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Unsweetened natural cocoa (UNCP) was evaluated for reproductive toxicity in rats. A preliminary genotoxic potential was evaluated by the DNA comet assay test using C57Bl/6 mice. Both therapeutic dose (TD; 900 mg/kg) and high dose (HD; 9000 mg/kg) of UNCP were used. White Wistar rats were used in two experimental groups. The females received UNCP 15 days before crossing with untreated males. The males received UNCP for 48 days before mating with untreated females. Subacute toxicity was observed during a 14-day oral administration of UNCP. Results show that a high tail DNA% was observed with methyl mesylate administration in all tissues analysed. The lowest tail DNA% value was observed in the liver (1.64 ± 0.26) and kidney (1.63 ± 0.30) during UNCP (TD) administration. UNCP did not induce observable physical congenital malformations on the pubs of treated female and male rats, lacks genotoxic potential, and did not adversely affect pregnancy index, pub weights, and survival index, but UNCP exhibited proimplantation potential ( p > 0.05 ).

Ethnopharmacological relevance Some local communities in Cote d'Ivoire use the mushroom Termitomyces schimperi combined with kaolin (TSK) to manage various cancers in patients. However, there is a paucity of data on toxicity, mutagenicity and trace metal constituent of TSK. Aim of the study We sought to investigate the acute and sub-chronic toxicities, mutagenic potential, and trace metal constituents of TSK. Materials and methods To assess acute toxicity, single doses (1000, 3000 and 5000 mg/kg) of aqueous extract of TSK were administrated per os to Sprague Dawley (SD) rats on Day 1. The rats were then monitored for 13 consecutive days. Sub-chronic toxicity was evaluated by daily administration of 200 and 500 mg/kg of the extract per os for 90 consecutive days. SD rats used as control received distilled water. Signs of toxicity, changes in body weight and mortality were monitored. After the aforementioned monitoring processes, rats were sacrificed and blood collected for full blood count and biochemistry analysis. Animal organs were also collected for histopathological examination. The mutagenic potential of the aqueous extract of TSK (10000 μg/mL) on TA98 Salmonella typhimurium was estimated. Additionally, energy-dispersive X-ray fluorescence (ED-XRF) method was employed to determine trace metal constituents of TSK. Results Single-dose administration of 5000 mg/kg of TSK did not cause any death in the SD rats; thus, LD50 was above 5000 mg/kg. Administration of 1000 and 3000 mg/kg of the aqueous extract of TSK did not cause any significant change in behaviour and body weight of SD rats during the 14-day monitoring period. However, the mean corpuscular volume and the mean corpuscular haemoglobin concentration increased significantly (p = 0.0122) in serum creatine level among rats administered 1000 and 3000 mg/kg of TSK extract compared with control. After 14 days, there were minimal changes with isolated organs of TSK-treated and control rats. Furthermore, 90-day treatment with extract of TSK caused no significant change in parameters assessed. TSK induced frameshift gene mutation in S. typhimurium before (p Conclusions The aqueous extract of TSK showed no toxicity (acute and sub-chronic) at doses tested. These findings are consistent with the absence of heavy metals (i.e., cadmium) and potentially toxic elements (i.e., uranium) in TSK samples analysed. TSK showed some level of mutagenic potential. Further mutagenic and chronic toxicity studies on TSK are required.

Background. There is considerable evidence that many patients concurrently administer dietary supplements with conventional drugs, creating a risk for potential drug-supplement interaction. The aim of this study was to determine the effect of Cellgevity® supplement on selected rat liver cytochrome P450 (CYP) enzymes. Also, based on our previous finding, we sought to determine the effect of Cellgevity® on the pharmacokinetics of carbamazepine, a CYP3A4 substrate. Methods. Male Sprague–Dawley (SD) rats were randomly put into 5 groups and administered either distilled water (negative control), Cellgevity® (3 separate doses), or phenobarbital (positive control), per os. Modulation of liver CYP enzyme activity was evaluated after 30 days of treatment, using probe substrates, spectroscopic, and high-performance liquid chromatographic methods. In the pharmacokinetic study, 12 SD rats were put into 2 groups and administered carbamazepine plus normal saline (group 1) or carbamazepine plus Cellgevity® (group 2), per os, both over a period of 14 days. Blood samples from rats in the same group were collected after treatment. Serum samples were prepared and pooled together at each specific sampling time point. Levels of carbamazepine were determined using a fluorescence polarization immunoassay. Results. Activities of rat liver CYP1A1/2, CYP2C9, and CYP2D6 were significantly increased by Cellgevity® after 30-day treatment. Pharmacokinetic parameters for rats administered carbamazepine with Cellgevity® vis-a-vis carbamazepine with normal saline were as follows: Cmax; 20 μmol/L vs 11 μmol/L, AUC0⟶24; 347 μmol h/L vs 170 μmol h/L, Ke; 0.28 h−1 vs 0.41 h−1, and t1/2; 2.3 h vs 1.7 h, respectively. Conclusions. Cellgevity® increased the activity of rat CYP1A1/2, CYP2C9, and CYP2D6 enzymes and was found to alter the pharmacokinetics of carbamazepine in rats.

