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    Comparative Clinical Study of Mist Amen Fevermix and Edhec Malacure: Two Polyherbal Products used for the Treatment of uncomplicated malaria in Ghana against Artemether/Lumefantrine
    (2020-11) Turkson, Bernard Kofi; https://orcid.org/0000-0002-4990-7725
    The use of herbal medicinal products for the treatment of malaria an infectious and a life threatening disease, has increased globally. However, inadequate scientific studies, questions about the quality, safety and efficacy of such herbal products have been raised. On the other hand, the reduced sensitivity of the malaria parasites to artemisinin-based combination therapies is also of concern. There is therefore the need for new antimalarial medications including those from alternative sources such as herbal medicinal products. In this study, methods for the quality control of Mist Amen Fevermix and Edhec Malacure, two polyherbal antimalarial products used in Ghana for the management of uncomplicated malaria was undertaken. The development of the quality parameters for the test samples was based on phytochemical, physicochemical, chromatographic and spectroscopic methods. The set parameters were found to be sufficient to evaluate Mist Amen Fevermix and Edhec Malacure, and can be used as reference standards for the quality control purposes. Qualitative phytochemical screening and fingerprinting were undertaken based on standard analytical methods. The antiplasmodial activity was assessed in vitro by using field isolates of Plasmodium falciparum with SYBR® Green assays to measure parasite growth inhibition. Thermo Elemental M5 Atomic Absorption Spectrophotometer (AAS) fitted with Graphite furnace and an auto sampler was used to determine the heavy metal contents of the herbal products. The herbal samples were evaluated for microbial load by using the appropriate culture media. In vivo antiparasitic activity in mice was assessed using the Rane’s curative method using ANKA strain of Plasmodium berghei parasites. A comparative clinical study was done to assess the safety and effectiveness of the test samples at the Tafo Government Hospital, Kumasi after Committee on Human Research, Publication and Ethics approval. Male and female patients aged 15-45 years with clinically established malaria were treated with Mist Amen Fevermix and Edhec Malacure, at the specified doses of 45 mls (0.1063 g) and 30 mls (0.0521 g) three times daily after meals for three days. Basic phytochemical screening of the two products indicated the presence of the following phytochemicals: alkaloids, saponins, tannins, phytosterols and flavonoids. From the data, it was established that Mist Amen Fevermix and Edhec Malacure complied with the pharmacopoeial standards after testing for microbes. The following heavy metals were present in Mist Amen Fevermix and Edhec Malacure: Fe, Ni, K, Zn, Hg, Cu, Mn, Cr, Cd, Pb, Fe, Cu, K and Na. Ni was below detectable limit in Edhec Malacure. The phytochemical screening of the products revealed the presence of alkaloid flavonoid, tannin, steroid and saponin. The HPLC method was validated for linearity, limits of detection and quantification, precision and accuracy. The test products were found not to have been adulterated with lumefantrine, artemether and quinine. The test herbal products showed in vitro and in vivo antiplasmodial activities against Plasmodium falciparum and Plasmodium berghei parasites. Inhibitory concentration (IC50) values for Edhec Malacure was 70.89 ng/ml and that of Mist Amen Fevermix was 112.5 ng/ml. Edhec Malacure suppressed 76.17% of parasitaemia while Mist Amen Fevermix suppressed 69.03% of parasitaemia. Edhec Malacure demonstrated curative chemo suppressive potentials of 80.93% at the dose of 2.234 mgkg-1 and Mist Amen Fevermix % suppression was 69.03% at a dose of 4.56mg/kg-1. Both products demonstrated antiplasmodial activity in human red blood cells. The clinical evaluation of the test samples showed that Mist Amen Fevermix exhibited a statistically significant difference between the mean malaria parasite load recorded at the first visit and those recorded at the second visit, t(23) = 4.59, p =0 .000. Similarly, there was a significant difference between the mean parasite count recorded on the second visit and the third visit, t(6) = 1.49, p =0 .187. No difference were recorded for the third and fourth visits t(3) = 1.00, p =0 .391. Edhec Malacure also exhibited a significant difference in efficacy between the mean malaria parasite count recorded at the first visit and those recorded at the second visit, t(26) =3.77, p =0 .001. Similarly, there is a statistically significant difference between malaria parasite count at the second visits and third visits, t(16) = 1.74, p =0 .100. This shows the significant effectiveness of the products. Kidney and liver panel as well as full blood count and vital signs were within normalviii reference range at the end of the 28-day study and thus established the safety of Mist Amen Fevermix and Edhec Malacure in the treatment of uncomplicated malaria. The results support claims that Mist Amen Fevermix and Edhec Malacure may be useful antimalarial agents. This study has demonstrated the in vitro and in vivo antiplasmodial activities of Mist Amen Fevermix and Edhec Malacure, and suggests that, the products have promising antimalarial activity. The in vivo findings showed that Mist Amen Fevermix and Edhec Malacure are relatively safe for oral administration at doses tested. In addition, the study supports the use of Mist Amen Fevermix and Edhec Malacure, two polyherbal products for the treatment of uncomplicated malaria. Both products achieved a comparable clinical treatment outcome to the reference control medication artemether/lumefantrine.
