The use of C-reactive protein to predict uncomplicated appendicitis and increase uptake of laparoscopy in low resource hospitals-A retrospective cohort study.

BACKGROUND: Acute appendicitis remains the most common surgical emergency worldwide. There has been a low uptake of laparoscopic appendicectomy in the South African public sector. Preoperative identification of cases of uncomplicated appendicitis that are amenable to a laparoscopic approach may facilitate the implementation of laparoscopic appendicectomy programs in training hospitals. With limited access to preoperative imaging, alternative strategies for this preoperative prediction are needed. METHODS: A retrospective audit of patients over the age of 12 years with a histologically confirmed diagnosis of acute appendicitis over a 5-year period was performed. Patients were categorized as uncomplicated or complicated appendicitis and C-reactive protein (CRP) and white cell count (WCC) reviewed. Receiver operating characteristics curves were constructed for these blood tests and acute appendicitis severity. Youden's J statistic was used to determine optimal cut off values for diagnosing complicated appendicitis. RESULTS: 358 patients had confirmed appendicitis and complete blood results. Of these, 189 (52.79%) had complicated appendicitis with a 40.22% perforation rate. Median CRP in uncomplicated and complicated groups were 68 mg/L (IQR 19-142) and 216 mg/L (IQR 103-313) with an area under the curve (AUC) of 0.75 (95% CI: 0.70-0.80). The median WCC in the two groups were 12.6 × 109 cells/L (IQR 9.9-15.6) and 14.4 × 109 cells/L (IQR 11.5-18.28) with an AUC of 0.61 (95% CI: 0.56-0.67). The optimal cut off value for CRP was found to be 110 mg/L with a sensitivity of 74.74% and specificity of 69.23%. CONCLUSION: A cutoff value of 110 mg/dl CRP can distinguish patients with early appendicitis from those with complicated disease and when used in conjunction with clinical assessment may help identify patients in whom laparoscopic appendicectomy is appropriate.

Protective Role of Phenolic Compounds from Whole Cardamom (Elettaria cardamomum (L.) Maton) against LPS-Induced Inflammation in Colon and Macrophage Cells.

The chemical profiling of phenolic and terpenoid compounds in whole cardamom, skin, and seeds (Elettaria cardamomum (L.) Maton) showed 11 phenolics and 16 terpenoids, many of which are reported for the first time. Herein, we report the anti-inflammatory properties of a methanolic extract of whole cardamom in colon and macrophage cells stimulated with an inflammatory bacteria lipopolysaccharide (LPS). The results show that cardamom extracts lowered the expression of pro-inflammatory genes NFkß, TNFα, IL-6, and COX2 in colon cells by reducing reactive oxygen species (ROS) while not affecting LXRα. In macrophages, cardamom extracts lowered the expression of pro-inflammatory genes NFkß, TNFα, IL-6, and COX2 and decreased NO levels through a reduction in ROS and enhanced gene expression of nuclear receptors LXRα and PPARγ. The cardamom extracts in a range of 200-800 µg/mL did not show toxicity effects in colon or macrophage cells. The whole-cardamom methanolic extracts contained high levels of phenolics compounds (e.g., protocatechuic acid, caffeic acid, syringic acid, and 5-O-caffeoylquinic acid, among others) and are likely responsible for the anti-inflammatory and multifunctional effects observed in this study. The generated information suggests that cardamom may play a protective role against low-grade inflammation that can be the basis of future in vivo studies using mice models of inflammation and associated chronic diseases.

Cross-Talk and Physiological Role of Jasmonic Acid, Ethylene, and Reactive Oxygen Species in Wound-Induced Phenolic Biosynthesis in Broccoli.

