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- ItemAnalgesic and Anti-Inflammatory constituents of annickia polycarpa stem and root barks and clausena anisata root.(AUGUST, 2016) Kumatia, Emmanuel KofiClausena 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.
- ItemAnalgesic and antispasmodic constituents, and standardization of the root bark of Cassia Sieberiana D. C. [Caesalpiniaceae](2013) Sam, George HenryMedicinal plants have been used by man for the treatment of various diseases however many of the products from these have not been scientifically standardized. Cassisa sieberiana D.C. (Caesalpiniaceae) is a tree whose root bark is traditionally used as an analgesic, especially for schistosomiasis in Ghana. This work was carried out to validate its analgesic and antispasmodic action, identify the isolates by guided fractionation and standardize the power root bark which is used in therapy...
- ItemAnti-inflammatory and anti-anaphylactic effects of trichilia monadelpha (thonn.) j. j. de wilde. ex oliv.pp (meliaceae) extracts in Rodents(2013-08-04) Okon, Inemesit BenTrichilia monadelpha (Meliaceae) in Ghana and other parts of Africa, is used locally to manage various inflammatory and pain conditions.Three extracts(petroleum ather extrct,PEE, etthyll acetate extract, EthE, and ethernal extarct, EAE) obtained from this plant and evaluated pharmacologically................
- ItemAnti-inflammatory and ethopharmacological effects of an ethanolic leaf extract of Palisota Hirsuta K. Schum. (Commelinaceae)(2009-08-09) Boakye-Gyasi, EricLeaves of Palisota hirsuta are used in Ghana and other West African states for various painful and inflammatory conditions. This study is aimed at evaluating the anti-inflammatory and ethopharmacological properties as well as toxicity profile of an ethanolic leaf extract of Palisota hirsuta using animal models. Preliminary phytochemical screening showed that the powdered leaves contained tannins, reducing sugars, flavonoids, steroids and terpenoids with traces of alkaloids. Effect of the extract on acute inflammation was assessed in the carrageenan- induced foot edema in 7-day-old chicks with diclofenac and dexamethasone as reference drugs. Pre treatment with the extract (30-300 mg kg-1; p.o.) significantly, inhibited foot edema in the chicks comparable to the NSAID diclofenac with maximal inhibition of 54.71±11.04%. Diclofenac and dexamethasone also dose- dependently inhibited carrageenan-induced foot edema. The anti-arthritic effect of the ethanolic leaf extract was assessed in the Freund’s adjuvant induced-arthritis model in rats. Palisota hirsuta extract (PHE) as well as dexamethasone and methotrexate, used as positive controls, showed significant dose-dependent anti-arthritic properties when administered prophylactically, curatively and also in combination therapy. PHE (30-300 mg kg-1) significantly reduced the arthritic edema in the ipsilateral paw with the highest dose used giving a maximum inhibition of 13.02±8.77%. PHE (300 mg kg-1) also significantly prevented the spread of the edema from the ipsilateral to the contralateral paw indicating inhibition of systemic spread. Dexamethasone (0.3-3 mg kg-1) and methotrexate (0.1-1.0 mg kg-1) significantly and in a dose dependent manner also inhibited polyarthritis edema. PHE in combination with methotrexate did not show significant effect. However there was a significant inhibition of arthritis in both the acute and the polyarthritic phases when PHE was combined with dexamethasone. Dexamethasone in combination with methotrexate caused the greatest inhibition of both phases with an extreme level of significance as expected. Overall, the present results demonstrate that PHE has anti-arthritic effect which could be similar to that exhibited by methotrexate. P. hirsuta (30-300 mg kg-1; p.o.) also dose-dependently decreased baker’s yeast induced fever in rats when Paracetamol (10-100 mg kg-1; p.o.) was used as the reference drug. The in vitro antioxidant properties of the extract were evaluated using the reducing power test; 2, 2-diphenyl-1-picrylhydrazyl hydrate (DPPH) radical scavenging assay and the lipid peroxidation assay. In all tests, n-propyl gallate was used as the reference antioxidant. The extract (0.1-3.0 mg ml -1) showed a reducing power potential (EC ; 133.7±7.59 mg ml-1) but was less than that of the reference antioxidant n-propyl gallate (EC 3.77±0.07 mg ml-1) in the reducing power test. The relative anti-oxidative activity in the DPPH de-colorization assay (defined by the EC ) was in the order: n-propyl gallate (8.02±0.01 × 10-4) > extract (1.77±0.40 × 10-1). The extract (0.1-1.0 mg ml-1) and n-propyl gallate (0.01-0.1 mg ml-1) exhibited a concentration dependent inhibition of lipid peroxidation. The rank order of potency (defined by ED in mg ml-1) was found to be: n-propyl gallate (1.31±3.00 × 10-2) > extract (4.29±0.95 × 10-1). These findings present the extract with potent antioxidant properties which may account in part for its anti-inflammatory and analgesic activities. In the analgesic assay, the leaf extract of P. hirsuta (PHE) (30, 100 and 300 mg kg-1 p.o) as well as morphine and diclofenac (positive controls), caused significant dose- dependent anti-nociceptive activity in all the pain models used. In the tail withdrawal test, PHE (300 mg kg-1) increased withdrawal latencies significantly by 43.83±11.62%. Also, PHE (300 mg kg-1) completely reversed the inflammatory- induced mechanical hyperalgesia with a maximum percentage effect of 154.79±15.84%. PHE significantly reduced the number of acetic acid induced writhing in mice. In the formalin test, PHE (10–300 mg kg-1, p.o.) caused a marked and dose-related inhibition of both phases of formalin-induced nociception. The anti-nociceptive effect exhibited by PHE in the formalin test was reversed by the systemic administration of the non-selective opioid antagonist, naloxone, the NO synthase inhibitor, NG-nitro-arginine methyl ester (L-NAME) and the ATP- sensitive K+ channel inhibitor, glibenclamide. However, theophylline a non- selective adenosine receptor antagonist did not reverse the effect. PHE, unlike morphine, did not induce tolerance to its anti-nociceptive effect in the formalin test after chronic administration and also morphine tolerance did not cross-generalize to PHE. Overall, the present results demonstrate that the central and peripheral anti-nociceptive of PHE may partially or wholly be due to the stimulation of peripheral and/or central opioid receptors through the activation of the nitric oxide-cyclic GMP- ATP-sensitive K+ (NO/cGMP/K+ATP)-channel pathway. As part of the present study, the ethopharmacological properties of the ethanolic leaf extract, in multiple behavioral paradigms of anxiety and depression— the open field test, the light/dark box, the elevated plus maze (EPM), the forced swimming test (FST) and tail suspension test (TST) was evaluated. P. hirsuta treated mice (30-300 mg kg-1) exhibited anxiolytic activity similar to diazepam in all the anxiety models used. PHE significantly increased the percentage number of center entries and the percentage time spent in the center of the open field. It also significantly increased the time spent in the lit area in relation to the time spent in the dark area of the light/dark box as well as significantly increasing open arm activity in the EPM. These effects were completely reversed in the presence of flumazenil (3 mg kg-1), a specific antagonist of the benzodiazepine site in the GABAA benzodiazepine receptor complex. The extract also dose-dependently reduced the duration of immobility in both the FST (ED50: 114.55±72.69 mg kg ) and TST (70.42±0.06 mg kg-1). Pretreatment with •-methyldopa (400 mg kg-1; 3 h; p.o.), to reduce brain NE and DA tissue content or reserpine (1 mg kg-1; 24 h; s.c.) for the disruption of vesicular storage of brain NE, DA and 5-HT tissue content or a combination of the two drugs to deplete both newly synthesized and vesicular components of NE and DA transmission attenuated the anti-immobility effects of both imipramime and the extract but not fluoxetine. Neither the extract nor the standard drugs used modified motor performance on the rota rod test at all doses tested. Collectively, these results suggest that the extract has anxiolytic and antidepressant-like effects in the models employed possibly by GABAergic activation and/or modification of monoamine transport and/or metabolism. In the toxicological study, there were no significant differences found in almost all of the hematological, serum biochemical parameters and organ/body weight ratio. No abnormality of any organ was found during histopathological examination. The results showed that the no-observed adverse- effect level (NOAEL) of P. hirsuta extract (PHE) was >3000 mg kg-1 body weight per day in rats, which can be regarded as virtually non-toxic. In conclusion, PHE had no overt organ specific toxicity and hence has a high safety profile in rats. Putting all together, these novel findings provide some pharmacological evidence and basis for the traditional use of the leaves of P. hirsuta in traditional medicine to manage various painful and inflammatory conditions.
