No description available.

No description available.

No description available.

No description available.

No description available.

The main objective of the study was to investigate the bioactive components, their effect on HL60 leukaemic and Caco-2 colon carcinoma cancer cell lines and their bioavailability from a decoction of a Chinese Herbal Remedy OldenIandia diffusa, using a Caco-2 monolayer as a mimic for intestinal absorption. The HPLC separation method was set up by investigating parameters such as column type, mobile phase, isoeratic/gradient elution, flow rate, pH of buffer and detection wavelength. Using this method eleven fractions (F1-F11) were collected. Results from a cytotoxicity investigation using the CyQUANT NF, trypan blue and neutral red uptake assays showed that the decoction has a cytotoxic effect on HL60 (V=13.5±4.3%, n=3) and Caco-2 (V=50.0±1.4%, n=3) cancer cell lines. The most cytotoxic active fraction was F9 (V=62.2±7.3% (HL60) and V=32.l±7.9%, n=3 (Caco-Zj). DAPI staining and Western blotting (detection of cleaved-PARP) studies on the decoction and F9 fraction indicated that mode of cell death was apoptosis, which was mediated by a caspase cascade. Fraction F9 was separated into eight further fractions (compound- fraction-l to 8) by optimisation of the HPLC method. By using liquid-liquid extraction with ethyl acetate, eleven sub fractions (compound-fraction-l to 8 and ethyl acetate fractions (FEA-l to 3) were collected purely. All collected fractions were analysed by high resolution-MS and their MWs were determined as 268.07,238.07,242.24,280.38,256.24,282.25, 284.27, 456.36,550.17,328.22 and 330.24. In the most cytotoxic fraction F9, oleanolic acid and ursolic acid were isolated and identified in concentrations of 0.068 mg/g and 0.166 mg/g, respectively. FEA-l to 3 also showed cytotoxic effects on these cancer cells (V=53.3±2.2%, 55.4±4.4% and 50.6±11.3% (HL60) and V=63.4±13.5%, 45.8±5.9%, and 59.5±9.7%, respectively (Caco-2), Fraction FEA-l was identified as E-6-0-p-coumaroyl seandoside methyl ester (MW 550.17). Cytotoxicity assessment results showed that it has a growth inhibition effect on both cancer cell lines. Bioavailability after ingestion of the decoction was studied using 21-day grown Caco-2 monolayers. The post absorption sample (PAS) of the decoction and fraction F9 were shown to have a good permeability (P[sub]app=3.575xlO[sup]-6 cm/s). The PAS also has a cytotoxic effect on cancer cells (V=67.0±7.5%, n=3). When analysed by LC-MS, the most of the compounds that had previously been seen in the original fractions were again observed.

Introduction. Giner, a traditional Chinese herbal medicine, is used to treat digestive disordrs in particular to alleviate symptoms of nausea and/or vomiting. Aims. Major aims were to measure the concentration of [6]-gingerol (6G), and elements in fresh ginger rhizome juice (GJ) by HPLC and ICP-AES and investigate their effects on gastrointestinal functions. Methods. Short circuit current (I[sub]sc) effects of 6G and the dietary phytochemicals quercetin and kaempferol (100 [mu]M) were measured in a Caco-2 cell monolayer. In vitro isometric recording was used to investigate GJ (50 [mu]L, 200 [mu]L), 6G [1.59x10[sup]-5M-1x10[sup]-4M), a selected combination of elements (K [4.6x10[sup]-2M], Mg [7.4x10[sup]-3M], Mn [8.3x10[sup]-4M], Na [1.1x10[sup]-3M, Ca [5.1x10[sup]-4M]), and a "faux" ginger juice on contractile activity of proximal and distal stomach and duodenum segments from 'Suncus murinus'. The effect of 6G (1x10[sup]-2Mx1x10[sup]-4M, po.) and a ginger capsule suspension on motion-induced emesis was investigated in vivo in 'Suncus'. Results. The concentration of 6G in GJ was 239.43 [plus or minus] 7.92 mg/L. 6G had no effect on I[sub]sc], however quercetin and kaempferol caused a significant increase on I[sub]sc and the ATP - induced chloride ion secretion. GJ (50 [mu]L and 200 [mu]L) caused a dose-realted biphasic effect resulting in an overall increase in tension on both regions of the stomach at 25 minutes and an inhibitory effect on duodenal contractions. "Faux" GJ (200 [mu]L) only partially accounted for the effects of GJ. 6G and a ginger capsule suspension had no antiemetic activity in vivo. Conclusions. Quercetin and kaempferol may be able to augment the signalling in the intestinal epithelia resulting in an increase in fluid secretion which could facilitate stool passage. "Faux" GJ did not fully account for the motility effects of GJ, indicating that there were ither bioactive constituents present in GJ (e.g. [6]-shogaol). GJ was most effective on the duodenum, suggesting this as a target for an enteric coated ginger capsule for gastrointestinal disorders.

