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Parkinson's disease (PD) is a neurodegenerative disorder characterized by nigrostriatal degeneration and the spreading of aggregated forms of the presynaptic protein α-synuclein (aSyn) throughout the brain. PD patients are currently only treated with symptomatic therapies, and strategies to slow or stop the progressive neurodegeneration underlying the disease's motor and cognitive symptoms are greatly needed. The time between the first neurobiochemical alterations and the initial presentation of symptoms is thought to span several years, and early neuroprotective dietary interventions could delay the disease onset or slow PD progression. In this study, we characterized the neuroprotective effects of isoflavones, a class of dietary polyphenols found in soy products and in the medicinal plant red clover (Trifolium pratense). We found that isoflavone-rich extracts and individual isoflavones rescued the loss of dopaminergic neurons and the shortening of neurites in primary mesencephalic cultures exposed to two PD-related insults, the environmental toxin rotenone and an adenovirus encoding the A53T aSyn mutant. The extracts and individual isoflavones also activated the Nrf2-mediated antioxidant response in astrocytes via a mechanism involving inhibition of the ubiquitin-proteasome system, and they alleviated deficits in mitochondrial respiration. Furthermore, an isoflavone-enriched soy extract reduced motor dysfunction exhibited by rats lesioned with the PD-related neurotoxin 6-OHDA. These findings suggest that plant-derived isoflavones could serve as dietary supplements to delay PD onset in at-risk individuals and mitigate neurodegeneration in the brains of patients.

The effects of protein carrier and drying technique on the concentration and bioaccessibility of lipophilic compounds (lutein, β-carotene, chlorophylls a and b) and hydrophilic flavonoids in freeze-dried (FD) or spray-dried (SD) spinach juice and protein-spinach particles were investigated. Carotenoid and chlorophyll contents were highest in FD spinach juice without protein (147 and 1355 mg/100 g, respectively). For both SD and FD protein-spinach particles, SPI best protected carotenoids and chlorophylls (123 and 1160 mg/g, respectively), although the bioaccessibility of lipophilic compounds in WPI particles was higher than SPI particles (p 0.05). For flavonoids, the drying technique was more important than the type of carrier, since FD particles had higher total flavonoids than SD. However, SD particles had higher bioaccessibility for most flavonoids (40-90 %) compared to FD (20 %). The drying method and protein carrier can be designed to produce protein-spinach ingredients with desired concentration of compounds and bioaccessibility.

Acai fruit is recognized for its health promoting properties. However, there is still a need to address the effects of industrial processing on this fruit. In this study, phenolic content, anti-inflammatory properties and dermal wound repair properties of 20 acai samples, before and after industrial processing, from various Amazon regions were investigated. Acai pulp was rich in total phenolics (18.9–58.8 mg g−1) and proanthocyanins (9.8–43.1 mg g−1), but contained trace anthocyanins (up to 0.1 mg g−1). Industrially processed samples lost substantial amounts of proanthocyanidins (up to 83.2%), while the anthocyanins inherently present were greatly enriched after processing (20-fold higher). Non-processed acai pulp extracts protected against early inflammation response which was correlated with proanthocyanidins, by significantly inhibiting nitric oxide production and suppressing pro-inflammatory gene expression including interleukin-1β, cyclooxygenase-2, nitric oxide synthase, and interleukin-6. The promotion of dermal wound repair of acai seed and pulp extracts was mainly contributed by anthocyanins and other bioactive compounds. The anti-inflammatory effect was diminished but wound healing effect was retained after pulp processing, suggesting the processing technology needs to be improved to maintain biological properties of acai fruit.

The World Health Organization estimates that 7 million people die every year due to pollution exposure. Among the different pollutants to which living organism are exposed, ozone (O3) represents one of the most toxic, because its location which is the skin is one of the direct tissues exposed to the outdoor environment. Chronic exposure to outdoor stressors can alter cutaneous redox state resulting in the activation of inflammatory pathways. Recently, a new player in the inflammation mechanism was discovered: the multiprotein complex NLRP1 inflammasome, which has been shown to be also expressed in the skin. The topical application of natural compounds has been studied for the last 40 years as a possible approach to prevent and eventually cure skin conditions. Recently, the possibility to use blueberry (BB) extract to prevent pollution-induced skin toxicity has been of great interest in the cosmeceutical industry. In the present study, we analyzed the cutaneous protective effect of BB extract in several skin models (2D, 3D, and human skin explants). Specifically, we observed that in the different skin models used, BB extracts were able to enhance keratinocyte wound closure and normalize proliferation and migration responses previously altered by O3. In addition, pretreatment with BB extracts was able to prevent ozone-induced ROS production and inflammasome activation measured as NRLP1-ASC scaffold formation and also prevent the transcripts of key inflammasome players such as CASP1 and IL-18, suggesting that this approach as a possible new technology to prevent cutaneous pollution damage. Our data support the hypothesis that BB extracts can effectively reduce skin inflammation and be a possible new technology against cutaneous pollution-induced damage.

The present study was designed to characterize the polyphenols isolated from Acacia mearnsii bark crude extract (B) and fractions (B1-B7) obtained by high-speed counter-current chromatography (HSCCC) and evaluate their anti-inflammatory and carbolytic enzymes (α-glucosidase and α-amylase) inhibitory activities. Fractions B4, B5, B6, B7 (total phenolics 850.3, 983.0, 843.9, and 572.5 mg·g-1, respectively; proanthocyanidins 75.7, 90.5, 95.0, and 44.8 mg·g-1, respectively) showed significant activities against reactive oxygen species (ROS), nitric oxide (NO) production, and expression of pro-inflammatory genes interleukin-1β (IL-1β) and inducible nitric oxide synthase (iNOS) in a lipopolysaccharide (LPS)-stimulated mouse macrophage cell line RAW 264.7. All the extracts suppressed α-glucosidase and α-amylase activities, two primary enzymes responsible for carbohydrate digestion. A. mearnsii bark samples possessed significantly stronger inhibitory effects against α-glucosidase enzyme (IC50 of 0.4-1.4 μg·mL-1) than the pharmaceutical acarbose (IC50 141.8 μg·mL-1). B6 and B7 (IC50 17.6 and 11.7 μg·mL-1, respectively) exhibited α-amylase inhibitory activity as efficacious as acarbose (IC50 15.4 μg·mL-1). Moreover, B extract, at 25 µg·mL-1, significantly decreased the non-mitochondrial oxidative burst that is often associated with inflammatory response in human monocytic macrophages.

