Department of Biochemistry, Cell and Molecular Biology
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Item Aflatoxin Contamination of Cassava Flour (Kokonte) Processed By Traditional Methods in Ghana(University of Ghana, 1978-12) Lokko, P.G.; Gyang, F.N.; Kordylas, M.J.; University of Ghana, College of Basic and Applied Sciences, School of Biological Sciences, Department of Biochemistry, Cell and Molecular BiologyAflatoxin contamination of cassava flour (kokonte) bought in the markets of Accra has been investigated. A survey of the methods of production and of processing of the cassava chips was conducted to determine the sources of contamination in the production of the flour. Samples of the flour were analysed for aflatoxin contamination and concentrations using Thin Layer Chromatography (T.L .C.). Known levels of aflatoxin in kokonte flour samples were fed to weanling rats to determine the effects on the rats. Serum alkaline phosphatase levels were determined in the serum samples of the individual rats. The study revealed that:- (1) Three types of kokonte flour are sold at the markets of Accra and these are named according to their colours. There are red, black and white kokonte types. (2) Methods used in the processing of the cassava chips exposed the fresh roots to mould growth and hence to aflatoxin contamination. (3) Twenty-two percent of the kokonte flour samples bought from the market were found to contain aflatoxin. The mean aflatoxin concentration was calculated to be 867pg/kg. (4) There were reduced rates of growth in the rats fed on aflatoxin-contaminated diets over a period of 25 days and their food intake was also found to have reduced when compared with those of the control group. (5) The rats on aflatoxin-contaminated diets had fatty livers and there was an evidence of a growth on one of the livers. (6) Serum alkaline phosphatase levels in rats fed aflatoxin-contaminated diets were found to be higher than those observed for the rats on control diets.Item Analysis of Antimicrobial Resistance in Candida Albicans Using Modulators of MDR/CDR Gene Expression(University of Ghana, 2018-07) Yeboah, R.Majority of life-threatening fungal infections in clinics are caused by Candida albicans. The emergence of azole resistance in fungi complicates patient management. In response to chemical stress, C. albicans make transient changes in the gene expression for survival. Notable among these is the upregulation of efflux pump which is known to be the main mechanism of antifungal resistance. Potent therapeutic agents targeting this resistance mechanism are urgently needed. Chemo-sensitization is postulated as one way to overcome antifungal resistance. Endophytic fungi produce bioactive metabolites which are used as chemotherapeutic agents. The aim of this study is to use modulators of CDR and MDRs genes as probes to study chemo-sensitization and resistance phenotypes. Also, fungal metabolites (alone and in combination with chemosensitizers) will be used to reverse antifungal resistance. On analysis of phenotypic switching of the fungal cells in the presence of efflux modulators and phenotypic modifiers, S. cerevisiae was frequently observed to switch phenotypes as compared to C. albicans. Chemical compounds, including, compounds PC04-10, PC04-11, PC04-16 and PC04-23, significantly modified the antimicrobial phenotypes of Candida albicans and Saccharomyces cerevisiae and could be considered for use as synergistic partners of antifungal drugs to overcome resistance. Also, it was realized that some compounds including rifampicin, estradiol, PC04-09 and PC04-14 caused resistance. A total of 40 out of 507 bioactive and 90 chemosensitizing extracts were identified from SBF and TEF fungal extracts. In the Rhodamine efflux assay, six compounds were found to inhibit S. cerevisiae efflux, these well trifluoperazine, trifluoprozerazine, thioridazine, chlorpromazine, deferasirox and ibuprofen, whereas in C. albicans only the last four out of the six compounds were active. Also, 13 out of 20 chemosensitizing extracts significantly inhibited efflux activity of C. albicans and S. cerevisiae. Thus, fungi are good sources of novel and potent antifungal and chemosensitizing compounds.Item Analysis of Histidine Rich Protein 2 And 3 Gene Deletion Polymorphisms in Northern Ghana(University of Ghana, 2017-07) Ayelazuno, F.A.The use of Plasmodium falciparum histidine rich protein 2 based (PfHRP2-based) Rapid Diagnostic Tests (RDTs) to accurately diagnose falciparum malaria from other febrile cases reporting to health facilities in Ghana plays a vital role in the control of malaria. However, false negatives due to deletion polymorphism in the pfhrp2 gene may lead to misdiagnosis, increased morbidity as a direct result of delayed treatment and ultimately high treatment cost. Therefore, determining the prevelance of parasites that carry these polymorphisms could be of relevance to National Malaria Control Programmes (NMCPs). The aim of this study was to determine the prevalence and geo-spatial distribution of P. falciparum histidine-rich protein 2 and 3 gene deletion polymorphism in the Kassena-Nankana Districts (KNDs). Patients reporting with fever or history of fever were recruited after informed consent was obtained. Thick and thin blood smears were used to assess malaria parasite species and density of parasitemia whilst filter paper dried blood spots (DBS) were made for parasite DNA extraction for detection of deletion polymorphism. DNA was extracted using Qiagen midi kit following the manufacturer’s instructions. The exon 1-2, exon 2 and the flanking genes of pfhrp2, the exon 2 of phrp3 were amplified using specific primer pairs. PCR products were resolved by 2.0 % agarose gel electrophoresis. A total of 197 samples were collected, out of which 99 were found to be positive for P. falciparum. The prevalence of parasites that were found to have deleted exon 1-2 of the pfhrp2 gene was 7.7% (7/99), whilst 14.1% (14/99) were detected to have deleted the upstream gene of pfhrp2 gene. I observed 4.0 % (4/99) of parasites that had deleted exon 2 of the pfhrp2 gene and only one parasite was found to have deleted the entire pfhrp3 gene. Geo-spatial analysis did not reveal location specific differences in prevalence of pfhrp2 deletion polymorphisms. Overall, this study shows a low prevalence pfhrp 2 and/or 3 gene delection in the KNDs of northern Ghana. Evidence that pfhrp2 based RDTsmay still be an effective tool for diagnosing malaria in this region.Item Analysis of Low Molecular Weight Compounds Produced by Indigenous Wood Decay Fungi(University of Ghana, 2011-03) Aboagye, S.Y.Over the years, natural products have played a major role in the search for novel drugs or drug candidates. Secondary metabolites from nature especially those of fungal origin exhibit unique biological activities and research continue to meet the keen interests which have potential pharmaceutical value. Fungi constitute an important source of secondary metabolites such as penicillin. The present study analyzed the biological activities of a variety of compound mixtures from wood decay fungi. A total of 54 wood decay fungi (WDF) were collected, with majority obtained from the University of Ghana campus and its surroundings. The WDF were cultured in potato dextrose broth (PDB) for 48 days and the time course analysis of two selected WDF were also performed to determine the profile of specific bioactivities. Cultures were terminated and extracted with ethyl acetate at the end of the 48th day of culture. WDF extracts were analyzed spectrophotometrically at wavelengths between 200 nm and 900 nm which showed high absorption in the UV region of the spectrum. TLC analyses of the fungal extracts were done using EtOAc: CH3CN: PetEth (7:2:1) as the solvent system and different classes of compounds were detected on TLC plates sprayed with Anisaldehyde reagent. UV detection of compounds on TLC showed unique band pattern for the different WDF. Assays for biological activities of the fungal extracts were performed against S. aureus ATCC.2, E. coli NMIMR.3, C. albicans KBTH.2 and A. niger ATCC.2 using disc diffusion assay method. From the primary screening of antimicrobial activity, a total of 40 WDF extracts were found to exhibit some form antimicrobial activity towards the test organism. Out of total 40 that had an activity, 10 of the WDF extracts were found to have biological activity selectively (SG+) against Staphylococcus aureus ATCC.2, and 13 extracts were also found to have a broad spectrum antimicrobial activity (BSAB) against Staphylococcus aureus ATCC.2 and Escherichia coli NMIMR.3. The number of extracts that had selective antifungal (SAF) activity towards Candida albicans KBTH.2 was found to be 3. The number of WDF that exhibited a non selective antimicrobial (NSAM) activity towards the three test organisms, S. aureus ATCC.2, E. coli NMIMR.3and C. albicans KBTH.2 were recorded to be 11. The time course analysis showed that fungal metabolites are produced as early as the 7th day of culture, however cultures that were harvested between 22nd and the 48th day of culture produced potent bioactive components. . After the secondary screening of 27 WDF, inhibitory activity against only S. aureus ATCC.2 was found to be possessed by 9 refermented extracts. Inhibitory activity towards both Gram positive and Gram negative bacteria tested was found in 6 extracts. Four (4) refermented extracts also exhibited inhibition towards S. aureus ATCC.2 and C. albicans KBTH.2. A shift in antimicrobial activity was observed after the secondary screen. Sephadex LH-20 fractions of the selected WDF extracts showed the broad spectrum activities of the individual fractions. All the extracts (A4, E2, E9 and F3) that showed broad spectrum activities against a Gram + and Gram – bacteria had common fraction/s possessing the biological activities. In those extracts (B6 and B7) that inhibited a Gram + bacteria and a fungus, the biological activities were seen in different fractions.Item Anti-Diabetic and Probiotic Effect of Kombucha on Alloxan-Induced Diabetic(University of Ghana, 2019-07) Adade, E.E.Diabetes mellitus, is a metabolic disorder caused by the inability of the beta pancreatic cells to adequately produce insulin or due to insulin resistance of cells. As a result of the increasingly high incidence of diabetes globally, the World Health Organization (WHO) has set timelines and guidelines for the reduction of the risk of mortalities and morbidities associated with non-communicable diseases including diabetes, by the year 2030. However, this agenda is hinged on the availability of affordable, safe and effective alternatives for the management and treatment of these diseases. Hence, there is a need to explore other alternatives to the conventional oral anti-hyperglycemic agents driven by factors such as patient’s preference, demand among others. Kombucha is tea fermented by a symbiotic culture of bacteria and yeasts (SCOBY). Consumers of Kombucha have reported several anecdotal evidences of its medicinal potential. This study seeks to investigate its anti-diabetic and probiotic effect on alloxan-induced diabetic rats. It was hypothesized that Kombucha, being a complex matrix of microorganisms and nutraceuticals, would play an essential role in the management of diabetes. Molecular characterization of the microbiome of Kombucha using shotgun metagenomics (Oxford Nanopore MINION sequencing technology) showed Brettanomyces bruxellensis CBS 2499 as the most abundant species within the microbial community accounting for about 51 % of all reads. Brettanomyces anomalus, Komagataeibacter xylinus NBRC 15237, Bacillus nealsonii AAU1, Zygosaccharomyces bailii CLIB 213, Acetobacter, Gluconobacter and over 300 other genera and species of microorganisms including archaea and viruses were also detected using a combination of REFSEQ and One Codex data bases (OXCDB). In-vivo experiment was used to evaluate the anti-diabetic property, safety and gut microbiome changes of Kombucha. Kombucha was found to perform better than the conventional antidiabetic drugs, metformin and glibenclamide in lowering the fasting blood glucose (FBG) of the diabetic rats. Daily administration of 25 mg/kg and 100 mg/kg of freeze-dried Kombucha tea demonstrated a 5 fold reduction in FBG (p<0.05) and 40% and 50% respective increases in body weight of the alloxan-induced diabetic rats compared to the diabetic control (DC). Histological analysis, shows Kombucha enhances pancreas regeneration and hence the concomitant increase in insulin secretion as demonstrated in the study. Serum lipid profiling showed 100mg/kg Kombucha treatment increases the levels of total cholesterol (16%), high density lipoproteins (HDL) (13%) and low-density lipoproteins (LDL) (10%) but conversely reduces triglyceride level (17%) compared to the DC (p>0.05). Further analyses demonstrated that Kombucha decreases the relative organ (liver and kidney) to body weight ratio in treated animals. In addition, Kombucha was able to reduce significantly the elevated levels of liver enzymes such as Alkaline phosphatase (ALP), Alanine transaminase (ALT) and Aspartate Aminotransferase (AST) as well as renal toxicity indices, creatinine and urea in treated animals. Histology of the kidney and liver also showed that Kombucha has no adverse effect on the morphology and cellular integrity of these organs suggesting its hepatoprotective and renal protective potentials. Urinanalysis also showed reduction of glucose in urine for the 100 mg/kg Kombucha-treated animals. Additionally, Kombucha protects the gut microbiome, most significantly by enhancing the Lactobaccillaceae family of bacteria within the gut and reduces the possibilities of colonization of the gut by other opportunistic bacterial species. The study demonstrated that Kombucha is enriched with diverse microbial population with probiotic value and daily intake of Kombucha may be potentially helpful in the management of diabetes, protection against renal and liver toxicity and offer gut microbiome protection.Item Anti-Diabetic and Probiotic Effect of Kombucha on Alloxan-Induced Diabetic Rats(University of Ghana, 2019-07) Adade, E.E.Diabetes mellitus, is a metabolic disorder caused by the inability of the beta pancreatic cells to adequately produce insulin or due to insulin resistance of cells. As a result of the increasingly high incidence of diabetes globally, the World Health Organization (WHO) has set timelines and guidelines for the reduction of the risk of mortalities and morbidities associated with non-communicable diseases including diabetes, by