Non‐alcoholic fatty liver diseases (NAFLD) is manifested in the absent of alcohol abuse. This disease is the major cause of liver failure and death among adults and children worldwide, including South Africa. Its increasing prevalence urges the need of therapeutic intervention. The main objectives of this study were to investigate the following: (1) The effect of 38.9% high fat diet (HFD)‐induced insulin resistance and fatty liver in male Wistar rats, (2) The efficacy of aqueous extracts from Sutherlandia frutescens leaves and Prunus africana bark and metformin in the treatment of HFDinduced insulin resistance and fatty liver. Male Wistar rats were fed on HFD (the HF group) or normal rat chow (the LF group) for 12 weeks. Even though the HFD‐fed rats had developed insulin resistance by week 12, fatty liver developed by week 16. After week 12, the HF group was divided into four groups of 6‐7 rats each and three of those groups were gavaged with either 0.125 mg P. africana extract/kg bwt/day (the HF+Pa group) or 50 mg S. frutescens extract kg bwt/day (the HF+Sf group) or 16 mg metformin/ kg bwt/day (HF+Met group), while kept on the same diet for an additional of 4 weeks, to investigate whether two medicinal plant extracts and metformin can prevent HFD to induce fatty liver or not. After 16 weeks, the liver histological images revealed that the HF group developed fatty liver in the form of both microsteatosis and macrosteatosis. Fatty liver was confirmed by significant increased liver total lipid (TL) and activities of glucose‐6‐phosphate dehydrogenase (cG6PD) and xanthine oxidase (XO), mitochondrial NADH oxidase (mNOX) and by a decrease (P<0.05) in the activities of the homogenate superoxide dismutase (hSOD) and mitochondrial complex II in the HF group, when compared to the LF group. Since the activities of mCS and cACL enzymes were not changed in the HF group, hence increased cG6PD activity in the HF group indicates that there was increased NADPH demand for lipid accumulation from activated NEFAs taken up by the liver from circulation and for maintenance of the NADPH‐dependent antioxidants and oxidants, respectively. The obtained data also show that mitochondria of the HFD‐fed rats adapted to an increase in energy availability, thereby compensation through decreasing complex II activity, to allow electron flux from β‐oxidation to respiratory chain in the HF group. Liver TL content was significantly decreased in the rats treated with metformin and P. africana extract, but not in the rats treated with S. frutescens when compared to the HF group (P < 0.05). However, the TL content remained >5% per liver weight in all treated groups. The present study demonstrates that these two plant extracts and metformin have different glucogenic and lipogenic effects from that presented by HFD alone when compared to the LFD alone. In conclusion, metformin and P. africana extract can attenuate HFD‐induced fatty liver without changing the dietary habits. Hence S. frutescens extract is less effective in the prevention of HFD‐induced fatty liver. A change in the dietary habits is recommended to be considered during the use of these three remedies in the treatment of HFD‐induced insulin resistance and fatty liver. All three treatments enhanced antioxidant capacity, and may improve insulin resistance and fatty liver mediated by the present HFD through different mechanism of actions in the liver.