Over 500 unique anthocyanins have been described to date, which vary in color, antioxidant, light-attenuating, and antimicrobial properties. Identification of anthocyanin chemical structure may therefore serve as an important clue to their in situ function in plants. We characterized the histological and chemical structures of anthocyanins associated with diverse leaf color patterns in the terrestrial orchid, Tipularia discolor, as a step towards understanding their ultimate function. Tipularia discolor produces a single wintergreen leaf in autumn, which is drab brown in color during expansion. Upper (adaxial) surfaces of fully-expanded leaves may be green, purple-spotted, or solid purple, while lower (abaxial) surfaces are bright magenta. Our results showed that the same three cyanidin 3,7,3'-triglucosides, in similar concentrations and proportions, accounted for coloration in each of these cases, and that different colors result from differences in histological location of anthocyanins (i.e. abaxial/adaxial epidermis, mesophyll). Anthocyanins with 3,7,3' linkage positions are rare in plants, occurring only within the orchid subfamily Epidendroideae, to which Tipularia belongs. These results are important to the discussion of anthocyanin structure-function because they serve as a reminder that 1) plants may employ the same anthocyanins in different anatomical locations to achieve a broad range of colors (and potentially adaptive functions), and 2) anthocyanin chemical structure and anatomical location are influenced by phylogenetic inertia, as well as natural selection.

Phytochemical and bioactivity analyses of pistachio hulls revealed the presence of anacardic acids (3198 mg/100 g), fatty acids (1500 mg/100 g), and phytosterols (192 mg/100 g) as major components. Carotenoids (4.93 mg/100 g), chlorophylls (10.27 mg/100 g), tocopherols (8.83 mg/100 g), and three triterpene acids (mangiferolic, isomangiferolic and mangiferonic acids) were characterized. A polar (P) extract contained quercetin-3- O -glucoside (6.27 mg/g), together with smaller concentrations of quercetin, myricetin and luteolin flavonoids, accounting for 5.53 mg/g. Gallotannins and other phenolic compounds esterified with a gallic acid moiety characterized the P extract. P extract potently inhibited the release of nitric oxide (NO) and reactive oxygen species (ROS) in lipopolysaccharide-stimulated RAW 264.7 macrophage cells. The mRNA expression levels of the anti-inflammatory cytokine COX-2 were significantly inhibited by fractions P2-P5, while IL-6 was only inhibited by fraction P3. Moreover, the P extract significantly decreased the non-mitochondrial oxidative burst associated with inflammatory response in macrophages.

Stability of protein-polyphenol aggregate particles, created by complexing polyphenols from blueberry and muscadine grape pomaces with a rice-pea protein isolate blend, was evaluated in an in vitro gastrointestinal model. Recovery index (RI; % total phenolics present post-digestion) was 69% and 62% from blueberry and muscadine grape protein-polyphenol particles, compared to 23% and 31% for the respective pomace extracts. Anthocyanins RI was 52% and 42% from particles (6% and 13% from pomace extracts), and proanthocyanidins RI was 77% and 73% from particles (25% and 14% from pomace extracts), from blueberry and grape, respectively. Protein-polyphenol particle digests retained 1.5 to 2-fold higher antioxidant capacity and suppressed the expression of pro-inflammatory cytokines, iNOS, IL6, and IL1β, compared to unmodified extract digests, which only suppressed IL6. Protein-polyphenol particles as a delivery vehicle in foods may confer better stability during gastrointestinal transit, allow protected polyphenols to reach the gut microbiota, and preserve polyphenol bioactivity.

A sensitive and straightforward LC-IT-TOF-MS method was validated for the profiling and simultaneous quantification of anthocyanins, flavan-3-ols, flavonols, phenolic acids, and resveratrol in blueberry genotypes with fruit color ranging from deep purple (Vaccinium angustifolium) to various shades of pink (crosses of V. corymbosum, V. darrowii, and V. ashei). Standard calibration curves were linear for all analytes with correlation coefficients >0.99. The relative standard deviation for intra- and inter-day precision was lower than 10%. The method allowed an easy and selective identification and quantification of phenolics in blueberries with divergent profiles. The in vitro antioxidant assay results were strongly correlated with total phenolics and total anthocyanin content. Lowbush blueberry extracts (50 μg/mL) reduced ROS and NO production, and inhibited the transcription of the proinflammatory cytokines IL-6β, COX2, iNOS, and IL-6 in the in vitro assays at much lower concentrations than pink fruited berries (250 μg/mL).

Particulate colloidal aggregate food ingredients were prepared by complexing wheat flour, chickpea flour, coconut flour and soy protein isolate with aqueous wild blueberry pomace extracts, then spray drying, freeze drying, or vacuum oven drying to prepare dry, flour-like matrices. Physico-chemical attributes, phytochemical content and stability during storage were compared. Eighteen anthocyanins peaks were identified for samples. Spray dried matrices produced with soy protein isolate had the highest concentration of polyphenols (156.2 mg GAE/g) and anthocyanins (13.4 mg/g) and the most potent DPPH scavenging activity (714.1 μmoles TE/g). Spray dried blueberry polyphenols complexed with protein were protected from degradation during 16 weeks at 4 °C and 20 °C. Soy protein isolate more efficiently captured and stabilized wild blueberry pomace phytochemicals than other protein sources. Overall, spray drying the blueberry extracts complexed with protein proved to be an environment-friendly strategy to produce stable functional ingredients with multiple applications for the food industry

Preparative isolation of complex mixtures of compounds from walnut polar extracts was established by a combination of high-speed counter-current chromatography (HSCCC) and electrospray ionization-ion trap-time of flight mass spectrometry (ESI-IT-TOF-MS). Compounds were isolated after a solvent optimisation selection based on solute distribution in a biphasic solvent system. Isolation was achieved through one or two successive HSCCC runs, and final purification on Sephadex LH-20. Isolated compounds included ellagitannins (1-11), gallic acid (12), dicarboxylic acid glucosides (13-15), hydrojuglone glucoside (16), catechin (17), procyanidin B2 (18), and megasterone glucosides (19-20). Praecoxin D (4) was isolated for the first time from walnut, while praecoxin A methyl ester (5) and glansreginin A n-butyl ester (14) are newly identified compounds. The purity and identity of isolated compounds were confirmed by NMR and HPLC-ESI-MS/MS. These results provided a foundation for in depth characterisation of walnut compounds and offered an efficient strategy for isolation of potentially health-relevant phytochemicals from walnuts.