the year 2030. However, this agenda is hinged on the availability of affordable, safe and effective alternatives for the management and treatment of these diseases. Hence, there is a need to explore other alternatives to the conventional oral anti-hyperglycemic agents driven by factors such as patient’s preference, demand among others. Kombucha is tea fermented by a symbiotic culture of bacteria and yeasts (SCOBY). Consumers of Kombucha have reported several anecdotal evidences of its medicinal potential. This study seeks to investigate its anti-diabetic and probiotic effect on alloxan-induced diabetic rats. It was hypothesized that Kombucha, being a complex matrix of microorganisms and nutraceuticals, would play an essential role in the management of diabetes. Molecular characterization of the microbiome of Kombucha using shotgun metagenomics (Oxford Nanopore MINION sequencing technology) showed Brettanomyces bruxellensis CBS 2499 as the most abundant species within the microbial community accounting for about 51 % of all reads. Brettanomyces anomalus, Komagataeibacter xylinus NBRC 15237, Bacillus nealsonii AAU1, Zygosaccharomyces bailii CLIB 213, Acetobacter, Gluconobacter and over 300 other genera and species of microorganisms including archaea and viruses were also detected using a combination of REFSEQ and One Codex data bases (OXCDB). In-vivo experiment was used to evaluate the anti-diabetic property, safety and gut microbiome changes of Kombucha. perform better than the conventional antidiabetic drugs, metformin and glibenclamide in lowering the fasting blood glucose (FBG) of the diabetic rats. Daily administration of 25 mg/kg and 100 mg/kg of freeze-dried Kombucha tea demonstrated a 5 fold reduction in FBG (p<0.05) and 40% and 50% respective increases in body weight of the alloxan-induced diabetic rats compared to the diabetic control (DC). Histological analysis, shows Kombucha enhances pancreas regeneration and hence the concomitant increase in insulin secretion as demonstrated in the study. Serum lipid profiling showed 100mg/kg Kombucha treatment increases the levels of total cholesterol (16%), high density lipoproteins (HDL) (13%) and low-density lipoproteins (LDL) (10%) but conversely reduces triglyceride level (17%) compared to the DC (p>0.05). Further analyses demonstrated that Kombucha decreases the relative organ (liver and kidney) to body weight ratio in treated animals. In addition, Kombucha was able to reduce significantly the elevated levels of liver enzymes such as Alkaline phosphatase (ALP), Alanine transaminase (ALT) and Aspartate Aminotransferase (AST) as well as renal toxicity indices, creatinine and urea in treated animals. Histology of the kidney and liver also showed that Kombucha has no adverse effect on the morphology and cellular integrity of these organs suggesting its hepatoprotective and renal protective potentials. Urinanalysis also showed reduction of glucose in urine for the 100 mg/kg Kombucha-treated animals. Additionally, Kombucha protects the gut microbiome, most significantly by enhancing the Lactobaccillaceae family of bacteria within the gut and reduces the possibilities of colonization of the gut by other opportunistic bacterial species. The study demonstrated that Kombucha is enriched with diverse microbial population with probiotic value and daily intake of Kombucha may be potentially helpful in the management of diabetes, protection against renal and liver toxicity and offer gut microbiome protection.Item Anti-Diabetic And Probiotic Effect Of Kombucha On Alloxan-Induced Diabetic Rats(2019-07) Adade, E.E.Diabetes mellitus, is a metabolic disorder caused by the inability of the beta pancreatic cells to adequately produce insulin or due to insulin resistance of cells. As a result of the increasingly high incidence of diabetes globally, the World Health Organization (WHO) has set timelines and guidelines for the reduction of the risk of mortalities and morbidities associated with non-communicable diseases including diabetes, by the year 2030. However, this agenda is hinged on the availability of affordable, safe and effective alternatives for the management and treatment of these diseases. Hence, there is a need to explore other alternatives to the conventional oral anti-hyperglycemic agents driven by factors such as patient’s preference, demand among others. Kombucha is tea fermented by a symbiotic culture of bacteria and yeasts (SCOBY). Consumers of Kombucha have reported several anecdotal evidences of its medicinal potential. This study seeks to investigate its anti-diabetic and probiotic effect on alloxan-induced diabetic rats. It was hypothesized that Kombucha, being a complex matrix of microorganisms and nutraceuticals, would play an essential role in the management of diabetes. Molecular characterization of the microbiome of Kombucha using shotgun metagenomics (Oxford Nanopore MINION sequencing technology) showed Brettanomyces bruxellensis CBS 2499 as the most abundant species within the microbial community accounting for about 51 % of all reads. Brettanomyces anomalus, Komagataeibacter xylinus NBRC 15237, Bacillus nealsonii AAU1, Zygosaccharomyces bailii CLIB 213, Acetobacter, Gluconobacter and over 300 other genera and species of microorganisms including archaea and viruses were also detected using a combination of REFSEQ and One Codex data bases (OXCDB). In-vivo experiment was used to evaluate the anti-diabetic property, safety and gut microbiome changes of Kombucha. Kombucha was found to perform better than the conventional antidiabetic drugs, metformin and glibenclamide in lowering the fasting blood glucose (FBG) of the diabetic rats. Daily administration of 25 mg/kg and 100 mg/kg of freeze-dried Kombucha tea demonstrated a 5 fold reduction in FBG (p<0.05) and 40% and 50% respective increases in body weight of the alloxan-induced diabetic rats compared to the diabetic control (DC). Histological analysis, shows Kombucha enhances pancreas regeneration and hence the concomitant increase in insulin secretion as demonstrated in the study. Serum lipid profiling showed 100mg/kg Kombucha treatment increases the levels of total cholesterol (16%), high density lipoproteins (HDL) (13%) and low-density lipoproteins (LDL) (10%) but conversely reduces triglyceride level (17%) compared to the DC (p>0.05). Further analyses demonstrated that Kombucha decreases the relative organ (liver and kidney) to body weight ratio in treated animals. In addition, Kombucha was able to reduce significantly the elevated levels of liver enzymes such as Alkaline phosphatase (ALP), Alanine transaminase (ALT) and Aspartate Aminotransferase (AST) as well as renal toxicity indices, creatinine and urea in treated animals. Histology of the kidney and liver also showed that Kombucha has no adverse effect on the morphology and cellular integrity of these organs suggesting its hepatoprotective and renal protective potentials. Urinanalysis also showed reduction of glucose in urine for the 100 mg/kg Kombucha-treated animals. Additionally, Kombucha protects the gut microbiome, most significantly by enhancing the Lactobaccillaceae family of bacteria within the gut and reduces the possibilities of colonization of the gut by other opportunistic bacterial species. The study demonstrated that Kombucha is enriched with diverse