Wounding induces phenolic biosynthesis in broccoli. However, there is scarce information about the physiological and molecular mechanisms governing this stress response. In the present study, a chemical-genetics approach was used to elucidate the role of reactive oxygen species (ROS), jasmonic acid (JA), and ethylene (ET) as stress-signaling molecules in the wound-induced phenolic biosynthesis in broccoli. Wounding activated the biosynthesis of ET and JA. Likewise, the wound-induced biosynthesis of ET and JA was regulated by ROS. JA activated primary metabolism, whereas the three signaling molecules activated phenylpropanoid metabolism. The signaling molecules inhibited the wound-induced activation of the hydroxycinnamoyl-CoA quinate hydroxycinnamoyl transferase (HQT) gene, which is involved in caffeoylquinic acids biosynthesis, and the main phenolics accumulated in wounded broccoli, suggesting that an alternative caffeoylquinic biosynthesis pathway is activated in the tissue due to wounding. ROS mediated the biosynthesis of most individual phenolic compounds evaluated. In conclusion, ROS, ET, and JA are essential in activating broccoli's primary and secondary metabolism, resulting in phenolic accumulation.

Extraction of phytochemicals from the pomegranate (Punica granatum L., Punicaceae) by reverse iontophoresis.

Plant metabolic profiling can provide a wealth of information regarding the biochemical status of the organism, but sample acquisition typically requires an invasive and/or destructive extraction process. Reverse iontophoresis (RI) imposes a small electric field across a biological membrane to substantially enhance the transport of charged and polar compounds and has been employed, in particular, to extract biomarkers of interest across human skin. The objective of this work was to examine the capability of RI to sample phytochemicals in a minimally invasive fashion in fructo (i.e., from the intact fruit). RI was principally used to extract a model, bioactive compound - specifically, ellagic acid - from the fruit peel of Punica granatum L. The RI sampling protocol was refined using isolated peel, and a number of experimental factors were examined and optimised, including preparation of the peel samples, the current intensity applied and the pH of the medium into which samples were collected. The most favourable conditions (3 mA current for a period of 1 hour, into a buffer at pH 7.4) were then applied to the successful RI extraction of ellagic acid from intact pomegranates. Multiple additional phytochemicals were also extracted and identified by liquid chromatography with tandem mass spectrometry (LC-MS/MS). A successful proof-of-concept has been achieved, demonstrating the capability to non-destructively extract phytochemicals of interest from intact fruit.

Non-destructive, reverse iontophoretic extraction of phytochemicals from Mangifera indica, Centella asiatica, Punica granatum, and Citrus sinensis.

For the commercial-scale isolation of phytochemicals, a suitable plant biomass source (including species, origin, growing season, etc.) must be identified, and frequent analytical verification is required to ensure that the phytochemicals are present at predefined minimum threshold concentrations. While the latter are typically assessed in the laboratory, a more efficient and less resource-intensive approach would involve non-destructive and environmentally friendly measurements in situ. Reverse iontophoretic (RI) sampling offers a potential solution to this challenge. OBJECTIVE: We aimed to demonstrate the non-destructive, RI sampling of phytochemicals of interest from biomass from four different sources. MATERIALS AND METHODS: RI experiments were performed in side-by-side diffusion cells using a current density of 0.5 mA/cm2 , for a predetermined time in a defined pH environment, using (1) fresh leaves from Mangifera indica and Centella asiatica and (2) isolated peel from Punica granatum and Citrus sinensis. RESULTS: Mangiferin, madecassoside, punicalagin, ellagic acid, and hesperidin were extracted from the different biomasses by RI. The amounts extracted ranged from 0.03 mg/100 mg of biomass for the cathodal extraction of madecassoside to 0.63 mg/100 mg of biomass for the anodal extraction of punicalagin. A linear relationship (r2  = 0.73) between the RI-extracted quantities of punicalagin and those determined using conventional methods was demonstrated. CONCLUSION: The non-destructive, in situ measurement of phytochemical levels by RI represents a feasible approach for timing the harvesting process.

Chemical Genetics Applied to Elucidate the Physiological Role of Stress-Signaling Molecules on the Wound-Induced Accumulation of Glucosinolates in Broccoli.