- ItemAnti-Nociceptive Properties of the Ethanolic Extract Of Fruits of Xylopia Aethiopica (Dunal) A. Rich (Annonaceae) and its Major Constituent, Xylopic Acid(2012) Ameyaw, Elvis OforiThe fruits of Xylopia aethiopica are traditionally used to treat malaria, fibroid (uterine), fungal infection, rheumatism, arthritis, amenorrhea, boil, haemorrhoids and flatulence whilst the crushed seeds are applied topically on the forehead in the treatment of headache and neuralgia. TLC and HPLC analyses of the extract revealed the presence of several compounds. The isolated xylopic acid produced a single spot in a number of solvent systems including petroleum ether: ethyl acetate (1:9) and hexane: ethyl acetate (1:9) and a single peak in HPLC analysis certifying the level of purity of the compound The extract and xylopic acid exhibited antinociception in all the pain models used. In the acetic acid-induced writhing test, the extract (30 – 300 mg kg-1) (F3,28=14.37, P<0.0001) and xylopic acid (10 – 100 mg kg-1) (F3,28=20.56, P<0.0001) significantly reduced abdominal writhes induced with acetic acid in mice with the highest dose of the extract inhibiting visceral nociception by 98.8 ± 0.8%. The highest dose of xylopic acid also inhibited visceral nociception by 93.8 ± 1.4%. Morphine (1 – 10 mg kg-1) (F3,28=9.77, P=0.00016) and diclofenac (1 - 10 mg kg-1) (F3,28=4.04, P=0.0165) used as controls in this model similarly exhibited significant antinociceptive activities in this test. The extract (30 – 300 mg kg-1) (F3,28=6.93, P=0.0012) significantly attenuated mechanical hyperalgesia in the Randall-Selitto test with maximum possible effect of 110 ± 16.17% at the highest dose used. Similarly xylopic acid (10 – 100 mg kg-1) significantly (F3,28=4.86, P=0.0076) attenuated mechanical hyperalgesia with a maximum possible effect of 94.58 ± 21.6% at the highest dose in the same test. The extract and xylopic acid were both relatively more effective in the Hargreaves thermal paw withdrawal test, (extract: F3,64= 8.10, P=0.0338, xylopic acid: F3,64= 7.11, P=0.03) compared to the tail flick test (extract: F3,64= 6.47, P=0.045, xylopic acid: F3,64= 19.5, P<0.0001). For the acute and chronic musculoskeletal pain tests morphine was most efficacious. The extract was also more efficacious than xylopic acid. Xylopic acid was however more potent in reducing both the chronic muscle and knee pain whereas the extract was more efficacious in the chronic skeletal pain model. The extract, xylopic acid and pregabalin ameliorated vincristine-induced neuropathic pain. Common symptoms experienced by patients with neuropathic pain such as mechanical hyperalgesia, tactile and cold allodynia were measured using Von Frey filaments and cold water. The extract (F3,28=5.12, P=0.006), xylopic acid (F3,28=3.72, P=0.0229) and pregabalin (F3,28=5.92, P=0.0029) produced tactile anti-allodynia. Similar effects were observed in the Von Frey intermediate and mechanical hyperalgesia as well as cold allodynia tests. In the formalin test the extract and xylopic acid inhibited both neurogenic and inflammatory phases of pain. The antinociception of the extract, xylopic acid and morphine involved the inhibition of opioid, NO-cGMP, 5-HT3, adenosine and muscarinic pathways Further determination of the mechanism of antinociception of xylopic acid carried using binding assay revealed the binding of xylopic acid to μ-opioid receptors with an enhancement of endogenous opioid binding. Capsaicin-sensitive C-fibres-glutamatergic-nociceptive pathway was found to participate in the antinociception of extract and xylopic acid. Tolerance to morphine antinociception on the opioid receptors developed after chronic treatment for eight days but failed to develop to the extract and xylopic acid. Also morphine tolerance did not cross-generalize to the extract and xylopic acid. In order to elucidate the drug—drug interaction between xylopic acid/morphine and xylopic acid/diclofenac in combination administrations, isobolographic analysis was performed. The experimental ED50’s (Zmix) of xylopic/morphine combination were smaller than their corresponding theoretical ED50’s (Zadd) in both phases of the formalin test indicating synergism. Isobolographic analysis of xylopic acid/diclofenac combination carried out also indicated potentiation of the combination as the experimental ED50 lay below the line of additivity. The degree of potentiation calculated as interaction index indicated that the combination synergized to produce antinociception. In summary these findings provide scientific data for the use of the fruit of X. aethiopica in the treatment of painful conditions and co- administration of xylopic acid/morphine and xylopic acid/diclofenac may be said to be beneficial as their various side effects may be reduced due to lower doses used with potentiation of their therapeutic effects.
- ItemAnticonvulsant and antidepressant effects of an ethanolic extract of the leaves of Pseudospondias microcarpa (Engl.) A. Rich. (Anacardiaceae) in animal models(2015-11-16) Adongo, Donatus WewuraThe search for novel pharmacotherapy from medicinal plants for central nervous system (CNS) disorders has become of importance since new agents with improved efficacy for more effective therapy are required. Pseudospondias microcapa has been extensively used in Ghana and other parts of Africa as medication for various diseases including CNS disorders. The present study examined the anticonvulsant, antidepressant, as well as some neurobehavioural properties of an ethanolic extract of the leaves of Pseudospondias microcarpa in animal models. Preliminary phytochemical screening of Pseudospondias microcarpa extract (PME) revealed the presence of saponins, tannins, glycosides, terpenoids, flavonoids and alkaloids. Neuropharmacological activities of the extract in mice was investigated. The extract produced sedation and analgesia in the Irwin test with an LD50 above 3000 mg kg-1. PME potentiated pentobarbitone sleeping time and was metabolised by hepatic enzymes. It however showed no effect on locomotor activity or motor coordination. Furthermore, the extract blocked convulsions induced by PTZ and showed analgesic activity in the tail withdrawal test. Based on the findings from the preliminary studies, the anticonvulsant activity of PME and possible mechanism(s) in animal models was further explored. The extract significantly delayed the onset as well as decreased the duration and frequency of pentylenetetrazole (PTZ)-, picrotoxin (PTX)- and strychnine (STN)-induced seizures. In addition, pretreatment of mice with PME before administration of 4-aminopyridine (4-AP) or isoniazid (INH) significantly increased the latency to seizures and reduced the incidence of mortality.The GABAA receptor antagonist, flumazenil, reversed the anticonvulsant effect of PME, further suggesting the possible involvement of the GABAergic system in its action. Furthermore, the extract protected against 6-Hz psychomotor seizures but had no effect in the MES test. Moreover, the anticonvulsant effect of PME (100 mg kg-1, p.o.) was prevented by pre-treatment with L-arginine or sildenafil. However, N-nitro-L-arginine methyl ester (L-NAME) or methylene blue (MB) augmented the anticonvulsant effect of PME. Acute treatment with PME reduced immobility of mice in the TST and FST. The antidepressant-like effect of PME (100 mg kg-1, p.o.) in the TST was blocked by p-Chlorophenylalanine and cyproheptadine but not prazosin, propranolol or yohimbine, thus suggesting 5-HT pathway may be involved in the action of PME. This is further confirmed by the potentiation of 5-hydroxytryptophan-induced head-twitch response by PME in mice. Pretreatment with a combination of reserpine and α-methyl-p-tyrosine (AMPT) completely prevented the behavioural effects of PME, fluoxetine and desipramine. Concomitant administration of D-cycloserine and the extract potentiated the anti-immobility effect. In contrast, D-serine a full agonist of glycine/NMDA receptors abolished the effects. In the repeated open-space swim test, mice showed a progressive decrease of active swimming and a corresponding increase in immobility that persisted for several days. However, the increased inactivity was reversed selectively by the extract and the classical antidepressant drugs, in that they increased distance swum and decreased immobility. Moreover, the depressive-behaviour induced by the repeated open-space swim test impaired spatial learning and memory performance in the Morris water maze (MWM) test. This was however reversed by the extract. The long-term effects of chronic mild stress (CMS) and PME treatment on depressive, anxiety-like behaviour and cognitive function were also investigated. Exposure of mice to the CMS paradigm displayed decreased sucrose intake, poor coat state, decreased grooming frequency (splash test), increased immobility, increased anxiety and impaired cognitive function. These effects were however reversed by chronic PME treatment. Anxiolytic-like effects of the extract in behavioural paradigms of anxiety was also evaluated. P. microcarpa treated mice (30-300 mg kg-1, p.o.) exhibited anxiolytic-like activity similar to diazepam in all the anxiety models used. PME increased open arm activity in the elevated plus maze (EPM) as well as increasing the time spent in the lit area in relation to the time spent in the dark area of the light/dark box. The extract also increased the number of central entries and time spent in the center of the open field. In addition, an increase in social interaction and decreased stress-induced hyperthermia was obseverd for PME-treated mice. Acute and subacute toxicity in rats did not show deaths after 14 days treatment with the extract (30–3000 mg kg-1). Haematological or serum biochemical parameters were not affected except decrease in lymphocytes (100−3000 mg kg-1), triglycerides (100 mg kg-1) and very low density lipoproteins (100 mg kg-1). Histopathological examination did not reveal toxic effect on the stomach, heart, liver, brain, kidney and spleen. Results of the present study suggests that PME possesses sedative, analgesic, anticonvulsant, antidepressant and anxiolytic effects without affecting motor coordination.