Overconsumption of energy dense foods and sedentary lifestyle are considered as major causes of obesity-associated insulin resistance and abnormal glucose metabolism. Results from both cohort studies and randomized trials suggested that anthocyanins from berries may lower metabolic risks, however these reports are equivocal. The present study was designed to examine effects of six berries with structurally diverse anthocyanin profiles (normalized to 400 µg/g total anthocyanin content) on development of metabolic risk factors in the C57BL/6 mouse model of polygenic obesity. Diets supplemented with blackberry (mono-glycosylated cyanidins), black raspberry (acylated mono-glycosylated cyanidins), blackcurrant (mono- and di-glycosylated cyanidins and delphinidins), maqui berry (di-glycosylated delphinidins), Concord grape (acylated mono-glycosylated delphinidins and petunidins), and blueberry (mono-glycosylated delphinidins, malvidins, and petunidins) showed a prominent discrepancy between biological activities of delphinidin/malvidin-versus cyanidin-type anthocyanins that could be explained by differences in their structure and metabolism in the gut. Consumption of berries also resulted in a strong shift in the gastrointestinal bacterial communities towards obligate anaerobes that correlated with decrease in the gastrointestinal luminal oxygen and oxidative stress. Further work is needed to understand mechanisms that lead to nearly anoxic conditions in the gut lumens, including the relative contributions of host, diet and/or microbial oxidative activity, and their implication to human health.

A comprehensive phytochemical analysis was conducted on pistachios to identify the differential contributions of skin and kernel phytochemicals to in vitro bioactivity. Qualitative and quantitative analyses of skin and kernel non-polar extracts (SNP and KNP, respectively) indicated that the major components are fatty acids (696.36 and 879.70 mg g−1), phytosterols (16.08 and 4.28 mg g−1), and γ-tocopherol (304.17 and 397.10 μg g−1). Analysis of the skin and kernel polar extracts (SP and KP, respectively) showed that skin accumulated higher levels of phenolic compounds, especially flavan-3-ols, compared to the kernel. An (epi)catechin hexoside was the major component in SP and KP (9.8 mg g−1 and 3.3 mg g−1, respectively). Flavan-3-ols with different degrees of polymerization were detected in SP, but only the monomers were identified in the KP. Quercetin glycosides were the major flavonols present in both SP and KP. Bioassays with 3T3L1 mouse adipocytes demonstrated that all extracts decreased lipid accumulation, with SNP demonstrating the highest activity (17% inhibition). Bioassay guided fractionation of SNP indicated that the lipolytic activity was highest in the fraction consisting of linoleic acid (20%), linolenic acid (10%), and β-sitosterol (50%). Radical scavenging assays indicated that all pistachio extracts significantly inhibited ROS, while SP was the most inhibiting to NO production in LPS-stimulated RAW 264.7 macrophages. Gene expression profiles associated with inflammation (IL6, iNOS, and COX2) were characterized in the LPS-stimulated RAW264.7 macrophages after treatment with pistachio extracts. SP and KP were the most potent to inhibit the expression of COX2. The SNP had the strongest effect in decreasing non-mitochondrial oxidative burst associated with inflammatory response in macrophages.

The present study demonstrated that defatted soybean flour (DSF) can sorb polyphenols from blueberry and cranberry juices while separating them from sugars. Depending on DSF concentration and juice dilution, the concentration of blueberry anthocyanins and total polyphenols sorbed to DSF ranged from 2–22 mg/g and 10–95 mg/g, respectively while the concentration of anthocyanins and proanthocyanidins in cranberry polyphenol-enriched DSF ranged from 2.5–17 mg/g and 21–101 mg/g, respectively. Blueberry polyphenols present in one serving of fresh blueberries (73 g) were delivered in just 1.4 g of blueberry polyphenol-enriched DSF. Similarly, one gram of cranberry polyphenol-enriched DSF delivered the amount of proanthocyanidins available in three 240 ml servings of cranberry juice cocktail. The concentration of blueberry anthocyanins and total polyphenols eluted from DSF remained constant after 22 weeks of incubation at 37 °C, demonstrating the high stability of the polyphenol–DSF matrix. LC–MS analysis of eluates confirmed that DSF retained major cranberry and blueberry polyphenols intact. Blueberry polyphenol-enriched DSF exhibited significant hypoglycaemic activities in C57bl/6J mice, and cranberry polyphenol-enriched DSF showed antimicrobial and anti-UTI activities in vitro , confirming its efficacy. The described sorption process provides a means to create protein-rich food ingredients containing concentrated plant bioactives without excess sugars, fats and water that can be incorporated in a variety of scientifically validated functional foods and dietary supplements.

The development of new antiplasmodial drugs is of primary importance due to the growing problem of multi-drug resistance of malaria parasites. Spilanthes acmella, a plant traditionally used for the treatment of toothache, was targeted as a lead for its potential antiplasmodial activity. A systematic approach for investigating a suitable centrifugal partition chromatography (CPC) solvent system for N-alkylamides separation was reported. The partition behavior of three N-alkylamides has been studied using several biphasic solvent mixtures in search of an adequate CPC solvent system for this class of compounds. Major N-alkylamides in S. acmella were isolated from a methanolic crude extract of flowers by CPC with the solvent system heptanes-ethyl acetate-methanol-water (3:2:3:2, v/v/v/v). Four N-alkylamides were purified and the structures were illustrated by electrospray ionization-ion trap-time of flight-mass spectrometry (ESI-IT-TOF-MS), ¹H nuclear magnetic resonance (¹H NMR) and ¹³C nuclear magnetic resonance (¹³C NMR). The CPC fractions, which contained natural mixtures of phytochemicals, demonstrated significantly higher antiplasmodial activity compared to corresponding purified N-alkylamides, thus suggesting that interactions between these N-alkylamides may potentiate antiplasmodial bioactivity.

Chromatographic fractionations of the toluene extract of the heartwood of Excoecaria parvifolia collected in Australia resulted in the isolation of 12 beyerane diterpenes (1–12), and the triterpene, lupeol. Four of the isolated diterpenoids (5–7 and 12) have unusual structures: ent-3-oxa-beyer-15-en-2-one, (5); ent-15,16-epoxy-2-hydroxy-19-norbeyer-1,4-dien-3-one (6); methyl ent-2,4-seco-15,16-epoxy-4-oxo-3,19-dinorbeyer-15-en-2-oate (7); and ent-2,17-dihydroxy-19-norbeyer-1,4,15-trien-3-one (12). The structures were established by spectroscopic analyses, NMR data comparisons with similar diterpenes, and chemical correlations. All the diterpenes are assumed to have the same absolute configuration as the co-occurring (+)-stachenol (4). Diosphenol 2 and nor-lactone 5 exhibited significant potency in bioassays for cytotoxic activity against leukemia cells (L1210). Plausible biosynthetic pathways are proposed to explain the origin of the diterpene metabolites.

