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Articles by J. Boateng
Total Records ( 24 ) for J. Boateng
  D.S. Williams , M. Verghese , L.T. Walker , J. Boateng , L.A. Shackelford , M. Guyton , J. Jones , J. Khatiwada and C.B. Chawan
  This study was designed to evaluate the anticarcinogenic effect of Flax Seed Meal (FSM) (10 and 20%) and Flax Seed Oil (FSO) (7 and 14%) on Azoxymethane (AOM)-induced colon tumors in Fisher 344 male rats during initiation (I), promotion (P) and Initiation + Promotion (I+P) stages of carcinogenesis. After an acclimatization period of 1 week, 14 groups of Fisher 344 male weanling rats, 3-4 week old (15 per group) were assigned to 2 control groups fed AIN 93G diet and AIN 93G + 14% soybean oil (SBO) (high fat control). The remaining 12 groups were assigned to 10 and 20% FSM (I, P and I+P) and 7 and 14% FSO (I, P and I+P). All rats received 16 mg kg-1 body weight of AOM at 7 and 8 week of age. At 20 weeks of age all animals were switched to AIN-93 M diets and at 45 week of age all rats were killed by CO2 asphyxiation. Tumor incidence (%) in colon of rats fed C (7 and 14%) was both 100. Tumor incidences for rats fed FSO (7 and 14%) at I, P and I+P were: 100, 100, 61, 60, 58 and 61, respectively and 80, 80, 66.6, 66.6, 66.6 and 31, respectively for rats fed FSM (10 and 20%) at I, P and I+P. Tumors per tumor-bearing ratios for groups fed C; 10 and 20% FSM (I, P and I + P) were 3.86, 1.28, 1.70, 1.75 and 1.0, 0.94, 0.64, respectively. In rats fed C (7 and 14%) and 7 and 14% FSO (I, P and I + P) T/TBR ratios were 3.86, 5.96; 1.4, 0.6, 0.6; 1.90, 0.8, 0.8, respectively. Glutathione-S-Transferase activity (a phase II detoxification enzyme) was significantly (p<0.05) higher in rats fed 10 and 20% FSM and 7 and 14% FSO compared to controls. The results of this study indicate that bioactive phytochemicals such as dietary fiber and lignans such as secoisolariciresinol diglycoside (SDG) found in flax seed meal and essential fatty acids such as α-linolenic acid (ALA) found in flax seed oil suppress colon tumors, particularly at the promotion stage and flax seed products may therefore be effective chemopreventive agents.
  J. Boateng , M. Verghese , L.T. Walker , L. Shackelford and C.B. Chawan
  The aim of this study was to examine antitumor effects of Kidney Beans (KB) on Azoxymethane (AOM) induced colon cancer and the cytotoxic effects of KB extracts on colon cancer cell lines (CaCO2). For ACF study, 12 Fisher 344 male rats were fed AIN-93G control diet (C) (n = 6) and 20% KB (n = 6), for 13 week. In the EPTM two groups of rats (n = 14) were fed AIN-93G control diet (C) and 20% KB. All rats received two s/c injections of AOM at 7 and 8 week of age at the rate of 16 mg kg-1 body weight in saline. At 17 week (ACF) and 45 week of age (End-point) all rats were killed by CO2 asphyxiation. For lactate dehydrogenase (LDH) assay, cells were incubated (24 and 48 h) with selected concentrations (0.5-25 mg/100 mL) of KB extract. Total number of ACF was 158 and 72 for groups fed C and 20% KB. Tumor size (mm) and TBR for C and 20% KB were 6.50; 1.16 and 3.8; 1.44, respectively. LDH release (%) in CaCO2 cells after 24 and 48 h incubation with KB extracts ranged from 13.8 to 62.8 and 23.5 to 84.1, respectively. Feeding KB significantly (p<0.05) reduced the incidence of AOM induced colon tumorigenesis and KB extracts demonstrated cytotoxic effects on colon cancer cell lines (CaCO2).
  R. Sunkara , M. Verghese , V. Panala , R. Field , J. Boateng , L.A. Shackelford and L.T. Walker
  In this study, the chemopreventive potential of Cranberry was analyzed in reducing the Aberrant Crypt Foci (ACF) induced by Azoxymethane (AOM) in Fisher 344 male rats. After 1 week period of acclimatization, rats were divided into five different groups. Cranberry meal was mixed in an AIN 93G based diet at 5 and 10% and juice was provided at 2.5 and 5%. Daily feed intake and weekly body weights were recorded. At 17 week of age, rats were killed and samples were collected. Number of ACF and number of cypts/foci were enumerated in the colon. There were no significant differences in feed intake, weight gain, cecal weight and cecal pH among all groups. Total ACF incidence (119) was significantly (p<0.05) higher in control group than in treatment groups. Reduction in total ACF induction was higher in rats fed 10% Cranberry (65.75%) compared to control. A two to six fold increase in selected hepatic enzymes activities (units/mg enzyme) were seen in rats fed 5 and 10% treatment diets compared to control. Results of this study showed that administration of Cranberry meal and juice resulted in significant (p<0.05) reductions in the incidence of ACF in azoxymethane induced preneoplastic lesions.