Wounding stress is an effective strategy to induce glucosinolate (GS) biosynthesis in broccoli. However, there is insufficient knowledge on the physiological and molecular mechanisms underlying this stress response. Herein, a chemical-genetic approach was applied to elucidate the role of jasmonic acid (JA), ethylene (ET), and reactive oxygen species (ROS) on the wound-induced biosynthesis of GS. Broccoli was processed into chops to induce wounding stress. Broccoli chops were treated with phenidone (PHEN) and diphenyleneiodonium chloride (DPI) as inhibitors of JA and ROS biosynthesis, respectively, whereas 1-methylcyclopropene (1-MCP) was applied as an inhibitor of ET action. Wounding stress induced the expression of genes related to the biosynthesis of indolic and aliphatic GS, which was correlated with the accumulation of GS and modulated by the inhibitors of signaling molecules applied. Results of gene expression analysis indicated that JA played a key role in the activation of most genes, followed by ROS. Furthermore, except for the CYP79B2 gene, PHEN and 1-MCP synergistically downregulated the expression of GS biosynthetic genes evaluated, showing that the interaction between JA and ET was fundamental to modulate GS biosynthesis. Results presented herein increased our knowledge of the physiological and molecular mechanisms governing the wound-induced biosynthesis of GS in broccoli.

UVC light modulates vitamin C and phenolic biosynthesis in acerola fruit: role of increased mitochondria activity and ROS production.

The effects of ultraviolet-C light (UVC) on vitamin C and phenolic compounds in acerola during postharvest storage were investigated in order to elucidate the mechanism inducing the antioxidant systems. The fruits, stored at 10 °C for 7 days after a hormetic UVC irradiation (two pulses of 0.3 J/cm2), showed significantly less degradation of vitamin C and phenolic compounds than the control without the UVC challenge. UVC activated the L-galactono-1,4-lactone dehydrogenase (GalDH), a key enzyme for vitamin C biosynthesis, and altered the composition of phenolic compounds, through phenolic biosynthesis, in acerola during postharvest storage. UVC also induced reactive oxygen species (ROS) productions at immediate (day 0) and late (day 7) times during postharvest storage through the mitochondrial electron transport chain and NADPH oxidase, respectively. Results suggest that UVC helps in the retention of vitamin C and phenolic content in acerola by altering ascorbic acid and phenolic metabolism through an increase in mitochondrial activity and a ROS-mediated mechanism. Data showed the beneficial effects of UVC on maintenance of nutraceutical quality in acerola during postharvest storage and supplied new insights into understanding the mechanism by which UVC irradiation enhance the antioxidant system in fruits.

Verrucosamide, a Cytotoxic 1,4-Thiazepane-Containing Thiodepsipeptide from a Marine-Derived Actinomycete.

A new cytotoxic thiodepsipeptide, verrucosamide (1), was isolated along with the known, related cyclic peptide thiocoraline, from the extract of a marine-derived actinomycete, a Verrucosispora sp., our strain CNX-026. The new peptide, which is composed of two rare seven-membered 1,4-thiazepane rings, was elucidated by a combination of spectral methods and the absolute configuration was determined by a single X-ray diffraction study. Verrucosamide (1) showed moderate cytotoxicity and selectivity in the NCI 60 cell line bioassay. The most susceptible cell lines were MDA-MB-468 breast carcinoma with an LD50 of 1.26 µM, and COLO 205 colon adenocarcinoma with an LD50 of 1.4 µM. Also isolated along with verrucosamide were three small 3-hydroxy(alkoxy)-quinaldic acid derivatives that appear to be products of the same biosynthetic pathway.

Sequential application of postharvest wounding stress and extrusion as an innovative tool to increase the concentration of free and bound phenolics in carrots.