Traditional medicine has become an important part of healthcare worldwide. It is estimated that about 25 percent of prescribed medicines contain plant products or active compounds derived from plants. In South Africa, traditional medicine forms part of the culture and tradition of most communities. Garlic compounds have been shown to have a variety of antimicrobial properties. Amongst these are antifungal, antibacterial, antiviral and anti protozoal activities. Allicin and its breakdown products have been shown to be the main active compounds which possess these properties. Tulbaghia violacea has been used for the treatment of a variety of illnesses including asthma, fever, oesophageal cancer, constipation and hypertension. This study investigated the antifungal nature of T.violacea on the morphology, spore germination and lipid synthesis of Aspergillus flavus and Aspergillus parasiticus. The results of this study showed that the plant extract inhibited A. flavus growth at a minimal inhibitory concentration of 15mg/ml and was fungicidal at 20mg/ml and above. A. parasiticus was not inhibited at 25mg/ml indicating resistance to the inhibitory component of the plant extract. A measure of metabolic activity using the XTT assay showed reduced metabolic activity in the presence of increasing concentrations of the plant extract. Higher extract concentrations resulted in higher percentage inhibition of fungal growth for both fungal species with up to 98 percent inhibition being observed for the highest extract concentrations for both fungi. Germination was also delayed in the presence of 15mg/ml plant extract concentration by up to 60hr for A. flavus and 48hr for A. parasititcus. The TEM results showed increased thickening of the cell wall with higher extract concentrations. The thickening was greater for A. flavus than for A. parasiticus. Cell wall thickening may be the reason for the delay in germination in both species. Lipid production was reduced in the presence of plant extracts when compared to the control. The plant extracts inhibited triglyceride production at 15mg/ml for both A. flavus and A. parasiticus. The results therefore indicate that T. violacea extracts are antifungal and probably affect germination through interactions with the cell wall. It is possible that the extract affects lipid production in Aspergillus species.

Anthracnose is one of the serious diseases of cowpea (Vigna unguiculata L. Walp) and common bean (Phaseolus vulgaris L.) caused by the Colletotrichum fungi. The disease is prevalent is small holder farmers’ fields due to the scarcity and high cost of the synthetic fungicides. This study was conducted with the main aim of improving food security and income of the smallholder farmers by increasing legumes, P. vulgaris and V. unguiculata thereby increasing production and improve food security and income of smallholder farmers. Investigations involved in vitro bioassaying for antifungal activities of the crude extracts on Colletotrichum lindemuthianum (Sacc. and Magn.) Bri. and Cav. and Colletotrichum dematium (Fr.) Grove var. truncata field isolates and evaluating the effect of crude plant extracts seed treatments on seed germination, emergence and control of anthracnose disease of common bean and cowpea. Furthermore, ultra-structural changes of plant extracts treated and efficacy of foliar application of extracts. The in vitro study showed that Allium sativum L., Agapanthus caulescens Spreng., Carica papaya L. and Syzygium cordatum Hochst.ex Krauss extracts have good antifungal activities against both C. lindemuthianum and C. dematium. The low concentrations (5 mg.ml-1) of Syzygium and Agapanthus water extracts and acetone extracts of Agapanthus and Carica gave a high percentage of bean seed germination, emergence, short mean emergence time (MET) and were effective in controlling the anthracnose disease. The treatment of Agapanthus (both water and acetone) extracts also increased the shoot length and dry weight of the seedlings. The Allium acetone extracts (5 mg.ml-1) was the only treatment that gave good results with respect to germination percentages, MET, shoot length, leaf area and dry mass of cowpea. Five mg.ml-1 concentrations of Syzygium and Agapanthus water extracts and acetone extracts of Agapanthus and Carica have potential as seed treatments on bean. Allium acetone extract (5 mg.ml-1) was the only potential cowpea seed treatment that could be recommended to farmers as an alternative to the synthetic fungicide. Electron microscopy revealed that principle differences were observed in the cotyledon-embryo connecting tissues of seeds treated with Agapanthus, which had few cristae in their mitochondria than