  R. Field , M. Verghese , L. T. Walker , V. Panala , L. Shackelford and J. Boateng
  The aim of present study was to determine the effects of feeding Wheat Germ Meal (WGM) at 5 and 10% and Wheat Germ Oil (WGO) at 7% (normal fat) and 14% (high fat) on azoxymethane (AOM) induced aberrant crypt foci in Fisher 344 male rats. Following a 1 week period of acclimatization, rats were assigned to 6 groups and fed American Institute of Nutrition 93-Growth diet (AIN-93G) with 7% (C1) and 14% (C2) soybean oil (SBO), AIN-93G diet with 7 and 14% WGO (instead of SBO) and Wheat Germ Meal (WGM) C1+ 5 and 10% WGM. At 7 and 8 week of age all rats received subcutaneous injections of azoxymethane (AOM) at 16 mg kg-1 body weight and were killed at 17 week of age by CO2 asphyxiation. AOM-induced Aberrant Crypt Foci (ACF) were counted. Results showed that ACF formation in distal colons in rats fed 7 SBO, 14 SBO, 7 WGO and 14% WGO (Means±SEM) were 99.2±15.9, 126.0±17.0, 68.2±0.4 and 79.7±1.9 respectively. In rats fed 5% WGM and 10% WGM the number of ACF were 23.75±1.11, 18.6±1.0, 75.2±2.39 and 49.6±2.2 in proximal and distal colon. Total ACF reductions (%) compared to control in rats fed 5% WGM and 10% WGM were 25% and 47%, respectively. Glutathione S-transferase (GST) activities were significantly (p<0.05) higher in treatment groups (5 and 10% WGM) compared to control (7 and 14% SBO). These results indicated that wheat germ and wheat germ oil significantly (p< 0.05) reduced AOM induced ACF. Regular consumption of wheat germ and wheat germ oil may have health implications.
  V. Panala , M. Verghese , J. Boateng , R. Field , L. Shackelford and L.T. Walker
  The objective of this study was to compare the inhibitory effects of Rice bran oil (RBO), Corn oil (CO) and Soybean oil (SBO) at 7% (normal fat level) and 14% (high fat level) on Azoxymethane (AOM) induced Aberrant Crypt Foci (ACF). The long term effect (End Point Tumor (EPT) study) of dietary fat from the above sources on colon cancer in Fisher 344 male rats was determined. In the ACF study 2 groups of F344 rats (4 weeks old) (n = 6) received AIN-93G Control (C) diet containing 7 and 14% Soybean oil (SBO). The remaining groups were assigned treatment diets consisting of 7 and 14% RBO and CO. The rats remained on their respective diets for 13 weeks. Rats in the EPT study were fed a control (AIN-93G) diet with 7% SBO, while the treatment groups were fed diets containing 7% RBO and CO, respectively. At 20 week of age rats in the EPT study were switched to AIN-93Maintenance (M) diets. All rats received 2 s/c injections of AOM at 7 and 8 week of age @ 16 mg kg-1 body weight in saline. At 17 and 45 week of age all rats were killed by CO2 asphyxiation. Total colonic ACF in the rats fed SBO, RBO and CO at 7 and 14% levels ranged from 101-189. In the EPT study, all the rats fed 7% SBO and CO developed tumors (100% tumor incidence) while tumor incidence in the groups fed RBO, was 54% while tumor size (mm) and tumor/Tumor Bearing Rat ratio (TBR) in the rats fed SBO, RBO and CO ranged from 1.3-6.86 and 1.83-5.86, respectively. Present results indicate that the type and constituents (such as n-3 PUFA, vitamin E, phytosterols) of dietary fat plays a significant role in the formation of AOM induced colonic ACF and tumors in Fisher 344 rats.
  D. Gajula , M. Verghese , J. Boateng , L.T. Walker , L. Shackelford , S.R. Mentreddy and S. Cedric
  Basil (Ocimum basilicum L. and Ocimum tenuiflorum L.) contains important phytochemicals that have been reported to afford protection against several chronic diseases due to their anti-inflammatory and antioxidant activities. The purpose of this study was to determine the effects of three accessions of Ocimum tenuiflorum (Holy Basil) Denmark (HBD), Cuba (HBC), India (HBI)) and one accession of Ocimum basilicum (Culinary Basil) (CB) at 1 and 2% levels on azoxymethane (AOM) induced Aberrant Crypt Foci (ACF) in Fisher 344 male rats and to determine the effect of oven drying on total phenolics, flavonoids and anthocyanins of Basil and antioxidative activity. Fifty four rats were divided into 9 groups (n = 6) after a 1 week period of acclimatization. Group 1 was fed a control (C) diet (AIN-93 G) and remaining groups were fed C+1 or 2% CB, HBD, HBC and HBI. All rats received s/c injections of AOM in saline at 16 mg kg-1 b.wt. at 7 and 8 week of age. Rats were killed by CO2 asphyxiation at 17 week of age. The ACF in rats fed C (158.1) was higher than in rats fed C+1% CB, HBD, HBC, HBI (77, 86, 76, 73) and C+2% CB, HBD, HBC, HBI (65, 78, 61, 67). The GST and CAT activities (╬╝mol mg-1) in rats fed C+1 and 2% CB, HBD, HBC and HBI were significantly (p<0.05) higher compared to C. Results showed that feeding culinary and Holy Basil leaves reduced the number of AOM-induced ACF and therefore may have implications in the food industry as a potential chemopreventive agent.