Quinoa (Chenopodium quinoa Willd.) contains high levels of biologically active phytoecdysteroids, which have been implicated in plant defense from insects, and have shown a range of beneficial pharmacological effects in mammals. We demonstrated that the most prevalent phytoecdysteroid, 20-hydroxyecdysone (20HE), was secreted (leached) from intact quinoa seeds into water during the initial stages of seed germination. Leaching efficiency was optimized by ethanol concentration (70% ethanol), temperature (80°C), time (4 h), and solvent ratio (5 ml/g seed). When compared to extraction of macerated seeds, the leaching procedure released essentially all the 20HE available in the seeds (491 μg/g seed). The optimized quinoa leachate (QL), containing 0.86% 20HE, 1.00% total phytoecdysteroids, 2.59% flavonoid glycosides, 11.9% oil, and 20.4% protein, significantly lowered fasting blood glucose in obese, hyperglycemic mice. Leaching effectively releases and concentrates bioactive phytochemicals from quinoa seeds, providing an efficient means to produce a food-grade mixture that may be useful for anti-diabetic applications.

We describe a series of four patients who presented with ‘high globus pharyngeus’, who all had an abnormally curled epiglottis tip touching and indenting the tongue base. The actual incidence of ‘curling epiglottis’, as well as the potential impact of this variation in persistent globus symptoms, is not known. We therefore describe for the first time a series of patients with this anatomical variant of the epiglottis, each of whom experienced unresolved globus symptoms despite receiving intensive medical treatment. In the literature, the success rate for improvement in symptoms following medical treatment ranges from 68 to 80 per cent.Following CO2 laser partial epiglottectomy, all four patients experienced complete relief of their symptoms.We advocate consideration of this treatment for high globus pharyngeus that fails to respond to conservative treatment, in cases with proven curled epiglottis on endoscopic examination.

Eryngium foetidum L. (Apiaceae) is a traditional herb that has been used for numerous medicinal applications, including as a treatment for parasitic infections, especially in the Neotropics from where it originates.This study evaluates the in vitro leishmanicidal and cytotoxicity activities of isolated compounds based on a bioassay-guided fractionation approach.Defatted aerial parts of E. foetidum were subjected to extraction with methanol followed by partitioning with n-hexane, ethyl acetate and 50% methanol. Then, the first two fractions were subsequently fractionated by column chromatography and HPLC. Compound identity was confirmed by mass spectrometry and NMR spectroscopy. Leishmania tarentolae (promastigotes) and L. donovani (amastigotes) were used as testing parasites. L6 rat myoblasts were used for cytotoxicity. All extracts and fractions were tested at 20 μg/mL.The initial methanol extract showed 20% growth inhibition of L. tarentolae. Then, the n-hexane and ethyl acetate fractions were also active showing approximately 40% growth inhibition. From these two fractions, the following compounds were isolated: lasidiol p-methoxybenzoate (1), a daucane sesquiterpene; and 4-hydroxy-1,1,5-trimethyl-2-formyl-cyclohexadien-(2,5)-[α-acetoxymethyl-cis-crotonate] (2), a terpene aldehyde ester derivative. Compound 1 inhibited the growth of both L. tarentolae and L. donovani with IC₅₀ values of 14.33 and 7.84 μM, respectively; and showed no cytotoxicity (IC₅₀ 50 μM). Compound 2 was inactive in the L. tarentolae assay (IC₅₀ 50 μM).This study presented the bioassay-guided fractionation with the leishmanicidal and cytotoxicity activities of two compounds isolated for the first time from an Eryngium species.

Sweetpotato phytochemical content was evaluated in four genotypes (NCPUR06-020, Covington, Yellow Covington, and NC07-847) at harvest and after curing/storage for 4 or 8 months. Curing and storage for up to 8 months did not significantly affect total phenolic content in Covington, Yellow Covington, and NC07-847, however for NCPUR06-020, a purple-fleshed selection, total phenolic content declined mainly due to anthocyanin degradation during storage. Covington had the highest carotenoid content at harvest time (281.9 μg/g DM), followed by NC07-847 (26.2 μg/g DM), and after 8 months, total carotenoids had increased by 25% and 50%, respectively. Antioxidant activity gradually declined during storage, and freshly harvested sweetpotatoes also demonstrated higher anti-inflammatory capacity as gauged by inhibition of lipopolysaccharide-induced reactive oxygen species (ROS) in SH-SY5Y cells. Gradual changes in sweetpotato phytochemical content and antioxidant and anti-inflammatory capacity were noted during normal long-term storage, but the specific effects were genotype-dependent.

The TNO intestinal model (TIM-1) of the human upper gastrointestinal tract was used to compare intestinal absorption/bioaccessibility of blueberry anthocyanins under different digestive conditions. Blueberry polyphenol-rich extract was delivered to TIM-1 in the absence or presence of a high-fat meal. HPLC analysis of seventeen anthocyanins showed that delphinidin-3-glucoside, delphinidin-3-galactoside, delphinidin-3-arabinoside and petunidin-3-arabinoside were twice as bioaccessible in fed state, whilst delphinidin-3-(6″-acetoyl)-glucoside and malvidin-3-arabinoside were twice as bioaccessible under fasted conditions, suggesting lipid-rich matrices selectively effect anthocyanin bioaccessibility. TIM-1 was fed blueberry juice (BBJ) or blueberry polyphenol-enriched defatted soybean flour (BB-DSF) containing equivalent amounts of free or DSF-sorbed anthocyanins, respectively. Anthocyanin bioaccessibility from BB-DSF (36.0 ± 10.4) was numerically, but not significantly, greater than that from BBJ (26.3 ± 10.3). Ileal efflux samples collected after digestion of BB-DSF contained 2.8-fold more anthocyanins than same from BBJ, suggesting that protein-rich DSF protects anthocyanins during transit through upper digestive tract for subsequent colonic delivery/metabolism.

The traditionally used antimalarial plant, Datisca glomerata (C. Presl) Baill, was subjected to antiplasmodial assay guided fractionation. This led to the isolation of seven cucurbitacin glycosides, datiscosides I–O, along with two known compounds, datiscoside and datiscoside B, from the aerial parts of D. glomerata . Their structures and relative stereochemistry were determined on the basis of mass spectrometry, 1D and 2D NMR spectroscopic data. Antiplasmodial IC 50 values were determined for all isolated compounds against a chloroquine sensitive strain of Plasmodium falciparum (D10), which were also evaluated in vitro for their antileishmanial activity against Leishmania tarentolae . Cytotoxicity was evaluated against rat skeletal muscle cells (L6) and Chinese ovarian hamster cells (CHO). The antiplasmodial activity of the compounds was moderate and ranged from 7.7 to 33.3 μM. None of the compounds showed appreciable antileishmanial activity. The compounds displayed cytotoxicity against L6 but not CHO mammalian cells.

