to consumers.

Bioactive Compounds, Antioxidants, and Health Benefits of Sweet Potato Leaves.

Sweet potato (Ipomoea batatas) is one of the most important food crops worldwide and its leaves provide a dietary source of nutrients and various bioactive compounds. These constituents of sweet potato leaves (SPL) vary among varieties and play important roles in treating and preventing various diseases. Recently, more attentions in health-promoting benefits have led to several in vitro and in vivo investigations, as well as the identification and quantification of bioactive compounds in SPL. Among them, many new compounds have been reported as the first identified compounds from SPL with their dominant bioactivities. This review summarizes the current knowledge of the bioactive compositions of SPL and their health benefits. Since SPL serve as a potential source of micronutrients and functional compounds, they can be further developed as a sustainable crop for food and medicinal industries.

Antioxidant activity in extracts of 27 indigenous Taiwanese vegetables.

The objectives of this study were to identify the antioxidants and antioxidant axtivity in 27 of Taiwan's indigenous vegetables. Lycium chinense (Lc), Lactuca indica (Li), and Perilla ocymoides (Po) contained abundant quercetin (Que), while Artemisia lactiflora (Al) and Gynura bicolor (Gb) were rich in morin and kaempferol, respectively. Additionally, Nymphoides cristata (Nc) and Sechium edule (Se)-yellow had significantly higher levels of myricetin (Myr) than other tested samples. Cyanidin (Cyan) and malvidin (Mal) were abundant in Gb, Abelmoschus esculentus Moench (Abe), Po, Anisogonium esculentum (Retz.) Presl (Ane), Ipomoea batatas (Ib)-purple, and Hemerocallis fulva (Hf)-bright orange. Relatively high levels of Trolox equivalent antioxidant capacity (TEAC), oxygen radical absorption capacity (ORAC), and 1,1-diphenyl-2-picryl-hydrazyl (DPPH) radical scavenger were generated from extracts of Toona sinensis (Ts) and Po. Significant and positive correlations between antioxidant activity and polyphenols, anthocyanidins, Que, Myr, and morin were observed, indicating that these phytochemicals were some of the main components responsible for the antioxidant activity of tested plants. The much higher antioxidant activity of Po, Ts, and Ib (purple leaf) may be related to their higher Cyan, Que, and polyphenol content.

Organ-specific distribution of chlorophyll-related compounds from dietary spinach in rabbits.

The distribution of chlorophyll-related compounds (CRCs) derived from dietary spinach was investigated in different organs the rabbits. The rabbits in the experimental group consumed 100 g of freeze-dried spinach powder after a 24 h fasting period and sacrificed 2, 4, 8, 12 and 24 h later and in the control group sacrificed after the 24 h fasting period. The main CRCs in the liver were found to be chlorophyll (Chl a) and b, chlorophyllide (Chlide) a and b, pheophytin (Phe) a and b and pheophorbide (Pho) a and b, which reached their peak values at 8 h post-feeding. The gallbladder contained mainly Chlide a and a', Pho a and a', Pho b and b', which peaked their values at 2 h post-feeding. Pho a and b were consistently observed in the blood and peaked at 12 h post-feeding. The earlier appearance of Chlide a', Pho a' and Pho b' in the gallbladder compared to the liver indicated that these CRCs were compartmentalized differently and might undergo the same type of vectorialized transport as characterized for the bile salts. Pho levels peaked later in the blood compared to the liver, suggesting that Pho might be released into the peripheral blood circulation from the liver. In conclusion, Chlide and Pho were the principal Chl metabolites in the rabbits. Our data may expand our understanding of the metabolism and biodistribution of CRCs in the human body. A number of biological functions, including anti-oxidation, anti-tumor and anti-aging have recently been attributed to CRCs, it will be interesting to explore, if the binding of Chlide and Pho to other nutrients or trace metal ions in the body mediate their biological functions.

Eliminating interference by anthocyanin in chlorophyll estimation of sweet potato (Ipomoea batatas L.) leaves.