  J. Boateng , M. Verghese , V. Panala , L.T. Walker and L. Shackelford
  In this study we examined the preventive properties of Rice Bran (RB) and germ on the incidence of azoxymethane induced colon tumorigenesis in Fisher 344 male rats. We also examined the cytotoxic and apoptotic properties of RB using an in vivo model. Tumor incidence (%) in C and RB 5% and RB 10% were 100, 55 and 64, respectively. Tumors/tumor Bearing Rats (TBR) were 3.8, 2 and 1.56 for C, RB 5% and RB 10%, respectively. Tumor size (mm) was larger in control (6.50) than in rats fed RB 5% and RB 10% (1.33 and 0.64). After 12, 24 and 48 h of incubation with RB extracts, LDH (%) release ranged from 2.25-46.79. Present results suggest that feeding RB at 5 and 10% levels significantly (p<0.05) reduced the incidence of AOM induced colon tumors in Fisher 344 male rats. We conclude that the protective effects of RB against colon tumorigenesis may possibly be attributed to the synergistic/additive actions of phytochemicals contained in RB.
  V.P. Gourineni , M. Verghese and J. Boateng
  Protective role of prebiotics such as inulin type fructans (Synergy 1®) in colon cancer prevention was demonstrated using animal models. In vitro and in vivo studies have shown the inhibitory effects of dietary fiber and isoflavones such as genistein against the development of cancer. However, the underlying mechanisms of these phytonutrients remain elusive. The aim of this study was to examine the anticancer effects of Synergy1® and soybean extracts in combinations on colon cancer cell line (Caco-2). Cells were incubated (12 and 24 h) with specific concentrations of Syn1®, SM, Syn1®+SM (25-200 μg mL-1), isoflavones (0.125-0.8 μg mL-1) and butyric acid (0.1-0.8 μg mL-1). Lactate dehydrogenase (LDH) release (%) was higher in cells at all concentrations following 24 h incubation with a mixture of Syn1® + SM compared to the cells treated with the extracts alone. LDH release (%) was increased by two fold in cells after incubation with genistein for 24 h compared to the LDH release after 12 h. Caspase-3 activity; a measure of apoptosis seems to be proportional to incubation time in cells treated with Syn1® and SM extracts. Histone associated DNA fragmentation and release of oligonucleotides was higher in cells incubated with a mixture of Syn1®+SM and butyric acid extracts. These results suggests anti-neoplastic properties of Synergy1® and Soybean in combinations by inducing cell toxicity and apoptosis (Caspase-3 and histone associated DNA fragmentation) as a result of synergism between bioactive components present in these functional ingredients. Phytochemical combinations tested at lowest and safer levels in this study may be effective in colon cancer prevention.
  A.A. Miller , M. Verghese , J. Boateng , L. Shackelford and L.T. Walker
  The aim of the study was to test the chemopreventive potential of almonds and pecans on (AOM) induced aberrant crypt foci (ACF). Following a 1 week period of acclimatization, 30 Fisher 344 male rats were randomly divided into 5 groups. One group was fed AIN93G (growth) diet as Control(C) and the other groups were fed almonds (A) and pecans (P) at 5% (5 g/100 g level) and 10% (10 g/100 g level). At 7 and 8 weeks of age rats received subcutaneous injections of Azoxymethane (AOM) at 16 mg kg-1 body weight and were killed by CO2 asphyxiation at 17 weeks of age. Selected enzyme activities such as, glutathione-s-transferase (GST), catalase (CAT) and superoxide dismutase (SOD) were determined. ACF incidence in rats fed Pecans and Almonds at 5 and 10% dose levels were significantly (p<0.05) lower than rats fed control. ACF was reduced by 46-61% in the treatment groups compared to the control. GST and CAT activities (μmol g-1) in rats fed Pecan and Almonds at 5 and 10% dose levels were significantly higher (p<0.05) compared to control fed rats and ranged from 22.03 to 26.44 and 2.38 to 2.98, respectively. We also noted a significant increase in SOD activity (μmol g-1) in rats fed the treatment diets compared to those fed control. Present data indicate that feeding almonds and pecans significantly (p<0.05) reduced incidence of AOM induced ACF which are precancerous lesions.