We used a murine model of type II diabetes, which reproduces the major features of the human disease, and a number of cellular models to study the antidiabetic effect of ANC, a standardised anthocyanin-rich formulation from maqui berry (Aristotelia chilensis). We also isolated delphinidin 3-sambubioside-5-glucoside (D3S5G), a characteristic anthocyanin from maqui berry, and studied its antidiabetic properties. We observed that oral administration of ANC improved fasting blood glucose levels and glucose tolerance in hyperglycaemic obese C57BL/6J mice fed a high fat diet. In H4IIE rat liver cells, ANC decreased glucose production and enhanced the insulin-stimulated down regulation of the gluconeogenic enzyme, glucose-6-phosphatase. In L6 myotubes ANC treatment increased both insulin and non-insulin mediated glucose uptake. As with the ACN, oral administration of pure D3S5G dose-dependently decreased fasting blood glucose levels in obese C57BL/6J mice, and decreased glucose production in rat liver cells. D3S5G also increased glucose uptake in L6 myotubes and is at least partially responsible for ANC's anti-diabetic properties.

Alkaloids from the stem bark of Geissospermum vellosii possess a variety of therapeutic properties including antimalarial activities, activity as a sexual stimulant and inhibition of the proliferation of HIV and herpes viruses. Methods currently used to isolate the active components from G. vellosii are time-consuming, labor intensive, and result in low recovery. In addition, there is a lack of sensitive and accurate analytical methods for the structural characterization and identification of alkaloid components in minor quantities. A combination of high performance counter-current chromatography and ESI tandem mass spectrometry (MS(n)) was established to isolate alkaloids from the stem bark of G. vellosii, and study their electrospray ionization mass spectrometry fragmentation behavior. Five indole alkaloids were successfully isolated and identified by nuclear magnetic resonance and mass spectrometry. The multi-stage tandem mass spectrometric data were used to study their fragmentation pattern and set a model for detailed structure characterization of related indole alkaloids. The presence of the even mass fragment ion suggestive of an odd number of nitrogen at m/z 144 corresponding to C(10)H(9)N was characteristic to indole alkaloids. The results of the experiments demonstrated that the combination of high performance counter current chromatography and ESI-MS(n) is a sensitive, selective and effective approach for rapid isolation and characterization of alkaloids from G. vellosii.

Objective:This study aimed to evaluate voice and quality of life after transoral laser resection of early glottic carcinoma.Methods:We studied 19 patients undergoing transoral laser resection of tumour stage (T) one or T2 glottic carcinoma. Laryngeal function was evaluated by video-stroboscopy, vocal function by the Voice Symptom Scale, the grade-roughness-breathiness-asthenia-strain scale and objective phoniatric assessment, and quality of life by the University of Washington Quality of Life questionnaire.Results:Patients’ glottic carcinoma tumour-node-metastasis (TNM) staging was T1 N0 M0 in 14 patients and T2 N0 M0 in five. Overall voice grade, roughness and breathiness were mild to moderate in 84 per cent; asthenia and voice strain were more uniformly distributed, with 15 per cent of patients having normal voice quality. Eight patients developed a glottic web post-operatively; anterior commissure web was significantly associated with worse voice grade (p = 0.05). Seven patients (47 per cent) had a ‘mucosal wave’ on the operated vocal fold; this was significantly associated with less strain on phonation (p = 0.05). Voice Symptom Scale score was low overall (15 patients (78.9 per cent) scored less than 30). The fundamental frequency and frequency irregularity were normal in nine patients (47.3 per cent); the closed quotient was normal in six (31.5 per cent). The averaged quality of life score was ≥90 in 14 patients (73.7 per cent); 18 (94.7 per cent) felt their health-related quality of life was either the same or better post-operatively; and overall quality of life was positive in all.Conclusion:Transoral laser resection of T1 and T2 glottic carcinoma enables adequate tumour tissue excision with preservation of acceptable vocal function. Most patients’ post-operative quality of life is very good. Anterior commissure web formation is associated with poorer vocal function.

From the methylene chloride extract of Ehretia ovalifolia leaves, a new flavone derivative named ovalifolin [3-(3-methyl-1-butenyl)-6-methoxy-5,7,4'-trihydroxy flavone] has been isolated and identified together with the known flavone aglycones apigenin, luteolin and the highly methoxylated flavanol araneosol. The structures of these metabolites have been established on the basis of chemical, chromatographic and spectral methods.

In comparison to the well-recognized adaptogenic herb Rhodiola rosea, phytochemical constituents of two other Rhodiola species (R. heterodonta and R. semenovii) were elucidated and characterized. Two major phytochemical groups; phenolic and/or cyanogenic glycosides and proanthocyanidins, were isolated and identified in the three species. Chemical similarities among the three species were observed; however, each species displayed differences in phytochemical constituents. R. heterodonta contained a newly detected phenylethanoid glycoside, heterodontoside, in addition to the known compounds tyrosol, viridoside, salidroside, and rhodiocyanoside A. Both R. heterodonta and R. rosea contained phenylethanoid/propanoid compounds that were not detected in R. semenovii. For R. semenovii, the cyanogenic glucosides rhodiocyanoside A and lotaustralin were detected. Although the three species have proanthocyanidins composed of (-)-epigallocatechin and its 3-O-gallate esters in common, the degree of polymerization greatly differed between them. In contrast to R. heterodonta and R. semenovii, R. rosea has higher molecular weight polymeric proanthocyanidins. This study resulted in the identification and isolation of phytochemical constituents for direct cross-comparison between three Rhodiola species of medicinal and pharmacological value.

The sensation of a foreign body in the throat when swallowing is a common and very discomforting symptom for those who experience it. Common causes are gastroesophageal reflux disease and possibly upper oesophageal dysmotility. This complaint may be dismissed as psychogenic when a cause for the problem is not readily apparent.We present two cases in which the patient had a foreign body sensation in the throat, associated with a dry swallow. The most helpful diagnostic procedure was careful examination of the hypopharynx and neck and flexible fibre-optic rhinolaryngoscopy. One case was treated with surgery of the neck and larynx to trim the medially projecting superior cornu of the thyroid cartilage. Surgery was successful in eliminating the symptoms.Although an uncommon finding, our experience is that symptoms due to a medially displaced thyroid cornu should be considered in the differential diagnosis as an important and surgically correctable cause of persistent foreign body sensation at the hyoid level in the neck.