- ItemAnticonvulsant, Antidepressant and Anxiolytic Effects of Mallotus Oppositifolius (Geiseler) Müll. Arg. (Euphorbiaceae)(2012-08-20) Kukuia, Kennedy Kwami EdemMallotus oppositifolius is used in Ghana for CNS disorders but very little scientific evidence exists to support its use. Thus central effects of 70% v/v hydroalcoholic extract of the leaves of Mallotus oppositifolius (MOE) was assessed. Anticonvulsant effects of the extract in acute and chronic seizure models were evaluated. The study also investigated the effect of the extract on animal models of depression and anxiety. In a preliminary screening of the central effects of the extract, oral dose of MOE induced sedation (1000 – 3000 mg kg-1); caused neuromuscular deficits in the rotarod test (300 – 3000 mg kg-1); reduced spontaneous locomotor activity in the activity cage; exhibited anticonvulsant effect (30 – 3000 mg kg-1) and central analgesic effect in the tail immersion test (100 – 3000 mg kg-1). The LD50 was approximately 6000 mg kg-1 in mice. M. oppositifolius (10 - 100 mg kg-1, p.o.) exhibited anticonvulsant effect in the picrotoxin and strychnine induced seizure tests. The extract significantly delayed onset of myoclonic jerks and clonic convulsions; decreased the frequency and duration of clonic convulsions in these models. In the MES test, the extract caused a significant and dose dependent decrease in the duration of tonic limb extensions. In the pilocarpine induced status epilepticus, MOE delayed the onset of clonic convulsions and decreased the duration of these seizures. Furthermore, the extract protected mice against mortality induced by 4-aminopyridine and delayed the onset of both clonic and tonic convulsions. Flumazenil, a GABAA/benzodiazepine antagonist, reversed the anticonvulsant effect of the extract in the PTZ-induced seizure test suggesting enhancement of GABAA neurotransmission is involved in the anticonvulsant effect of the extract. Isobolographic analysis of the combination of diazepam and extract showed a synergistic effect but the mode of action of this effect may not be dependent on enhancement of GABAA neurotransmission since flumazenil failed to reverse their anticonvulsant effect. Oral doses of MOE (10 - 100 mg kg-1), fluoxetine (3 - 30 mg kg-1) and imipramine (3 - 30 mg kg-1) decreased the frequency of immobility and immobility periods of mice in both the FST and TST when compared to control group, indicating significant antidepressant activity. In the open space swim test, a chronic depression model, MOE demonstrated antidepressant-like effect on the first day of treatment and sustained it throughout the period of drug treatment. MOE decreased immobility time while increasing the distanced travelled by the mice. The depression induced in this model induced significant impairment in spatial learning and memory in the Morris water maze—this was reversed by the extract and fluoxetine but not imipramine. Extract, fluoxetine and imipramine treatments did not have significant effects on weight variation. A day after the 14th day of drug treatment, the antidepressant effect was still significant. A 3-day subcutaneous pretreatment with 200 mg kg-1 para-chlorophenylalanine (pCPA), reversed the antidepressant effect of MOE and fluoxetine but not imipramine, suggesting that serotoninergic enhancement may be involved in the behavioural effect of the extract. This was confirmed by the ability of the extract to potentiate the head twitch responses induced by 5-hydroxytryptophan in mice, a model sensitive to 5-HT2A receptor activation. Pretreatment with α-methyldopa (400 mg kg-1) however, failed to reverse the behavioural effect of the extract and fluoxetine treatments in the forced swim test. The same result as above was observed for extract and fluoxetine treatments when mice were pretreated with reserpine (1 mg kg-1) or a combination of α-methyldopa (200 mg kg-1) and reserpine (1 mg kg-1). This suggests that the antidepressant effect of the extract may not be dependent on central noradrenergic mechanisms. Administration of D-serine (600 mg kg-1), a full agonist on the glycine site of the NMDA receptors, reversed the antidepressant effect of the extract, fluoxetine and desipramine in both the TST and FST. D-cycloserine (2.5 mg kg-1), a partial agonist potentiated this behavioural effect in both extract and fluoxetine treated mice but not desipramine in both the TST and FST. This suggests possible involvement of glycine/NMDA receptor or pathway antagonism in the antidepressant effect of the extract. MOE slightly increased curling score in the tail suspension test and this was significantly potentiated by D-cycloserine, suggestive of possible opioidergic activity. MOE (10 - 100 mg kg-1, p.o.) showed anxiolytic effect in the three anxiety models used namely; elevated plus maze, light-dark box and open field tests. M. oppositifolius treatment significantly increased the percentage of centre entries and the percentage time spent in the centre of the open field. M. oppositifolius also increased the time spent in the lit area and the latency to leave the lit area in light/dark box. In the EPM, it significantly increased open arm activities by increasing percentage open arm entries and duration. MOE also decreased risk assessment behaviours such as the head dips, stretch-attend postures and rearing. Acute and subacute toxicity in rats did show deaths after 14 day treatment with the extract (30 – 3000 mg kg-1). Extract treatment did not affect weight of rats or the relative organ weights. Haematological or serum biochemical parameters were not affected except increases in serum bilirubin (300 and 3000 mg kg-1), urea and creatinine (30 and 100 mg kg-1). Histopathological examination did not reveal toxic effect on the stomach, heart, liver and spleen. There were however some morphological changes of the kidney at 30 mg kg-1. These results suggest that the extract has anticonvulsant effect possibly through enhancement of GABAergic, glycinergic and potassium channel activation or increased potassium conductance. Possible inhibition of muscarinic and glutamatergic transmission may also be involved. The antidepressant-like effects of the extract may be due to the interplay of serotoninergic, glycine/NMDA and opioidergic pathways. The extract also demonstrated anxiolytic-like effects possibly by the involvement of GABAergic and serotoninergic mechanisms.
- ItemAntiplasmodial compounds from Ghanaian medicinal plants(December 2015) Komlaga, GustavMalaria 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.