  D. Asiamah , M. Verghese , J. Boateng , B. Kanda , L. Shackelford and L.T. Walker
  Bitter Melon, (BM) Momordica charantia is known in Asia for regulation of blood glucose levels and as a treatment for Diabetes mellitus. However, studies on its effect in preventing cancer are scarce. This study was conducted to investigate the chemopreventive properties of bitter melon on azoxymethane (AOM)-induced Aberrant Crypt Foci (ACF) in Fisher 344 male rats and determine its effects on selected hepatic detoxification and antioxidant enzymes. Rats were fed AIN93G as control (C) and treatment diets containing 2 and 4% BM. At 7 and 8 weeks of age, ACF was induced by administering 2 s/c injections of AOM at 16 mg kg-1 b.wt. and killed by CO2 asphyxiation at 17 weeks of age. Total ACF in proximal and distal colons were 35, 26 and 21; 116, 54 and 38 in rats fed C, 2% BM and 4% BM, respectively. Catalase and SOD activities (μmol mg-1) were higher in rats fed treatment diets (0.293 and 0.03; 1.82 and 0.27 for rats fed 2 and 4% BM, respectively) compared to control (0.04). Glutathione-S-Transferase (GST) (μmol mg-1) activity and Glutathione (GSH) levels were higher in treatment groups compared to control. Physicochemical analysis was conducted on bitter melon fruit (total phenolics, free radical scavenging activity (DPPH) and Ferric Reducing Antioxidant Power (FRAP)). Total phenolic content was 34.31 mg/GAE/g fresh fruit, DPPH activity was 2.99% and FRAP was 0.652 μmol Fe2+/g/mL. Results indicate bitter melon reduced incidence of AOM induced ACF. Thus inclusion of BM in diets may have possible implications in reducing the risk of colon cancer.
  B. Kanda , J. Boateng , L. Shackelford , S. Appiah , K. Campbell L. T. Walker and M. Verghese
  Peach is an important fruit consumed worldwide. This study was conducted to investigate the effects of processed peaches (dried, frozen and juice) on Azoxymethane-induced (AOM) Aberrant Crypt Foci (ACF) in Fisher 344 male rats, their effect on selected hepatic enzyme activities and to determine the effect of processing (Blanching+Freezing, Blanching+Freeze drying) on total phenolic content and antioxidant potential (DPPH and FRAP). Rats were fed AIN93G as control (CON), treatment diets containing 2 and 4% peaches, and pasteurized juice (PJ) (2 and 4%). Rats received AOM injections s/c at 16 mg kg-1 b wt. during the 7th and 8th wk of age and killed by CO2 asphyxiation at 17 weeks of age. ACF incidence was higher in rats fed CON compared to treatment diets. Rats fed PJ had the lowest ACF (100 and 59 for 2 and 4%) compared to the treatment groups. Blanching (STB+BWB) influenced the incidence of ACF with STB having lower ACF numbers compared to BWB. GST, SOD and CAT (mmol/mg) activities, (9.56-1.43, 1.98-0.49 and 0.53-0.33), respectively were higher in rats fed treatment diets, compared to CON. Total phenolics, ranged from 98.09-120.38, 111.05-120.19, 104.76-122.09 (mg/GAE/g fwb) in FU, FRB and FDB peaches, respectively. FRAP (μg g-1) in FU, FRB and FDB peaches ranged from 0.028-0.042 while (%) DPPH (T30) ranged from 21.588-73.449. Results indicate processing of fruits may impact its role in chemoprevention, total phenolics and antioxidant activity, since fruits are highly perishable; optimum technique is required to minimize losses during and after processing.
  R.L. Miller-Cebert , J. Boateng , L. Shackelford , E. Cebert , L.T. Walker and M. Verghese
  Cruciferous vegetables contain naturally occurring substances that are beneficial to health. The purpose of this study was to investigate the effects of selected cruciferous vegetables (Collard Green (CG), cabbage (CB), turnip green (TG) and canola green (CN) on Azoxymethane (AOM)-induced Aberrant Crypt Foci (ACF) on Fisher 344 male rats. Thirty-six rats were randomly assigned to 9 groups (n = 4). The control group was fed AIN-93G diet and the other eight groups were fed AIN-93G along with the selected vegetables at 5 and 10%. The rats were administered subcutaneous injections of AOM at 7 and 8 weeks of age at 16 mg kg-1 body weight. At 17 weeks of age, rats were killed by CO2 asphyxiation. Total ACF numbers in rats fed CB, TG, CG and CN at 5% were 55, 41, 47 and 59, while at the 10% level, ACF numbers were 54, 63, 54 and 46, respectively. Total ACF in rats fed 5 and 10% cruciferous vegetables were significantly (p<0.05) lower (41-63) than rats fed the control diet (151). Findings indicate that cruciferous vegetables, including canola reduced the incidence of ACF and could potentially be used as a dietary chemopreventive agent against colon cancer.
