Antioxidant Activity In Vitro and Hepatoprotective Effect of Phlomis Maximowiczii In Vivo

Background

A number of medicinal plants and there compounds played a major role in the treatment of hepatic disorders. They were widely used for the treatment of these disorders, and oxidant stress injury was one of the liver injury mechanisms. The present study evaluated the antioxidant activity and the hepatoprotective effect of each extracts of Phlomis maximowiczii.

Materials and Methods

The antioxidant activity was assayed by the methods of ABTS, FRAP and DPPH in vitro. Hepatoprotective effect of P. maximowiczii extracts was examined using carbon tetrachloride-induced acute liver injury in mice.

Results

P. maximowiczii n-butanol (PMBU) extract, ABTS (IC₅₀=18.96 µg/mL), DPPH (IC₅₀=25.15 µg/mL), and FRAP (RACT₅₀=2775.6±144.18 µmol/g), showed higher scavenging capacity than that of P. maximowiczii ethyl acetate (PMEA). The n-butanol extract could significantly reduce the level of GPT, GOT and MDA (P<0.05, P<0.001 and P<0.001, respectively) and increase the level of SOD (P<0.001), respectively.

Conclusion

The antioxidant activity of n-butanol extract in vitro was related with the level of MDA and SOD in vivo, and hepatoprotective effect of n-butanol extract also had relationship with its antioxidant activity in vivo.     

In vitro susceptibility of Plasmodium falciparum to monodesethylamodiaquine, quinine, mefloquine and halofantrine in Abidjan (Côte d'Ivoire)

Background

Malaria is the primary cause of hospitalization in Côte d''Ivoire. Early treatment is one of the strategies to control this illness. However, the spread of resistance of Plasmodium falciparum to antimalarial drugs can seriously compromise this strategy.

Objectives

The aim of this study was to assess the in vitro susceptibility of P. falciparum to monodesethylamodiaquine and aminoalcohols in Abidjan (Côte d''Ivoire).

Methods

We assessed the in vitro susceptibility of isolates collected from patients with uncomplicated malaria by using the WHO optical microtest technique.

Results

The proportions of resistance to monodesethylamodiaquine, méfloquine and halofantrine were 12.5%, 15.6% and 25.9%, respectively. For quinine, none of isolates showed evidence of in vitro resistance. However, two isolates (6.1%) had IC₅₀ values above 300 nM. The IC₅₀ of each drug was positively and significantly correlated to that of the other three drugs, and the correlation was higher between halofantrine and mefloquine.

Conclusions

Our results showed that the in vitro chloroquine resistance reported in previous studies has been extended to other antimalarial drugs investigated in this study except for quinine. Therefore, it is necessary to implement a long-term monitoring system of antimalarial drug resistance.     

Antioxidant Activities In Vitro and Hepatoprotective Effects of Nelumbo Nucifera Leaves In Vivo

Background

Herbal medicines played a major role in the treatment of hepatic disorders, and a number of medicinal plants and their compounds were widely used for the treatment of these disorders, and oxidant stress injury was one of the mechanism of liver injury.

Materials and Methods

Antioxidant activity of Nelumbo nucifera leaves (NU) extracts was assayed by the methods of scavenging 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2′-azino-bis (3-ethylbenzo-thiazoline-6-sulfonicacid) (ABTS) radical and ferric reducing antioxidant power (FRAP) in vitro. By intraperitoneal injection carbon tetrachloride (CCl₄) to establish acute liver injury model in mice, the levels of Glutamic-pyruvic transaminase (GPT), glutamic-oxaloacetic transaminase (GOT), superoxide dismutase (SOD) and the content of and maleicdialdehyde (MDA) were detected to evaluate hepatoprotective effect of NU using corresponding test kit.