- ItemAntiplasmodial evaluation of extracts of selected Ghanaian medicinal plants and other bioactivities of isolates of polyalthia longifolia var. pendula (annonaceae)(2014) Gbedema, Stephen YaoMalaria is one of the most important tropical infectious diseases. About half of the world’s population is at risk of developing malaria. It is a major public health challenge as well as a significant economic burden on many developing countries, especially in the African region. Most malaria patients are usually anaemic and also experience reduced or compromised immune systems which therefore predispose them to secondary bacterial and fungal infections. People living in extreme poverty are the most vulnerable to these infectious diseases simultaneously. This affects their ability to make a living and move out of poverty. Many of these people rely on herbs for treatment of the disease due to lack of access to efficacious antimalarial drugs against, especially the fast spreading multidrug-resistant Plasmodium falciparum. However, very few of these folklore herbs have been scientifically investigated and authenticated for used in the management of malaria and other infectious diseases. This study therefore sought to investigate and validate or otherwise the traditional uses of some Ghanaian plants for treating malaria. The ethanolic extracts of ten plant species which were selected based on their traditional medical application were screened against the multidrug resistant P. falciparum (K1 strain) by the parasite lactate dehydrogenase (pLDH) assay. Seven of the plant species (Adenia cissampeloides Planch. ex Hook. Anthocleista nobilis G. Don, Elaeis guineensis Jacq., Entandrophragma angolense (Welw.) CDC, Mallotus oppositifolius (Geisel.) Müll. Arg., Sarcocephalus latifolius (J.E.Sm) E. A Bruce and Polyalthia longifolia var. pendula) showed potent antiplasmodial activity with IC50 < 50 μg/ml. The extracts P. longifolia exhibited the most potent activity (IC50 < 23 μg/ml). Bioassay guided fractionation of P. longifolia extracts yielded four clerodane diterpenes [16- hydroxycleroda-3,13-dien-16,15-olide (1) and its acetylated derivative acetyl-16-oxycleroda- 3,13-dien-16,15-olide (1a), 16-oxocleroda-3,13E-dien-15-oic acid (2) and 3,16- dihydroxycleroda-4(18),13(14)Z-dien-15,16-olide (3)], a steroid [β-Stigmasterol (4)] and two vii alkaloids [Darienine (5) and L-Stepholidine (6)]. Even though the steroid and the alkaloids showed weak antiplasmodial activity (IC50: 22 -105 μg/ml), the clerodane diterpenes exhibited significantly (p < 0.005) potent blood schizonticidal activity (IC50: 3 - 6 μg/ml) against the multidrug resistant malaria parasite. Although the isolated clerodane diterpenes 1a, 2 and 3 are known compounds, this is the first report of their antiplasmodial activity especially against the multi-drug resistant Plasmodium falciparum. The clerodane diterpenes also displayed potent antibacterial and antifungal activities (MIC ranged between 3.13 and 50.00 μg/ml) against Staphylococcus aureus (ATCC 25923), Streptococcus pneumoniae (clinical isolate), Bacillus subtilis (NCTC 10073), Pseudomonas aeruginosa (ATCC 4853), Salmonella typhi (NCTC 8385), Escherichia coli (NCTC 25922) and Candida albicans (NCPF 3179). The study therefore appeared to lend support to the traditional uses of these plants as remedies for malaria in Ghana and formulations containing especially P. longifolia extracts will be highly beneficial in treating mixed infections associated with malaria.
- ItemAssessment of the analgesic properties of maerua angolensis dc (capparidaceae) in animal models of pain(OCTOBER, 2015) Iliya, Hosea AziMaerua angolensis is a medicinal plant used traditionally to relieve pain. Although the leaves, roots and stem barks are used, their efficacy, and safety have not been proven scientifically. This study therefore assessed the antinociceptive properties of Maerua angolensis. The antinociceptive activity on hydroethanolic extracts of the leaf, root and stem bark (30 – 300 mg/kg, p.o.) in the writhing test showed significant (P<0.0002) reduction of pain induced by acetic acid with the stem bark being more potent. Subsequently, the stem bark was extracted with petroleum ether, ethyl acetate or hydroethanol to obtain three (3) extracts which at 3 - 30 mg/kg, p.o. significantly (P<0.0006) reduced pain in both neurogenic and inflammatory phases of the formalin test in rats with the petroleum ether extract being more potent in neurogenic while ethyl acetate was more potent in inflammatory phase. Phytochemical results of the three solvent extracts shows presence of saponins, steroids, tannins, terpenoids, alkaloids, glycosides, flavonoids, oils and fats. The two most potent extracts were combined and subsequently referred to as MAE, fractionated in column to two (2) fractions F1 and F32 and purified leading to isolation of four (4) compounds C1, C2, C3 and C5 identified and characterized by 1H-NMR, GCMS and IR spectroscopy to be fatty acid and fatty acid esters namely octadecanoic acid methyl ester, bis (2-ethylhexyl) phthalate, octadecanoic acid and oleic acid methyl ester, respectively. MAE and fractions (3 – 30 mg/kg, p.o.) produced significant (P<0.05) antinociceptive effects in writhing, formalin, prostaglandin E2-induced mechanical hyperalgesia, bradykinin- and epinephrine-induced thermal hyperalgesia, tail-flick and paw withdrawal tests exhibiting both peripheral and central analgesic action. MAE and fractions reduced the number of jumps (intensity of withdrawal syndrome of morphine dependence) by mice but their effect was blocked by bicuculline and aminophylline. MAE and fractions suppress morphine withdrawal syndrome via stimulation of GABAergic and adenosinergic transmission. MAE and fractions (3 and 10 mg/kg) did not compromise the motor coordination of mice in the rotarod test, suggesting lack of central depressant effect in their antinociceptive effect. MAE and fractions effects on mechanical hyperalgesia, tactile and cold allodynia measured with Von Frey filaments and cold water in a mouse model of vincristine-induced neuropathy showed inhibition of pain suggesting their analgesic effects in cancer patients with vincristine-induced neuropathy. Theophylline, L-NAME, atropine, glibenclamide and yohimbine reversed MAE and fractions antinociception in writhing test while naloxone and ondansetron additionally reversed it in the tail-flick test. MAE and fractions inhibited capsaicin- and glutamate-induced nociception implying involvement of TRPV1 and glutamate receptors. Peripheral analgesic action of MAE and fractions involved ATP sensitive K+ channels, adenosinergic, muscarinic, α2 adrenergic and NO-cGMP paths while central action in addition involved 5-HT3 and opioid receptors. The compounds (1 – 10 mg/kg, p.o.) in writhing and wiping tests in mice reduced pain suggesting their peripheral and central analgesic action. PCPA, ondansetron, capsazepine and naloxone reversed antinociception of compounds in wiping test indicating involvement of 5-HT3, TRPV1 and opioid receptors. The compounds (3 - 30 µg/ml) reduced locomotor activity of zebrafish larvae exposed to acetic acid. Acute and sub-acute toxicity tests of MAE in rats revealed LD50 above 3000 mg/kg orally with no significant changes in body weight, relative organ weights, haematological and serum biochemical indices but significant histological changes in livers at 1000 and 3000 mg/kg MAE, showing its relative safety at therapeutic dose. Collectively, this study provides scientific data for the use of Maerua angolensis in the treatment of pains and contribute to the analgesic knowledge of this species.
- ItemBioactive constituents from the Ghanaian medicinal plant Chlorophora regia and its root endophytic fungus JK10.( AUGUST, 2016 ) Kyekyeku, James OppongThe extracts of the Ghanaian medicinal plant Chlorophora regia A. Chev (Moraceae) has been used for the treatment of various ailments in traditional medicine including burns, wounds, snake bite, wasp bite. A search through the literature, however, revealed there were no available data on the phytochemical composition of the plant. Therefore, the main objective of this study was to isolate, characterize and evaluate some biological activities of secondary metabolites from the stem bark of the plant and further investigate the endophytic community harbored in the inner tissues of the plant. Extensive phytochemical investigation of the stem bark resulted in the isolation and characterization of four new metabolites, regiafuran A–C and 6–prenylated–3,5,7,4ʹ–tetrahydroxy–2ʹ–methoxyflavonol in addition to fifteen known compounds. The isolated compounds were tested for their free radical scavenging activities. Regiafuran A–B, mulberrofuran Y, kuwanol E and 5,7,4ʹ–trihydroxy–2ʹ–methoxyflavanone demonstrated significant free radical scavenging activities with IC50 values of 1.9 μg/ml, 2.4 μg/ml, 2.2 μg/ml, 2.1 μg/ml and 1.8 μg/ml respectively. An unidentified endophytic fungus, JK10, was isolated from the root of C. regia. Twelve compounds including seven new 7–desmethyl derivatives of fusarin C and five known compounds were isolated from the endophytic fungus, JK10. The planar and relative configurations of the new compounds were elucidated by combined spectroscopic analyses of their UV, IR, HRESI–MSn, ECD and NMR data. The absolute configuration of solaniol was established for the first time by X–ray diffraction analysis of a single crystal. The antibacterial activities of the isolated compounds were evaluated. 7–desmethyl fusarin C–22/23 and 7–desmethyl fusarin C–25 exhibited remarkable activity at concentrations of 10.0 μg/mL against the soil bacterium Acinetobacter sp. BD4 comparable to the reference standard streptomycin. All the tested compounds demonstrated activity against the environmental strain of E. coli. Based on the results it could be proposed that the endophytic fungus, whose origin is from the roots, contributes a chemical–mediated defensive mechanism to the host plant against iii invading specific soil and environmental bacterial pathogens. This may confirm the existence of a unique cost–benefit endophyte–plant association. The spatial distribution of three kaurane diterpenes, xylopic acid, ent–kaur–16–en–19–oic acid and 15–oxo–ent–kaur–16–en–19–oic acid, in the fruits of Xylopia aethiopica (Dunal) A. Rich (Annonaceae) were visualized by MALDI–HRMS imaging techniques. The distribution of the compounds was predominantly in the pericarp region of the fruit with non-detectable levels in the seed.