  S. Appiah , M. Verghese , J. Boateng , L.A. Shackelford , B. Kanda , J. Patterson and L.T. Walker
  Beets (Beta vulgaris) have been reported to be a very nutritious vegetable which may provide health benefits against chronic diseases such as colon cancer due to phytochemicals present. The aim of this study was to investigate the effect of feeding Freeze Dried (FD), Cabinet Dried (CD) and pasteurized juice at 2 and 4% levels on Azoxymethane (AOM) induced Aberrant Crypt Foci (ACF) in Fisher 344 male rats and to determine the total phenolics and flavonoids content and antioxidants activity measured using FRAP and DPPH in steam blanched (SB), frozen, “individually quick frozen” (IQF), freeze dried and cabinet dried compared to fresh beets. Twenty eight rats were divided into 7 groups at 4 rats per group. The groups were fed control (C) diet (AIN-93G), C+2 and 4% FD, CD and beet juice. All rats received s/c injections of AOM in saline at 16 mg kg-1 b.wt. at 7 and 8 weeks. Rats were killed by CO2 asphyxiation at the 17 week of age. Number of ACF, crypts/foci and total crypts were enumerated in the colon. Total phenolics, flavonoids and monomeric anthocyanin content as well antioxidant activity (using DPPH and FRAP) of beets (freeze dried, cabinet dried, “individually quick frozen”, conventionally frozen steam blanched and fresh) were also determined. The total ACF incidence was significantly (p<0.05) higher in the control compared to beet fed groups. ACF reductions ranged from a low of 63% in rat fed 2% freeze dried beets to a high of 80% in rats fed 4% freeze dried beets. Total phenolics and flavonoids were significantly (p<0.05) higher in cabinet dried and freeze dried beets and higher antioxidant activity compared to the others (fresh, steam blanched, IQF and frozen). The results from the experiment indicates that feeding beets(cabinet dried, freeze dried or juice) reduced the incidence of AOM-induced ACF and therefore may be explored for its chemopreventive potential and other health benefits by the food industry.
  K. Busambwa , R.L. Miller-Cebert , L. Aboagye , L. Dalrymple , J. Boateng , L. Shackelford , L.T. Walker and M. Verghese
  Peas (yellow and green) and lentils contain bioactive compounds with protective properties. Sprouting of grains has shown to improve nutritional value due to activation of hydrolytic enzymes. The aim of this study was to determine anticarcinogenic potential of sprouted and non-sprouted green-split and yellow peas and lentils at 5 and 10% on Azoxymethane (AOM)-induced Aberrant Crypt Foci (ACF) in Fisher 344 male rats. Seventy-eight rats were randomly assigned to 13 groups: Control (C) received AIN-93G diet; treatment diets included Lentils sprouted and non-sprouted (LS, LNS), Green peas sprouted and non-sprouted (GS, GNS), Yellow peas sprouted and non-sprouted (YS, YNS) at 5 and 10% levels in an AIN 93 based diet. At 7 and 8 week, rats received 2 subcutaneous injections of AOM at 16 mg kg-1 b.wt. Rats were euthanized at 17 week by CO2 Asphyxiation. Total ACF ranged from 88 to 181.33 in rats fed C, LS and LNS, GS and GNS, YS and YNS. ACF reductions (%) compared to C were 68.56; 63.25 and 51.46 in rats fed LS at 10%, LNS at 10% and YNS at 10%, respectively. Glutathione content, Glutathione-S-transferase and Catalase activities were significantly higher (p<0.05) in rats fed treatment diets compared to C. Findings from this study indicate that sprouted and non sprouted green split and yellow peas and lentils reduced AOM-induced aberrant crypt foci in F344 male rats with the greatest reduction in rats fed Lentil sprouted (10%). Selected legumes appear to have beneficial potential in reducing colon cancer.
  R. Offei- Oknye , J.L. Patterson , L.T. Walker , J. Boateng and M. Verghese
  Phytochemicals are known to have antioxidative, antibacterial, antifungal, anti-inflammatory and anticancer properties. In this study, HepG2 hepatoma cells were incubated after 12 and 24 h with 80% methanol extracts of processed ginger: Oven dried, sun dried, freeze dried and fresh ginger (control). The objectives of this study were to investigate the chemopreventive potential of ginger extracts in hepatoma cell lines (induction of cytotoxicity and apoptosis) and to determine the effects of ginger extracts on selected detoxification and anti-oxidative enzymes: Glutathione-S-transferase, Glutathione reductase, Glutathione peroxidase, Superoxide dismutase, Catalase activity and Glutathione in cell lines (hepatoma). Processed ginger extracts induced higher cytotoxicity (%) in HepG2 (SD 60 μg mL-1 [92.89]). Induction of cytotoxicity (%) was higher after 24 h of incubation compared to 12 h. Histone related DNA fragmentation (enrichment factor), an indicator of apoptosis was higher after 12 h compared 24 h of incubation except with cell incubated with FD (240 μg mL-1). Detoxification and antioxidative enzymes studied generally showed a higher induction after 24 h compared to 12 h of incubation in SOD, GR, CAT, GPx and GSH (antioxidant substrate). These findings indicate that ginger may exerts its chemopreventive effects by its cytotoxic activity and the induction of detoxification and antioxidative enzymes.