The outer coat (exine) of plant pollen grains and spores is composed of the material sporopollenin. Sporopollenin is exceptionally resistant to chemical and physical attack and is perhaps one of the most chemically resistant, naturally occurring organic materials known. Its stability to chemical and physical stress is illustrated by its survival in ancient sedimentary rock of around 3.7 billion years old. Sporopollenin from Lycopodium clavatum has been functionalised in the past for use in the preparation of peptides and for use in ion and ligand exchange. It was found that sporopollenin. had a number of advantages over the synthetic resins commonly used in solid phase synthesis which included a constant chemical structure, constant pore size, chemical and physical stability and commercial availability. Investigations into the isolation of sporopollenin using various procedures outlined in the literature were reviewed in order to find the most efficient protocol for sporopollenin preparation. Detailed studies of the sporopollenin were undertaken to establish its chemical and physical stability, swelling, filtering and hydration characteristics. Results indicated that sporopollenin would make a good alternative to other solid supports as it showed little swelling in organic solvents, was easily filtered through grade 3-4 sinters and the size of the sporopollenin was constant. Also, it could be dried efficiently and showed good chemical and physical stability, e. g. it can be refluxed for several days in strong acid and base and a variety of organic solvents without decomposition. It also can be stirred for several days with little degradation. In contrast, a parallel study with a commercial PS-DVB resin showed it to be considerably less stable, e. g. stirring for one day resulted in severe degradation of the resin. Also, most of the commercially available resins are found to have a variable particle size and variable loadings. An investigation into the types of functional groups present on the sporopollenin surface was undertaken to explain the types of functionalities present on sporopollenin and examine their reactivity. The objective was to assess the potential of sporopollenin for use as a solid phase support for synthesis and drug delivery. Studies indicated that ketone, acid and hydroxyl groups were present. Thus, in order to obtain uniformity of functionality sporopollenin was reduced with the objective of obtaining solely hydroxyl groups on its surface. Suprisingly the loading of hydroxyl groups varied between one batch to another (0.40-0.92 mmol/g). Attempts to halogenate the hydroxy groups with PCl5 and SOCl2 or condense them with amino acids to form ester linkages was only moderately successful. In contrast, chloromethylation of sporopollenin using dirnethoxymethane and SOCl2 with ZnCl2 as a catalyst gave consistent levels of chloride loading from one batch to another (1.01-1.28 mmol/g). Such a form of chloromethylated sporopollenin could be compared to Merrifield resin. Preliminary attempts to substitute the chloride proved difficult due presumably to the attachment of chloromethyl groups to aliphatic sites rather than aromatic sites. Amination of different batches of sporopollenin with primary amines, e. g. n-butylamine gave loadings of around 1-2 mmol/g. An extensive study was undertaken to assess the stability of the amine attachment to acid, bases and organic solvents. In all of these a relatively robust attachment was found with no less than 70% of the amine still attached. This study encourages the use of diamines as linker groups with the intention that one primary amine would attach to the sporopollenin with the other being available for further substitution to attach either synthons or drugs.

