Inhibitory potential of honey on the enzymatic activity of Helicobacter pylori urease
- Authors: Matongo, Fredrick
- Date: 2012
- Subjects: Honey , Helicobacter pylori infections , Enzyme inhibitors , Traditional medicine , Antifungal agents
- Language: English
- Type: Thesis , Masters , MSc (Biochemistry)
- Identifier: vital:11253 , http://hdl.handle.net/10353/431 , Honey , Helicobacter pylori infections , Enzyme inhibitors , Traditional medicine , Antifungal agents
- Description: Urease of Helicobacter pylori is an important virulence factor implicated in the pathogenesis of many clinical conditions, such as chronic gastritis, peptic ulceration, and gastric cancer. Many urease inhibitors have been discovered, like phosphorodiamidates, hydroxamic acid derivatives, and imidazoles. Despite good activities at the enzyme level and excellent kinetic properties most of them have not been used as therapeutic agents in vivo because of their side effects, toxicity and instability. This has led to much attention to focus on exploring the novel urease inhibitory activities of natural products because of their low toxicity and good bioavailability. Honey, a natural product has been used in folk medicine due to its antitumor, antioxidant, antimicrobial and anti-inflammatory properties. The aims of this study were to isolate, characterise, purify urease produced by H. pylori and investigate the inhibitory effects of solvent honey extracts on its enzymatic activity. Urease was found to be both surface-associated and cytoplasmic. Maximum cytoplasmic urease activity was found to occur after 72 hr whereas maximum extracellular urease activities were found to occur after 96 hr. Characterization of the crude cytoplasmic urease revealed optimal activity at a pH of 7.5 and temperature of 40°C. The kinetic parameters Vmax and Km were 45.32 U ml-1 and 61.11 mM respectively.The honey extracts inhibited the activity of the crude urease in a concentration dependent manner. The Lineweaver-Burk plots indicated a non-competitive type of inhibition against H. pylori urease. The two honey extracts gave promising inhibitory activities against urease of H. pylori. Thus the results of this study delineates that inhibition of urease can ease development in therapeutic and preventative approaches based on the enzymatic activity of this Helicobacter protein.
- Full Text:
- Date Issued: 2012
- Authors: Matongo, Fredrick
- Date: 2012
- Subjects: Honey , Helicobacter pylori infections , Enzyme inhibitors , Traditional medicine , Antifungal agents
- Language: English
- Type: Thesis , Masters , MSc (Biochemistry)
- Identifier: vital:11253 , http://hdl.handle.net/10353/431 , Honey , Helicobacter pylori infections , Enzyme inhibitors , Traditional medicine , Antifungal agents
- Description: Urease of Helicobacter pylori is an important virulence factor implicated in the pathogenesis of many clinical conditions, such as chronic gastritis, peptic ulceration, and gastric cancer. Many urease inhibitors have been discovered, like phosphorodiamidates, hydroxamic acid derivatives, and imidazoles. Despite good activities at the enzyme level and excellent kinetic properties most of them have not been used as therapeutic agents in vivo because of their side effects, toxicity and instability. This has led to much attention to focus on exploring the novel urease inhibitory activities of natural products because of their low toxicity and good bioavailability. Honey, a natural product has been used in folk medicine due to its antitumor, antioxidant, antimicrobial and anti-inflammatory properties. The aims of this study were to isolate, characterise, purify urease produced by H. pylori and investigate the inhibitory effects of solvent honey extracts on its enzymatic activity. Urease was found to be both surface-associated and cytoplasmic. Maximum cytoplasmic urease activity was found to occur after 72 hr whereas maximum extracellular urease activities were found to occur after 96 hr. Characterization of the crude cytoplasmic urease revealed optimal activity at a pH of 7.5 and temperature of 40°C. The kinetic parameters Vmax and Km were 45.32 U ml-1 and 61.11 mM respectively.The honey extracts inhibited the activity of the crude urease in a concentration dependent manner. The Lineweaver-Burk plots indicated a non-competitive type of inhibition against H. pylori urease. The two honey extracts gave promising inhibitory activities against urease of H. pylori. Thus the results of this study delineates that inhibition of urease can ease development in therapeutic and preventative approaches based on the enzymatic activity of this Helicobacter protein.