the cells from other treatments. The embryonic root cells of bean seeds treated with Agapanthus had coalescing protein bodies. The embryonic root cells of cowpea and bean treated with Syzygium had fewer lipid bodies as compared to the control and the Agapanthus treated seeds. Bean plants that were foliar treated with the 15 mg.ml-1 concentrations of Allium water, Agapanthus water, Carica water, Agapanthus acetone, Carica 5 and 15 mg.ml-1 acetone, Syzygium 5 mg.ml-1 acetone extracts and the combinations (2.5 mg.ml-1 + 2.5 mg.ml-1) of Allium + Agapanthus, Allium + Carica, Agapanthus + Syzygium and Carica + Syzygium extracts registered low anthracnose (C. lindemuthianum,) disease severity and high leaf area. The cowpea plants treated with 15 mg.ml-1 water extracts of Agapanthus and the combinations of Allium + Agapanthus, Agapanthus + Carica and Agapanthus + Syzygium extracts recorded low cowpea anthracnose (C. dematium) disease severity, highest leaf area and dry mass. The study revealed that A. sativum, Agapanthus, C. papaya and S. cordatum plant extracts have antifungal activities and can be used as alternative seed treatments and foliar fungicides against the anthracnose diseases of legumes (cowpea and common bean) instead of synthetic fungicides without causing any negative effect on seed germination, emergence, ultra-structure of seeds and plant growth. Copyright / Dissertation (MSc)--University of Pretoria, 2012. / Microbiology and Plant Pathology / Unrestricted

Cannabis sativa has a complex history of classification and traditional use. Cannabis sativa ssp. sativa and spp. indica are the two major lineages of cannabis and, through artificial selection, many strains or cultivars are found within each group are bred together to yield hybrid plants. New methods of classification based on Δ⁹-Tetrahydrocannbinol (THC) and Cannabidiol (CBD) content as well as minor cannabinoids and terpenes have emerged as a more effective classification of cannabis. However, the fidelity of cannabis varieties relative to their respective strain names and lineages (indica, sativa and hybrid) based on chemistry has been brought into question. THC and more recently CBD are collectively responsible for the psychoactive and therapeutic effects of cannabis and minor cannabinoids and terpenes are emerging as having their own unique bioactivity or synergistic effects in vitro. Considering the variation in cannabis chemical profile and infidelity to strain names or lineages, we investigated the cannabinoid, terpene and metabolomic profiles of 33 THC-dominant strains (113 samples) to evaluate existing and alternative chemistry-based classification systems using multivariate analyses. Here, we conclude that Indica-Sativa-Hybrid designations are insufficient in describing variation in cannabinoid, terpene and metabolomic data, and that a terpene-based profile classification revealed robust groupings in cannabinoid-terpene data. However, terpene profiles were not discernable in metabolomic data. To investigate how chemical complexity and variability impacts bioactivity, we compared the activity of cannabis extracts to that of pure THC (and CBD) to determine if THC alone is driving activity. THC, CBD, and plant extracts were tested in vitro for cytotoxicity in BEAS-2Bs cells and for cannabinoid receptor signalling activity using a human CB₁-HEK293 cell model. THC did not completely dictate 24-hour toxicity in BEAS-2B cells suggesting that other extract components (beyond THC and CBD) are contributing to cytotoxicity. While CBD alone was 2x more toxic than THC alone, THC:CBD do no predict toxic concentration. THC within extracts appeared to drive efficacy at CB₁ receptors by reducing intracellular cAMP accumulation but did not dictate variation in EC₅₀. THC in extract also appeared to increase percent cAMP reduction in cells regardless of low CBD content but a 1:1 balanced THC/CBD extract revealed reduced percent cAMP reduction. Pure compounds compared to extracts of the same THC/CBD ratio performed very similarly at CB1 receptors besides 1:1 extract preparation having reduced % reduction of cAMP compared to 1:1 pure compounds suggestive of NAM by extract components. Regression modelling of THC within extract revealed a significant positive relationship in % cAMP reduction (Emax) but no significance in TC₅₀ and EC₅₀. This work demonstrates the importance of rigorous analysis of cannabis chemistry as well as evaluation of extracts in bioactivity assays.

This document only includes an excerpt of the corresponding thesis or dissertation. To request a digital scan of the full text, please contact the Ruth Lilly Medical Library's Interlibrary Loan Department (rlmlill@iu.edu).