BACKGROUND: Spectral reflectance was evaluated for its usefulness as a nondestructive estimation of chlorophyll (Chl) content from three cultivars of sweet potato (Ipomoea batatas L.) with green, yellow, and purple leaves grown in a greenhouse for 22 days. While the green and yellow leaves contain variant amount of photosynthetic pigments without or with little level of anthocyanins, the purple leaves, except large amount of photosynthetic pigments, have high quantity of anthocyanins. RESULTS: For green and yellow leaves, the reciprocal reflectance (R-1) and derived indices incorporating near infrared (NIR) reflectance, [(Rλ)-1 - (RNIR)-1] and [(RNIR/Rλ) - 1], in the green and red edge spectral ranges were shown to be strongly correlated (r2 = 0.8 ~ 0.9) with the chlorophyll content. The root mean square error (RMSE) of the chlorophyll content estimation using these indices was < 50 mg m-2. However, when purple leaves containing high levels of anthocyanins were included in the sample, R-1 in the green spectral range and the above-mentioned indices displayed much weaker correlations with the chlorophyll content. The RMSE of chlorophyll estimation using these indices in the green spectral range sharply increased to > 110 mg m-2 when the sample included purple leaves. The new index, [1 - (Rλ/RNIR)], was therefore inferred and developed to eliminate the distorting effect of anthocyanins on chlorophyll content estimation using reflectance in the green spectral range. For leaves with high levels of anthocyanins, the correlation between [1 - (Rλ/RNIR)] and the chlorophyll content remained strong (r2 = 0.8 ~ 0.9) in the green spectral range, and the RMSE was minimal. CONCLUSION: The reflectance index, [1 - (Rλ/RNIR)], therefore represents a new and useful parameter for estimating leaf chlorophyll content in leaves with any level of anthocyanins such as purple rice leaf.

Antioxidant activity of herbaceous plant extracts protect against hydrogen peroxide-induced DNA damage in human lymphocytes.

BACKGROUND: Herbaceous plants containing antioxidants can protect against DNA damage. The purpose of this study was to evaluate the antioxidant substances, antioxidant activity, and protection of DNA from oxidative damage in human lymphocytes induced by hydrogen peroxide (H2O2). Our methods used acidic methanol and water extractions from six herbaceous plants, including Bidens alba (BA), Lycium chinense (LC), Mentha arvensis (MA), Plantago asiatica (PA), Houttuynia cordata (HC), and Centella asiatica (CA). METHODS: Antioxidant compounds such as flavonol and polyphenol were analyzed. Antioxidant activity was determined by the inhibition percentage of conjugated diene formation in a linoleic acid emulsion system and by trolox-equivalent antioxidant capacity (TEAC) assay. Their antioxidative capacities for protecting human lymphocyte DNA from H2O2-induced strand breaks was evaluated by comet assay. RESULTS: The studied plants were found to be rich in flavonols, especially myricetin in BA, morin in MA, quercetin in HC, and kaemperol in CA. In addition, polyphenol abounded in BA and CA. The best conjugated diene formation inhibition percentage was found in the acidic methanolic extract of PA. Regarding TEAC, the best antioxidant activity was generated from the acidic methanolic extract of HC. Water and acidic methanolic extracts of MA and HC both had better inhibition percentages of tail DNA% and tail moment as compared to the rest of the tested extracts, and significantly suppressed oxidative damage to lymphocyte DNA. CONCLUSION: Quercetin and morin are important for preventing peroxidation and oxidative damage to DNA, and the leaves of MA and HC extracts may have excellent potential as functional ingredients representing potential sources of natural antioxidants.

Chlorophyll-related compounds inhibit cell adhesion and inflammation in human aortic cells.