- Full Text:
- Date Issued: 2012
The allelopathic potential of Arctotis Arctotoides (L.f.) O. Hoffm on some vegetables
- Authors: Badmus, Abimbola Adesile
- Date: 2012
- Subjects: Allelopathy , Allelopathic agents , Vegetables -- Microbiology , Enzyme inhibitors
- Language: English
- Type: Thesis , Doctoral , PhD (Botany)
- Identifier: vital:11306 , http://hdl.handle.net/10353/454 , Allelopathy , Allelopathic agents , Vegetables -- Microbiology , Enzyme inhibitors
- Description: Laboratory and greenhouse experiments were conducted to evaluate the allelopathic effects of the extracts and residue Arctotis arctotoides (L.f.) O. Hoffm on selected vegetable crops. The study aimed to address the following specific objectives to (i) examine the ultra structures of the leaf of A. arctotoides using the Scanning Electron Microscope (SEM), (ii) carry out comprehensive qualitative and quantitative phytochemical analysis of the root and shoot materials of the plant, (iii) investigate the allelopathic activities of the root and shoot aqueous extracts of A. arctotoides at concentrations of 10, 8, 6, 4 and 2 mg/ml on germination, radicle and plumule growth of cabbage, carrot, tomato and spinach, (iv) evaluate the inhibitory effects of the dried shoot residue of the plant at 10, 20 and 40 g kg-3 of soil (treatments B, C and D) and the control (treatment A) on the morphology, growth and chlorophyll pigment content of tomato and cabbage transplants at 1, 2, 3 and 4 weeks after transplanting and (v) assess the effects of the dried shoot residue of A. arctotoides on the yield, nutrient uptake by the leaves of tomato and cabbage at 4 and 12 weeks after transplanting. Finally, to analyze the residual mineral content of the soils with tomato and cabbage transplants at 12 weeks after transplanting. The the SEM revealed that anisocytic stomata and glandular trichomes (GTs) were numerous on the abaxial than the adaxial surfaces of A. arctotoides. The non glandular trichomes (NGTs) were also present on both surfaces but lesser on the abaxial. Morphologically, the GTs were peltate, uniseriate and globular head while the NGTs were cylindrical and filamentous with variable number of cells at the basal portion. The energy dispersive X-ray spectroscopy of some crystals showed that Na+ Mg2+ and Ca2+ were the major constituents of the crystal deposit found around the GTs and stomata. The results of the phytochemical composition of the root and shoot extracts of A. arctotoides confirmed the occurrence of alkaloids, flavonoids, phenolics, saponnins, tannins and triterpenes as the common constituents. In addition, cardiac glycosides and steroids were also detected in the shoot of the A. arctotoides. Quantitative estimation of the chemical constituents of the crude extracts further revealed that the alkaloid content in the root higher (0.97 percent) than the shoot (0.64 percent). The quantity of flavonoids detected in the shoot (1.02 percent) was more than that observed in the root (0.35 percent). Others (phenolics and tannins) were marginal in the two plant parts. The results of the inhibitory effects of the root and shoot aqueous extract at the varying concentrations showed that root extract at 10 mg/ml considerably reduced the germination of cabbage, carrot, tomato and spinach seeds by 84.0, 83.2, 72.8 and 37.4 percent respectively. Incubation of the shoot extract at the same concentration resulted in 100 percent inhibition of cabbage and carrot seed germination whereas those of tomato and spinach were suppressed by 91.5 and 61.2 percent respectively. The two extracts at the varying concentrations also had a significant reduction on the radicle and plumule growth of the four vegetables. Addition of the shoot residue to the soil showed massive chlorosis, necrotic lesions and wilting of tomato and cabbage leaves under treatments C and D at 2 weeks after transplanting. The number of leaves, leaf area, dry shoot and root weight of the two