microbial population with probiotic value and daily intake of Kombucha may be potentially helpful in the management of diabetes, protection against renal and liver toxicity and offer gut microbiome protection.Item Anti-Diabetic and Probiotic Effect of Kombucha on Alloxan-Induced Diabetic Rats(University of Ghana, 2019-07) Adade, E.E.Diabetes mellitus, is a metabolic disorder caused by the inability of the beta pancreatic cells to adequately produce insulin or due to insulin resistance of cells. As a result of the increasingly high incidence of diabetes globally, the World Health Organization (WHO) has set timelines and guidelines for the reduction of the risk of mortalities and morbidities associated with non-communicable diseases including diabetes, by the year 2030. However, this agenda is hinged on the availability of affordable, safe and effective alternatives for the management and treatment of these diseases. Hence, there is a need to explore other alternatives to the conventional oral anti-hyperglycemic agents driven by factors such as patient’s preference, demand among others. Kombucha is tea fermented by a symbiotic culture of bacteria and yeasts (SCOBY). Consumers of Kombucha have reported several anecdotal evidences of its medicinal potential. This study seeks to investigate its anti-diabetic and probiotic effect on alloxan-induced diabetic rats. It was hypothesized that Kombucha, being a complex matrix of microorganisms and nutraceuticals, would play an essential role in the management of diabetes. Molecular characterization of the microbiome of Kombucha using shotgun metagenomics (Oxford Nanopore MINION sequencing technology) showed Brettanomyces bruxellensis CBS 2499 as the most abundant species within the microbial community accounting for about 51 % of all reads. Brettanomyces anomalus, Komagataeibacter xylinus NBRC 15237, Bacillus nealsonii AAU1, Zygosaccharomyces bailii CLIB 213, Acetobacter, Gluconobacter and over 300 other genera and species of microorganisms including archaea and viruses were also detected using a combination of REFSEQ and One Codex data bases (OXCDB). In-vivo experiment was used to evaluate the anti-diabetic property, safety and gut microbiome changes of Kombucha. Kombucha was found to perform better than the conventional antidiabetic drugs, metformin and glibenclamide in lowering the fasting blood glucose (FBG) of the diabetic rats. Daily administration of 25 mg/kg and 100 mg/kg of freeze-dried Kombucha tea demonstrated a 5 fold reduction in FBG (p<0.05) and 40% and 50% respective increases in body weight of the alloxan-induced diabetic rats compared to the diabetic control (DC). Histological analysis, shows Kombucha enhances pancreas regeneration and hence the concomitant increase in insulin secretion as demonstrated in the study. Serum lipid profiling showed 100mg/kg Kombucha treatment increases the levels of total cholesterol (16%), high density lipoproteins (HDL) (13%) and low-density lipoproteins (LDL) (10%) but conversely reduces triglyceride level (17%) compared to the DC (p>0.05). Further analyses demonstrated that Kombucha decreases the relative organ (liver and kidney) to body weight ratio in treated animals. In addition, Kombucha was able to reduce significantly the elevated levels of liver enzymes such as Alkaline phosphatase (ALP), Alanine transaminase (ALT) and Aspartate Aminotransferase (AST) as well as renal toxicity indices, creatinine and urea in treated animals. Histology of the kidney and liver also showed that Kombucha has no adverse effect on the morphology and cellular integrity of these organs suggesting its hepatoprotective and renal protective potentials. Urinanalysis also showed reduction of glucose in urine for the 100 mg/kg Kombucha-treated animals. Additionally, Kombucha protects the gut microbiome, most significantly by enhancing the Lactobaccillaceae family of bacteria within the gut and reduces the possibilities of colonization of the gut by other opportunistic bacterial species. The study demonstrated that Kombucha is enriched with diverse microbial population with probiotic value and daily intake of Kombucha may be potentially helpful in the management of diabetes, protection against renal and liver toxicity and offer gut microbiome protection.Item Anti-Inflammatory Activity And The Mechanism Of Action Of Morinda Lucida Benth(University of Ghana, 2016-12) Ayertey, F.A commonly used medicinal plant in African folk medicine for the treatment of various diseases including inflammation is Morinda lucida Benth. However, scientific data supporting its antiinflammatory activity is scarce. In the current study, the anti-inflammatory activity of the hydroethanolic leaf extract of Morinda lucida Benth (HEML) was assessed using carrageenaninduced paw edema in female Sprague-Dawley rats (SDRs). The potential mode of action of HEML was determined by assessing its effect on the levels of PGE2, NO, IL-1β, TNF-α and IL- 10, and also expressional levels of COX-2 and iNOS in RAW 264.7 cells in vitro. Its phytochemical constituents were determined by standard methods, and its antioxidant activity was investigated by DPPH free radical scavenging assay. The ability of HEML to significantly reduce rat paw edema caused by carrageenan demonstrated its anti-inflammatory activity. HEML also inhibited paw edema induced by histamine or serotonin, and further suppressed LPSinduced fever in the SDRs, which indicates that both early and late phases of acute inflammation were affected by the extract. Persistence of the late phase mediators may plunge the organism into a chronic state of inflammation. Results obtained through the determination of levels of PGE2, NO, IL-1β, TNF-α and IL-10 in culture supernatant of LPS-activated RAW 264.7 cells showed that HEML reduced NO and PGE2 concentrations by downregulating iNOS expression but not COX-2. It also suppressed the levels of pro-inflammatory cytokines IL-1β and TNF-α possibly by boosting levels of the anti-inflammatory cytokine IL-10. HEML contained saponins, reducing sugars, polyphenolics including flavonoids. It possesses antioxidant activity, which may be due to its polyphenolic content, particularly flavonoids. The current findings provide evidence-based scientific data to support the anecdotal use of M. lucida as treatment agent for inflammation.Item Anti-Inflammatory Medicinal Plants as Anti-Oxidants and Inhibitors of Proinflammatory Eicosanoid Biosynthesis(University of Ghana, 2001-05) Amponsah-Manager, K.; Addy, M.E.; Nyarko, A.K.; University of Ghana, College of Basic and Applied Sciences, School of Biological Sciences, Department of Biochemistry, Cell and Molecular BiologyArachidonic acid (AA) metabolism that leads to the production of both anti- and pro-inflammatory eicosanoids is a standard assay used to investigate the basis for the therapeutic action of anti-inflammatory medicinal plants. Earlier investigations have established the efficacy of some herbal preparations in terms of their ability to increase the amounts of anti-inflammatory eicosanoids. Desmodium adscendens and Parkitina sp. ('Tina A' ) increased PGE2 and PGI2 synthesis and inhibited phospholipase A2 activity while Thonningia sanguinea decreased the release of histamine and slow reacting substances of anaphylaxis (SRA-S). So far none of these extracts