ETHNOPHARMACOLOGICAL RELEVANCE Advancement in cancer therapy has improved survival among patients. However, use of anticancer drugs like anthracyclines (e.g., doxorubicin) is not without adverse effects. Notable among adverse effects of doxorubicin (DOX) is cardiotoxicity, which ranges from mild transient blood pressure changes to potentially serious heart failure. Anecdotal reports suggest that Kalanchoe integra (KI) may have cardio-protective potential. AIMS OF THE STUDY This study sought to determine the cardio-protective potential of KI against doxorubicin-induced cardiotoxicity and also examined any possible genotoxic potential of KI in selected organs. Additionally, the nitric oxide modulatory potential of KI was assessed. MATERIALS AND METHODS The leaves of KI were collected, air-dried, pulverised and extracted using 70% ethanol. High-performance liquid chromatography (HPLC) fingerprinting was done for KI. Also, the single-cell gel electrophoresis assay (Comet assay) was employed to ascertain the genotoxic potential of KI. In assessment of cardio-protective potential of KI against doxorubicin-induced cardiotoxicity, a total of 42 female Sprague-Dawley rats were put into 7 groups (n = 6). Group I: vehicle control, received normal saline (1 mL/kg p.o) for 30 days. Group II: toxic control, received DOX (20 mg/kg i.p.) once on the 29th day. Group III: KI control, received KI (300 mg/kg p.o) for 30 days. Group IV: vitamin E control, received vitamin E (100 mg/kg p.o) for 30 days. Group V: KI treated-1, received KI (300 mg/kg p.o) for 30 days and DOX (20 mg/kg i.p) on the 29th day. Group VI: KI treated-2, received KI (600 mg/kg p.o) for 30 days and DOX (20 mg/kg i.p) on the 29th day. Group VII: vitamin E treated, received vitamin E (100 mg/kg p.o) for 30 days and DOX (20 mg/kg i.p) on the 29th day. Thirty-six (36) hours after last administration, rats were sacrificed. Blood samples were taken via cardiac puncture to determine levels of aspartate aminotransferase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), creatine kinase (CK), lactate dehydrogenase (LDH), enzymatic antioxidants such as glutathione (GSH), superoxide dismutase (SOD), and catalase (CAT). Nitric oxide level was also determined. Hearts of rats in each group were excised and taken through histopathological examination. RESULTS In the HPLC fingerprint analysis, 13 peaks were identified, and peak with retention time of 24.0 min had the highest peak area (3.223 x104 mAU). Comet assay showed that the KI extract was non-genotoxic. Pretreatment with KI protected rats against doxorubicin-induced cardiotoxicity as evidenced by the low levels of AST, ALT, ALP, CK and LDH compared with the controls (p

Trichilia monadelpha is a common medicinal plant used traditionally in treating central nervous system conditions such as epilepsy, depression, pain, and psychosis. In this study, the antidepressant-like effect of crude extracts of the stem bark of T. monadelpha was investigated using two classical murine models, forced swimming test (FST) and tail suspension test (TST). The extracts, petroleum ether, ethyl acetate, and hydroethanolic extracts (30–300 mg/kg, p.o.), standard drug (imipramine; fluoxetine, 3–30 mg/kg, p.o.), and saline (vehicle) were given to mice one hour prior to the acute study. In a separate experiment the components (flavonoids, saponins, alkaloids, tannins, and terpenoids; 30–300 mg/kg, p.o.) from the most efficacious extract fraction were screened to ascertain which components possessed the antidepressant effect. All the extracts and components significantly induced a decline in immobility in the FST and TST, indicative of an antidepressant-like activity. The extracts and some components showed increase in swimming and climbing in the FST as well as a significant enhancement in swinging and/or curling scores in the TST, suggesting a possible involvement of monoaminergic and/or opioidergic activity. This study reveals the antidepressant-like potential of the stem bark extracts and components of T. monadelpha.