Postharvest wounding stress in carrots induces the accumulation of phenolics, whereas extrusion generates modifications in the nutritional profiles of food matrixes. In the present study, the sequential application of wounding stress and extrusion on total free and bound phenolics as well as on carotenoid profiles of carrots was evaluated. Wounding was applied by shredding carrots and storing the tissue (48 h, 15 °C). The stressed-tissue was dehydrated and extruded at 63 °C or 109 °C and at continuous or expansion screw configurations. Extrudates were milled and sieved before phytochemical analysis. Wounding increased total free (288.1%) and bound (407.6%) phenolic content, whereas the carotenoid content was unaltered. The free and bound phenolics that showed the highest increase due to wounding were the chlorogenic (579.8%) and p-coumaric (390.9%) acids. Extrusion, at 109 °C under expansion screw configuration, further increased the wound-induced accumulation of total free (296.6%) and bound (22.1%) phenolics and induced trans-cis isomerization of ß-carotene.

Wounding and UVB Light Synergistically Induce the Biosynthesis of Phenolic Compounds and Ascorbic Acid in Red Prickly Pears (Opuntia ficus-indica cv. Rojo Vigor).

The present study evaluated the effects of ultraviolet B (UVB) radiation and wounding stress, applied alone or combined, on the biosynthesis of phenolic compounds and ascorbic acid in the peel and pulp of red prickly pear (Opuntia ficus-indica cv. Rojo Vigor). Whole and wounded-fruit samples were treated with UVB radiation (6.4 W·m-2) for 0 and 15 min, and stored for 24 h at 16 °C. Phytochemical analyses were performed separately in the peel and pulp. The highest phenolic accumulation occurred after storage of the whole tissue treated with UVB, where the main phenolic compounds accumulated in the peel and pulp were quercetin, sinapic acid, kaempferol, rosmarinic acid, and sinapoyl malate, showing increases of 709.8%, 570.2%, 442.8%, 439.9%, and 186.2%, respectively, as compared with the control before storage. Phenylalanine ammonia-lyase (PAL) activity was increased after storage of the whole and wounded tissue treated with UVB light, and this increase in PAL activity was associated to phenolic accumulation. On the other hand, l-galactono-γ-lactone dehydrogenase (GalLDH) activity and ascorbic acid biosynthesis was enhanced due to UVB radiation, and the effect was increased when UVB was applied in the wounded tissue showing 125.1% and 94.1% higher vitamin C content after storage when compared with the control. Respiration rate was increased due to wounding stress, whereas ethylene production was increased by wounding and UVB radiation in prickly pears. Results allowed the generation of a physiological model explaining the UVB and wound-induced accumulation of phenolic compounds and ascorbic acid in prickly pears, where wounding facilitates UVB to access the underlying tissue and enhances an apparent synergistic response.

UVA, UVB Light, and Methyl Jasmonate, Alone or Combined, Redirect the Biosynthesis of Glucosinolates, Phenolics, Carotenoids, and Chlorophylls in Broccoli Sprouts.

Broccoli sprouts contain health-promoting phytochemicals that can be enhanced by applying ultraviolet light (UV) or phytohormones. The separate and combined effects of methyl jasmonate (MJ), UVA, or UVB lights on glucosinolate, phenolic, carotenoid, and chlorophyll profiles were assessed in broccoli sprouts. Seven-day-old broccoli sprouts were exposed to UVA (9.47 W/m²) or UVB (7.16 W/m²) radiation for 120 min alone or in combination with a 25 µM MJ solution, also applied to sprouts without UV supplementation. UVA + MJ and UVB + MJ treatments increased the total glucosinolate content by ~154% and ~148%, respectively. MJ induced the biosynthesis of indole glucosinolates, especially neoglucobrassicin (~538%), showing a synergistic effect with UVA stress. UVB increased the content of aliphatic and indole glucosinolates, such as glucoraphanin (~78%) and 4-methoxy-glucobrassicin (~177%). UVA increased several phenolics such as gallic acid (~57%) and a kaempferol glucoside (~25.4%). MJ treatment decreased most phenolic levels but greatly induced accumulation of 5-sinapoylquinic acid (~239%). MJ treatments also reduced carotenoid and chlorophyll content, while UVA increased lutein (~23%), chlorophyll b (~31%), neoxanthin (~34%), and chlorophyll a (~67%). Results indicated that UV- and/or MJ-treated broccoli sprouts redirect the carbon flux to the biosynthesis of specific glucosinolates, phenolics, carotenoids, and chlorophylls depending on the type of stress applied.