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    Analgesic and Anti-Inflammatory constituents of annickia polycarpa stem and root barks and clausena anisata root.
    (AUGUST, 2016) Kumatia, Emmanuel Kofi
    Clausena anisata and Annickia polycarpa are medicinal plants used to treat various painful and inflammatory disorders among other ailments in traditional medicine. The aim of this study was to investigate the analgesic/antinociceptive and anti-inflammatory activities of the ethanol extracts of C. anisata root (CRE), A. polycarpa stem (ASE) and root barks (AR) in order to provide scientific justification for their use as anti-inflammatory and analgesic agents. Analgesic activity was evaluated using the hot plate and the acetic acid induced writhing assays. The mechanism of antinociception was evaluated by employing pharmacological antagonism assays at the opioid and cholinergic receptors in the hot plate and the writhing assays. Anti-inflammatory activity was also evaluated by carrageenan induced edema in rats’ paw assay. The compounds were isolated using bioassay-guided fractionation and their structures identified by spectroscopic methods. CRE at 1000 mg/kg p.o. produce significant (p < 0.001) analgesic activity of 72.15 and 48.05 % in the hot plate and writhing assays respectively and significant (p < 0.01) anti-inflammatory activity of 27.53 %. ASE also produced significant (p < 0.001) analgesic activity of 82.54 and 44.03 % in the hot plate and writhing assays respectively and significant anti-inflammatory activity of 69.64 %. Furthermore, the results also showed that the petroleum ether (pet ether) fraction (PEF) of C. anisata root extract and the chloroform fraction (AC) of A. polycarpa stem bark extract were the most active fractions among the petroleum ether, chloroform and aqueous fractions of these extracts. A total of seven (7) compounds were isolated. Four (4) coumarins, namely, anisocoumarin B, osthol, imperatorin and xanthotoxol in addition to a carbazole alkaloid, heptaphyline were isolated from PEF. Two (2) protoberberine alkaloids namely jatrorrhizine and palmatine were also isolated from AC. Palmatine was further isolated from the chloroform fraction of A. polycarpa root bark. The seven isolated compounds were tested for analgesic activity in the writhing test. Six of them at 6 mg/kg p.o., produced significant analgesic activity of 38.13 to 47.28 %. One of the isolates (xantothoxol) was inactive. Analgesic activity of diclofenac in the writhing test was 32.92 % at 6 mg/kg p.o. Four of the isolates were also tested for analgesic activity in the hot plate assay. These isolates at 9 mg/kg p.o. produced immence analgesic effect of 30.13 to 93.87 %. The analgesic effect of tramadol 9 mg/kg p.o. was 27.13 % in the hot plate test.
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    Risk factors and human coronaviruses associated with upper respiratory tract infections in three rural areas of Ghana
    (May 2014) Owusu, Michael
    Acute respiratory tract infections (ARI) are the leading cause of morbidity and mortality in developing countries, especially in Africa. In spite of its importance, information on the viral aetiology and risk factors associated with ARI are limited in Ghana. Even though human coronaviruses (HCoVs) are known to be associated with respiratory disease outbreaks and severe infections in some developed countries, their epidemiological role is understudied in many African countries including Ghana. It is therefore not known whether HCoVs are pathogenic viruses associated with ARI or only exist as normal commensals of the upper respiratory tract. The aim of this study was to find the association between HCoVs and ARIs, describe the sero-molecular epidemiology of HCoVs and identify the risk factors associated with upper respiratory tract infection. An unmatched case control study was conducted in Buoyem, Forikrom and Kwamang communities of Ghana. Subjects were interviewed on various socio-demographic factors and hygienic practices using structured questionnaires. Nasal/Nasopharyngeal swabs were taken from older children and adults, and tested for Middle East respiratory syndrome coronavirus (MERS-CoV), HCoV-229E, HCoV-OC43, HCoV-NL63 and HCoV-HKU1 using Reverse Transcriptase Real-Time Polymerase Chain Reaction. A total of 1272 subjects were recruited comprising of 662 (52%) controls and 610 (48%) cases. Risk factors associated with upper respiratory tract infections were school attendance to the level of Senior High and tertiary education, and being a health worker. Out of 322 subset of cases interviewed, 212 (66%) covered their nose with handkerchiefs when they sneezed, 52 (16%) covered with their hands upon xix sneezing and 79 (25%) sneezed in the open. Self-administered drugs such as herbs (2%), analgesics (25%) and antibiotics (16%) were used to manage upper respiratory tract infections. Out of the 1,272 subjects recruited, nasal swabs were taken from 1,213. Of the 1,213, 150 (12.4%) subjects were positive for one or more viruses. Of