- ItemBiochemical Markers of Oxidative Stress as Indices of HIV/AIDS Progression(2009-07-12) Obirikorang, ChristianReactive Oxygen Species (ROS) has been implicated in the pathogenesis and the progression of HIV infection. This study was aimed at investigating the levels of oxidative stress and their probable relationship as markers of HIV disease progression in HIV positive patients in two established HIV/ART centres in Ghana. In all two hundred and twenty eight (228) confirmed People Living with HIV/AIDS (PLWHAs) were included in the study. The subjects were recruited from the Central Regional Hospital (CRH) and Bolgatanga Regional Hospital (BRH). One hundred and forty-three (143; 62.72%) PLWHAs were recruited from CRH and they comprised of eighty one (81; 56.64%) males and sixty two (62; 43.36%) females whilst eighty five (85; 37.28%) PLWHAs were recruited from BRH comprising of forty three (43; 50.59%) males and forty two (42; 49.41%) females. These two hospitals were chosen to give a fair representation of PLWHAs in Ghana. Another 100 sex- and age-matched healthy, HIV-seronegative individuals were studied in parallel as controls. Ethical clearance was obtained from Committee on Human Research, Publications and Ethics (CHRPE), School of Medical Sciences, Kwame Nkrumah University of Science & Technology (KNUST), Kumasi. All subjects gave informed consent to take part in the study after verbal and written explanation of the methods and risks involved had been given. Venous blood samples were taken and assayed for the haematological parameters (haemoglobin (Hb), haematocrit (HCT) and total white blood cell count (WBC), biochemical assays (total Serum Protein, Serum Albumin, Triglycerides, Total Cholesterol, HDL-cholesterol and LDL-cholesterol and markers of oxidative stress (Serum Malondialdehyde (MDA), Ferric Reducing Ability of Plasma (FRAP) and serum Vitamin C and E, Superoxide Dismutase (SOD) Glutathione Peroxidase (GPx). The subjects with the mean age of 36.9±10.9 years which was not significantly different from that of the control group of 39.4+13.4 years. The PLWHAs subjects were grouped per the criteria of the Center for Disease Control (CDC) as: CD4 count (1) ≥ 500 mm-3; (2) 200 – 499 mm-3; and (3) < 200 mm-3. Forty three (43; 18.86%) patients (38.7+10.94 years) had a CD4+ count ≥ 500 mm-3, sixty three (63; 27.63%) patients (37.4+9.54 years) had a CD4+ count between 200 – 499 mm-3 and one hundred and twenty two (122; 53.51%) patients (36.7+10.93 years) had a CD4+ count < 200 mm-3. Markers of oxidative stress revealed significant differences between the patients and control subjects. Malondialdehyde (p<0.001) in the patients was markedly increased as compared to the control group. This suggests increased lipid peroxidation in HIV infected individuals and this increased with the progression of the infection. Ferric reducing ability of Plasma (p<0.0001), Glutathione peroxidase (p<0.0001), Superoxide Dismutase (p<0.0001) were decreased in the patients compared to the control group indicating an increase in free radicals product. Vitamin C (p<0.0001) and E (p<0.0001) were reduced in the patients compared to the control group suggesting a decrease in the antioxidant level as the HIV infection progressed in the patients. Results from the haematological assay revealed a significant decrease in the mean blood haemoglobin levels of the HIV positive patients compared to the control subjects. The significant positive correlation (p<0.0001) between Hb and CD4 count highlights its usefulness in the progression of HIV infection. The haematocrit result pattern showed a consistency with the Hb. There was a significant positive correlation between HCT and CD4 counts (p<0.0001). No significant difference was observed in the total white blood cell count (WBC) of HIV positive patients and the control group (p=0.1830). Apart from serum total Protein (p<0.0001) and Triglycerides (p<0.001), which showed significant increase as compared to the control group, serum albumin (p=0.0106), Total cholesterol (p<0.0001), HDL-cholesterol (p<0.0001) and LDL cholesterol (p<0.0001) showed a significant decrease as compared to the control group. From the correlation analysis, serum Albumin(R=0.41), Total cholesterol (R=0.67), HDL-cholesterol (R=0.27) and LDL cholesterol (R=0.27) showed a positive significant correlation in relation to the CD4 count. However serum total Protein (R=-0.36), Potassium (R=-0.67) and Triglycerides (R=-0.27) did show a negative correlation in relation to the CD4 count. Altogether the findings of this study have revealed that oxidative stress increases as HIV infection progressed. We propose from the study that the interaction of the mechanism underlying oxidative stress and HIV progression and subsequent apoptosis is a receptor-mediated process. During the early phase of HIV infection, generation of ROS has been known to activate HIV replication in vivo through the activation of a factor that binds to a DNA-binding protein, NF-kappa B which is a known activator of HIV replication. This leads to increase in the disturbance in the antioxidant system leading to an increase in ROS production with concomitant biochemical and haematological derangement as observed in the study. Therefore an early intervention with antioxidant supplements in the early phase of HIV infection is likely to reduce HIV progression and improve the immune function.
- ItemThe biological validity and feasibility of the community based questionnaire approach in determining the epidemiological status of disease and insect vector nuisance(1998) Lale, Von LouisIn the present study, the community-based questionnaire approach using an existing disease surveillance network and the school system was compared and validated with standardized biomedical and other techniques for guineaworm infection, schistosomiasis morbidity and blackfly nuisance monitoring. In all, 707 community key informants and 6,020 children were interviewed in 194 communities (70 with schools and 124 without schools) in East Mamprusi District of the Northern Region of Ghana. The communities/schools were randomly allocated to be interviewed during the rainy season or dry season. Children in ninety-nine communities were interviewed in the rainy season and those in the remaining ninety-five communities interviewed in the dry season. All questionnaires were returned within four weeks. Reagent strip tests for haematuria, urine filtration and microscopy were used for validating schistosomiasis, whilst re-interviews by research team for consistency and checks on evidence of infection were used to validate guineaworm and indirect validation for blackfly nuisance was done with checks on the probability of occurrence of the flies at the time of the survey, using Atlas GIS computer software. The sensitivity, specificity, and predictive values of questionnaires in identifying guineaworm were very high. The kappa statistic (K) was very high (0.92), indicating almost complete agreement between questionnaire responses and guineaworm validation. The prevalence of guineaworm of 1.2% from questionnaires is in line with previous epidemiological findings of guineaworm infection in the study area. In this study, the 3+ positives of the investigators’ and interviewers’ reagent strip testing indicated good agreement (K 083 and 0.77 respectively) with microscopy and is therefore a better indicator of positive status of schistosomiasis in the study area. The validity of the questionnaires was less than satisfactory, with very low sensitivity and positive predictive value (4.2%) and very poor agreement with microscopy (K = 0.07). The specificity and negative predictive value were high as generally expected in low prevalence areas. The low prevalence of schistosomiasis in the study area (<1%) complements the key informants’ low ranking for ‘blood in urine’. Questionnaires were fairly sensitive and specific in a high prevalence school (Buipe) but not in the low prevalence schools (New Buipe and Daboya). The use of colour charts for recall of urine colour as opposed to direct questioning, significantly improved the ,c sensitivity, and predictive values in both low and high prevalence schools. Both the school teachers and guineaworm volunteers performed well in respect of questionnaire administration and reagent strip testing. The use of questionnaires cost two times less than reagent strip testing and 25 times less than microscopy in communities with and without schools, whilst the combined use of questionnaires and reagent strips was 12 times and 14 times less in communities with and without schools respectively. About 93% of the children interviewed experienced blackfly bites in the rainy season. The key informant respondents also indicated rainy season as most important for fly nuisance. The 50% or more bites had the highest K for fly nuisance and is thus the best community cut-off for blackfly nuisance monitoring in the study area. There was no significant difference in blackfly nuisance with respect to proximity of communities to potential breeding sites (P>0.05, Mann-Whitney U-test, 95% Cl). The cost of US$ 125 per surveyed community or school under general expenditure represents the logistical expenses for all the diseases and insect vectors included in the questionnaires and this underscores the cost-effectiveness and need to integrate guineaworm surveillance with the monitoring of other health conditions, insect vectors as well as relevant occurrences at the community level.
