Clinical Pathology Department

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    N-Substituted Phenylhydrazones Kill the Ring Stage of Plasmodium falciparum
    (BioMed Research International, 2024) Gyan, P.; Amengor, C. D. K.; Biniyam, P. D.
    Antimalarial resistance has hampered the effective treatment of malaria, a parasitic disease caused by Plasmodium species. As part of our campaign on phenotypic screening of phenylhydrazones, a library of six phenylhydrazones was reconstructed and evaluated for their in vitro antimalarial and in silico receptor binding and pharmacokinetic properties. The structures of the phenylhydrazone hybrids were largely confirmed using nuclear magnetic resonance techniques. We identified two compounds which exhibited significant antimalarial potential against the ring stage (trophozoite) of 3D7 chloroquine-sensitive (CS) strain and DD2 chloroquine-resistant (CR) strains of Plasmodium falciparum with monosubstituted analogs bearing meta or para electron-donating groups showing significant activity in the single-digit micromolar range. Structure activity relationship is presented showing that electron-donating groups on the substituent hydrophobic pharmacophore are required for antimalarial activity. Compounds PHN6 and PHN3 were found to be the most potent with pIC50s (calculated form in vitro IC50s) of 5.37 and 5.18 against 3D7 CS and DD2 CR strains, respectively. Our selected ligands (PHN3 and PHN6) performed better when compared to chloroquine regarding binding affinity and molecular stability with the regulatory proteins of Plasmodium falciparum, hence predicted to be largely responsible for their in vitro activity. Pharmacokinetic prediction demonstrated that the phenylhydrazones may not cross the blood-brain barrier and are not P-glycoprotein (P-gp) substrates, a good absorption of 62% to 69%, and classified as a category IV compound based on toxicity grading.
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    Assessment Of Diesel Fuel Quality
    (Heliyon, 2024) Kwao-Boateng, E.; Ankudey, E.G.; Danquah, K.O.; Darkwah, L.
    Diesel is an essential energy source in the transportation and industrial sectors worldwide; hence, The quality of this commodity is crucial. This study compares various fuel samples to understand the quality of the fuels in terms of sulphur content, density, surface tension, viscosity, and calorific value. The properties of diesel fuel samples from eight (8) Filling Stations (marketing) Companies (MC) were examined and compared with GSA 141:2022 and ISO 8217:2017 standards. Fuel from two companies, MC-A and MC-G, had slightly lower densities than the standard. indicative of a possible contamination with lower-density fuels such as kerosene. The surface tension of all samples, except one, was within the standard range. The only sample with the lower than the standard value also displayed high sulphur content. Although all the fuel samples met the minimum requirement for calorific value, the viscosities of the fuels from three companies were slightly higher than the specified standard value, which can potentially result in higher emissions. In the case of sulphur content, fuel samples from only three companies were in compliance with the maximum 50 ppm standard. This means 62.5 % of the diesel fuel within the study area at the time contained more than the acceptable amount of sulphur. The findings in this Research highlights the need to re-examine the quality of fuels along the distribution chain.
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    In vitro antiplasmodial activity, LC-MS analysis, and molecular docking studies of bioactive compounds from Tetrapleura tetraptera (Fabaceae) fruits
    (Scientific African, 2023) Hamidu, S.; Adams, L.; Oduro, P.K.; et al.
    Malaria continues to be a major public health concern, particularly for children and pregnant women in areas where the disease is endemic. Developing safe and efficient antimalarial therapies to fight the disease is essential. Medicinal plants represent a potential source for the development of new antimalarial drugs. Tetrapleura tetraptera is a plant native to West Africa and traditionally used to treat several diseases including Malaria. Here, we investigated the antiplasmodial ac tivities of T. tetraptera fruit extracts against the chloroquine-sensitive (Pf3D7) and chloroquine resistant (PfDD2) strains of Plasmodium falciparum in vitro using SYBR green assay. In addition, the antioxidant potential of the fruit extracts was also determined. LC-MS analysis was carried out to identify the bioactive compounds in the extracts. Molecular docking studies provide significant prima facie evidence of inhibition hence, to evaluate the potential inhibition of Plasmodium fal ciparum dihydroorotate dehydrogenase (PfDHODH), a validated malaria drug target, the identi fied compounds were docked against PfDHODH. Strong antiplasmodial activities were demonstrated by the ethyl acetate and ethanolic extracts of T. tetraptera fruit, with IC50 values of 16.12 ± 0.04 µg/mL and 2.06 ± 0.02 µg/mL against the Pf3D7 strain, respectively. In the DPPH radical scavenging experiment, the ethanolic extract revealed considerable antioxidant activity with an EC50 value of 0.21 ± 0.82 mg/mL. Seven bioactive compounds were identified in the extract using LC-MS analysis. N-Methyl-1H-indole-3-propanamide (I), Tazolol (II), and Isopentyl salicylate (III) were identified as potential inhibitors of PfDHODH with high binding affinities ranging from -32.08 to -30.69 kcal/mol. The potential lead compounds also interacted with Gly181, Leu531, and Arg265, which are critical amino acid residues in the catalytic activity of PfDHODH. These findings support the traditional use of T. tetraptera fruit extracts for the treat ment of malaria and as promising avenues for antimalarial drug development.