these, single virus detections occurred in 146 subjects (12.0%) and multiple detections occurred in 4 (0.3%). Compared with control subjects, infections with HCoV-229E (OR = 5.15, 95% CI = 2.24 – 11.78), HCoV-OC43 (OR = 6.16, 95% CI = 1.77 – 21.65) and combine HCoVs (OR = 2.36, 95% CI = 1.5 = 3.72) were associated with upper respiratory tract infections. Significant median virus concentration difference was observed for only HCoV- NL63 (cases: 2.41 x 106 copies per PCR reaction; IQR = 1.96 x 104 - 2.3 x 106 vrs controls: 1876.5 copies per PCR reaction; IQR =387.2 – 8.6 x 104, P=0.003) and the clinically relevant cut-off viral concentration was determined to be 7,510 copies per PCR reaction. HCoVs were found to be seasonally dependent with high proportions identified in the harmattan season (54/215, 25.1%) compared to the wet (80/516, 15.5%) seasons. The most frequent viruses detected in the harmattan and wet seasons were HCoV-229E and HCoV-NL63 respectively. HCoV-OC43 and HCoV-HKU1 were almost distributed equally throughout the year. Sequencing of the partial spike region was successful for 53 out of 146 samples (36.3%). Of the 53, 12 (22.6%) were HCoV-OC43, 14 (26.4%) were HCoV-NL63, 24 (45.3%) were HCoV-229E and 3 (5.7%) were HCoV-HKU1. A comparison of the obtained sequences resulted in no differences to sequences already published in GenBank. xx This study has identified risk factors of URTI and also demonstrated that HCoVs could play significant role in causing upper respiratory tract infections among adults and older children in rural arrears of Ghana. This information could be useful to policy makers, public health practitioners and other stakeholders in Ghana.
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    Antiplasmodial compounds from Ghanaian medicinal plants
    (December 2015) Komlaga, Gustav
    Malaria is a major public health challenge in Ghana, and many indigenes employ medicinal plants, beside orthodox medicines, to treat the disease. An ethnobotanical survey was performed in the Bosomtwi and Sekyere East Districts of Ghana to identify plants used locally to manage malaria. This was done in comparison with the plant ingredients in marketed herbal antimalarial remedies in the Kumasi metropolis. The survey inventoried ninety-eight (98) plant species; twelve (12; 12.2%) reported for the first time globally, and twenty (20; 20.4%) others for the first time in Ghana for the treatment of malaria. Twenty-three (23) locally available finished, often multi/polyherbal antimalarial products examined contained aerial or underground parts of twenty-nine (29) of the plants cited in the survey as ingredients. Twenty-two (22) of these products were registered by the Ghana Food and Drugs Authority; four (4) were included in the Ghana Health Service recommended herbal medicine list for treating malaria in Ghana. The aqueous as well as serially extracted organic solvents (petroleum ether, ethyl acetate, and methanol) extracts of five plants parts, selected based on their importance in the traditional treatment of malaria and lack of the appropriate data in the literature, were studied against the chloroquine-sensitive 3D7 and chloroquine-resistant W2 P. falciparum parasite in vitro. The plant materials included the whole of Phyllanthus fraternus, leaves of Tectona grandis, Terminalia ivorensis and Bambusa vulgaris, and root of Senna siamea. All the aqueous extracts showed notable antiplasmodial activity (IC50 < 10 µg/mL), except that of S. siamea, against 3D7 P. falciparum. Only T. ivorensis and S. siamea extracts showed activity against W2 P. falciparum (IC50 < 50 µg/mL). The extracts demonstrated high selectivity index (SI) for 3D7 P. falciparum (SI > 3.5) but very low SI for W2 P. falciparum. Resistance index (RI) was largely under 20. The organic fractions were equally active (IC50 < 50 µg/mL; 3D7 P. falciparum). The methanol extracts of the two most potent plant materials, the whole of P. fraternus and leaf of B. vulgaris, were subjected to phytochemical study to isolate and elucidate the chemical constituents, which were then assayed for antiplasmodial activity. The phytochemical study of the v methanol extract of P. fraternus yielded six compounds; Pf 1 to Pf 6 identified as the lignan, phyllanthin, and five securinega alkaloids namely nirurine, ent-norsecurinine, allo-norsecurinine, bubbialine and epibubbialine. This is the first isolation of allo - norsecurinine from a natural source and bubbialine from the Phyllanthus genus. The compounds displayed significant antiplasmodial activity against both 3D7 and W2 P. falciparum (1.14 ± 0.32 µM ≤ IC50 ≤ 59.00 ± 5.43 µM); ent-norsecurinine being the most active (IC50=1.14± 0.32 µM) and against the W2 P. falciparum. Only Pf2 (nirurine) and Pf1 (phyllanthin) displayed cytotoxicity (CC50 < 100 μM; HUVECs). This is the first report of the antiplasmodial activity of these compounds. Similar study of the methanol extract of B. vulgaris yielded 6 compounds, Bv1 to Bv6, identified as p-coumaric acid [(E)-3-(4-hydroxyphenyl) acrylic