Two samples, water extract and blended whole leaves, of freshKalanchoe integraleaves (Crassulaceae), a traditional antihypertensive medicine used in Ghana, were analyzed with Energy Dispersive X-Ray Fluorescence spectroscopy (EDXRF). Analysis revealed 12 macro and 26 micro elements in both extracts. Further quantitative assessment of the results for amounts of elements that are pharmacologically significant revealed that the amounts of calcium, potassium, and magnesium present in the extracts could be correlated to its traditional usage in managing hypertension and arrhythmias. However, heavy metals (lead and inorganic arsenic) detected in the extracts may pose a threat at doses normally used traditionally for the treatment of hypertension.

Kalanchoe intergra (Ki) leaf extract is an orally administered multipurpose plant medicine in Ghana and other parts of the world for the treatment of ulcers, pain and adenoma of the prostate gland. There is paucity of information concerning its short-term usage. The present study is aimed at conducting histopathological and biochemical studies in a 14-day sub-acute toxicity studies using female Sprague-Dawley rats.Crude extract of Ki leaves was prepared and freeze-dried. A 14-day sub-acute toxicity studies was conducted using 2 week old nulliparous and non-pregnant female Sprague-Dawley rats (120-150g). Reconstituted Ki was administered at a dosage of 900mgkg(-1) (high dose), 300mgkg(-1) with a control group receiving an equivalent volume of distilled water (as vehicle) by gastric lavage. Histopathological studies of major organs and blood chemistry analysis were performed on blood obtained via cardiac puncture into EDTA tubes after euthanisation.There was a significant decrease in urea (p0.016) and creatinine levels (p0.001) in both the high and low dose groups. There was an increase in ALP levels (P=0.01) in both the high and low dose groups. ALT and AST rather decreased significantly in both the high and low dose groups (p0.0001). Histopathological results did not show any abnormalities in all the HE stained paraffin sections. Thus the photomicrographs of the liver, kidney and heart were within histopathological limits.Ki leaf extract is non-toxic when administered by the oral route over a time period of 14 days at the above doses.

Objective: Kalanchoe integra is known to possess antihistaminic and mast cell stabilizing effect. Inflammatory mediators, such as histamine and eicosanoids have been implicated in the pathophysiology of allergen-induced asthma including bronchospasm, vasodilation, increased vascular permeability, perivascular and peribronchial oedema, acute functional changes in the lungs and diarrhea due to increased intestinal motility. This study is to Original Research Article European Journal of Medicinal Plants, 4(5): 542-562, 2014 543 ascertain the anti-inflammatory effect of Kalanchoe integra aqueous leaf extract (KILE) on bronchial hyper-responsiveness in ovalbumin-sensitized guinea-pigs. Method: Bronchial asthma was induced in guinea pigs using Ovalbumin. In vivo skin tests were carried out on all guinea pigs using ovalbumin and histamine as allergens. The diameters of wheals were recorded and the means determined. OA-sensitized guineapigs were challenged with 2% OA aerosols after 1 hour per os of drugs (KILE or prednisolonefor the treated groups) for two weeks. A piece of excised trachea was suspended in a tissue bath and challenged with histamine in the presence and absence of KILE, as well as Prednisolone (2.5 mg/kg). The results were reported as mean ±S.E.M. Statistical analysis was performed using one-way ANOVA and Bonferroni’s post hoc test. Results: Biological assaying of KILE showed significant dose-dependent reduction in histamine induced vasodilation of cutaneal blood vessels (P

Background: Kalanchoe integra is widely used in folklore medicine as an antiasthmaticagent. Previous studies have shown the ameliorating effect of Kalanchoe integra leaf extract [KILE] on bronchial hyperesponsiveness and inflammation.Further, the stabilizing effect of Kalanchoe sp on mast cell degranulation,suggests that Kalanchoe species are suitable candidates for allergicasthma therapy. This study is designed to investigate the anti-asthmaticpotential of KILE and monitor the accompanying histopathological andimmunobiochemical changes that occur in an animal model of bronchial asthmausing ovalbumin sensitized guinea pigs. Method: Thirty male guinea pigs weredivided into five groups of six animals each. Bronchial asthma was simulated inguinea pigs using ovalbumin. Both low dose (300 mg/kg) and high dose extract(900 mg/kg) were administered daily for 42 days. Prednisolone (2.5 mg/kg) wasthe standard drug used. Results: Guinea pigs in all KILE treated groups maintained the integrity of their airwaystructures: bronchial folds and walls, alveoli, alveolar ductsand sacs. KILE and prednisolone caused a reduction in immune parameters (p 0.001), extent of bronchoconstriction, bronchialwall thickness and goblet cell accumulation in the sensitized guineapigs. Conclusion: This study demonstrates the anti-asthmatic potential of KILE duringprolonged administration by the oral route.