has shown any significant effect on thromboxane (TX) synthesis. The present study was conducted to evaluate the efficacy of four plant extracts, D. adscendens, Tina A, T. sanguinea and L. multi flora in decreasing the amounts of the pro-inflammatory eicosanoids, TXB2 and cysteinyl leukotrienes (cyst.LT), and their antioxidant properties to act as anti-oxidants. Isolated guinea pig lungs perfused via the trachea were used as a model to study the release and inhibition of the release of the TXB2 and cyst. LTs by the extracts. The organs were perfused with Kreb's solution with and without the plant extracts and TXB2 and cyst.LT released were estimated by an ELISA. There were significant decreases in the amounts of TX B2 and cyst.LT released from lungs perfused with the extract of 'Tina A' or T. sanguinea and lungs from animals treated with 'Tina A'. Generally, the treatment of the animals with the extract decreased the release of TX more than LTs, while perfusion of the isolated lungs (i.e. short term treatment) had more effect on cyst.LT than TX. The effects of D. adscendens, 'Tina A’, L. multiflora and T. sanguinea on the in vitro synthesis of TXB2 using blood platelet microsomes were investigated. Microsomes for this study were prepared from blood platelets. Except L. multiflora, for which higher concentration showed minimum effects on TXB2 synthesis, there was a concentration-dependent inhibition of TXB2 by all the extracts. The effect of T. sanguinea became significant at a relatively higher concentration compared to that of D. adscendens or 'Tina A'. At the highest concentration of 100ug/ml, D. adscendens, 'Tina A’ and T. sanguinea caused 81%, 81.6% and 87.5% decreases respectively in the amounts of TXB2 synthesized. The effects of the extracts on hydroxyl radical generation and total and watersoluble phenolic content were determined. There was a concentration-dependent inhibition of hydroxyl generation by all the extracts. Large amounts of phenolic compounds were identified in all the extracts. For 'Tina A', 74% of the total phenols was water soluble while T. sanguinea which gave the highest amount of total phenols had 31% being water soluble. There was a positive correlation between total phenolic content and inhibition of hydroxyl radical generation. T. sanguinea and D. adscendens which had the highest and lowest amounts of total phenols respectively, showed the highest and lowest inhibition of hydroxyl radical generations respectively at all concentrations studied. These findings suggest that the inhibition of both synthesis/release of pro-inflammatory eicosanoids and generation of reactive oxygen species by the plant extracts studied validates their use in folk medicine in the management of asthma and other inflammatory disorders.Item Application of Competitive Binding Assay Methods to Serum Vitamin D Levels in Health And Disease(University of Ghana, 1979) Ahene, I.S.; Chaplin,M.F.; University of Ghana, College of Basic and Applied Sciences, School of Biological Sciences, Department of Biochemistry, Cell and Molecular Biology.Competitive Binding Assay methods have been used to determine the serum levels of 25- hydroxyvitamin D^ in healthy Ghanaians and patients suffering from cirrhosis of the liver and chronic renal failure. Although the assay followed fairly standard methods as used by several workers, an improvement in stability and sensitivity was obtained by the use of gelatin in the buffer; the final assay buffer being 0.02M phosphate buffer, pH 7 ,C with 0.1% gelatin. Though rachitic rat serum was used for the main part of the work, it was confirmed that normal rat serum contained the binding protein at a suitable titre for the assay as has been reported in the literature. The normal circulating levels in 47 healthy Ghanaians was 129.0 + 55.7 ng/ml sermn, 46.0 + 16.1 ng/ml*?-.t^) for 11 patients with cirrhosis of the liver and 83.5 + 82.7 ng/ml for 8 patients suffering from chronic renal failure. Both patients'lavels were significantly different from the normal mean value ( P < 0 . 0 5 ) .Item Assessment of Sensitivity of P. Falciparum to Antifolate Antimalarials In Southern Ghana By Polymerase Chain Reaction (PCR) Assay Systems(University of Ghana, 1996-07) Mak-Mensah, E. E.Drug treatment of malaria is being hampered by the emergence of drug resistant strains of malaria parasites. Rapid methods for monitoring patients' response to treatment, applicable to field conditions, would aid management and control of drug resistant P. falciparum infections. In this study, mutation specific polymerase chain reaction assays using 3'-mismatched oligonucleotide primers which annealed to the wild or mutated parasites gene encoding dihydrofolate reductase-thymidylate synthase (DHFR-TS) were used to survey P.falciparum strains in two Regions of Southern Ghana (Volta and Greater Accra Regions). Mutations were identified directly from blood samples obtained from patients attending Out Patients Departments in Hospitals. Amplified P.falciparum DHFR-TS nucleotide sequences of 337bps were obtained when reaction products were analyzed by electrophoresis. Of 151 smear positive samples analysed, 13 (8.6%) contained the Asn-10S codon AAC that confers pyrimethamine resistance, 133 (88%) samples contained only the wild type Ser-10S codon AGC and none contained the Val-16 codon GTA found in cycloguanil-resistant pyrimethamine sensitive parasites. Out of the 13 pyrimethamine resistant cases, 10 were found in samples obtained from the Volta Region while the rest (3) were from Korle-Bu Teaching Hospital in the Greater Accra Region. No PCR product was found in 5 (3.3%) of the 151 samples. After second (nested) PCR, only one sample showed amplified product: contamination was negligible. PCR amplification of the DHFR-TS presents a rapid alternative to in vitro drug testing for monitoring the resistance of P.falciparum to antifolate antimalarials.Item Assessment of the Effects of Cocoa and Tea Kombucha on Asexual Stages of Plasmodium Falciparum(University of Ghana, 2017-07) Agbemafle, S.Tea and Cocoa Kombucha are fermented beverages produced by the action of a symbiotic culture of bacteria and yeasts. Their health benefits have been hailed and staunchly declared by Kombucha drinkers worldwide. However, there is minimal scientific evidence on their anti-malarial properties. This research aimed to further investigate the anti-plasmodial properties and possible mechanism of action of tea Kombucha and cocoa Kombucha by determining the inhibitory concentrations, evaluate the stage specificity, assess the inhibition of Plasmodium falciparum erythrocyte invasion and determine the antioxidant and cytotoxic activities of Kombucha extracts and fractions. Kombucha black and green tea and Kombucha cocoa were prepared, neutralized and fractionated using three different solvents (hexane, ethyl acetate and water) to obtain the respective extracts for in-vitro assays. The growth inhibition effects of the various extracts on in vitro P. falciparum were assessed using the SYBR Green I assay. Ring stage survival assays and schizont stage specific assays were evaluated after 72 hours using flow cytometry. Inhibition of P. falciparum invasion of human red blood cells (RBCs) treated with Kombucha extracts/fractions were also assessed using flow cytometry. The 