  K. Busambwa , R. Sunkara , N. Diby , R. Offei- Okyne , J. Boateng and M. Verghese
  Legumes such as lentils, green and yellow split-peas have been reported to provide health benefits against colon cancer due to nutrients and non-nutrient phytochemical compounds present in them. The aim of this study was to investigate the effect of selected legumes in inducing cytotoxicity, apoptosis and/or necrosis in human epithelial colorectal adenocarcinoma cells. Methanolic extracts of sprouted and non-sprouted legumes (Lentils, green peas, yellow peas) were prepared. Caco-2, human colonic carcinoma cell line was obtained from the American Type Culture Collection (ATCC, Manassas, VA). Cells were incubated with legume extracts at selected concentrations (1.5, 3, 4.5, 6, 7.5 and 10 mg mL–1) for determination of lactate dehydrogenase (LDH) release (cytotoxicity %). The ability of legume extracts in inducing cell apoptosis was analyzed at selected concentrations of 1.5, 3, 6 and 12 mg mL–1 by quantifying DNA fragmentation. Morphological changes in cells were observed microscopically. The activity of caspase-3 enzyme was also determined. Results showed that legume extracts induced cytotoxicity in caco-2 cells in a dose-dependent manner. A 20-80% increase in release of LDH was observed in cells incubated with extracts compared to untreated cells. Total fragmented DNA (%) was higher in cells treated with sprouted legume extracts compared to their non-sprouted extracts. More than 40% of total DNA fragmentation was observed in cells treated with sprouted legumes. Caspase-3 activity was highest in cells incubated with non-sprouted yellow split peas at 6 mg mL–1. Specific morphological changes related to apoptosis such as: surface blebbing, formation of apoptotic bodies, blisters and echinoid spikes and cell membrane destabilization were observed in cells incubated with extracts. These results suggested the cytotoxic and apoptosis inducing potential of legume extracts. Bioactive components in these selected legumes may be used as chemopreventive agents.
  J. Boateng , R. Miller-Cebert , L. Shackelford and M. Verghese
  Background and Objectives: There is limited information on the effect of pistachio nuts in cancer prevention. Since pistachios contain several protective compounds with antioxidants properties, the aim of this study is to determine if pistachios can reduce precancerous colon cancer lesions in rats by affecting several biomarkers of oxidative stress including induction of endogenous antioxidant enzymes. Methodology: Thirty Fisher 344 male rats were randomly assigned to 5 groups. Rats in groups 1 and 2 were fed AIN-93G as positive (CON+) and negative control (CON-), while rats in groups 3-5 were assigned AIN-93G with 5, 10 and 15% pistachio meal (PM). Rats (except group 2), received AOM injections at 7 and 8 weeks of age and killed at 17 weeks of age by CO2 asphyxiation. Results: Total ACF and crypt multiplicity, antioxidant enzymes: Glutathione-s-transferase (GST), glutathione (GSH), glutathione peroxidase (GPx), catalase (CAT), glutathione reductase (GR) and superoxide dismutase (SOD) were determined. Total serum cholesterol (TC) and triglycerides (TG) were also assessed. Results showed significant (p<0.05) reductions in ACF in all treatment groups compared to the rats fed CON+(158). The ACF ranged from 72-119 in rats fed 5, 10 and 15% PM. Rats in group 2 developed no ACF. Significant (p<0.05) increase in antioxidant enzyme activities (protein μmol mg–1) was observed compared to CON+. The GST (5.86-10.84), GR (4.91-7.21), CAT (1.66-3.38), GPx (4.25-7.61) and SOD (protein U mg–1) (1.02-1.73). The TC and TG were significantly (p<0.05) decreased in the treatment groups compared to CON+. Data indicated PM reduced ACF by enhancing phase II and antioxidant enzyme activities and reducing serum lipids. Conclusion: The PM could be investigated as putative dietary chemoprevention in colon cancer therapy.
  J. Boateng and M. Verghese
  Consumption of fruits and the other dietary antioxidants are considered beneficial due to the protection they afford in the pathogenesis associated with oxidative stress. The aim of this study was to evaluate the antioxidative effects of selected fruit extracts (Plums, Apples, Grapes and Cranberries) on human lung fibroblasts (CCD-25LU) exposed to tert-butyl hydroperoxide (tBHP) oxidative stress. Lactate Dehydrogenase (LDH) was used to assess cytotoxicity (cell integrity) and antioxidant enzymes catalase (CAT), glutathione-s-transferase (GST), glutathione peroxidases (GPx) and concentrations of reduced glutathione (GSH) were determined. Results showed that LDH release by cells pretreated with fruits extracts were significantly (p<0.05) lower compared to cells treated with tBHP alone. Antioxidant enzymes (CAT, GST and GPx) in cells pretreated with fruit extracts were increased by 2-4 folds compared to cell exposed to tBHP alone. GSH levels which were significantly (p<0.05) reduced after exposure to tBHP were restored by pretreatment with fruit extracts. Fruits extracts used in this study protected CCD-25LU against oxidative stress induced by tBHP and reduced cell damage. Consumption of fruits may therefore play a significant role in protection against oxidative induced lung diseases.