Results

EtOAC (NUEA) and n-BuOH extracts (NUBU) of N. nucifera leaves had good scavenging DPPH and ABTS radical activity and ferric reducing antioxidant power in vitro. DPPH radical scavenging activity and ferric reducing antioxidant power of NUEA (IC₅₀= 6.68±0.29 µg/mL, RACT₅₀=1749.82±67.03 µmol/g) and NUBU (IC₅₀= 4.61±0.01 µg/mL, RACT₅₀=1995.27±135.71 µmol/g ) were higher than that of BHT (IC₅₀=8.76±0.20 µg/mL, RACT₅₀=1581.68±97.41 µmol/g) and Dangfeiliganning (IC₅₀=28.06±0.17 µg/mL, RACT₅₀=1028.55±3.28 µmol/g). ABTS radical scavenging activity of NUEA (IC₅₀= 5.32±0.12 µg/mL) and NUBU (IC₅₀= 8.16±0.27 µg/mL) were higher than that of Dangfeiliganning (IC₅₀= 9.76±0.16 µg/mL). Thus, hepatoprotective effect of NUEA and NUBU was evaluated on CCl₄-induced acute liver injury mice. The results showed that the levels of GOT and GPT in each treatment group significantly decreased (p<0.001 and p<0.01, p<0.05, respectively) except for the group of NUEA (130.8 mg/kg) (p>0.05). The contents of malondialdehyde (MDA) in liver in groups of NUEA (523 mg/kg), NUBU (840.5 and 420.5 mg/kg, repectively) had significant decrease (p<0.001 and p<0.05, respectively), and the level of SOD in liver for each treatment group could significantly decrease (p<0.001, p<0.05, respectively).

Conclusion

NUEA and NUBU had significantly hepatoprotective effect for Calcium tetrachloride CCl₄-induced liver injury, which might be attributable to its antioxidant activity.     

Experimental Study on the Optimization of Extraction Process of Garlic Oil and its Antibacterial Effects

Background

Garlic oil which is the main active constituent of garlic has a wide range of pharmacological activities, and a broad antibacterial spectrum. It also has a strong anti-cancer activity, and can significantly inhibit a variety of tumors such as liver cancer, gastric cancer and colon cancer. The objective is to study the extraction process of garlic oil and its antibacterial effects.

Materials and Methods

CO₂ Supercritical extraction was used to investigate the optimal processing conditions for garlic oil extraction; filter paper test and suspension dilution test were applied to determine the bacteriostatic action of garlic oil.

Results

In the CO₂ supercritical extraction experiment, factors influencing the yield of garlic oil were: extraction pressure > extraction temperature > extraction time in descending order. Range analysis showed that the optimal experimental conditions for CO₂ supercritical extraction of garlic oil were extraction pressure of 15 Mpa, temperature of 40 °C, and duration of 1 h. Different concentrations of garlic oil could all inhibit the growth of Staphylococcus aureus, Escherichia coli and Bacillus subtilis, suggesting that garlic oil has an antibacterial effect.

Conclusion

The optimal experimental conditions for CO₂ supercritical extraction of garlic oil were: extraction pressure of 15 Mpa, temperature of 40 °C, and duration of 1 h; garlic oil has an antibacterial effect.     

The Antibacterial Activity of Traditionally Used Salvadora Persica L. (Miswak) and Commiphora Gileadensis (Palsam) in Saudi Arabia

Background

Nowadays there is a need to find naturally occurring substances from plants with antimicrobial activity as an alternative to available used antibiotics.

Materials and Methods

Salvadora persica (miswak) and Commiphora gileadensis were collected, dried and extracted with either methanol or warm water and the obtained extracts were assessed for their antibacterial activity against 5 different genera of bacteria using agar well diffusion method. The tested bacteria included some human pathogens.

Results

The obtained extracts exhibited considerable inhibitory effects against all the tested bacteria with various degrees of growth inhibition. It was shown that methanol extract was more effective compared to water extracts. The minimum inhibitory concentrations (MIC) of the methanol extracts ranged from 50–100 °g/ml. No toxicity was found using Artimia salina as test organism and no antitumor activity against Ehrlich ascites carcinoma.

Conclusion

S. persica and C. gileadensis showed moderate to high inhibitory activity on pathogenic bacteria with no toxicity and can be used traditionally as alternative medicine     

Pigmentation and Dermal Conservative Effects of the Astonishing Algae Sargassum Polycystum and Padina Tenuis on Guinea Pigs,Human Epidermal Melanocytes (HEM) and Chang Cells

Background

The preference for a fairer skin-tone has become a common trend among both men and women around the world. In this study, seaweeds Sargassum polycystum and Padina tenuis were investigated for their in vitro and in vivo potentials in working as skin whitening agents. Seaweed has been used as a revolutionary skin repairing agent in both traditional and modern preparations. The high antioxidant content is one of the prime reasons for its potent action. It has been employed in traditional Chinese and Japanese medicine. For centuries, most medical practitioners in the Asian cultures have known seaweed as an organic source of vitamins, minerals, fatty acids like omega-3 and omega-6 and antioxidants. The present objective of the study was to evaluate the potent dermal protective effect of the two seaweeds Sargassum polycystum and Padina tenuis on human cell lines and guinea pigs.