vegetables grown in the amended soils were also drastically reduced. The inhibition percentages due to the addition of the three concentrations of A. arctotoides dried shoot residue on the dry shoot weight at 4 weeks after transplanting were 38.6, 45.5 and 70.3. for tomato and 57.5, 73.3 and 87.5 percent for cabbage. Similarly, the declines in the dry root weight of 61.3, 82.9.4 and 83.4 percent for tomato as well as 53.1, 54.7 and 67.2 percent for cabbages were recorded for the two vegetables under treatment B, C and D during the period. The results further showed that the dry fruit yield and shoot weight of tomato under the treatments B, C and D decreased with increase in shoot residue concentrations of A. arctotoides. Relative to treatment A, no significant differences were recorded in the dry head weight of cabbage under the residue treated groups. The reductions in the fruit yield and fresh head weight caused by treatments C and D were 37.2 and 84.8 percent for tomato and 30.9 and 72.4 percent for cabbage. The findings on the mineral contents in the leaves of the two vegetables revealed significant differences in the uptake of N, Mg, Na, Cu and Fe by tomato leaves. The concentrations of N, K, Na and Zn in cabbage leaves also differed. However, the P content was relatively constant in the leaves of the two vegetables at 4 and 12 weeks after transplanting. At 12 weeks after transplanting, the Fe content in soils with tomato and cabbage treatments C and D was greatly enhanced in comparison with the other nutrients. The residual N, P and Zn detected in soils planted to cabbage were similarly equal among all the groups including the control. Thus, under the greenhouse experiment, Arctotis arctotoides (L.f) O. Hoffm has been shown to contain some phytotoxic chemical compounds in its root and shoot materials. The compounds either singly or collectively have demonstrated some inhibitory potentials on the germination, growth and yields of cabbage, carrot, tomato and spinach evaluated in this study.
- Full Text:
- Date Issued: 2012
- Authors: Badmus, Abimbola Adesile
- Date: 2012
- Subjects: Allelopathy , Allelopathic agents , Vegetables -- Microbiology , Enzyme inhibitors
- Language: English
- Type: Thesis , Doctoral , PhD (Botany)
- Identifier: vital:11306 , http://hdl.handle.net/10353/454 , Allelopathy , Allelopathic agents , Vegetables -- Microbiology , Enzyme inhibitors
- Description: Laboratory and greenhouse experiments were conducted to evaluate the allelopathic effects of the extracts and residue Arctotis arctotoides (L.f.) O. Hoffm on selected vegetable crops. The study aimed to address the following specific objectives to (i) examine the ultra structures of the leaf of A. arctotoides using the Scanning Electron Microscope (SEM), (ii) carry out comprehensive qualitative and quantitative phytochemical analysis of the root and shoot materials of the plant, (iii) investigate the allelopathic activities of the root and shoot aqueous extracts of A. arctotoides at concentrations of 10, 8, 6, 4 and 2 mg/ml on germination, radicle and plumule growth of cabbage, carrot, tomato and spinach, (iv) evaluate the inhibitory effects of the dried shoot residue of the plant at 10, 20 and 40 g kg-3 of soil (treatments B, C and D) and the control (treatment A) on the morphology, growth and chlorophyll pigment content of tomato and cabbage transplants at 1, 2, 3 and 4 weeks after transplanting and (v) assess the effects of the dried shoot residue of A. arctotoides on the yield, nutrient uptake by the leaves of tomato and cabbage at 4 and 12 weeks after transplanting. Finally, to analyze the residual mineral content of the soils with tomato and cabbage transplants at 12 weeks after transplanting. The the SEM revealed that anisocytic stomata and glandular trichomes (GTs) were numerous on the abaxial than the adaxial surfaces of A. arctotoides. The non glandular trichomes (NGTs) were also present on both surfaces but