UVA, UVB Light Doses and Harvesting Time Differentially Tailor Glucosinolate and Phenolic Profiles in Broccoli Sprouts.

Broccoli sprouts contain health-promoting glucosinolate and phenolic compounds that can be enhanced by applying ultraviolet light (UV). Here, the effect of UVA or UVB radiation on glucosinolate and phenolic profiles was assessed in broccoli sprouts. Sprouts were exposed for 120 min to low intensity and high intensity UVA (UVAL, UVAH) or UVB (UVBL, UVBH) with UV intensity values of 3.16, 4.05, 2.28 and 3.34 W/m², respectively. Harvest occurred 2 or 24 h post-treatment; and methanol/water or ethanol/water (70%, v/v) extracts were prepared. Seven glucosinolates and 22 phenolics were identified. Ethanol extracts showed higher levels of certain glucosinolates such as glucoraphanin, whereas methanol extracts showed slight higher levels of phenolics. The highest glucosinolate accumulation occurred 24 h after UVBH treatment, increasing 4-methoxy-glucobrassicin, glucobrassicin and glucoraphanin by ~170, 78 and 73%, respectively. Furthermore, UVAL radiation and harvest 2 h afterwards accumulated gallic acid hexoside I (~14%), 4-O-caffeoylquinic acid (~42%), gallic acid derivative (~48%) and 1-sinapoyl-2,2-diferulolyl-gentiobiose (~61%). Increases in sinapoyl malate (~12%), gallotannic acid (~48%) and 5-sinapoyl-quinic acid (~121%) were observed with UVBH Results indicate that UV-irradiated broccoli sprouts could be exploited as a functional food for fresh consumption or as a source of bioactive phytochemicals with potential industrial applications.

Stability of Bioactive Compounds in Broccoli as Affected by Cutting Styles and Storage Time.

Broccoli contains bioactive molecules and thus its consumption is related with the prevention of chronic and degenerative diseases. The application of wounding stress to horticultural crops is a common practice, since it is the basis for the fresh-cut produce industry. In this study, the effect of four different cutting styles (CSs) (florets (CS1), florets cut into two even pieces (CS2), florets cut into four even pieces (CS3), and florets processed into chops (CS4)) and storage time (0 and 24 h at 20 °C) on the content of bioactive compounds in broccoli was evaluated. Immediately after cutting, 5-O-caffeoylquinic acid and caffeic acid content increased by 122.4% and 41.6% in CS4 and CS2, respectively. Likewise, after storage, 3-O-caffeoylquinic acid and 5-O-caffeoylquinic acid increased by 46.7% and 98.2%, respectively in CS1. Glucoerucin and gluconasturtiin content decreased by 62% and 50%, respectively in CS3; whereas after storage most glucosinolates increased in CS1. Total isothiocyanates, increased by 133% immediately in CS4, and after storage CS1 showed 65% higher levels of sulforaphane. Total ascorbic acid increased 35% after cutting in CS2, and remained stable after storage. Results presented herein would allow broccoli producers to select proper cutting styles that preserve or increase the content of bioactive molecules.

Marine Bacteria, XLVII - Psychrotolerant Bacteria from Extreme Antarctic Habitats as Producers of Rare Bis- and Trisindole Alkaloids.