1,1-diphenyl-2-picrylhydrazyl (DPPH) method was used to assess antioxidant activity of Kombucha on human peripheral blood mononuclear cells (PBMCs). The MTT (3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide) method was used to assess cytotoxicity profiles of Kombucha extracts and their fractions. The neutralized ethyl acetate fraction of cocoa Kombucha exhibited over 20 times (20x) the inhibitory activity of the corresponding unneutralized ethyl acetate fraction on Dd2 P. falciparum, and on NF54, the activity of neutral cocoa was almost 6 times (5.8x) its unneutralized counterpart. The ethyl acetate fraction of the neutralized GTK was 6x less potent than the non-neutralized ethyl acetate fractionof green tea Kombucha in inhibiting the ring stage of P. falciparum. The ethyl acetate fraction of green tea extract demonstrated schizonticidal activity at concentrations above its IC50(2mg/ml). The aqueous CTK extract displayed more potency in inhibiting invasion (46.6%) of P. falciparum Dd2 at its IC50 (280 μg/ml). The neutral cocoa Kombucha (NCTK) was 66% more effective at radical scavenging than the fermented CTK extract. The ethyl acetate NCTK fraction displayed the highest selective index (4.21). Generally, ethyl acetate selectively extracted bioactive components of Kombucha that exhibited anti-plasmodial and antioxidant activities coupled with good selectivity index compared to all other extracts and fractions. Cocoa Kombucha and its fractions exhibited the most active anti-plasmodial and stage specific activity on P. falciparum and could be explored as a food supplement for malaria prophylaxis.Item Association between Haptoglobin Phenotypes and Dyslipidemia as a Risk Factor of Cardiovascular Disease Among Ghanaian Hiv/Aids Patients(University of Ghana, 2016-07) Tagoe, I.N.A.; Amanquah, S.; Quay, O.; University of Ghana, College of Basic and Applied Sciences, School of Biological Sciences, Department of Biochemistry, Cell and Molecular BiologyCardiovascular disease (CVD) is one of the major complications in patient with human immunodeficiency virus (HIV) infection. The development of dyslipidemia as primary risk factor of CVD has been attributed to the contribution of oxidative stress. Studies have reported that these complications can be averted in HIV/AIDS with an effective antioxidant system. The association between haptoglobin phenotypes and dyslipidemia as risk factor to the development of cardiovascular disease in Ghanaian HIV/AIDS patients was studied in this cross-sectional study. A total of 180 HIV/AIDS patients were recruited from the Fevers unit of the Korle-Bu Teaching Hospital, Ghana. Patients on HAART were 105 (58.7%), was made up of 46 (43.8%) males and 59 (56.2%) females. HAARTNaïve patients were 75 (41.6%) comprising of 19 (25.3%) males and 56 (74.6%) females. The control group (100) was made up of 39 (39 %) males and 61 (61%) females. In this study, biochemical parameters such as total cholesterol (TC), triglycerides (TG), LDL and HDL were measured. The HIV viral RNA and cluster of differentiation (CD4) count were also measured among HIV patients. Haptoglobin phenotype was determined on polyacrylamide gel electrophoresis. The mean diastolic blood pressure (DBP), body mass index (BMI), visceral fat and body fat of patients compared with apparently health controls were not statistically significant (p > 0.05). However, the mean systolic blood pressure was significantly elevated in patients on HAART compared to those naïve of antiretroviral treatment. Mean hemoglobin level in HAART-naïve patients was significantly low (p < 0.05) when compared to mean hemoglobin of HAART treated patients and control group. However, lactate dehydrogenase (LDH) activity was significantly raised in both HAART and HAART-naïve patients than the controls (p < 0.0001). The CD4 count in the patients on treatment was significantly elevated than the HAART-naïve group (p < 0.0001). The mean HIV viral RNA level in HAART-naïve patients compared to those on treatment was significantly higher (p < 0.01). Malondialdehyde (MDA) was significantly elevated in both HAART and HAART-naïve patients compared to control group. The study participants had varied but significant mean MDA in relation to stratified CD4 counts. Generally, lipid parameters were altered among the study population. Total cholesterol, triglycerides, LDL, TC/HDL, LDL/HDL ratios and atherogenic index of plasma (AIP) were elevated in HIV patients (p < 0.0001) with significant variations between HAART and HAART-naïve groups when compared to the control group. The mean HDL level was significantly low in HIV groups whiles it was increased in the control group. The odds for developing hypercholesterolemia was 3.54 times more in patients on HAART compared to HAART-naïve patients. Again, HDL was significantly lower in HAART patients compared to HAART-naïve patients (OR = 0.22, p < 0.001). The CD4 count was negatively correlated with AIP. Elevated MDA was shown to occur in subjects with Hp2-2 compared to the other phenotypes. Patients with HIV expressing Hp2-2 were 1.29 times at risk of developing CVD although not significant compared to the other phenotypes. Haptoglobin phenotype therefore may not be a risk factor for the development of CVD in Ghanaian HIV/AIDS patients.Item Autopsy Characterization of Lung Microbiome of Hiv-Positive Patients in A Tertiary Referral Hospital in Ghana(University of Ghana, 2017-07) Badu, P.Pulmonary infections are the underlying cause of high morbidity and mortality among Human Immunodeficiency Virus (HIV) infected persons. Notwithstanding, there is limited data on pulmonary co-infecting pathogens and their susceptibility to commonly used antibiotics. Knowledge on these pathogens is therefore critical for implementing effective interventions and treatment guidelines especially, in highly burdened countries with scarce diagnostic facilities. The aim of this study was to characterize the lung microbiome of post-mortem biopsy samples of HIV/AIDS (Human Immunodeficiency Virus/Acquired Immunodeficiency Syndrome) patients in Ghana. A total of 102 lung biopsies from HIV/AIDS decedents from the Korle-Bu Teaching Hospital in Ghana were examined for mycobacteria, other bacteria, fungi and viruses. The techniques utilized included; culture, Gram/Ziehl Neelsen (ZN) staining, Matrix Assisted Laser Desorption/Ionization Time of Flight (MALDI-TOF) mass spectrometry and Polymerase Chain Reaction (PCR). The drug susceptibility pattern of Mycobacterium tuberculosis complex (MTBC) isolates and other bacteria were subsequently determined using Genotype MTBDRplus and disc diffusion assays respectively. Clinical data and autopsy causes of death were also retrieved for 86 cases and inferences were made by correlating the data to that of the laboratory investigations. From the clinical data, 42 (48.8%) of the cases were males and 42 (48.8%) were females with the mean ages of 40.4 (±10.6) and 37.1 (±11.5) respectively. HIV type was defined in 39 (45.3%) cases and co-infections with tuberculosis (TB), pneumonia, oesophageal candidiasis and/or cryptococcal disease occurred in 12 (14.3%) cases. Characterization of mycobacterial isolates yielded, 25 MTBC and 5 M. abscessus. In addition, an acid-fast isolate was identified as Norcardia farcinica. Drug susceptibility testing of the MTBC