acid], cinnamic acid, dehydrovomifoliol [(E)-4-hydroxy-3,5,5-trimethyl-4-(3-oxobut-1-en-1-yl)cyclohex- 2-en-1-one], 3-oxo-α-ionol [9-hydroxy megastigma-4, 7-dien-3-one], loliolide [6- hydroxy-4, 4, 7a-trimethyl-5, 6, 7, 7a-tetrahydrobenzofuran-2(4H)-one] and tricin [5,7,4’-trihydroxy-3’,5’-dimethoxyflavone]. The six compounds are the first everreported isolations from B. vulgaris. All the compounds from B. vulgaris displayed significant activity against 3D7 (IC50 < 5 μΜ and W2 strains of P. falciparum (IC50 < 7 μM). Bv1 (p-coumaric acid) was the most active against 3D7 P. falciparum (IC50: 0.84 ± 0.90 μM) and Bv2 (cinnamic acid) the most active against W2 P. falciparum (IC50: 1.41 ± 0.38 μM). The compounds displayed no cytotoxicity (CC50 > 100 μM; HUVECs). This is the first report of the antiplasmodial activity of the six compounds. These twelve (12) compounds with remarkable antiplasmodial activity add to the library of natural compounds with antiplasmodial activity. This study has illustrated the potentials of Ghanaian medicinal plants as source of natural antiplasmodial compounds, and has justified the use of the plants in traditional treatment of malaria.
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    Evaluating the role of the Ghanaian pharmacist in medication safety strategies in hospitals
    (2015-11-04) Acheampong, Frankline
    Healthcare has always been a risky venture with a lot of harm associated with it. Medicines form a major and vital part of the healthcare delivery system. The World Health Organisation and many national safety organisations have created a lot of awareness about the importance of enhancing patient safety in the healthcare system. Evidence available suggests that a lot of adverse drug events especially medication administration errors occur during the medication use process. Consequently, it is important to detect and prevent errors at the drug administration stage, since it is also the last step before these errors could reach patients. In Ghana, little is known of the prevalence and contributory factors of medication administration errors. Current literature suggests the existence of many medication safety strategies that are being employed. Pharmacists in particular, have been identified to contribute immensely to the safe use of medicines globally. The aim of this study was to determine the existence of adverse drug events with emphasis on medication administration errors, explore the perceived roles and documented evidences of pharmacists’ roles in the safe use of medicines and understand the experiences and expectations of doctors and nurses on such roles. Methods The methods used were the following: • A direct non-participatory observation of medication administration by nurses followed by face-to-face interview with a sample of these nurses at the Surgical Medical Emergency Department of Korle Bu Teaching Hospital. • Survey of pharmacists working in Ghanaian hospitals across the country using a structured questionnaire. • Retrospective evaluation of documented clinical intervention reports followed by key informant interviews of pharmacists involved in the reporting. • Open and close-ended questionnaires administered to a conveniently sampled doctors and nurses at the hospital to explore their perceptions and expectations. Key findings • Medication administration errors occurred at a rate of 27.2% at the emergency setting. It was also shown that most of the causes of the errors were related to staff and environmental factors such as workload, and lack of adequate knowledge about medication and their use. • Pharmacists in Ghanaian hospitals perceived their services to be useful in preventing adverse drug events. They indicated that they spent more time on activities with perceived greatest impact on patient care such as reviewing pharmacotherapies, monitoring adverse drug reactions and counselling patients on medication use. • Documented evidence of Pharmacist’s clinical interventions activities revealed that 24 pharmacists made 1019 clinical interventions in 448 handwritten reports. Majority (76.1%) of the interventions related to drug therapy changes. The pharmacists reported that the major barrier to their medication safety roles was the perceived discrepant attitude of doctors and nurses. • In contrast, doctors and nurses indicated that they interacted frequently with pharmacists and acknowledged their roles to be useful in contributing to medication safety. Conclusions and Recommendations • Medication administration errors were observed in over a quarter of the activities of the nurses involved in the study. • There was an overwhelming evidence of the strategic role of hospital pharmacists in identifying and preventing adverse drug events. • Unlike the perception that pharmacists had about the discrepant attitudes of doctors and nurses, the clinicians acknowledged and appreciated the role of the pharmacist in medication safety. • The clinical role of pharmacist in hospitals should be intensified to enhance safety and patient care.