From the gastrointestinal tract of a fish dredged near the South Orkney Islands in Antarctica, we isolated the psychrotolerant bacterial strain T262, which belongs to the species Vibrio splendidus. Investigation of this strain led to the isolation of a series of 15 bis- and trisindole derivatives. Among them, six new indole alkaloids, namely, turbomycin C [4'-n-butoxyphenyl-bis(1H-indol-3-yl)methane, 1a], turbomycin D [4'-n-propoxyphenyl-bis(1H-indol-3-yl)methane, 1b], turbomycin E [4'-ethoxyphenyl-bis(1H-indol-3-yl)methane, 1c], turbomycin F [4'-methoxy-3',5'-dinitrophenyl-bis(1H-indol-3-yl)methane, 2], trisindolal (3a), and 4-(1H-indol-3-yl-sulfanyl)phenol (4). Another new bisindole derivative elucidated as 2-(indol-3-ylmethyl)-indol-3-ylethanol (7a) was obtained together with six known compounds from the psychrotolerant Arthrobacter psychrochitiniphilus strain T406, isolated from the excrement of penguins. Some of the isolated compounds showed activity against both gram-positive and gram-negative bacteria at 10 µg/paper disk. Trisindolal (3a) was active against the peronosporomycetes Botrytis cinerea and Phytophthora infestans, and some of the indole derivatives indicated promising cytotoxicity towards human tumor cell lines. By exhibiting a mean IC50 of 0.45 µg/mL (1.17 µM), trisindolal (3a) showed pronounced potency and selectivity in a panel of 11 human tumor cell lines derived from 10 different tumor histotypes.

Protective role of terpenes and polyphenols from three species of Oregano (Lippia graveolens, Lippia palmeri and Hedeoma patens) on the suppression of lipopolysaccharide-induced inflammation in RAW 264.7 macrophage cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Mexican oregano infusions have been traditionally used in México for the treatment of inflammation-related diseases, such as respiratory and digestive disorders, headaches and rheumatism, among others. Nevertheless, there is limited information regarding the phenolic compounds, terpenes and composition as well as biological activity of Mexican oregano. AIM OF THE STUDY: To determine the phenolic and terpene composition and to evaluate the anti-inflammatory potential of three species of Mexican oregano (Lippia graveolens (LG), Lippia palmeri (LP) and Hedeoma patens (HP)) in order to provide a scientific basis for their use. MATERIALS AND METHODS: We obtained methanol and chloroform extracts from dried oregano leaves of each species. We used LC-DAD-ESI-MS/MS and GC-MS to determine the phenolic and terpene profiles of the leaves, respectively. We evaluated anti-inflammatory potential by measuring the effect of Mexican oregano extracts on some pro-inflammatory mediators, such as nitric oxide (NO) and reactive oxygen species (ROS) using lipopolysaccharide(LPS)-stimulated RAW 264.7 macrophage cells and evaluating cyclooxygenase activity (COX-1, COX-2). RESULTS: Nine phenolic compounds (phenolic acids and flavonoids) and 22 terpenes (monoterpenes and sesquiterpenes) were detected in LG, LP and HP. We studied extracts from LG, LP and HP, and fractions from LG and LP in order to know their effect on some pro-inflammatory mediators. The phenolic and terpene extracts from LG, LP and HP exhibited significant inhibitory effect on ROS and NO production and mitochondrial activity in LPS-induced inflammation in RAW 264.7 macrophage cells. Nitric oxide production was also diminished by the terpene LG fraction LGF2 and the LP fractions LPF1, LPF2 and LPF3, confirming that both monoterpenes and sesquiterpenes are active compounds of oregano. Furthermore, the total extracts of LG, LP and HP exhibited non-selective inhibitions against the activity of the cyclooxygenases COX-1 and COX-2. CONCLUSIONS: Our results suggest that Lippia graveolens, Lippia palmeri and Hedeoma patens extracts have the potential to treat inflammatory diseases; their activity is mediated by polyphenols and terpenes. These findings support the claim for their traditional use in the treatment of inflammation-related diseases.