The objectives of this study were to investigate the effects of chlorophyll-related compounds (CRCs) and chlorophyll (Chl) a+b on inflammation in human aortic endothelial cells. Adhesion molecule expression and interleukin (IL)-8, nuclear factor (NF)-κB p65 protein, and NF-κB and activator protein (AP)-1 DNA binding were assessed. The effects of CRCs on inflammatory signaling pathways of signal transducers and activators of transcription 3 (STAT3) and mothers against decapentaplegic homolog 4, respectively induced by IL-6 and transforming growth factor (TGF)-ß, in human aortic smooth muscle cells cultured in vitro were also investigated. HAECs were pretreated with 10 µM of CRCs, Chl a+b, and aspirin (Asp) for 18 h followed by tumor necrosis factor (TNF)-α (2 ng/mL) for 6 h, and U937 cell adhesion was determined. TNF-α-induced monocyte-endothelial cell adhesion was significantly inhibited by CRCs. Moreover, CRCs and Chl a+b significantly attenuated vascular cell adhesion molecule-1, intercellular adhesion molecule-1, and IL-8 expressions. Treatments also significantly decreased in NF-κB expression, DNA binding, and AP-1 DNA binding by CRCs and Asp. Thus, CRCs exert anti-inflammatory effects through modulation of NF-κB and AP-1 signaling. Ten micromoles of CRCs and Asp upregulated the expression of mothers against decapentaplegic homolog 4 (Drosophila) (SMAD4) in the TGF-ß receptor signaling pathway, and SMAD3/4 transcription activity was also increased. Ten micromoles of CRCs were able to potently inhibit STAT3-binding activity by repressing IL-6-induced STAT3 expression. Our results provide a potential mechanism that explains the anti-inflammatory activities of these CRCs.

Inhibitive effect of purple sweet potato leaf extract and its components on cell adhesion and inflammatory response in human aortic endothelial cells.

This study investigated the effects of purple sweet potato leaf extract (PSPLE) and its components, cyanidin and quercetin, on human aortic endothelial cells (HAECs) during the inflammatory process. HAECs were pretreated with 100 µg/mL PSPLE or 10 µM quercetin, cyanidin or aspirin for 18 h followed by TNF-α (2 ng/mL) for 6 h, and U937 cell adhesion was determined. Adhesion molecule expression and CD40 were evaluated; NFκB p65 protein localization and DNA binding were assessed. PSPLE, aspirin, cyanidin and quercetin significantly inhibited TNF-α-induced monocyte-endothelial cell adhesion (p < 0.05). Cyanidin, quercetin and PSPLE also significantly attenuated VCAM-1, IL-8 and CD40 expression, and quercetin significantly attenuated ICAM-1 and E-selectin expression (p < 0.05). Significant reductions in NFκB expression and DNA binding by aspirin, cyanidin and quercetin were also observed in addition to decreased expression of ERK1, ERK2 and p38 MAPK (p < 0.05). Thus, PSPLE and its components, cyanidin and quercetin, have anti-inflammatory effects through modulation of NFκB and MAPK signaling. Further in vivo studies are necessary to explore the possible therapeutic effects of PSPLE on atherosclerosis.

Naturally occurring chlorophyll derivatives inhibit aflatoxin B1-DNA adduct formation in hepatoma cells.

The inhibitory effects of four chlorophyll derivatives (chlorophyllide [Chlide] a and b and pheophorbide [Pho] a and b) on aflatoxin B1 (AFB1)-DNA adduct formation, and on the modulation of hepatic glutathione S-transferase (GST) were evaluated in murine hepatoma (Hepa-1) cells. Enzyme-linked immunosorbent assay showed that pretreatment with Chlide or Pho significantly reduced the formation of AFB1-DNA adducts, and that Pho was the most potent inhibitor. However, wash-out prior to adding AFB1 totally eliminated inhibition by Childe and partially eliminated inhibition by Pho, indicating that the inhibitory effect of Chlide, and to some extent Pho, was mediated through direct trapping of AFB1. Furthermore, spectrophotometric analysis showed that Pho treatment could increase GST activity in Hepa-1 cells. These observations indicate that the chlorophyll derivatives studied may attenuate AFB1-induced DNA damage in the Hepa-1 cell by direct trapping of AFB1. Pho provided additional protection not only by direct trapping, but also by increasing GST activity against hepatic AFB1 metabolites.

Effects of chlorophyll-related compounds on hydrogen peroxide induced DNA damage within human lymphocytes.