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    Antioxidant and Anticancer Potential of Natural Cocoa Extract
    (IGI Global Gateway, 2023) Ayembilla, J.A.; Otu, P.N.Y.; Dotse, E.A.; et al.
    The aim of this study was to investigate antioxidant and anticancer potentials of natural cocoa extracts. Antioxidant properties of the extracts were evaluated by DPPH assay, total phenolic content by Folin-Ciocalteau and reduced glutathione content by O-phthalaldehyde assays. Antiproliferative activity was also evaluated on leukaemia and prostate cancer cell lines using MTT assay. The hydroethanolic extracts of cocoa roots and beans and aqueous cocoa leaves showed high total antioxidant activities with the root extract having the strongest scavenging effect on DPPH and the highest total phenolics content. The hydroethanolic cocoa bean crude extract showed the highest anticancer activity against LNCaP cells, whiles the ethyl acetate fraction of hydroethanolic cocoa leaf exhibited the highest anticancer against PC3. Jurkat and PC3 cells had similar susceptibilities to the natural cocoa extracts. Thus, cocoa has antioxidant and anticancer activity, and the specific bioactive compounds responsible for these activities need to be elucidated in future research.
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    Comparison of Modified Manual Acid-Phenol Chloroform Method and Commercial RNA Extraction Kits for Resource Limited Laboratories
    (Hindawi Limited, 2023) Danquah, K.O.; Sakyi, S. A.; Efah, A.; et al.
    Background and Aim. RNA extraction is a commonly used technique in molecular biology. In recent years, commercially available RNA extraction kits have largely replaced conventional approaches. However, these commercial kits are expensive and are not readily available in many resource-constrained institutions and laboratories. Tis study therefore compared the performance of the conventional acid guanidinium thiocyanate-phenol-chloroform (AGPC) extraction method to QIAamp® Viral RNA Mini Kit (QIAGEN, Cat. No. 52906) and OxGEn RNA Kit (OxGEn Molecular Solutions, GE-009) to build an in-house RNA extraction technique from blood and oral swab samples. Method. In a comparative experimental cross-sectional study, RNA was extracted from oral swabs and blood samples from 25 healthy individuals at the Department of Molecular Medicine, KNUST. RNA was extracted by the manual AGPC extraction method and commercial RNA extraction kits. Te quantity (ng/μl) and purities (260/ 280 nm) of the extracted RNA were measured spectrophotometrically using the IMPLEN NanoPhotometer® N60. Te presence of RNA in the extracts was confrmed using 2% agarose gel electrophoresis. Statistical analyses were conducted using R language. Results. Te yield of RNA extracted from blood and oral swab samples using modifed AGPC was signifcantly higher compared to the commercial methods (p < 0.0001). However, the purity of RNA extracted by the manual AGPC method from blood was signifcantly lower than the commercial methods (p < 0.0001). Moreover, the purity from oral swabs using the manual AGPC method was signifcantly lower compared to QIAamp (p < 0.0001) and the OxGEn kits method (p < 0.001). Conclusion. Te modifed manual AGPC method has a very high yield of RNA extracts using blood samples, which could serve as an alternate costefective method for RNA extraction in resource-limited laboratories; however, its purity may not be suitable for downstream Hindawi International Journal of Clinical Practice Volume 2023, Article ID 9593796, 8 pages https://doi.org/10.1155/2023/9593796 processes. Moreover, the manual AGPC method may not be suitable for extracting RNA from oral swab samples. Future investigation is needed to improve the purity of the manual AGPC RNA extraction method and also confrmation of the obtained results by PCR amplifcation and RNA purity verifcation by sequencing
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    Investigating potential sand fy vectors after the frst reported outbreak of cutaneous leishmaniasis in Ghana
    (Parasites & Vectors, 2023) de Souza, D.K.; Addo, S.O.; Desewu, K.; et al.