- ItemCellular immune response to mycobacterium ulcerans infection(2015-07-12) Frimpong, MichaelBackground: M. ulcerans infection (Buruli ulcer) is a disease of the skin and soft tissue endemic in sub-Sahara Africa, with the major burden in West Africa. It belongs to a large group of environmental mycobacteria. Generally, protection against mycobacterial infection is thought to be based on cell-mediated immunity, specifically Th-1 type cellular immune responses are essential for control of mycobacterial infections, while humoral response have little benefit. The reasons why only some individuals in endemic areas who are exposed to M. ulcerans develop lesions are not known but are likely to reflect individual differences in the immune response to infections with this mycobacterium. Aim of study: These study aims at describing the cellular immune response to Mycobacterium ulcerans infection (Buruli ulcer) associated with protection and determine the efficacy of two subunit proteins (MUL 4978 and MUL 3720) as potential candidates for vaccine against Buruli ulcer. And finally investigate the effect of co-infection with M. perstans on BU susceptibility. Methods: All clinically suspected cases of Buruli ulcer were confirmed by standard PCR and ZN microscopy. Interferon gamma (IFN-γ) secretion following stimulation with mycobacterial antigens of peripheral blood mononuclear cells (PBMC) and whole blood from Buruli ulcer confirmed patients and household contacts were investigated using IFN-γ ELISA. The effectiveness of two subunit protein vaccines (MUL 3720 and MUL 4978) as potential vaccine candidates was tested using the method mention above. Also CD4+, CD8+ T-cell profiles and CD19 + cell populations in patients with Buruli ulcer were determined by flow cytometry analysis. A case control of 66 M. ulcerans (Mu) disease patients and 30 household contacts were investigated for Mansonella perstans (Mp) co-infection. Patients confirmed to have Mu disease vi by PCR were given the WHO recommended combination of rifampicin 10mg/kg and streptomycin 15mg/kg for 8 weeks. Ivermectin 150ug/kg and doxycycline 200mg were administered for 6 weeks as treatment of Mp infection when present. Results: The results showed that following stimulation with M. ulcerans antigens, PBMC from Buruli ulcer patients and their household contacts mounted high IFN-γ response. Also the Buruli ulcer patients with ulcerative lesions produced more IFN-γ than those with pre-ulcerative lesions (p = 0.026). IFN-γ secretion increased with treatment, with significant difference (p = 0.0078) at 6 weeks compared to baseline (pre-treatment), corresponding to a decrease in patients’ lesion sizes. Patients with severe forms of Buruli lesions (Category II and III) had a significantly decreased CD4+ T-cell population compared with healthy contacts (p = 0.0395). There were no statistically significant differences in the populations of CD8+ T-cell and B cell (CD3-CD19+) populations. There were high IFN-γ responses to subunit protein vaccines and IFN-γ levels in both candidates vaccine antigens (MUL 3720 and MUL 4978) were significantly high after 6 weeks of treatment compared to before treatment (p = 0.03 and 0.005) respectively. Fifteen out of 66 (23%) patients with Mu disease were co-infected with Mp while 4 out of 30 (13%) of the household contacts had infection with Mp. While filarial infection was more common among Buruli ulcer patients than household contacts it did not influence healing time of Buruli lesions (p = 0.93) or predispose patients to more severe forms of the disease. Conclusions: These findings suggest that T helper cell-1 (Th-1) immune response to M. ulcerans may play a protective role in the control of the disease. Also patients with severe forms of Buruli ulcer had depleted CD4+ T-lymphocyte populations. Furthermore, the results indicate that subunit protein vaccines MUL 3720 and MUL 4978 are immunogenic in human ex-vivo assays. This study also provided clear evidence of M. perstans co-infection in Buruli ulcer patients.
- ItemChemical constituent(s), anti-inflammatory, anti-oxidant and anti-nociceptive activities of the roots Of Palisota Hirsuta K.Schum (Commelinaceae).(February, 2012 ) Sarpong, Francis MainooThe roots of Palisota hirsuta are used in Ghana and other West African countries where the plant grows for the treatment of various disease conditions such as rheumatoid arthritis (other painful and inflammatory conditions), infertility in females, anaemia, dysentery etc. The current study seeks to evaluate the anti-inflammatory, analgesic and anti-oxidant properties of the ethanolic root extract of P.hirsuta, the methanolic and petroleum ether fractions thereof and the constituents isolated from these fractions using animal models. The carrageenan-induced foot oedema test in 7-day old chicks was used to assess the anti-inflammatory effect of the ethanolic extract, methanolic and petroleum ether fractions and all the isolated constituents. Diclofenac and dexamethasone were used as reference drugs for these assessments. The effect before the induction of inflammation (pre-emptive) paradigm was used for the assessment of inflammation. The analgesic (anti-nociceptive) effect of the ethanolic extract methanolic and petroleum ether fractions as well as the isolated constituents was assessed using the formalin-induced nociception test in the mice paw. The in vitro antioxidant properties of the isolates PH I, II, III, IV, V and VI were evaluated by using the lipid peroxidation assay, 2,2-diphenyl-1-picrylhydrazylhydrate (DPPH) radical scavenging assay and reducing power test. The standard antioxidant n-propyl gallate was used as the reference antioxidant in all the assays (the anti-oxidant assays were carried on the isolates following qualitative tests with DPPH). As part of the current study, the structures of the isolated constituents were evaluated using Gas Chromatography coupled with Mass Spectrometer (GC/MS) and Nuclear Magnetic Resonance. The crude extract PHC, methanolic fraction PHM and the petroleum ether fraction PHE of P.hirsuta (30-300mg/kg, p.o) dose dependently reduced oedema with maximal effects of 64.01±10.90%, 72.91±4.06%, and 57.86±11.89% (prophylactic) respectively. The extract PHC (30-300mg kg-1, p.o) caused a dose-related inhibition of both phases (i.e. phase 1 and phase 2) of the formalin-induced nociception. The highest dose caused maximal inhibitions of 40.89±24.10% and 66.25±32.08%. The methanolic fraction; PHM (30-300mgkg-1, p.o), also produced maximal inhibitions of both phases by 72.00±15% and 61.00±41.97%. PHE, which is the petroleum ether fraction dose-dependently, inhibited both phases of the formalin-induced nociception. Maximal inhibitions of 45.02±29.81% and 80.50±4.62% were respectively produced by PHE. For the isolates, PH I, II, III, IV, V and VI, the maximal inhibitions produced are as follows; PH I: 55.08±14.90% and 63.57±25.91%, PH II: 90.86±6.407% and 67.47±20.84%, PH III: 57.87±8.43% and 79.90±7.05%, PH IV: 71.39±9.19% and 89.19±3.81%, PH V: 61.34±11.07% and 82.89±3.97%, PH VI: 70.14±8.60% and 86.18±7.42% of the licking time in the early and late phases. Morphine as the reference drug (positive control) reduced the duration of formalin evoked nociceptive behaviour by a maximum percentage of 92.49±4.679% in the early phase and 95.16±5.49% in the late phase of the test. The isolates (0.5-3mgml-1) exhibited a lipid peroxidation activity as per the IC50 in the following corresponding manner; PH I 0.260, PH II 0.295, PH III 1.545, PH IV 0.007, PH V 1.769 and PHVI 1.453.The n-propyl gallate (0.001-0.3mgml-1) showed a lipid peroxidation activity by an IC50 of 0.015. The relative anti-oxidative activity in the DPPH decolourization assay (DPPH scavenging activity) of isolates as defined by the IC50 produced by the isolates (0.5-3mgml-1) were correspondingly PH I—3.304; PH II—3.636; PH III—0.241; PH IV—0.880, PH V--- 4.414 and PH VI—0.163. For n-propyl gallate (0.01-0.3mgml-1) the IC50 was found to be 0.058 in the DPPH decolourization assay. The isolates showed reducing power potential by their EC50 values corresponding to PH I—5.874; PH II —5.122; PH III—5.238; PHIV—0.524, PH V 4.501 and PH VI—1.723. The reference antioxidant n-propyl gallate showed a reducing power by an EC50 of 1.723. The findings in all three antioxidant activity assays show that the isolates have potent antioxidant properties which might partly account for the anti-inflammatory and anti-nociceptive/analgesic activities. Spectral analyses resolved PH IV to be 20-Hydroxyecdysone, PH I to contain the fatty acids eicosanoic acid and ethyl stearate, PH II to be a mixture of trimethyl benzene, cymene and ethyl hexadecanoate (ethyl palmitate) and PH III to be a mixture of methyl palmitate and octadecanoic acid. The roots extract PHC, the methanolic fraction PHM and ether fraction PHE exhibited anti-inflammatory activity as well as anti-nociceptive activity. The isolates PH I, II, III, IV, V and VI showed both anti-inflammatory and anti-nociceptive activities. The isolates also showed anti-oxidant action. 20-Hydroxyecdysone was isolated for the first time from the roots of Palisota hirsuta.