The severity and number of people affected by cancer is a worldwide problem with millions of people affected annually. The search for treatment and cures of cancer continues to be a global effort. As part of this global effort, many natural products have been tested against cancer cell lines, most from plants located in tropical regions. However, this study reports that extracts of Atriplex confertifolia, a native North American plant, has significant bioactivity against human breast cancer cell lines MCF-7, 435, and 231, and HeLa cells (cervical cancer cells). The bioactivity of A. confertifolia extracts of these cells lines was compared to an FDA-approved cancer drug and an industry-standard leukocyte control cell line. Active portions of the extracts were found primarily in the polar fractions of the plant. A dose-response curve of the extracts clearly showed significant cell death similar to the FDA-approved drug. The plant extracts did not inhibit the viability of the leukocyte cell line. Cancer cell death was followed as a function of time and concentration. Cell death appears to be a result of apoptosis.

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Blackberries are rich in polyphenols including anthocyanins. Polyphenols are hypothesized to have biological activities that impact positively on human health. The purpose of these studies was to develop phenolic extracts from selected cultivars of blackberries currently grown in Kentucky as potential Botanical Drug Products for the treatment and prevention of cancer and inflammatory diseases. An ultrasound-assisted ethanol extraction method was employed to obtain anthocyanin-containing extracts (ACEs) from puree or powder (lyophilized puree) of blackberries. ACEs were analyzed for total anthocyanin and phenolics content, polymeric color, and total antioxidant capacity (TAC). The influence of water content in the extraction system was evaluated. A 90 day stability study of the extract and a 48 h stability study of the extract in biologically relevant buffers were completed. HPLC-MS results showed the anthocyanins in ACE were mainly cyanidin-based. As compared to powder-derived ACEs, puree-derived ACEs contained similar amounts of anthocyanins, but greater levels of phenolics and increased TAC. The in vitro antiproliferative effects of ACEs were evaluated in human leukemia (HL- 60), colon (HT-29), and breast (MCF-7) cancer cells. The anticancer mechanism involving reactive oxygen species (ROS) generation was investigated. It was found puree-derived ACEs significantly enhanced production of H2O2 and cytotoxicity in all cell lines as compared to powder-derived ACEs. Cyanidin 3-glucoside exerted anticancer effect by acting synergistically or additively with other active components in the extracts. Furthermore, the phenolic-enriched fractions were separated from non-phenolic fractions in ACEs and found to have potent antioxidant and antiproliferative activities. Pureederived ACE and corresponding phenolic-enriched methanol fraction (MF) induced cell death through ROS-independent caspase 3 pathway whereas the cytotoxicity induced by powder-derived ACE and corresponding MF is related to ROS mechanisms. The in vitro anti-inflammatory studies showed ACEs inhibited Lipid A-induced Interleukin-12 (IL-12) release from mouse dendritic cells, and modulated lipopolysaccharide (LPS)-induced secretion of tumor necrosis factor-α (TNF-α) and Interleukin-6 (IL-6) from murine macrophages. These studies have important implications for the potential use of blackberry extracts for the treatment and prevention of cancer and inflammation diseases and provide essential information for the development of Botanical Drug Products from extracts derived from blackberries and other fruits.