Plants as Biofactories: Postharvest Stress-Induced Accumulation of Phenolic Compounds and Glucosinolates in Broccoli Subjected to Wounding Stress and Exogenous Phytohormones.

Broccoli contains high levels of bioactive molecules and is considered a functional food. In this study, postharvest treatments to enhance the concentration of glucosinolates and phenolic compounds were evaluated. Broccoli whole heads were wounded to obtain florets and wounded florets (florets cut into four even pieces) and stored for 24 h at 20 °C with or without exogenous ethylene (ET, 1000 ppm) or methyl jasmonate (MeJA, 250 ppm). Whole heads were used as a control for wounding treatments. Regarding glucosinolate accumulation, ET selectively induced the 4-hydroxylation of glucobrassicin in whole heads, resulting in ∼223% higher 4-hydroxyglucobrassicin than time 0 h samples. Additionally, glucoraphanin was increased by ∼53% in whole heads treated with ET, while neoglucobrassicin was greatly accumulated in wounded florets treated with ET or MeJA, showing increases of ∼193 and ∼286%, respectively. On the other hand, although only whole heads stored without phytohormones showed higher concentrations of phenolic compounds, which was reflected in ∼33, ∼30, and ∼46% higher levels of 1,2,2-trisinapoylgentiobose, 1,2-diferulolylgentiobiose, and 1,2-disinapoyl-2-ferulolylgentiobiose, respectively; broccoli florets stored under air control conditions showed enhanced concentrations of 3-O-caffeoylquinic acid, 1,2-disinapoylgentiobiose, and 1,2-disinapoyl-2-ferulolylgentiobiose (∼22, ∼185, and ∼65% more, respectively). Furthermore, exogenous ET and MeJA impeded individual phenolics accumulation. Results allowed the elucidation of simple and effective postharvest treatment to enhance the content of individual glucosinolates and phenolic compounds in broccoli. The stressed-broccoli tissue could be subjected to downstream processing in order to extract and purify bioactive molecules with applications in the dietary supplements, agrochemical and cosmetics markets.

Protective Role of Flavonoids and Lipophilic Compounds from Jatropha platyphylla on the Suppression of Lipopolysaccharide (LPS)-Induced Inflammation in Macrophage Cells.

Seventeen polyphenols (e.g, apigenin, genistein, and luteolin glycosides) and 11 lipophilic compounds (e.g., fatty acids, sterols, and terpenes) were detected by LC-MS/MS-ESI and GC-MS, respectively, in Jatropha platyphylla. Extracts from pulp, kernel, and leaves and fractions were studied to know their effect on some pro-inflammatory mediators. Phenolic and lipophilic extracts showed significant inhibitory effects on ROS and NO production while not affecting mitochondrial activity or superoxide generation rate in lipopolysaccharide (LPS)-induced inflammation in RAW 264.7 macrophage cells. In addition, NO production was also diminished by lipophilic leaf fractions F1 and F2 with the latter fraction showing a greater effect and composed mainly of sterols and terpene. Furthermore, total extracts showed nonselective inhibitions against cyclooxygenase COX-1 and COX-2 activities. All together, these results suggest that J. platyphylla extracts have potential in treating inflammatory diseases and their activity is mediated by flavonoids and lipophilic compounds.

Combined effect of water loss and wounding stress on gene activation of metabolic pathways associated with phenolic biosynthesis in carrot.