Extraction of Petalostigma pubescens heartwood followed by chromatographic purifications and crystallizations afforded five tricyclic diterpenes: 5,9-syn-rosanes petalostigmones A and B (1 and 2), the erythroxylane petalostigmone C (3), the norditerpene lactone pubescenone (4), and the known ent-cleistanthane diterpene (−)-sonderianol (5). The structures and relative stereochemistry were elucidated by means of spectroscopic methods, chemical correlations, and, in the cases of 1 and 4, by X-ray crystallographic analyses. The new isolates 1–4 are assumed to belong to the same absolute configurational family (9αCH3) of ent-pimarane-derived diterpenes as the known co-occurring (−)-5 (10αCH3). Biogenetic schemes originating from a common ent-copalyl diphosphate intermediate are presented to rationalize the structures of these natural products. A novel ring contraction–ring expansion mechanism is suggested to account for the 7-membered B ring of pubescenone. Compounds 1–5 were evaluated for their cytotoxicity; sonderianol (5) showed the highest activity against mouse leukemia cell lines L1210, P388 and mouse liver cancer cells HEPA1c1c7.

Cystic lesions of the larynx are most commonly benign. However, we report a case of non-Hodgkin’s lymphoma (NHL) of the larynx which presented as a supraglottic cyst. This form of presentation has not been previously described in the medical literature.

A 50 year old woman with a 20 year history of Crohn's disease underwent laparotomy which revealed extensive disease in the small and large bowel, and this was resected. Gross examination of the resected bowel showed features of Crohn's disease as well as a polypoid tumour in the caecum. Histopathological examination of the tumour showed it to be an infiltrating mixed adenocarcinoma/carcinoid tumour arising in a tubulovillous adenoma. Random sampling of the rest of the bowel affected by Crohn's disease also showed a focus of dysplasia and adenomatous change. It is suggested that Crohn's disease may have played a part in the pathogenesis of the tumour.

A new eudesmanolide, 1beta,9alpha-dihydroxyeudesm-3-en-5beta,6alpha,7alpha,11alphaH-12,6-olide (1), was isolated from Crataegus flava fruits. Its structure was determined by spectroscopic methods.

The aerial parts of the Egyptian herb Cornulaca monacantha have yielded two new galloyltannin analogs (named monacanthin A and monacanthin B) characterized by protoctechuoyl moiety at C-6, and also a new flavonol glycoside, quercetin-4′- O -β- d -galactoside. The structures of the new compounds were established by conventional analytical methods and confirmed by spectral analyses. Two known flavonoids, luteolin-7- O -rhamnoside, luteolin-7- O -glucoside, and two known gallotannins, penta- O -galloyl-β- d -glucose and 1,2,3,6-tetra- O -galloyl-β- d -glucose were also isolated.

Eversion of the rectum during restorative proctocolectomy with stapled ileal pouch-anal anastomosis (IPAA) remains a controversial surgical manoeuvre because of concern that it may impair anal sphincter function and adversely affect outcome. We have reviewed the long-term results in 41 patients whose operation included formation of a 20 cm J-pouch with stapled IPAA by the technique of rectal eversion. At median follow-up of 4 years (range 1–6 years), 4 pouches (10%) had been removed (2 for pelvic sepsis, 1 for rectovaginal fistula and 1 for Crohn's disease). In 34 patients with functioning pouches in situ, median stool frequency was 5 per 24 h (range 2–10). 11 patients (33%) regularly had to evacuate their pouch at night and 4 (12%) used antidiarrhoeal medication. No patients reported major incontinence; 2 (6%) had minor leakage, and in another 2 minor leakage had now ceased. 4 patients had had episodes of pouchitis. These favourable results offer no support for the contention that rectal eversion substantially worsens the long-term results of restorative proctocolectomy.

EDITOR,—T C B Dehn has commented on the problems of accruing patients into randomised controlled trials, noting the understandable reluctance of patients to enter some trials (“I want the best treatment, doctor”) in which the patient's outcome (for example, time to death) is the main end point and, consequently, the patient is unable to benefit from thestudy directly.1 This raises a fundamental issue concerning the usefulness of randomised controlled trials in reliably identifying new clinical knowledge. Less recognised, but equally important, is inconsistent clinical enthusiasm to offer the …

In our ongoing investigation of new compounds with activity against malaria parasites, we tested the in vitro antiplasmodial activity of fractions and purified compounds from Cassia fistula L., a plant traditionally used by native populations of Tanzania, Zimbabwe, Mozambique and Brazil to treat malaria or symptoms associated with this disease. Crude extracts from leaves, bark and fruits were tested for their antiplasmodial activity against the chloroquine-sensitive strain of Plasmodium falciparum(D10), where leaf extracts showed the highest activity. The chloroform extract of the leaves was further bioassay-guided fractionated using a combination of centrifugal partition chromatography and flash column chromatography. Three main antiplasmodial principles, phytol (1) (IC50 18.9 ± 0.60 μM), lutein (2) (IC50 12.5 ± 0.35 μM), and di-lineolylgalactopyranosyl-glycerol (DLGG) (IC50 5.8 ± 0.27 μM) (3), were isolated and identified using spectroscopic methods. When the three active principles were tested for their cytotoxicity using a Chinese Hamster Ovarian (CHO) cell line, compound 3 showed very weak toxicity (IC50 75.9 ± 0.28 μM), while the other two compounds were nontoxic, even at the highest concentration tested. The study provides evidence to support the use of Cassia fistula as an antimalarial remedy and describes the antiplasmodial constituents from the leaves.

PURPOSE: The aim of our study was to determine the five-year survival of patients with colorectal carcinoma whose Dukes classification had changed following fat clearance of the mesocolon or mesorectum. METHODS: One hundred three patients with colorectal carcinoma were followed up at a special clinic for at least five years after surgery. The tumors from these patients had previously been given a Dukes classification before and after fat clearance. RESULTS: Four of the five patients whose Dukes status changed from B to C as a result of fat clearance died of malignant disease during the five-year follow-up period. After fat clearance it was apparent that Dukes B patients survived, on average, 11 months longer than Dukes C patients. A significantly increased mean number of positive nodes was found after fat clearance in Dukes C cases, both in those who were alive at five years and those who died of their malignant disease. CONCLUSIONS: The fat clearance technique is a useful aid to improving the accuracy of the Dukes classification and has prognostic significance. It should be used in specimens of colorectal carcinoma, which on initial examination appear to be Dukes B cases.