Obesity is a disease that has reached epidemic proportions across the world. Many types of treatments have been used to combat obesity including using synthetic drugs such as sibutramine and orlistat. However, the high cost and potentially hazardous side-effects of anti-obesity drugs have led many researchers to turn to naturally occurring compounds obtained from fruits, vegetables, herbs and plants for the treatment of obesity. In our study, the anti-obesity effects of S. crispus crude extract (SCE) and individual polyphenols (EGCG, Resveratrol, Phloridzin, Quercetin and Verbascoside) found in fruits, vegetables and herbs have been investigated. The effects of S. crispus extract (SCE) in vivo were tested on high fat-induced obese LDLr KO mice maintained on high fat diet (HFD) or switched to low fat diet (LFD). All mice were HFD for 25 weeks to induce obesity, after which half were maintained on the HFD and half switched to LFD. At the same time, mice were given normal water or 0.1% (w/v) SCE in water at Week 0-4 which was increased to 1% (w/v) at Week 5-10. Oxygen consumption (VO2), CO2 production (VCO2), RER, locomotor activity (LMA) and heat production (HP) were measured at Week 0, 5 and 10. Food intake, water intake and body weight was measured weekly. Plasma glycerol (PG) and abdominal adipose tissue (AAT) weight were determined at Week 10. Mice switched to LFD lost weight (p< 0.001), mainly due to decreased energy intake (p<0.001). They also had lower AAT weight and PG concentration (all p<0.001). SCE had no effect at either dose on body weight, VO2, VCO2 or LMA, but significantly reduced respiratory exchange ratio (RER) (p=0.034) and increased HP at Week 4 (P=0.048), without altering food or water intake (p=0.1, p=0.222). PG concentration were also increased in SCE treated mice (p=0.032). The effects of SCE and individual polyphenols in vitro were tested on rat epididymal and human omental adipose tissue explants and results were compared with with the results from the pig perirenal adipose tissue explants. SCE does not appear to have any direct effect on lipolysis in the rat epididymal adipose tissue explants and human omental adipose tissue explants. EGCG was found to consistently inhibit lipolysis in rat, human and pig adipose tissue explants and the effects were greatest at 100µM. The effects of Phloridzin in human, rat and pig fat explants were inconsistent as it was found to either increase or decrease lipolysis with different treatments. In all experiments, when Isoproterenol (IP) was present Resveratrol inhibited lipolysis and was independent of adenosine deaminase (ADA), with greater inhibition found at 100µM compared with 50µM Resveratrol. The effects of Resveratrol on lipolysis in the human adipose tissue explants was found to be different when compared with the effects found in the pig and rat adipose tissue explants when incubated for 24 and 26hr. The effects of Resveratrol on lipolysis in the human adipose tissue itself are also dependent on the presence and absence of ADA and IP. Subsequent experiments were carried out where basal lipolysis and effects of the presence and absence of ADA were also investigated. Basal lipolysis was found to be higher in pig adipose tissue explants (Headland, 2007) than in human adipose tissue explants, but lower than rat adipose tissue explants. This is also true for the IP stimulated lipolysis in pig perirenal adipose tissue explants, but not in the pig subcutaneous adipose tissue explants, where IP stimulated lipolysis was similar to that in human omental adipose tissue. As expected, the presence of adenosine does have an effect on the lipolysis rate in rat, human and pig adipose tissue explants, since the addition of ADA (to metabolise/remove adenosine) increased basal lipolysis. However, only in the pig perirenal adipose tissue explants was IP stimulated lipolysis found to be increased with ADA. In the human omental adipose tissue explants, we also found that although BMI and age had weak negative correlations with lipolysis, these were not statistically significant (P=0.097 for BMI, P=0.48 for age). However, the trend suggests that IP stimulated lipolysis decreased with increased BMI. Thus, SCE appeared to induce lipolysis and body fat oxidation in vivo but no direct effect on lipolysis were found in vitro. Resveratrol is the most promising polyphenol to induce lipolysis based on the studies across the rat, human and pig species compared with quercetin, EGCG and Phloridzin. The consistent effects of EGCG on lipolysis inhibition however, might also be an anti-obesity effect through the mechanism of adipocyte apoptosis which requires further study.

The methanolic crude extract of aerial parts of the plant Scrophularia nodosa was shown to have potent DPPH radical scavenging activity (IC50 = 48.75 ± 7.00 μg/ml). Activity-guided fractionation resulted in the isolation of three principal antioxidant compounds; acteoside, angoroside C and angoroside A. Acteoside (yield = 1.21%, IC50 = 15.2 μM) appeared to be the most abundant and most antioxidant-active. The potent antioxidant activity is in support of the traditional use of the plant for wound healing and anti-inflammatory conditions. The methanolic extract of aerial parts of Tanacetum vulgare has potent DPPH radical scavenging activity (IC50 = 37.00 ± 1.20 μg/ml). Activity-guided fractionation on the methanolic extract of T. vulgare resulted in the isolation of 3,5-di-caffeoylquinic acid (3,5-DCQA), axillarin and luteolin. 3,5-DCQA appeared to be the most abundant and most antioxidant-active compound (yield = 7.28%, IC50 = 9.7 μM). The potent antioxidant activity is in support of the traditional use of the herb for fever, rheumatic conditions and anti-inflammatory conditions. The methanolic crude extract of Cassia auriculata and its fractions were shown to have potent scavenging activity on DPPH, hydroxyl and hydroperoxide radicals, moderate superoxide radical scavenging activity and potent ion(III) reducing power. The activity-directed studies resulted in the isolation of kaempferol-3-0-β-D-rutinoside, kaempferol, luteolin, quercetin and unknown antioxidant inactive compound. The previously reported pharmacological aspects of the above flavonoids and flavonoid glycosides (anti-carcinogenic, anti-inflammatory, hyperglycaemic, antidiabetic) along with the shown antioxidant behaviour explain the traditional medicinal values of the plant. Cassia alata L crude extract and its fractions showed potent radical scavenging activity against formation of lipid peroxide radicals. The activity directed isolations resulted in the isolation of kaempferol along with p-hydroxybenzoic acid. These may contribute towards the traditional medicinal values of the plant as an antidiabetic, anti-microbial and for skin diseases.