The application of postharvest abiotic stresses is an effective strategy to activate the primary and secondary metabolism of plants inducing the accumulation of antioxidant phenolic compounds. In the present study, the effect of water stress applied alone and in combination with wounding stress on the activation of primary (shikimic acid) and secondary (phenylpropanoid) metabolic pathways related with the accumulation of phenolic compound in plants was evaluated. Carrot (Daucus carota) was used as model system for this study, and the effect of abiotic stresses was evaluated at the gene expression level and on the accumulation of metabolites. As control of the study, whole carrots were stored under the same conditions. Results demonstrated that water stress activated the primary and secondary metabolism of carrots, favoring the lignification process. Likewise, wounding stress induced higher activation of the primary and secondary metabolism of carrots as compared to water stress alone, leading to higher accumulation of shikimic acid, phenolic compounds, and lignin. Additional water stress applied on wounded carrots exerted a synergistic effect on the wound-response at the gene expression level. For instance, when wounded carrots were treated with water stress, the tissue showed 20- and 14-fold increases in the relative expression of 3-deoxy-D-arabino-heptulosanate synthase and phenylalanine ammonia-lyase genes, respectively. However, since lignification was increased, lower accumulation of phenolic compounds was detected. Indicatively, at 48 h of storage, wounded carrots treated with water stress showed ~31% lower levels of phenolic compounds and ~23% higher lignin content as compared with wounded controls. In the present study, it was demonstrated that water stress is one of the pivotal mechanism of the wound-response in carrot. Results allowed the elucidation of strategies to induce the accumulation of specific primary or secondary metabolites when plants are treated with water stress alone or when additional water stress is applied on wounded tissue. If the accumulation of a specific primary or secondary metabolite were desirable, it would be recommended to apply both stresses to accelerate their biosynthesis. However, strategies such as the use of enzymatic inhibitors to block the carbon flux and enhance the accumulation of specific compounds should be designed.

Protective Role of Ternatin Anthocyanins and Quercetin Glycosides from Butterfly Pea (Clitoria ternatea Leguminosae) Blue Flower Petals against Lipopolysaccharide (LPS)-Induced Inflammation in Macrophage Cells.

Twelve phenolic metabolites (nine ternatin anthocyanins and three glycosylated quercetins) were identified from the blue flowers of Clitoria ternatea by high-performance liquid chromatography diode array detection and electrospray ionization/mass spectrometry (HPLC-DAD-ESI/MS(n)). Three anthocyanins not reported in this species before show fragmentation pattern of the ternatin class. Extracts were fractionated in fractions containing flavonols (F3) and ternatin anthocyanins (F4). In general, C. ternatea polyphenols showed anti-inflammatory properties in lipopolysaccharide (LPS)-induced inflammation in RAW 264.7 macrophage cells with distinct molecular targets. Flavonols (F3) showed strong inhibition of COX-2 activity and partial ROS suppression. On the other hand, the ternatin anthocyanins (F4) inhibited nuclear NF-κB translocation, iNOS protein expression, and NO production through a non-ROS suppression mechanism. Accordingly, quercetin glycosides and ternatin anthocyanins from the blue flower petals of C. ternatea may be useful in developing drugs or nutraceuticals for protection against chronic inflammatory diseases by suppressing the excessive production of pro-inflammatory mediators from macrophage cells.

Rare prenylated isoflavones from Tephrosia calophylla.

Chemical re-examination of the roots of T. calophylla led to the isolation of four isoflavones, namely 2',4',5-trihydroxy-6,7-(2",2"-dimethylchromene)-8-gamma,gamma-dimethylallylisoflavone (1a, auriculatin), 2',5-dihydroxy-6,7-(2",2"-dimethylchromene)-4'-gamma,gamma-dimethylallyloxyisoflavone (2a, isoauriculatin), 3',4',5-trihydroxy-6,7-(2",2"-dimethylchromene)-8-gamma,gamma-dimethylallylisoflavone (1b, auriculasin), and 3',5-dihydroxy-6,7-(2",2"-dimethylchromene)-4'-gamma,gamma-dimethylallyl-oxyisoflavone (2b, isoauriculasin). Auriculasin and isoauriculasin are reported for the first time with full NMR assignments. All metabolites were fully characterized by 1D and 2D NMR techniques including COSY, HSQC, HMBC experiments, mass spectra and in part on the basis of X-ray diffraction. Accurate 'invariom' refinements with aspherical scattering factors were carried out for 1a, 1b and 2a.