  M. Verghese , J.E. Richardson , J. Boateng , L.A. Shackelford , C. Howard , L.T. Walker and C.B. Chawan
  The aim of the present study was to determine the effect of lycopene on hypercholesterolemia in New Zealand male rabbits. Rabbits (n = 25) were divided into five groups (n = 5) and fed a normal laboratory diet (NC), a high cholesterol (HC) (0.5 g/100 g) diet, a high cholesterol diet with 42.6 ppm (HC-LYC1), 85.2 ppm (HC-LYC2) and 127.8 ppm (HC-LYC3) lycopene, respectively. Rabbits were fed their respective diets for 12 weeks. Serum was analyzed for lipids: total cholesterol (TC), (triacylglycerol) TG, LDL-C (low density lipoprotein-cholesterol), High density lipoprotein-Cholesterol (HDL-C) and apolipoproteins A1 (Apo A1) and B (Apo B). Livers were collected for measurement of HMG-CoA reductase and ACAT activities and atherosclerotic plaque formation were assessed in aorta. Results showed that rabbits fed HC-LYC1, HC-LYC2 and HC-LYC3 had significantly (p≤0.05) lower serum TC, LDL, TG, ApoB and significantly higher HDL and Apo A1 levels compared to the control-HC. HDL-C (mmol L-1) in HC-LYC1, HC-LYC2 and HC-LYC3 were 2.81, 2.99 and 3.71, respectively compared to 1.93 in the control-HC group. There was a highly significant (p≤0.0001) reduction in hepatic HMG-CoA reductase levels in the groups fed the lycopene diets (HC-LYC1, HC-LYC2 and HC-LYC3) compared to the control-HC. A significant (p≤0.05) reduction in ACAT activity was seen as lycopene levels increased. The percent of atherosclerotic plaque in the aorta of the rabbits fed the control-HC diets (41.5%) was reduced to 14.8 ppm when lycopene was added to the diet at 127.8 ppm. The results of this study demonstrated that lycopene might play a significant role in the prevention of cardiovascular diseases.
  A. Clisby , M. Verghese , E. Cebert , L.T. Walker , R. Field , L. Shackelford , J. Boateng and C.B. Chawan
  The aim of the current study was to determine the efficacy of selected canola or mustard seeds (S) and canola sprouts (SP) cultivars (5 and 10% levels) on azoxymethane (AOM) induced Aberrant Crypt Foci (ACF) in Fisher 344 male rats. Following a one week period of acclimatization Fisher 344 male rats were randomly assigned to groups (n = 6). At seven and eight weeks of age rats received two successive s/c injections of AOM in saline at 16 mg kg-1 body weight. Rats were killed by CO2 asphyxiation at 17 weeks of age. The number of ACF in the proximal and distal colon in the treatment groups ranged from 6.33 (5% Pacific gold) to 15.55 (5% Jetton) and 21.33 (10% Pacific gold) to 52.0 (5% Idagold). Glutathione-S-transferase activity (μmol mg-1) was significantly (p<0.05) higher in the groups fed canola or mustard seeds compared to the control. Among the treatment groups the highest GST activity (μmol mg-1) was observed in the rats fed 10% Pacific gold group (32.50). The results from this study show that canola or mustard sprouts and seeds reduced the incidence of AOM-induced ACF and may possibly prevent the incidence of colon cancer.
  M. Yang , R. Hardin , S. Ogutu , M. Verghese and J. Boateng
  Background and Objective: Basil and ginger possess various beneficial effects such as antioxidant, anti-inflammatory and antimicrobial. However, studies on their bioefficacy do not consider changes to composition and chemical structure from processes such as digestion which may alter their bioactivity. The objective of this study was to evaluate the bioefficacy of bioaccessible fractions of basil and ginger in a HepG2 cell model before and after simulated in vitro digestion. Methodology: Digested and non-digested basil (BD and BND) and ginger (GD and GND) extracts were prepared and used for the determinations of Total Phenolic Content (TPC), Total Flavonoid Content (TFC), Ferric Reducing Antioxidant Power (FRAP), Oxygen Radical Absorbance Capacity (ORAC) Assay and Trolox Equivalent Antioxidant Capacity (TEAC). Lactate dehydrogenase (LDH) and 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) were used as measures of efficacy. Glutathione (GSH) and Glutathione-S-Transferase (GST) were also determined. Results: Results showed that TFC was significantly (p≤0.05) increased after in vitro digestion while, a 3 fold decrease was noted in TPC. Antioxidant activities were also decreased after in vitro digestion. The BD extracts exerted significant (p≤0.05) cytotoxicity (LDH) and reduced (p≤0.05) cell viability (MTT) compared to cells treated with BND extracts. The reverse was however, observed in cells treated with GD and GND. LDH in cells treated with GND ranged from 5-53% and 6-67%, respectively for 12 and 24 h compared to cells treated with GD which ranged from 4-18% and 9-28%, respectively for 12 and 24 h. The GSH levels and GST activities were significantly (p≤0.05) higher in cells treated with BND extracts compared to cells treated with BD extracts. However, results varied with ginger extracts. Conclusion: Although, the results indicated that digested and non-digested extracts of basil and ginger induced cytotoxicity and reduced cell viability in HepG2 cells, the distinct differences in the level of efficacy may reflect alterations in the polyphenolic composition caused by the digestion process.