Chlorophylls (Chl's) are the most abundant natural plant pigments. Four chlorophyll-related compounds (CRCs), including chlorophyllide a and b (Chlide a and b) and pheophorbide a and b (Pho a and b), were investigated for their antioxidative capacities to protect human lymphocyte DNA from hydrogen peroxide (H(2)O(2)) induced strand breaks and oxidative damage ex vivo. Lymphocytes exposed to H(2)O(2) at concentrations of 10 and 50 microM revealed an increased frequency of DNA single-strand breaks (ssb's; as measured by the comet assay) and also an increased level of oxidized nucleoside (as measured by 8-hydroxydeoxyguanosine, 8-OHdG). All Chl's reduced the level of DNA ssb's and 8-OHdG within human lymphocytes following exposure to 10 microM H(2)O(2). Only Pho a and b were able to decrease DNA ssb's and 8-OHdG following treatment of lymphocytes with 50 microM H(2)O(2), in a concentration-dependent fashion. It was demonstrated herein that Pho a and b were more antioxidative than others. We applied DPPH free-radical scavenge assays in vitro, and got similar results. Pho a and b had higher ability in scavenging capacities than others. We conclude that water-extract Chl's are able to enhance the ability of human lymphocytes to resist H(2)O(2)-induced oxidative damage, especially for Pho a and b.

Effects of frying oil and Houttuynia cordata thunb on xenobiotic-metabolizing enzyme system of rodents.

AIM: To evaluate the effects of frying oil and Houttuynia cordata Thunb (H. cordata), a vegetable traditionally consumed in Taiwan, on the xenobiotic-metabolizing enzyme system of rodents. METHODS: Forty-eight Sprague-Dawley rats were fed with a diet containing 0%, 2% or 5% H. cordata powder and 15% fresh soybean oil or 24-h oxidized frying oil (OFO) for 28 d respectively. The level of microsomal protein, total cytochrome 450 content (CYP450) and enzyme activities including NADPH reductase, ethoxyresorufin O-deethylase (EROD), pentoxyresorufin O-dealkylase (PROD), aniline hydroxylase (ANH), aminopyrine demethylase (AMD), and quinone reductase (QR) were determined. QR represented phase II enzymes, the rest of the enzymes tested represented phase I enzymes. RESULTS: The oxidized frying oil feeding produced a significant increase in phase I and II enzyme systems, including the content of CYP450 and microsomal protein, and the activities of NADPH reductase, EROD, PROD, ANH, AMD and QR in rats (P<0.05). In addition, the activities of EROD, ANH and AMD decreased and QR increased after feeding with H. cordata in OFO-fed group (P<0.05). The feeding with 2% H. cordata diet showed the most significant effect. CONCLUSION: The OFO diet induces phases I and II enzyme activity, and the 2% H. cordata diet resulted in a better regulation of the xenobiotic-metabolizing enzyme system.

A study of the antioxidative and antimutagenic effects of Houttuynia cordata Thunb. using an oxidized frying oil-fed model.

The aims of this study were to evaluate the antioxidative effect of a traditional Taiwanese vegetable, Houttuynia cordata Thunb. (H. cordata), by subjecting rodents to oxidized frying oil-induced oxidative stress, and to examine the antimutagenic effects of H. cordata using the Ames test. Forty-eight Sprague-Dawley rats were fed a diet of 0, 2, or 5% H. cordata and 15% fresh oil or oxidized frying oil (OFO) for 28 d. Levels of polyphenol in the feces, plasma, and liver were determined. The LDL lag time, plasma total antioxidant status (TAS), and levels of thiobarbituric acid-reactive substances (TBARS) were used as antioxidative indices, and the protein carbonyl group was used as an oxidative index. The results showed that the polyphenol content decreased in the plasma and increased in the feces when administering OFO, and the apparent absorption of polyphenol also decreased. The polyphenol content in plasma increased when giving H. cordata. There was a higher polyphenol concentration in the water extracts of H. cordata than in the methanol extracts. The OFO-fed groups had higher plasma TBARS and hepatic protein carbonyl group concentrations and shorter LDL lag times than those of the control group. The total TAS was elevated and the LDL lag time was prolonged when fed with H. cordata. In addition, both water and methanol extracts of H. cordata had an antimutagenic effect on benzo(a)pyrene, aflatoxin B1. and OFO, and showed a dose-dependent response using the Ames test. The antimutagenic ability of water extracts was higher than that of the methanol extracts. In conclusion, the polyphenol in H. cordata is easily absorbed and metabolized by rodents. H. cordata showed both antioxidative and antimutagenic properties under OFO feeding-induced oxidative stress.