lesser on the abaxial. Morphologically, the GTs were peltate, uniseriate and globular head while the NGTs were cylindrical and filamentous with variable number of cells at the basal portion. The energy dispersive X-ray spectroscopy of some crystals showed that Na+ Mg2+ and Ca2+ were the major constituents of the crystal deposit found around the GTs and stomata. The results of the phytochemical composition of the root and shoot extracts of A. arctotoides confirmed the occurrence of alkaloids, flavonoids, phenolics, saponnins, tannins and triterpenes as the common constituents. In addition, cardiac glycosides and steroids were also detected in the shoot of the A. arctotoides. Quantitative estimation of the chemical constituents of the crude extracts further revealed that the alkaloid content in the root higher (0.97 percent) than the shoot (0.64 percent). The quantity of flavonoids detected in the shoot (1.02 percent) was more than that observed in the root (0.35 percent). Others (phenolics and tannins) were marginal in the two plant parts. The results of the inhibitory effects of the root and shoot aqueous extract at the varying concentrations showed that root extract at 10 mg/ml considerably reduced the germination of cabbage, carrot, tomato and spinach seeds by 84.0, 83.2, 72.8 and 37.4 percent respectively. Incubation of the shoot extract at the same concentration resulted in 100 percent inhibition of cabbage and carrot seed germination whereas those of tomato and spinach were suppressed by 91.5 and 61.2 percent respectively. The two extracts at the varying concentrations also had a significant reduction on the radicle and plumule growth of the four vegetables. Addition of the shoot residue to the soil showed massive chlorosis, necrotic lesions and wilting of tomato and cabbage leaves under treatments C and D at 2 weeks after transplanting. The number of leaves, leaf area, dry shoot and root weight of the two vegetables grown in the amended soils were also drastically reduced. The inhibition percentages due to the addition of the three concentrations of A. arctotoides dried shoot residue on the dry shoot weight at 4 weeks after transplanting were 38.6, 45.5 and 70.3. for tomato and 57.5, 73.3 and 87.5 percent for cabbage. Similarly, the declines in the dry root weight of 61.3, 82.9.4 and 83.4 percent for tomato as well as 53.1, 54.7 and 67.2 percent for cabbages were recorded for the two vegetables under treatment B, C and D during the period. The results further showed that the dry fruit yield and shoot weight of tomato under the treatments B, C and D decreased with increase in shoot residue concentrations of A. arctotoides. Relative to treatment A, no significant differences were recorded in the dry head weight of cabbage under the residue treated groups. The reductions in the fruit yield and fresh head weight caused by treatments C and D were 37.2 and 84.8 percent for tomato and 30.9 and 72.4 percent for cabbage. The findings on the mineral contents in the leaves of the two vegetables revealed significant differences in the uptake of N, Mg, Na, Cu and Fe by tomato leaves. The concentrations of N, K, Na and Zn in cabbage leaves also differed. However, the P content was relatively constant in the leaves of the two vegetables at 4 and 12 weeks after transplanting. At 12 weeks after transplanting, the Fe content in soils with tomato and cabbage treatments C and D was greatly enhanced in comparison with the other nutrients. The residual N, P and Zn detected in soils planted to cabbage were similarly equal among all the groups including the control. Thus, under the greenhouse experiment, Arctotis arctotoides (L.f) O. Hoffm has been shown to contain some phytotoxic chemical compounds in its root and shoot materials. The compounds either singly or collectively have demonstrated some inhibitory potentials on the germination, growth and yields of cabbage, carrot, tomato and spinach evaluated in this study.