About half the patients treated with curative resection for colorectal cancer do not survive long-term. Adjuvant chemotherapy given during and after surgery may prevent hepatic metastases and improve patient survival. In patients with colorectal cancer, we have done a multicentre, randomised controlled trial comparing five-year survival after intraportal infusion of fluorouracil (1 g per day) plus heparin (10,000 U per day) (130 patients) or heparin alone (123) during curative resection and for 7 days thereafter, or after resection alone (145). There was no reduction in liver metastasis or increased overall survival advantage in either active-treatment arm of the study. However, patients who had stage III, Dukes' C (lymph-node-positive) tumours resected and were treated with fluorouracil plus heparin had a significant (p less than 0.03) survival advantage of about 16% compared with surgery-only controls. Further study of intraportal infusion of chemotherapeutic agent as adjuvant treatment to surgery in patients with colorectal cancer appears worthwhile.

Twenty four primary stromal tumours of the stomach and small intestine were investigated by silver staining of interphase nucleolar organiser regions (AgNORs) in an attempt to obtain an objective criterion for prediction of malignant tumour behaviour. Malignant tumours tended to have higher AgNOR counts than their benign counterparts, but this increase was small and there was some overlap between the two groups. There was a correlation between the mean AgNOR count and the mitotic count. There was no correlation between tumour size and these two measurements. This study suggests that in these stromal tumours the AgNOR count is not a useful independent predictor of malignant behaviour.

Summary The rectal mucosa-associated flora (MAF) of patients with ulcerative colitis has been studied in 25 patients with newly diagnosed disease, 20 with relapse of existing disease, and 44 who were in remission. Patients with active disease were re-examined twice during treatment. The MAF was simpler and less dense than the microflora of faeces. Obligate anaerobes usually predominated in the MAF although the ratio of obligate anaerobes to facultative species was lower than that found in faeces. Viable counts of the total flora and of its constituent genera varied considerably between patients. Counts of the total flora, of obligate anaerobes (including bifidobacteria, eubacteria and clostridia), and facultative organisms and micro-aerobes (enterobacteria and lactobacilli) were reduced in patients with active disease compared with those with inactive disease; corresponding carriage rates were also lower. Counts and carriage rates increased during treatment and approached those found in quiescent disease. The alterations in the MAF were especially marked in patients experiencing their first attack of ulcerative colitis. The relationship between these alterations and the aetiology and pathogenesis of this disease remains unclear.