isolates showed 1 isoniazid mono-resistance and 3 rifampicin mono-resistance. Other bacteria isolated were 217 (83.8%) with Enterococcus species (61), Staphylococcus species (35), Escherichia coli (28) and Klebsiella pneumoniae (23) dominating. Of the 217, 75 Gram-negatives and 117 Gram-positives were profiled for drug sensitivity. Gram-negative isolates were most susceptible to cefoxitin and gentamicin (45.3% each) but highly resistant to cefuroxime sodium (84.0%). The Gram-positives were also fairly susceptible to levofloxacin (58.0%) but highly resistant to oxacillin (81.2%) and flucloxacillin (75.2%). Also, nine (9) culturable fungi; Candida species (6), Cryptococcus neoformans (1), Pichia occidentalis (1) and Yarrowia lipolytica (1) were identified whereas PCR detected Pneumocystis jiroveci in 43 samples. Viruses detected in the samples included Cytomegalovirus (67); a DNA virus and RNA viruses; Parainfluenza-2 (1) and Enterovirus (1). Findings from this study calls for the introduction of comprehensive definitive laboratory investigation rather than empirical clinical diagnosis to identifying the exact underlying pathogens that cause pulmonary infections in managing HIV patients in Ghana.Item Baseline Assessment Of Biomarkers For Crude Oil Pollutants: DNA Adducts In Fish And Polycyclic Aromatic Hydrocarbons In Some Environmental Samples In The Western Coast Of Ghana(University Of Ghana, 2015-07) Osei-Yeboah, E.The industrialization of our society, including drilling for oil, mining of coal and minerals has led to increasing production of xenobiotic and natural chemical substances. Continual releases of polycyclic aromatic hydrocarbons (PAHs) from mining and drilling activities leads to their deposition in coastal environments and ultimately bioaccumulation in plants and animals, creating the danger of toxicity. PAH are deemed to be carcinogenic, mutagenic and teratogenic. In Ghana, oil drilling is ongoing at the Jubilee oil fields in the Western region, however, onshore baseline environmental assessment of pollutants such as PAHs have not yet been performed. In this study, an environmental assessment of some communities bordering the oil drilling fields was performed to establish (1) levels of fish DNA adduct formation of fishes, (2) levels of polycyclic aromatic hydrocarbons in soil, plants, water and fishes. Examination of blood smears of several fish species showed the presence of micronuclei in fish from three of the study areas although their mean micronucleated frequencies were low and below the threshold frequency of 15%. There was no statistical difference between their mean frequencies upon one way ANOVA analysis with a p value < 0.05 except the mean of the fish Chloroscombrus chrysurus which showed a significant difference. Reverse phase HPLC analysis of fish, water, plant and soil samples collected from six study sites were done. Only two fish species Pomadasys incises at Aboadze and Thunnus alalunga from Dixcove recorded four and one PAH compounds respectively with concentrations above the maximum contaminant levels of 30 μg/Kg set by the USEPA. Mean concentration of PAHs in water samples were in the concentration range 1.4 to 1255 μg/L in water samples from two of the four study areas where water was found. All samples recorded concentrations above the threshold limit value of 50 ng/L set by the World Health Organization. Sixty plant samples were collected across the six study areas and only Erythrina senegalensis and Ficus umbellata recorded the presence of PAHs with concentrations in the range of 0.15-4.70 mg/Kg. Soil samples were collected from depths of 0-15 cm and 15-30 cm. The mean concentration of PAHs in surface soils (0-15cm) ranged from 0.1 to 95 mg/Kg with that at 15-30 cm ranging from 0.12 to 105 mg/Kg. The PAH composition profile in all the samples were similar with 2–3 ring PAHs being dominant which is suggestive of petrogenic source.Item Biochemical Toxicology of Desmodium Adscendens(University of Ghana, 2001-09) Quaye, O.; Addy, M.E.; Nyarko, A.K.; Okine, L.K.N.; University of Ghana, College of Basic and Applied Sciences, School of Biological Sciences, Department of Biochemistry, Cell and Molecular BiologyAqueous crude extract of Desmodium adscendens, as dispensed at the Centre for Scientific Research into Plant Medicine (CSRPM), was freeze-dried into a powder and used in this study to evaluate its toxicological effects in rats. For acute toxicity studies, high doses of the freeze-dried extract, were administered orally as single doses to rats, and the behaviours of the animals and organ toxicity were noted. Low, medium and high doses of the plant extract, representing 3, 10, and 30% of the dose that caused 50% lethality over the period of observation were administered to another group of rats in subchronic toxicity studies, and certain biochemical parameters measured as indices of liver and kidney toxicity. In order to investigate possible drug-drug interaction with other drugs, the effects of the plant material on zoxazolamine paralysis time, thiopentone sleeping time and the induction/inhibition of isozymes of the cytochrome P450 (CYP) enzyme systems were studied. From the results of the acute toxicity studies, the dose of the plant extract that caused 50% lethality was estimated to be 1125mg/kg body weight and this is about 456 times the prescribed dose in humans. When the extract was chronically administered, no significant differences were observed in levels of γ-glutamyltransferase activity, total protein, total bilirubin, creatinine and blood urea nitrogen concentrations in sera of test and control animals, irrespective of the doses of the extract administered. There were however increases in alanine and aspartate aminotransferase activities, and direct bilirubin concentration with increasing dose levels of the plant extract. Suggesting a possible hepatocellular damage, and hepatic excretory dysfunction or increased RBC haemolysis respectively. The plant extract caused a decrease in zoxazolamine paralysis time and prevented thiopentone from causing sleep in test animals as compared to controls. Results of the studies on the hepatic microsomal CYP isozymes suggest that CYP2B1/2B2 are involed in the metabolism of zoxazolamine and thiopentone. This study has indicated that the doses that caused lethality are much higher than the effective therapeutic dose of the extract of D. adscendens. Chronic administration also showed minimal signs of hepatotoxicity at doses 15-150 times higher than the effective therapeutic dose, suggesting that the plant extract may be safe at the therapeutic dose. Thus although D. adscendens may be safe, its induction of CYP2B1/2B2 and inhibition of CYP2E suggest that when the plant extract is co-administered with other drugs, may lead to possible drug interaction which may cause increase in /loss of therapeutic effectiveness or toxicity of the co-administered drug.Item The Carotenoid Biosynthesis Pathway In The Asexual Intraerythrocytic Stages Of Plasmodium Falciparum: In Vitro Inhibition Assays, Hplc Profiling Of Carotenoids And Bioinformatics Analysis(University of Ghana, 2016-03) Nortey-Mensah, R.; Teye, M. A.; Ofori, M. F.; University of Ghana, College of Basic and Applied Sciences, School of Biological Sciences, Department of Biochemistry, Cell and Molecular