  J. Patterson , J. Boateng , L.T. Walker and M. Verghese
  N-nitrosamines are considered human carcinogens and have been found in cured meats, seafood, vegetables, apples, beer, drinking water, waste water, tobacco products and rubber products. Limited studies exist on the effects of low dose exposure to multiple N-nitrosamines compounds. The objectives of this study were to investigate the cellular mechanisms of action by which N-nitrosamines exhibit toxicity resulting in liver tumors and other effects. Hep2G human liver cells (ATCC HTB-37) was obtained from the American Type Culture Collection (ATCC, Manassas, VA). For assay, 5x104 Hep2G cells/well were seeded in a 24 well culture plate and incubated at 37°C and 7% CO2 until development of a monolayer. Cells were incubated with a combination of selected N-nitrosamines at selected concentrations (0, 4, 8, 16, 32, 64 mM) for 12 and 24 h. Lactate dehydrogenase (LDH) release (% cytotoxicity), histone-related DNA fragmentation and detoxification enzymes were determined. After 12 and 24 h incubation with N-nitrosamines,% cytotoxicity in Hep2G cells displayed a dose-dependent relationship at concentrations of 4, 8 and 16 mM. Cytotoxicity peaked at 16 mM for both time periods and then decreased with increasing concentration (64 mM) to 19.46 (12 h) and 55.73 (24 h). Overall, levels of glutathione-S-transferase (GST), glutathione peroxidase (GPx), Glutathione Reductase (GR) and Superoxide Dismutase (SOD) were higher with control compared to N-nitrosamines-treated cells. Histone-related DNA fragmentation was highest in cells treated with 8 mM (24 h). Possible mechanisms of action may be due to lower detoxification enzymes and/or an increase in H2O2 production, leading to cell death.
  K. Busambwa , M. Verghse , R.M. Cebert , L. Dalrymple , J. Allen , J. Boateng , L. Shackelford and L.T. Walker
  Lentils, green and yellow split-peas have been reported to provide health benefits against colon cancer due to the amount of nutrients and non-nutrient phytochemical compounds present. The aim of this study was to investigate and compare the chemopreventive potential of sprouted and non-sprouted lentils (LS, LNS), green (GS, GNS) and yellow split-peas (YS, YNS) on azoxymethane (AOM)-induced colon cancer. Following a 1 week acclimatization period, 42 Fisher-344 male rats were randomly assigned to 6 groups (n = 6). Five groups were fed treatment diets consisting of the selected legumes (Sprouted and non-sprouted), while the control group (C) was fed AIN-93 growth and maintenance of diet. Colon tumors were induced by administration of AOM at 7 and 8 weeks of age. Rats were killed by CO2 asphyxiation at age 46 weeks. Results showed lower tumor incidence in treatment groups at 66.7% in GS compared to 100% in LNS and the control. Rats fed control diet had higher Tumors/Tumor Bearing Rat (TBR) ratio (4.33) compared to those in treatment groups (1.2-2). Cecal pH was significantly higher in control (7.81) compared to the treatment diets. Glutathione-S-Transferase (GST) activity was significantly higher in sprouted legumes (8.55-14.04 μM min–1 mL–1) compared to non-sprouted legumes (4.53-5.67 μM min–1 mL–1). Glutathione concentration (GSH) ranged from a low of 636.34 μM in rats fed GNS to a high of 791.07 μM in rats fed YNS. Selected legumes were effective in reducing incidence of AOM-induced colon tumors in Fisher-344 male rats (2.1-4.3 times) and may be promoted for consumption as part of healthy eating habits to prevent chronic diseases such as cancer.
  S. Willis , J. Boateng , K. Busambwa , L. Shackelford and M. Verghese
  Background and Objective: Demand for plant preparations is on the rise urging the need to investigate their potential for toxicity. The aim of this study was to evaluate the effect of oral supplementation of bitter melon (BMLT) and blueberry leaf teas (BLUT) on redox and drug metabolizing enzymes: Glutathione (GSH) levels, Glutathione-S-Transferase (GST), superoxide dismutase (SOD), catalase (CAT) activities were determined. Methodology: Rats (n = 5) were administered once daily by gavages 0, 1000, 2000 and 4000 mg kg–1 b.wt., of BMLT, BLUT for 28 days. Day 29, rats were killed by CO2 asphyxiation and tissue and blood samples were collected for analysis. Results: Results also showed GST, CAT and SOD (μmol mg–1 protein) activities in experimental groups ranged from 0.89-2.14, 0.93-1.38 and 0.35-0.55, respectively. There were no significant differences in GSH (μmol/mg protein) levels between the control and treatment groups. Conclusion: Results indicated BMLT and BLUT prevented oxidative stress by improving the activity of antioxidant enzymes. Results also showed no apparent toxicity in rats administered LHT.
 
 
 
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