Rhodiola spp. (Crassulaceae) have been known for centuries as medicinal herbs in Eastern Europe and Asia. R. rosea has been popular as an adaptogenic plant [1-5]. The term adaptogen refers to substances which help the body and brain cells resist physical, chemical, and environmental stresses. Repeated administration of adaptogens leads to a stress-protective effect in a way similar to that produced by repeated physical exercise. Adaptogens have a broad range of therapeutic effects without causing any disturbance to the normal functioning of the organism [6-9]. An important direction in modern pharmacy today is to search for drugs that increase working capacity, stability under stressful conditions, and ability to adapt to rapidly-changing industrial and environmental conditions. The increase in body resistance to external stress conditions, like hypoxia, is considered an indication of adaptogenic activity [10,11]. Previously, a series of in vivo experiments to evaluate the increase in endurance of animals under intense physical loads was established as a means to gauge the ability of ingested substances to act as natural adaptogens [12]. Wild species of Rhodiola have been used in traditional medicine systems [13,14], including R. heterodonta, a species endemic to the mountains of Central Asia. Rhodiola species were recently reported to contain phenylethanoid/ propanoid compounds along with other biologically active constituents [15]. Analytical methods to determine salidroside and tyrosol have been reported [16-20], but methods to determine the presence of other co-occurring constituents are very limited. To standardize a pharmaceutical preparation, it is essential to fully characterize and quantify the constituents of the preparation. In this study we used simple and efficient analytical methods for characterization and quantification of multiple components in a R. heterodonta standardized ethanolic preparation that demonstrated an in-vivo antihypoxic efficacy. Our results showed that administration of the 80% EtOH extract of R. heterodonta (40 mg/kg) increased the life span of the treated mice under hypoxic conditions by 1.9 times compared with the untreated control (Table 1). This increase in the mice tolerance to hypoxia is evidence of the drug’s adaptogenic activity [10]. Our findings are consistent with the published data on epigalokhin, a polymeric proanthocyanidin isolated from a related species, R. semenovii, which exhibited pronounced antihypoxic effect in both in-vivo and in-vitro experiments [21]. Additionally, the most abundant phenylethanoid glycoside, salidroside, was reported to exert a protective effect on PC12 (neuronal pheochromocytoma cells), where it prevented the morphological changes of these cells under conditions of chemical hypoxia induced by Na2S2O4 [22]. Table 1 Antihypoxic activity of the R. heterodonta 80% EtOH extract, intra-peritoneally administered to mice. R. heterodonta rhizome contains a wide range of secondary metabolites including polymeric proanthocyanidins [15], which interfere with direct HPLC analysis. The presence of proanthocyanidin compounds along with phenylethanoids in the EtOH extract made both qualitative and quantitative analysis of both components difficult, for two reasons: the presence of a polymeric proanthocyanidin spectral hump, and overlapping of the peaks from the two groups because of similarity in UV wavelength absorption. An HPLC-PDA analysis of the 80% EtOH extract acquired at 225 nm and 280 nm displayed a complex hump (Figure 1). This problem was reported before, for the analysis of 5 marker compounds in R. rosea [23]. In this study, we separated these compounds on a Sephadex LH-20 column, to two major groups, phenylethanoids (fraction A) and proanthocyanidins (fraction B). This separation facilitated characterization of individual components. Also, partition of fraction B with EtOAc allowed for the separation of three major oligomeric proanthocyanidins (sub-fraction C) from the more complex polymers (sub-fraction D). This separation allowed accurate quantification of oligomeric proanthocyanidins and overcame the chromatographic hump characteristic for polymeric proanthocyanidin mixtures, which would inevitably affect the quantification of other constituents. Figure 1 HPLC chromatogram (from LC-MS) for R. heterodonta 80% EtOH extract measured at 225 nm (A), maximum absorption of phenylethanoids, and at 280 nm (B), maximum absorption of proanthocyanidins. Phenylethanoids isolated from fraction A included tyrosol (1), viridoside (2), salidroside (3), tyrosol methyl ether (4), heterodontoide (5), and mongrhoside (6), and the cyanogenic glucoside rhodiocyanoside A (7) (Figure 2) [15]. Tyrosol methyl ether and mongrhoside were characterized for the first time in this study. Mongrhoside, the arabinose derivative of viridoside, was recently identified in R. quadrifida and R. rosea [24, 25]. 1H NMR and 13C NMR spectra were identical to the data recently reported for monrhoside [25]. Figure 2 Compounds isolated and quantified in R. heterodonta HPLC-PDA quantification of fraction A (Figure 3), using the above isolated compounds as external standards, revealed that heterodontoside was the most abundant phenylethanoid in the 80% EtOH dry extract of R. heterodonta (53.2 mg/g), followed by tyrosol methyl ether (41.6 mg/g), salidroside (25.8 mg/g), viridoside (22.6 mg/g), and mongrhoside (18.3 mg/g), with tyrosol and rhodiocyanoside A as minor components (5.2 and 7.1 mg/g respectively). Collectively, the concentration of the seven compounds was 173.8 mg/g (17.4%) in the 80% EtOH extract of R. heterodonta. When completely dried, R. heterodonta fresh rhizomes contained 38.2% ± 0.7% dry matter (n = 3), of which 37.5% ± 2.9% constituted the 80% EtOH extract. The 80% EtOH extract constituted 14.3% ± 1.0% (n = 3) of the rhizome fresh weight. The total phenylethanoid/cyanogenic compounds comprised 6.4% and 2.4% of the dry and fresh rhizomes, respectively (Table 2). Figure 3 RP-HPLC chromatogram for phenylethanoids and cyanogenic glycosides in R. heterodonta (fraction A, UV 225 nm). Table 2 Concentrations of the phenylethanoid and cyanogenic glycosides in R. heterodonta 80% EtOH extract and the corresponding percentages in the rhizome dry and fresh tissues. Fraction B contained a large number of oligomeric and polymeric proanthocyanidins, as indicated by its LC-ESI-MS profile. HPLC-ESI-MS of sub-fraction C (the EtOAc partition from fraction B) revealed that there were three main components with m/z of 763, 915 and 459 [M+H]+ in the positive ion mode. The three components were isolated utilizing semi-preparative HPLC and identified as: epigallocatechinepigallocatechin-,3-O-gallate (EGC-EGCG, 9), 3-O-galloyl-epigallocatechin-epigallocatechin-3-O-gallate (EGCG-EGCG, 10), and epigallocatechin gallate (EGCG, 8). EGCG was identified by comparing its physical and spectral data with a standard compound. EGC-EGCG (9) and EGCG-EGCG (10) were identified by comparing their physical and NMR characteristics with published data [26]. Compounds 8, 9 & 10 were used as external standards for the HPLC quantification of oligomeric proanthocyanidins in sub-fraction C (Figure 4). The concentrations of EGC-EGCG, EGCG-EGCG, and EGCG in the 80% EtOH extract were 4.35, 4.68, and 8.51 mg/g, respectively. Collectively, the three measured oligomers composed 17.5 ± 0.82 mg/g (n = 3) or 1.75% of the 80% EtOH extract. The three oligomers represented 0.65% and 0.25% of the dry and fresh rhizomes, respectively (Table 3). Sub-fraction D contained a mixture of several complex polymeric compounds and, therefore, it was not analyzed further. This is the first report to quantify oligomeric proanthocyanidins in Rhodiola plants. Figure 4 RP-HPLC chromatogram for oligomeric proanthocyanidins in R. heterodonta (sub-fraction C, UV 280 nm). EGC-EGCG = epigallocatechin-epigallocatechin-3-O-gallate, EGCG-EGCG = 3-O-galloyl-epigallocatechin-epigallocatechin-3-O-gallate, EGCG = epigallocatechin ... Table 3 Concentrations of oligomeric proanthocyanidins in R. heterodonta 80% EtOH extract and the corresponding percentages in the rhizome dry and fresh tissues. Data on the concentration of phenylethanoids in Rhodiola species has rarely been reported outside of publications in Russia, China, and other Central Asian countries. In one study it was reported that the concentrations of salidroside and tyrosol ranged from 1.3-11.1 mg/g and 0.3- 2.2 mg/g, respectively in dry Rhodiola rosea rhizomes collected from various parts of China [22]. In another study, the concentration of salidroside in a selection of dried R. rosea samples ranged from 0.4 to 2.7 mg/g [23]. In our study, the concentrations of salidroside and tyrosol were 1.0 and 0.2 mg/g dry weight, respectively, while the methyl ether of tyrosol showed higher concentration (1.5 mg/g dry weight).

Bioassay-guided fractionation of extracts from Acacia burkittii and A. acuminata heartwoods against an L1210 (mouse lymphoma) cell line led to the isolation of two flavan-3,4-diols, melacacidin (1) and isomelacacidin (2), and three flavonoids, 3,7,8,3′,4′-pentahydroxyflavone (3), 7,8,3′,4′-tetrahydroxyflavanone (4) and 3,7,8,3′,4′-pentahydroxyflavanone (5). HPLC analyses (280 nm) of the 70% acetone extracts of the two plants showed different profiles in terms of the relative concentration of the five identified compounds. When tested against L1210, compounds 1 and 2 were the most active, giving ID50 values of 2.5 and 4.5 μg/mL, respectively. The lower activity of the other isolated compounds indicated the importance of the 3,4-hydroxyl groups for the cytotoxic activity of these flavonoids. The isolated compounds were either non-toxic or had very low toxicities against the “normal” CV-1 cell line (green monkey kidney cells). The O-methyl and O-acetyl derivatives of these compounds were inactive. Ten commercially available phenolic compounds (6-15) were also tested for their activity against both CV-1 and L1210 cell lines. Flavan-3,4-diols 1 and 2 were more cytotoxic to L1210 cells than all tested compounds, including catechin and epicatechin, the most abundant flavan-3-ols in many fruits and vegetable.

An association between non-suppurative otitis media (NSOM) and symptoms of dysequilibrium has been observed but not previously quantified in children (Gates, 1980;Busis, 1983; Blayney and Coleman, 1984). This study compared the incidence of balance related problems in 154 children with surgically proven glue ear and 51 children with normal ear function. Symptoms ranging from true vertigo to mild ataxia were discovered in 22 per cent of the children with NSOM but in none of those within the control group (p>0.001). Periods of dysequilibrium were associated with episodes of otalgia in 64 per cent of the children but were not increased in those with unilateral compared to those with bilateral effusions. Complete resolution of symptoms occurred in 85 per cent of children following the insertion of grommets. A history of balance disturbance should be actively sought in all children with otitis media with effusion, and when present, provides a strong indication for early operative intervention.