    Background Leishmaniasis is a parasitic disease caused by species of the genus Leishmania, which are transmitted through the bite of infected female sand flies. Since the first reported outbreak of cutaneous leishmaniasis in Ghana, in 1999, there has been limited published information on its vectors and reservoir hosts there. Previous studies have shown strong dominance of the sand fy genus Sergentomyia over the genus Phlebotomus in Ghana. Thus the aim of this study was to determine the possible sand fy vector species in Ghana, as well as their human-feeding behavior, from the time of the first reported outbreak of CL in the country. Methods Sand flies were collected from randomly selected houses in three communities. They were identified and used for blood meal source identification and the detection of Leishmania infection using molecular methods. Results A total of 1051 female sand flies were morphologically identified, of which Sergentomyia africana africana (29%) was the predominant species. Among the 275 female sand flies that had blood-fed, the identified blood meal sources included chicken (33.8%) and goat (12.4%); the percentage of human blood meals was 32%. Single-source and mixed-source blood meals were identified in Sergentomyia africana africana (11.6%), Sergentomyia ingrami (14.9%) and Sergentomyia simillima (20%), with S. simillima having the highest proportion of blood meals that included human blood (14.6%). Using molecular methods, unfed sand flies and identified human-feeding species were examined for the presence of Leishmania DNA. Pool screening analysis revealed three pools of S. ingrami positive for Leishmania major DNA, with an infection rate of 1.27% (95% confidence interval 2.467–3.647). Conclusions The findings suggest that some Sergentomyia species may be involved in the transmission of cutaneous leishmaniasis in Ghana. However, the role of S. ingrami as a vector of leishmaniasis in Ghana needs to be conclusively validated by isolating the parasite from this species and through experimental transmission studies.
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    In vitro antileishmanial activity and molecular docking studies of lupeol and monostearin, isolated from Parkia biglobosa
    (Scientific African, 2023) Osafo, T.; Philips, T.J.; Adomako, A.K.; Borquaye, L.S.; Ekuadzi, E.; Appiah-Opong, R.; Dickson, R.A.
    Parkia biglobosa (Leguminosae) is used traditionally for managing leishmaniasis. However, there are no reports of the antileishmanial constituents from the plant. This study is aimed at isolating antileishmanial compounds, from the stem bark of P. biglobosa, and assessing their mechanism of action using molecular docking studies. Bioassay-guided fractionation led to the isolation of two known compounds, lupeol and monostearin. Their structures were elucidated using their mass spectra. Using the 3-(4,5-dimethylthiazol-2- yl)-5-(3-carboxymathoxyphenyl)-2-(4-sulfophenyl)-2H-tretrazolium (MTS) assay, the evaluated antileishmanial effects of lupeol (1) and monostearin (2) showed IC50 values of 164.42 and 151.99 μg/mL respectively. The antileishmanial effects of the crude extract, non-polar, mid-polar and polar fractions were determined to be 64.43, 126.25, 725.65 and 167.52 μg/mL respectively. Molecular docking studies of the two isolates within the active sites of two validated targets in the Leishmania parasite: trypanothione reductase (TR) and pteridine reductase 1 (PTR1), showed that both compounds established important interactions with key amino acid residues in both proteins. Lupeol exhibited binding affinities of -7.1 kcal/mol and -10.2 kcal/mol for TR and PTR1, respectively, better than monostearin in both cases. Our findings back the claim that P. biglobosa stem bark possesses antileishmanial effects. Furthermore, the isolates lupeol and monostearin may be partly responsible for the observed activities.
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    Anti-plasmodial, Cytotoxic and Antioxidant Activities of Selected Ghanaian Medicinal Plants
    (SAGE, 2022) Appiah-Opong, R.; Agyemang, K.; Dotse, E.; Atchoglo, P.; Owusu, K.B.; Aning, A.; Sakyiamah, M.; Adegle, R.; Ayertey, F.; Appiah, A.A.; Nyarko, A.K.
    Malaria affects about half of the world’s population. The sub-Saharan African region is the most affected. Plant natural products have been a major source of antimalarial drugs; the first (quinine) and present (artemisinin) antimalarials are of natural product origin. Some secondary metabolites demonstrate adjuvant antioxidant effects and selective activity. The focus of this study was to investigate the anti-plasmodial activity, cytotoxicities and antioxidant properties of eight (8) Ghanaian medicinal plants. The anti-plasmodial activity was determined using the SYBR green assay and the tetrazolium-based colorimetric assay (MTT) was employed to assess cytotoxicity of extracts to human RBCs and HL-60 cells. Antioxidant potential of plant extracts was evaluated using Folin-Ciocalteu and superoxide dismutase assays. Phytochemical contstituents of the plant extracts were also assessed. All the extracts demonstrated anti-plasmodial activities at concentrations <50 μg/ml. Parkia clappertoniana and Terminalia ivorensis elicited the strongest anti-plasmodial activities with 50% inhibitory concentrations (IC50) of 1.13 μg/ml and 0.95 μg/ml, respectively. This is the first report on anti-plasmodial activities of Baphia nitida, Tabernaemontana crassa and Treculia Africana. T. Africana showed moderate anti-plasmodial activity with IC50 value of 6.62 µg/mL. Extracts of P. clappertoniana, T. Africana and T. ivorensis (0.4 mg/mL) showed >50% antioxidant effect (SOD). The extracts were not cytotoxicity towards RBCs at the concentration tested (200 μg/ml) but were weakly cytotoxic to HL-60 cell. Selectivity indices of most of the extracts were greater than 10. Our results suggest that most of the plant extracts have strong anti-plasmodial activity and antioxidant activity which warrants further investigations.