Croton membranaceus Mull.Arg. (Euphorbiaceae) is used for benign prostate hyperplasia (BPH) treatment. The study aimed at investigating organs that the aqueous root extracts of C. membranaceus (CMARE) target, which is absent in literature. Twenty-four male Sprague-Dawley rats (100-140 g) were randomly divided into 4 groups. Group 1, the control group received distilled water. Groups 2, 3 and 4 received 30, 150 and 300 mg kg(-1) b.wt CMARE respectively (oral gavage). Rats fed 90 days the standard chow diet ad libitum. Upon sacrifice, major organs were histologically examined and serum prostate-specific antigen (PSA) biochemically determined. Only the prostate was abnormal. Histologically, H&E staining revealed thickness and infoldings of the epithelial cells shrinking with increasing dose. The 30 mg kg(-1) group showed low columnar or flattened epithelium cells, whereas the columnar epithelium infoldings of the 150 mg kg(-1) b.wt and 300 mg kg(-1) b.wt groups were virtually nonexistent. The acini of the control, 30 mg kg(-1) b.wt group and the 150 mg kg(-1) b.wt groups showed clear pinkish secretion. However, secretion of the high-dose group appeared light pink in colour and the stroma cells appeared much darker than all the treated and control group. C. membranaceus targets the prostate with significant PSA reduction (P < 0.01).

Background : Conflicting reports about the toxicity of Synedrella nodiflora (L) Gaertn (family Asteraceae), a plant traditionally used in Ghana for the management of epilepsy, abound in literature. The present study evaluates the effect of a 90-day continuous oral administration of a hydro-ethanolic whole plant extract of Synedrella nodiflora (SNE) in male Sprague-Dawley rats. Methods : The toxicological evaluation of the extract (100, 300 and 1000 mgkg-1) was focused on haematological, serum biochemical parameters and histopathological changes of some isolated organs. Results : The extract produced no mortality in the rats treated during the study period. Only SNE 100 mgkg-1 produced significant decrease in white blood cell and neutrophil counts and an increase in albumin, globulin, total bilirubin, total protein and potassium levels. The higher doses (SNE 300 and 1000 mgkg-1) had no significant effect on all the haematological and biochemical parameters measured. Histopathological assessment of the liver, kidney and heart revealed no abnormalities in rats treated with the extracts. Only the SNE 1000 mgkg-1 produced distortions of the branching arrangements of the myocardial fibres and a congested vessel which indicates a healed infarction. Conclusions : The findings suggest hydro-ethanolic extract of Synedrella nodiflora (L) Gaertn generally has a low toxicity profile following a 90-day continuous oral administration in male Sprague-Dawley rats under the present laboratory conditions. However patients with renal or cardiac problems should use the plant with caution. Funding : Jointly supported by the International Foundation for Science, Stockholm, Sweden, through a grant (# F/5191-1) to Dr. Patrick Amoateng and the Office of Research, Innovation and Development (ORID), University of Ghana, Accra, Ghana, grant awarded to Dr. Patrick Amoateng (reference number: URF/6/ILG-002/2012-2013) Keywords : : Synedrella nodiflora, Sprague-Dawley rats, histopathological, haematological

Synedrella nodiflora (L) Gaertn (family Asteraceae), a common weed in Ghana, is traditionally used for the management of epilepsy, hiccup and threatened abortion. To further promote the ethno-pharmacological uses of the plant, an acute toxicity of a hydro-ethanolic whole plant extract was assessed in male Sprague-Dawley rats. The lethal dose (LD 50 ) and effects of a single oral admin-istration of the extract (1600, 3200 and 6400 mg kg -1 ) on haematological and serum biochemical parameters were measured. The extract produced no mortality in the rats treated during a 48-hour examination and after a subsequent 12-day assessment. Thus the LD 50 was indicated as being greater than 6400 mg kg -1 . The extract also did not significantly affect any of the haematological and serum biochemical indices. This result suggests that acute oral administration of the hydro-ethanolic extract of Synedrella nodiflora is virtually non-toxic in male Sprague-Dawley rats under normal laboratory conditions. Keywords: Synedrella nodiflora, acute toxicity, Sprague-Dawley rats, haematology, biochemical analysis