- Full Text:
- Date Issued: 2012
Studies towards the development of novel HIV-1 integrase inhibitors
- Authors: Lee, Yi-Chen
- Date: 2010
- Subjects: HIV infections -- Treatment , HIV (Viruses) , AIDS (Disease) -- Treatment , Nuclear magnetic resonance , Heterocyclic compounds -- Derivatives , Enzyme inhibitors , Chemical inhibitors , Quinoline
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4357 , http://hdl.handle.net/10962/d1005022 , HIV infections -- Treatment , HIV (Viruses) , AIDS (Disease) -- Treatment , Nuclear magnetic resonance , Heterocyclic compounds -- Derivatives , Enzyme inhibitors , Chemical inhibitors , Quinoline
- Description: The project has focused on the preparation of several series of compounds designed as potential HIV-1 integrase inhibitors. Various 2-nitrobenzaldehydes have been reacted with two activated alkenes, methyl vinyl ketone (MVK) and methyl acrylate, under Baylis-Hillman conditions to afford α-methylene-β-hydroxylalkyl derivatives in moderate to excellent yields. The reactions were conducted using the tertiary amine catalysts, 1,4-diazabicyclo[2.2.2]octane(DABCO) or 3-hydroxyquinuclidine (3-HQ) with chloroform as solvent, and yields were optimised by varying the catalyst, reagent concentrations and the reaction time. Reductive cyclization of the Baylis-Hillman adducts via catalytic hydrogenation, using 10% palladiumon-carbon catalyst in ethanol, afforded quinoline and quinoline N-oxide derivatives. In some cases “acyclic” reduction products were also isolated. Reaction of the Baylis-Hillman MVK adducts with HCl, has resulted in effective nucleophilic (SN’) displacement of the hydroxyl group to afford allylic chloride derivatives. Direct substitution of these chloro derivatives by secondary or primary amines, followed by catalytic hydrogenation gave quinoline derivatives containing a 3-aminomethyl substituent. The Baylis-Hillman ester adducts obtained from reaction with methyl acrylate were treated directly with various amines to give diastereomeric conjugate addition products. Reactions with piperazine gave N,N’-disubstituted piperazine products. The piperidine derivatives have been dehydrated to give cinnamate esters in moderate yields. The products, which have all been satisfactorily characterised by elemental (HRMS) and spectroscopic (1- and 2-D NMR) analysis, constitute a “library” of compounds for in silico and in vitro studies as potential HIV integrase inhibitors.
- Full Text:
- Date Issued: 2010
- Authors: Lee, Yi-Chen
- Date: 2010
- Subjects: HIV infections -- Treatment , HIV (Viruses) , AIDS (Disease) -- Treatment , Nuclear magnetic resonance , Heterocyclic compounds -- Derivatives , Enzyme inhibitors , Chemical inhibitors , Quinoline
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4357 , http://hdl.handle.net/10962/d1005022 , HIV infections -- Treatment , HIV (Viruses) , AIDS (Disease) -- Treatment , Nuclear magnetic resonance , Heterocyclic compounds -- Derivatives , Enzyme inhibitors , Chemical inhibitors , Quinoline
- Description: The project has focused on the preparation of several series of compounds designed as potential HIV-1 integrase inhibitors. Various 2-nitrobenzaldehydes have been reacted with two activated alkenes, methyl vinyl ketone (MVK) and methyl acrylate, under Baylis-Hillman conditions to afford α-methylene-β-hydroxylalkyl derivatives in moderate to excellent yields. The reactions were conducted using the tertiary amine catalysts, 1,4-diazabicyclo[2.2.2]octane(DABCO) or 3-hydroxyquinuclidine (3-HQ) with chloroform as solvent, and yields were optimised by varying the catalyst, reagent concentrations and the reaction time. Reductive cyclization of the Baylis-Hillman adducts via catalytic hydrogenation, using 10% palladiumon-carbon catalyst in ethanol, afforded quinoline and quinoline N-oxide derivatives. In some cases “acyclic” reduction products were also isolated. Reaction of the Baylis-Hillman MVK adducts with HCl, has resulted in effective nucleophilic (SN’) displacement of the hydroxyl group to afford allylic chloride derivatives. Direct substitution of these chloro derivatives by secondary or primary amines, followed by catalytic hydrogenation gave quinoline derivatives containing a 3-aminomethyl substituent. The Baylis-Hillman ester adducts obtained from reaction with methyl acrylate were treated directly with various amines to give diastereomeric conjugate addition products. Reactions with piperazine gave N,N’-disubstituted piperazine products. The piperidine derivatives have been dehydrated to give cinnamate esters in moderate yields. The products, which have all been satisfactorily characterised by elemental (HRMS) and spectroscopic (1- and 2-D NMR) analysis, constitute a “library” of compounds for in silico and in vitro studies as potential HIV integrase inhibitors.
- Full Text:
- Date Issued: 2010
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