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    Effect of Anthonotha macrophyla (P. Beauv) leaf extract on carrageenan‑induced paw oedema, oxidative stress makers and hyperalgesia in murine models
    (Springer, 2021) Kumatia, E.K.; Ayertey, F.; Appiah‑Opong, R.; Ofori‑Attah, E.; Bolah, P.; Antwi, S.; Appiah, A.A.; Ocloo, A.
    Anthonotha macrophylla leaf (AML) is employed in the management of pain, infammation, and some diseases associated with oxidative stress in African traditional medicine. This study investigated the anti-infammatory and analgesic activities, the acute toxicity and anti-oxidant properties of AML, characterised the phytochemical constituents, and quantified its total phenolic and favonoid contents. Anti-infammatory and analgesic activities of AML extract were evaluated using carrageenan-induced oedema assay in rats’ paw and acetic acid-induced writhing assays respectively. Anti-oxidant efect of the extract was assessed in vivo by measuring liver antioxidant enzymes and in vitro by using DPPH radical scavenging assay and the acute toxicity of the extract was investigated in vivo in mice. The phytochemicals were characterised using basic phytochemical screening and total phenolics and favonoids were quantified using established methods. The AML extract exhibited signifcantly high anti-infammatory activity and moderate analgesic activity compared to diclofenac sodium. The extract exhibited moderate DPPH scavenging activity, signifcantly increased Superoxide Dismutase (SOD) and Glutathione peroxidase (GPX) activities, and did not have any effect on lipid peroxidation. AML was found to contain high phenolic and low favonoids. The LD50 of the extract was above 5000 mg/kg body weight. In conclusion, the AML extract possessed signifcant anti-infammatory and moderate analgesic activities, which may be mediated through its strong antioxidant prop erties due to its phenolic contents. The LD50 value showed that the extract was safe in the short-term utilization. The extract may therefore serve as a source of anti-infammatory, analgesic and antioxidant agents.
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    The Hydroethanolic Stem Bark Extract of Tieghemella heckelii (A.Chev.) Pierre ex Dubard (Sapotaceae) Produced N-Methyl-D Aspartate (NMDA) Receptor-Dependent Analgesia and Attenuates Acute Inflammatory Pain via Disruption of Oxidative Stress
    (Hindawi, 2021) Kumatia, E.K.; Appiah-Opong, R.
    Background. Tieghemella heckelii stem bark is used in African traditional medicine to treat inflammatory pain conditions. However, these biological actions of the plant have not been proven. )is study investigates the phytochemical composition and the mechanisms of analgesic and anti-inflammatory actions of the hydroethanolic stem bark extract of T. heckelii (THBE). Methods. Phytochemical composition of THBE was investigated using qualitative and quantitative phytochemical analyses. Anti inflammatory activity was evaluated using the carrageenan-induced paw oedema assay. Analgesic activity was evaluated using hot plate and acetic acid-induced writhing assays. Mechanism of analgesic action was determined using pharmacological antagonist such as naloxone, atropine, flumazenil, nifedipine, or ketamine. Test agents were administered orally as follows: Tween 80 (5%) (control), diclofenac sodium (DS) 10/tramadol 9 mg/kg (standard), or THBE 10, 100, and 450 mg/kg. Glutathione peroxidase (GPx), superoxide dismutase (SOD), and lipid peroxidation levels were also measured. Results. THBE which contained 58.45% saponins, 229.04 ± 0.049 GAE mg/g phenolic compounds,and 0.482 ± 0.0028 QE mg/g flavonoids produced (p < 0.5) anti inflammatory effect of 56.22% and analgesia of 330 ± 72% and 50.4% in the hot plate and writhing assays, respectively, at 10 mg/ kg and inhibited oxidative stress by GPx and SOD elevation in rats during inflammation. Ketamine significantly blocked the analgesia of THBE, indicating NMDA receptor-dependent analgesic action. Whereas, naloxone, atropine, nifedipine, and flumazenil could not antagonize the analgesic action of THBE. Conclusion. )ese results show that THBE produced potent anti-inflammatory effect via disruption of oxidative stress and also generated NMDA receptor-dependent analgesia.