- ItemChemical constituents, anti-inflammatory, anti-oxidant and antimicrobial activities of the stem bark and leaves of Ficus Exasperata (VAHL)(2012) Amponsah, Isaac KingsleyThe work presented in this thesis involves the scientific investigation of the traditional uses of the leaves and stem bark of Ficus exasperata Vahl (Moraceae) as an anti-inflammatory, analgesic and wound healing agent. It also describes the isolation and characterization of the active principles from the Ficus exasperata. The petroleum ether, chloroform, ethyl acetate and 70% ethanolic extracts of the leaves and stem bark were assessed for anti-inflammatory, antioxidant and anti-microbial activities. The anti-inflammatory activities of the extracts and isolates were investigated using the carrageenan – induced foot pad eodema model in seven – day old chicks. The extracts were given orally to the chicks at 30, 100 and 300 mg/kg body weight, 1 hour after induction of oedema with carrageenan. Diclofenac and dexamethasone were used as reference drugs and the foot volume measured by water displacement plethysmography for five hours. All extracts exhibited anti – inflammatory effect with the stem bark showing the highest activity (ED50 = 50.65± 0.012). Antioxidant properties of the extracts were investigated using five assays; total antioxidant capacity, total phenolic content, DPPH scavenging activity, reducing power and lipid peroxidation activity. The most active antioxidant extract was the stem bark with IC50 values of 42.27±0.012, 20.09±0.001 and 61.80±0.001 µg/ml for the lipid peroxidation, DPPH scavenging and reducing power assays. The respective values for the standard antioxidant compound n-propyl gallate were 73.54±0.014, 10.8±0.002 and 66.88±0.002 µg/ml. Antimicrobial evaluation of extracts at concentrations of 10 mg/ ml was done using the agar well diffusion and micro-dilution assays. Seven organisms; P. aeruginosa, S. typhi, K. pneumoniae, E. coli, B. subtlilis, S. aureus and C. albicans were used. The chloroform extract of the stem bark was the most active with MIC of 1000 µg/ml against P. aeruginosa and S. aureus. Bergapten, oxypeucedanin hydrate and sitosterol-3-O-β-D-glucopyranoside were isolated from the bioactive chloroform extract of the stem bark whereas β-sitosterol and sitosterol-3-O-β-D-glucopyranoside were isolated from the pet-ether and ethyl acetate extracts of the leaves. To the best of our knowledge, this is the first report of the isolation of these compounds in Ficus exasperata. They exhibited dose-dependent anti-inflammatory activities with ED50 values of 101.6 ± 0.003, 126.4 ± 0.011, 275.9 ± 0.012 and 123.4 ± 0.033 mg/kg body weight for bergapten, oxypeucedanin hydrate, sitosterol-3-O-β-D-glucopyranoside and β-sitosterol respectively. They also showed significant DPPH scavenging effect with IC50 values of 63.38 ± 0.010, 46.63 ± 0.011, 220.3 ± 0.031 and >1000 µg/ml for the respective compounds. In the antimicrobial assay, β-sitosterol and its glucoside were inactive against all the organisms. Bergapten and oxypeucedanin hydrate gave MIC’s >1000 µg/ml against all susceptible organisms. They were the most active anti-inflammatory, anti-oxidant and antimicrobial compounds. The results of these studies have demonstrated that extracts of the leaves and stem bark of F. exasperata possess anti-inflammatory activity and also display antioxidant and antimicrobial activities. These findings provide scientific justification for the use of the stem bark and leaves of F. exasperata Vahl, in various traditional medicines, for the treatment of inflammatory and infectious conditions.
- ItemThe Chemical Pathology of Leiomyoma(2009-10-11) Dapilah, TheophilusUterine leiomyomas or fibroids are a major public health problem among women, occurring more frequently in women of reproductive age and are associated with diverse symptoms. In spite of the frequency with which fibroids occur, their biology is poorly understood. Studies indicate that oestrogen and progesterone play a role in the regulation of tumour growth, and there is increasing evidence that this response may be mediated via a number of growth factors. The literature regarding predisposing risk factors for development of myomas in Ghanaian women is very limited by the paucity of studies available. This is against the background that Ghana is thought to have a high prevalence of fibroids due to its indigenous black population, since uterine fibroids are more prevalent in black women. Almost all the studies done so far have been on women in the U.S.A or some other developed country where the environmental factors that are thought to influence the development of fibroids are very different from what pertains in the under developed world, particularly, Africa. Against this background, there is the need to investigate the risk factors that are believed to influence the development of fibroids in Ghanaian women. Therefore, the chemical pathology of fibroids was studied to bridge the gap in information on fibroids between the developed and the under developed world. The specific objectives of this study were to: 1) establish the demographic characteristics of women with fibroids in Ghana. 2) assess the association between fibroid development and growth, and the gynaecologic history of patients. 3) assess the relationship between the life styles of patients and the risk of developing fibroids among Ghanaian women. 4) determine the haematological profile of Ghanaian women with fibroids in relation to the tumour growth and development. 5) assess the impact of fibroids on the renal and liver functions of Ghanaian women with fibroids. 6) assess the association of oxidative stress with the development and growth of fibroids among Ghanaian women. A consecutive study of 200 women with fibroids between the ages of 20 to 40 was done in which questionnaires were designed to elicit information on their socioeconomic background and gynaecologic history. Anthropometric features were also taken and their blood samples analyzed for oxidative stress markers, biochemical, and haematological profiles. Women with obvious hormonal imbalances and chronic or malignant diseases were excluded. Control subjects recruited had similar age distribution as the patients and had been examined to exclude fibroids. Significant difference exist between the patients and the controls in terms of socio-economic characteristics most especially education and income earnings. Abortion, nulliparity, early onset of menarche and history of sexually transmitted diseases were observed to be strongly associated positively with the risk of fibroid development. BMI and waist-to-hip ratios of the patients were significantly higher than those of the controls. The results of the liver and renal function tests of patients were not significantly different from those of the controls and generally showed that both patients and controls had normal liver and renal functions. The patients had higher serum malondialdehyde and lower vitamin C levels compared to the controls. It was observed that most red blood cell indices were higher in the patients compared to the controls. However all other haematological parameters of the patients were not significantly different from those of the controls. Though generally, this study’s findings were similar to what has been observed by previous studies, the observation that higher incidence of abortions, PIDs, and STIs in patients may interplay through mediating hormonal and probably growth factors leading to the development of fibroids is quite novel.