BiologyPlasmodium falciparum, like other Apicomplexans, has retained a relict plastid known as the apicoplast. This organelle represents a new and exciting target for the chemotherapeutic management of malaria because it houses metabolic pathways that are unique to the parasite such as isoprenoid biosynthesis. The phytoene synthase (PSY) gene, has been demonstrated to be very important in carotenogenesis, however, little is known about the evolutionary relatedness of this gene in P. falciparum and other Apicomplexans. This study therefore aimed at profiling the carotenoids synthesized in the asexual intraerythrocytic stages of P. falciparum and to determine the evolutionary history and relatedness of the PSY gene in Apicomplexans and other organisms. In vitro inhibition assays were performed on the asexual intraerythrocytic stages of P. falciparum using fluridone to determine its IC50 and effect on parasite population. HPLC was used to profile carotenoids synthesized at the asexual stages and to determine the evolutionary history and relatedness of the PSY gene in Apicomplexans and other organisms, an unrooted phylogenetic tree was generated using MEGA 6. A dose-dependent inhibition of parasite population was observed with fluridone treatment on all the asexual stages, with the ring stages being the most susceptible. The carotenoid profiles showed that synthesis of carotenoids in P. falciparum is cumulative through the asexual intraerythrocytic stages with carotenoids such lycopene, α-, β-carotene among others being synthesized. An exciting novel finding of this study was the discovery of relatively high amounts of abscisic acid (ABA) in the schizont stages and not in the other stages. This is the first time ABA has been demonstrated to be synthesized by P. falciparum and it would be pioneering to further investigate the specific role of ABA in P. falciparum schizont stages. The phylogenetic analysis showed that the P. falciparum PSY was most related to P. University of Ghana http://ugspace.ug.edu.gh xii reichenowi, the chimpanzee strain of the malaria causing parasites further lending support to the proposed origin of malaria species in humans.Item Characterization of Liver Function, Immune Response And Hepatitis B Virus (Hbv) Genotypes in Pregnant Women With Malaria-Hbv Co-Infection in Northern Ghana(University of Ghana, 2017-07) Anabire, N.G.Background: The overlap of malaria and chronic hepatitis B (CHB) is common in endemic regions, however, the impact of this co-infection on liver function and immune responses is unknown. This study sought to investigate these interactions in pregnant women reporting to antenatal clinic in the Northern Region of Ghana. Methodology: Levels of malaria parasitemia, hepatitis B viremia, liver biochemical parameters and inflammatory cytokines were assayed and compared across four categories of pregnant women: uninfected, infected with Plasmodium falciparum alone (Malaria group), infected with hepatitis B virus (HBV) alone (CHB group) and co-infected with P. falciparum and hepatitis B virus (Malaria+CHB group). Circulating HBV genotypes were determined by nested PCR. Results: the prevalence of HBV genotypes were: 91.6% genotype E, 5.2% mixed genotypes E/A, and 3.2% mixed genotypes E/D. Relative to the CHB group, the Malaria+CHB group had lower viremia levels (P=0.0157). However, levels of malaria parasitemia were similar in women in the Malaria and Malaria+CHB groups (P=0.3038). Furthermore, levels of markers for liver injury/damage, including alanine aminotransferase (P <0.0001), aspartate aminotransferase (P <0.0001) and total bilirubin (P <0.0001) were elevated in women in the Malaria+CHB group relative to those in the other groups. Similarly, proinflammatory cytokines, including tumour necrosis factor alpha (TNF-α) (P <0.0001), interleukin (IL)-1β (P =0.0270), and IL-6 (P =0.0106) were higher in women with Malaria+CHB compared to those in the other categories. For anti-inflammatory cytokines, including IL-10 (P <0.0001) and IL-4 (P =0.0007), the pattern was exactly the opposite of that for the pro-inflammatory cytokines. Conclusions: The study reports HBV genotype E to be dominant among pregnant women in Northern Ghana. Our findings showed that malaria/CHB co-infection in pregnancy appeared to exacerbate the release of biomarkers for liver damage and inflammatory mediators while reducing immune-modulatory mediators. Recommendation: Further longitudinal studies to investigatechanges in the levels of these biomarkers after the malaria infection is cleared will be important to confirm the findings of this study.Item Characterizing Pathogenic Risk Factors Associated with Breast Cancer in Ghanaian Women(University of Ghana, 2019-07) Buckman, M.A.Globally, the second leading cause of cancer death in women is breast cancer. It is responsible for 16% of all cancer cases and the most common cancer in Ghana. Breast cancer pathogenesis has not been fully elucidated but is however known to have a complex aetiology involving both genetic and environmental factors. Pathogens are one of the environmental factors that have been linked with breast cancer development. Epstein-Barr Virus (EBV), Human Papilloma Virus (HPV) and Mouse Mammary Tumour virus (MMTV) have been implicated in breast cancer. Research has showed that the local microbiome of the host could modulate breast cancer risk, yet it is unknown what microbes (pathogenic or probiotic) inhabit breast tumour tissues in Ghana. The objectives of this study were to detect the presence of HPV, EBV, MMTV and bacteria in breast tumour tissues from Ghanaian women and determine the effect of bacteria on DNA in HeLa and MCF-7 cells. Formalin-Fixed Paraffin-Embedded (FFPE) tissues and fresh breast cancer tissues were obtained from 204 breast cancer patients at the Department of Surgery, Korle-bu Teaching Hospital. Nucleic acid was extracted from the samples and amplified using standard Polymerase Chain Reaction (PCR) to detect the viruses. HPV-positive tumours were sequenced using the Sanger sequencing method. EBV typing of EBV positive samples was done by a nested PCR reaction using EBV-1 and EBV-2 specific primers. Bacteria from fresh breast cancer tissues were obtained and identified using basic biochemical tests and Sanger sequencing. The DNA damage potential of the isolates was tested on HeLa and MCF-7 cells using the histone-2AX phosphorylation assay. HPV was detected in 27 (13.2%) of the 204 cases analyzed. EBV was identified in 66 (32.4%) of the samples. Out of the EBV positive cases, 2 (1%) were positive for EBV-1, 30 (14.7%) were positive for EBV-2, 26 (12.7%) were positive for both EBV-1 and EBV-2 and 8 (3.9%) could not be genotyped using available methods. Co-infection of both HPV and EBV was detected in 11 (5.4%) of the cases. MMTV was not detected in any of the samples analyzed. Microbiome analysis showed relatively high evidence of bacteria belonging to the Staphylococcus and Bacillus species. Staphylococcus sciuri, Staphylococcus epidermidis, Staphylococcus lugdunensis obtained from breast tumours induced DNA double-strand breaks in MCF-7 cells. The diversity and the probable role of the microbiome in breast cancer were also identified. Therefore, the presence of these pathogens showed probable involvement in breast cancer carcinogenesis.