Bronchospasm is an important characteristic of allergen-induced airway inflammation.Taraxacum officinale herb is believed to have several therapeutic effects including anti-inflammatory properties. The anticholinergic effect of the T. officinale leaves extract (TOLE) was determined in ovalbumin (OA)-sensitized guinea-pig trachea. The test drugs (TOLE or prednisolone) were administered orally 1 h prior to aerosolized 2% ovalbumin challenge. Isolated ovalbumin-sensitized guinea-pig tracheal was challenged with acetylcholine and OA in the absence and presence of TOLE. The blood samples were collected and neutrophils, lymphocytes and monocytes (eosinophils, basophils and macrophages) counts assayed. Prednisolone (2.5 mg/kg body weight (BW) orally) was used as reference standard. TOLE application showed significant antagonistic effect on contraction of trachea to both acetylcholine (35.10 ± 0.04) and OA (18.6 ± 1.5). Also, the extract reduced monocytes (0.62 ± 0.23), lymphocytes (3.4 ± 0.59) and neutrophils (3.65 ± 0.20) counts in OA-sensitized guinea-pigs. Thus, TOLE possesses anticholinergic and reduces neutrophil, eosinophil and basophil counts in ovalbumin-sensitized guinea-pigs. Key words: Taraxacum officinale, inflammation, blood cell counts, anticholinergic.

Objectives: This study presents the antispasmodic and antibacterial properties of an ethanol extract and fractions the of stem bark of Piliostigma reticulatum. Materials and Methods: The antispasmodic effects of the extract and its fractions were performed on isolated rabbit duodenum. The antibacterial properties were determined as minimal inhibitory and bactericidal concentration of the extract and fractions of P. reticulatum on susceptible and resistant strains of Escherichia coli, Vibrio cholerae, Staphylococcus aureus, Shigella dysenteriae and Salmonella tiphymurium. Results: The ethanol extract of P. reticulatum and fractions (except for heptane) produced concentration-dependent relaxant effects on isolated duodenum preparations. The IC 50 of the extract and dichloromethane, ethyl acetate, butanol and aqueous fractions are 0.88452, 0.2453, 0.2909, 0.3946 and 0.3231 mg/ml respectively. The extract was found to significantly antagonize acetylcholine-induced contraction. The susceptible strains E. coli and V. cholerae were the most inhibited by the dichloromethane fraction at 60 mg/mL, as shown by their diameter of inhibition of 13.2 ± 0.76 and 13.3 ± 0.67 mm respectively. Conversely, the dichloromethane fraction, the most active antibacterial fraction, did not inhibit the resistant strains S. dysenteriae and S. tiphymurium. Conclusion: The results showed that P. reticulatum stem bark possesses spasmolytic and antibacterial properties and this may contribute to its traditional medicinal use for the treatment of diarrhea.

Unsweetened natural cocoa powder is enriched with nutraceutical abundance of anti-asthmatic compounds theobromine and theophylline. Cocoa powder, which is prepared after removal of the cocoa butter, contains about 1.9% theobromine and 0.21% caffeine. Anecdotal reports indicate that regular consumption of unsweetened natural cocoa powder (UNCP), a common practice in Ghana, West Africa, has the potential to reduce the tendency of asthmatic episodes. In the present paper we studied the effect of regular ingestion of aqueous extract of UNCP on hematological and histopathological changes that occur in ovalbumin (OVA)-sensitized guinea pigs. OVA-sensitized guinea pigs were challenged with aerosolized OVA 1 hour after ingestion of 300 mg/kg (low dose) or 600 mg/kg (high dose) of UNCP for 35 consecutive days. Histopathological and haematological changes in the OVA-sensitized guinea pigs were evaluated. Both negative and positive controls with distilled water and prednisolone, respectively, were used. OVA-sensitized guinea pigs demonstrated concentration-independent reduction in immune response to aerosolized OVA. There were no histo-architectural changes in the bronchiolar smooth muscles of the treated groups. Unsweetened natural cocoa powder has potential anti-asthmatic properties when administered orally at the doses tested.