- ItemClinical evaluation and monograph development for a Ghanaian polyherbal product (Eaf-2011) used in the management of superficial mycoses(2015-07-11) Kwesi Prah, ThomfordHerbal medicines are the most accessible form of healthcare product for majority of the world’s population and have been used over time to address the health needs of several societies. In the present study, a Ghanaian polyherbal product (ointment) from the Centre for Plant Medicine Research comprising: Alchornea cordifolia, Eugenia caryophyllata, Psidium guajava, Zanthoxylum zanthoxyloides and Tridax procumbens, coded EAF-2011 and used in the management of superficial fungal skin diseases was assessed for its quality, safety and effectiveness. Qualitative chemical fingerprinting of the ointment indicated the presence of phytochemicals including alkaloids, phenols, triterpenes and flavonoids. Thin layer chromatograms also produced three marker spots whose properties make them suitable for use as analytical markers. Quantitative chemical assay of three flavonoid compounds in the product EAF-2011 using High Performance Liquid Chromatography (HPLC) showed the presence of 8.6810% (w/w) of rutin (RU), 0.2670% (w/w) of quercetin (QE) and 0.0610% (w/w) of kaempferol (KA). A twelve (12) month stability study that assessed the product for its pharmaceutical quality using organoleptic and physicochemical tests, Thin Layer Chromatography (TLC), HPLC and an antimicrobial assay during the storage of the product under ambient conditions revealed marginal changes in chemical constituents with one of the spots obtained on thin layer analysis undergoing a colour change to purple compared to the baseline colour of brown. The concentration of quercetin was also undetectable after the 6th month of assay during the HPLC analysis. This change in chemistry was however considered insignificant as the biological activity of the product remained unaffected over the period of study based on the results of the antimicrobial assay. An in-vivo chronic toxicity and skin sensitisation test using male Sprague-Dawley rats showed that three concentrations of the herbal extracts [i.e. 2% (w/w), 5% (w/w) and 10% (w/w)] had no adverse effect on the haematological, biochemical and urine analytical iv parameters of the animals used. The ointment did not induce any histological changes in skin, liver, kidney and spleen of the animals used. In the clinical study involving 84 participants diagnosed with superficial mycoses, the 10% (w/w) concentration of the herbal product was most efficacious with 91.3% of participants randomised to the group achieving the primary outcome of complete cure compared to 30.0% achieved with the standard treatment of Whitfield ointment after 3 months. The efficacy of two (2) other concentrations of the herbal product tested [2% (w/w) and 5% (w/w)] was also comparable to Whitfield ointment. The products tested were also safe for human use as haematological, biochemical and urine biochemistry parameters were normal at the end of the study for all the treatment groups. Re-evaluation of the component raw materials of the product using a combination and interactive study to establish their contribution to the overall activity of the product showed that Eugenia caryophyllata, Alchornea cordifolia and Zanthoxylum zanthoxyloides had better activity individually than the total crude extract of five plants used in the formulation of the original product. The combination of Eugenia caryophyllata 60% (w/w) and Alchornea cordifolia 40% (w/w) was selected after further screening and analysis using the fractional inhibitory concentration (FIC) and an isobolographic analysis. A new product (RF-2013) formulated using this combination as the recipe at a concentration of 5% (w/w) was clinically evaluated against superficial fungal skin infections in another human trial. This product was subjected to a randomised controlled single blind study in 15 participants with the 10% (w/w) EAF-2011 as the control treatment. Primary outcome was achieved by all participants receiving the control treatment compared to the 60% attained in the 5% (w/w) RF-2013. Based on the number of participants and their time taken to achieve the primary outcome, the 10% (w/w) EAF-2011 which is the original formulation was proposed as the preferred treatment in the management of superficial fungal skin infections.
- ItemClinical, Metabolic and Immunological Characteristics of Ghanaian Patients with Diabetes Mellitus(Febuary, 2009) Titty, Felix-Val KwakuObjectives: This study investigated the clinical characteristics of Ghanaian diabetic patients; prevalence of metabolic syndrome and its components in Ghanaian diabetic patients; and determinants of the metabolic syndrome diagnosed from Ghanaian diabetic patients. Further, association of metabolic syndrome with poor glycaemic control and occurrence and features of autoimmune diabetes and autoantibody-negative type 2 diabetes in Ghanaian diabetic patients were investigated. Research design and methods: This research was a prospective study covering a period of four years, from August 2004 to June 2008. The study was carried out at the Komfo Anokye Teaching Hospital and the Kwame Nkrumah University of Science and Technology, both in Kumasi on two study populations. For the first population, Ghanaian diabetic patients diagnosed by the WHO criteria were consecutively selected. The sample size was 456. Controls included 120 age- and sex-matched nondiabetic controls. For the second population, recently diagnosed (<1 year) Ghanaian diabetic patients were consecutively selected. The sample size was 120. Controls included 60 age- and sex-matched healthy nondiabetic controls. Socio-demographic and clinical characteristics of subjects were investigated using a standardized questionnaire. Blood pressure and anthropometric measurements (height, weight, waist circumference) of subjects were measured using mercury sphygmomanometer, physician standiometer and scale and a plastic tape respectively. Blood and serum samples from subjects were analysed for relevant biochemical indices using enzymatic methods and ATAC® 8000 Random Access Chemistry Analyzer and its reagent kits. Metabolic syndrome was diagnosed using the National Cholesterol Education Programme Adult Treatment Panel III (NCEP ATP III) criteria. HbA1C was analysed using an inhibition of latex agglutination test, DCA® 2000+ analyzer (Bayer model, USA) and its reagent kits. Insulin and autoantibodies were tested using an Enzyme linked immunosorbent assay (ELISA) technique, DRG International Inc. USA EIA reagent kits, ELISA reader (Tipo model, Italy) and washer (Murex, Great Britain). iii Results: The sex distribution of the Ghanaian diabetic patients of the first study population was 30.9% males and 69.1% females. Insulin-requiring diabetics were 24.6% and non-insulin requiring diabetics 75.4%. The mean age was 55.8 ± 12.3 years; 90.6% were ≥40 years of age. The mean age of onset was 49.7 ± 12.5 years; 89.5% had an age of onset ≥35 years or late onset diabetes. The mean diabetes duration was 6.0 ± 5.4 years and 60.5% had diabetes duration 1 – 9 years. Mean preprandial (fasting) glucose level was 9.4 ± 4.5 mmol/L with 62.1% having high preprandial glucose >7.2 mmol/L. The mean BMI was 25.1 ± 4.8 kg/m2; 44.5% were overweight and obese or had a BMI ≥25.0 kg/m2. The mean waist circumference was 87.0 ± 13.7 cm; 43.6% had central obesity. Diabetics with confirmed hypertension were 40.1%; hypertensives with inadequate blood pressure control ≥130/80 mmHg were 91.3%. The prevalence of the metabolic syndrome was 55.9% in the first population; prevalence in females (66.0%) was higher than males (33.3%). Low HDL cholesterol was the commonest component (47.4%) of the metabolic syndrome in the first population, followed by hypertension (46.9%). In females central obesity (57.1%) was the commonest component, followed by low HDL cholesterol (53.0%); in males, hypertension (39.7%) was the commonest component, followed by hypertriglyceridaemia (36.2%). Central obesity, hypertension and low HDL cholesterol prevalence was higher in females than males, while hypertriglyceridaemia prevalence was comparable in females and males. Female diabetics individually carried more metabolic syndrome factors than males. The major determinant of the metabolic syndrome diagnosed from Ghanaian diabetics was central obesity (69.4%) followed by hypertension (67.5%). In females the major determinant was central obesity (76.9%), followed by hypertension (67.3%), while in males the major determinant was hypertriglyceridaemia (74.5%), followed by hypertension (68.1%). The most frequent combination of different components was hyperglycaemia, central obesity and hypertension (46.3%). For the second population, the prevalence of the metabolic syndrome in patients with poor glycaemic control was 50.0% and patients with good glycaemic control 33.3%. The prevalence of autoimmunty in the insulin-requiring recently diagnosed diabetic patients was 35.3% and in the non-insulin requiring patients 16.5%. The prevalence of LADA in the non-insulin requiring diabetic patients was iv 13.5%. The prevalence of LADA and autoimmune type 1 diabetes in the second population were 11.7% and 7.5% respectively, giving total autoimmune diabetes prevalence of 19.2%; single autoantibody positivity was 77.3% and multiple autoantibody positivity 22.7% in the autoimmune diabetic patients. Most of the clinical and metabolic parameters of autoimmune diabetes and type 2 diabetes did not differ. The exceptions were hypertension and central obesity which were more likely in type 2 diabetes than autoimmune diabetes and HbA1C which was higher in autoimmune diabetes than type 2 diabetes. Conclusion: There were more Ghanaian females with diabetes mellitus than males and more non-insulin requiring than insulin requiring patients. Majority of the diabetic patients were 40 years and above and had late onset diabetes. More than half had diabetes disease duration of 1-9 years and carry clinical modifiable risk factors for microvascular and macrovascular disease. Less than half of the subjects had clinical modifiable risk factors for cardiovascular disease. The metabolic syndrome was frequent in Ghanaian diabetic patients, especially females, and was present at an prevalence slightly less than that in developed countries. Future prevention and control strategies should not overlook the importance of metabolic disease risk factors in Ghana. Interventions that address obesity and hypertension in females and hypertriglyceridaemia and hypertension in males and reduce waist circumference, blood pressure and hypertriglyceridaemia, may reduce the prevalence of the metabolic syndrome, and hence cardiovascular disease in Ghanaian diabetic patients. The major determinants of the metabolic syndrome were not necessarily the commonest metabolic syndrome components of the population under consideration. The metabolic syndrome was associated with poor glycaemic control. However, metabolic syndrome and poor glycaemic control are independent risk factors for cardiovascular disease. Autoimmune diabetes, including autoimmune type 1 diabetes and LADA, occurs in recently diagnosed Ghanaian diabetic patients. Both ICA and GAD autoantibody tests are required to identify autoimmune diabetes, and distinguish it from autoantibody-negative type 2 diabetes. Clinical and metabolic markers cannot be used for this purpose.
- ItemComparative 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/0000-0002-4990-7725The 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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