This study aimed to evaluate the antimicrobial efficacy of medicinal hemp plant extracts to determine the antibacterial effects of indigenous Sansevieria species and exotic Cannabis sativa phytotherapy varieties. This study also assessed whether aqueous o

Because of global warming the energy production development has progressed towards more renewable energy sources. Biomass has great potential in this matter and pellet is already a big market that has increased seven times the past decade. A periodically strained woodchip resource market and statements of short supply in the future has got actors exploring opportunities with other commodities. Grasses such as Canary grass has shown great potential in this matter and in this study a wetland grass is tested as an additive, 0,5, 1,0, 1,5, and 1,9%, with spruce woodchips. The test production series was performed at a production unit located at the department of environmental and energy system at Karlstad University, Karlstad. Quality was controlled accordingly to the European standard and parameters such as energy consumption, moisture content, mechanical durability and bulk density was tested. For comparison, a sample with only spruce wood chips was produced, and a sample containing 1% of a commonly used additive, potato starch. The results showed that a decrease in energy consumption with 14% when 2% wetland grass was added, part of the decline may be due to the increased production flow compared with the reference sample. The positive effects on decrease in energy consumption, that 1% potato starch results in, is equal to reults from 1% wetlandgrass. This indicates lubricating properties in wetlandgrass. This is attributed to that herbaceous plants have a high content of extracts such as waxes and that they cause less friction in the press. Tests also showed that pellet with wetland grass did not qualify the European standard in terms of mechanical durability. Extracts can form a weak boundary layer in the pellet and cause this. A possible trend shows a better mechanical durability with more grass in pellets. The presence of different size of particles can be a reason. Moisture content qualifies according to the European standard but is below optimum 8%. This despite to relatively high moisture content in the mixer. Higher moisture content in the press would certainly result in a generally higher quality. Suggestions for future studies are to produce pellets with greater distribution on the wetland grass added, to easier interpret a connection. Also examine the extracts behavior with different moisture content. For a sustainable development accordingly renewable energy it is important to ensure the future commodity market for pellets. Further studies should be performed to help the development of alternative raw materials in conjunction with pellet production. / Den globala uppvärmningen är en påverkande faktor gällande dagens energiutveckling. Europeiska unionens krav på mer förnyelsebar energi är därtill en anledning till det ökade intresset för bioenergi. Pelletsmarknaden har sjufaldigt ökat globalt det senaste decenniet med tidvis ansträngd resursmarknad som följd. Träspån anses komma bli en bristvara i framtiden och aktörer ser sig redan om efter alternativa råvaror. Olika typer av gräs som till exempel rörflen har visat sig ha stor potential. I denna studie testas ett våtmarksgräs som additiv i bränslepellets, med 0,5, 1, 1,5, och 1,9% inblandning. Pelletsproduktion samt tester gjordes i pilotanläggningen på Karlstads universitet, avdelningen för miljö-, och energisystem.  Energiförbrukning, fukthalt, hållfasthet och bulkdensitet är några av de variabler som testades. Pellets med våtmarksgräs jämfördes med ett referensprov med endast granspån samt ett prov med ett vanligt förekommande additiv (potatisstärkelse 1%). Kvalitén på pellets bedömdes utifrån den europeiska standarden. Resultatet visade att den specifika elförbrukningen minskade med 14% när 2% våtmarksgräs adderades, en del av den minskningen kan bero på det ökade produktionsflödet jämfört med referensprovet. De positiva effekter på specifik elförbrukning som 1% potatisstärkelse resulterade i, visade våtmarksgräs 1% liknande effekt. Detta indikerar smörjande egenskaper hos våtmarksgräset. En orsak kan vara den höga halt extrakt, såsom vaxer, som många örtartade växter består av. Dessa har i tidigare studier setts minska friktionen i pressen vilket reducerar elförbrukningen. Pellets med våtmarksgräs som additiv klarade inte de Europeiska kraven på hållfasthet. En låg fukthalt och närvaron av extrakt kan ha medfört svaga gränslager i pellets och därmed påverkat hållfastheten negativt. En svag trend visade på förbättring av hållfastheten vid högre andelar våtmarksgräs. Den varierande storleken på partiklar som sampelleteras kan ha bidragit till detta. Fukthalten är godkänd enligt den europeiska standarden men under det optimala 8%. Detta trots en relativt hög uppmätt fukthalt innan pressen. En högre fukthalt i samband med pressning hade troligen höjt kvaliteten generellt. Förslag till vidare studier är att utföra tester med större variation på mängden våtmarksgräs, för att få ett tydligare resultat. Extraktivens beteende i samband med andra varierande parametrar som exempelvis fukthalt, borde studeras närmare för att kunna bedöma våtmarksgräsets möjligheter som additiv. För en hållbar utveckling av förnybar energi är det viktigt att säkerställa den framtida råvarumarknaden för pellets. Fortsatta studier bör göras för att bidra till utvecklingen av alternativa råvaror i samband med pelletsproduktion.