Material and Methods

Seaweeds were extracted with ethanol and further fractionated with hexane, ethyl acetate and water. The extracts were tested for mushroom tyrosinase inhibitory activity, cytotoxicity in human epidermal melanocyte (HEM), and Chang cells. Extracts with potent melanocytotoxicity were formulated into cosmetic cream and tested on guinea pigs in dermal irritation tests and de-pigmentation assessments.

Results

Both Sargassum polycystum and Padina tenuis seaweeds showed significant inhibitory effect on mushroom tyrosinase in the concentration tested. SPEt showed most potent cytotoxicity on HEM (IC₅₀ of 36µg/ml), followed by SPHF (65µg/ml), and PTHF (78.5µg/ml). SPHF and SPEt reduced melanin content in skin of guinea pigs when assessed histologically.

Conclusion

SPEt, SPHF and PTHF were able to inhibit HEM proliferation in vitro, with SPHF being most potent and did not cause any dermal irritation in guinea pigs. The results obtained indicate that SPHF is a promising pharmacological or cosmetic agent.     

Evaluation of Antioxidant and Antimicrobial Potential of Two Endangered Plant Species Atropa Belladonna and Matricaria Chamomilla

Background

Plants are the natural source of antioxidants as well as antimicrobial compounds that has great potentials in pharmaceutical industry. In the present study, two medicinal plants Atropa belladonna and Matricaria chamomilla were collected from Northern areas of Pakistan.

Materials and Methods

The extracts of the collected plants were obtained by microwave assisted extraction (MAE) with changing parameters, power level and time; methanol and ethanol were solvents used during extraction. The extracts of plants were tested against different bacterial strains.

Results

It was observed that ethanolic extracts of Atropa belladonna has more significant antimicrobial activity against S.aureus than E.coli. In parallel, methanolic extract of Matricaria chamomilla showed greater significant antibacterial activity against S.aureus when compared with E.coli. In comparison, ethanolic extracts of Matricaria chamomilla has shown more significant results against S. aureus than E.coli (p≤0.05). Both plants had no antibacterial activity against S.typhi. The free radical scavenging activity observed by DPPH assay, indicate that both plants have antioxidant activity at all levels of concentrations in solvent tested during the present work. However, methanolic extracts had greater antioxidant activity when compared with ethanolic extracts.

Conclusion

Present study is thus helpful in highlighting present potentials for antioxidant and antimicrobial properties in the selected plants.     

The Immunostimulatory and Antimicrobial Property of Two Herbal Decoctions Used in the Management of HIV/AIDS in Ghana

Background

HIV/AIDS is a pandemic retarding economic growth and destroying human capital globally. This study therefore investigated the perceived efficacy of Betula alba (BA) and Sutherlandia frutescens (SF) decoctions used in the management of HIV/AIDS in Ghana.

Materials and Methods

A study on the records of HIV/AIDS patients attending Habibi Herbal Clinic, Kumasi, Ghana, was conducted to obtain information on the initial viral load presented during their maiden visit and results after treatment with the herbal decoctions. The decoctions were assessed for immunostimulatory property in cyclophosphamide-induced immunosuppressed ICR mice. Total white blood cell count, as well as lymphocyte and neutrophil counts were determined and their effects compared with Levamisole. The decoctions were also screened for antimicrobial activity by the micro dilution method.

Results

The two herbal decoctions used significantly reduced (P ≤ 0.001) the patients'' viral loads (47.42 ± 17.28 % to 13.69 ± 12.42 %; n=16). BA (1, 2, and 4 mg/kg) and SF (0.4, 0.8, and 1.6 mg/kg) caused significant increment (P ≤ 0.001) in total WBC and lymphocyte count in mice comparable to that produced by 2.5 mg/kg Levamisole. The decoctions also exhibited antimicrobial activity against gram negative and gram positive bacteria as well as Candida albicans (MIC 0.607 to 3.062 mg/ml). Phytochemicals present in both plants include saponins, terpenoids and coumarins.

Conclusion

The Betula alba and Sutherlandia frutescens decoctions have interesting immunostimulatory and antimicrobial properties and hence could be useful in the management of HIV/AIDS and associated opportunistic infections.     

Hepatoprotective Properties of Aqueous Leaf Extract of Persea Americana,Mill (Lauraceae) ‘Avocado’ Against CCL4-Induced Damage in Rats

Background

Natural products from plants have received considerable attention in recent years due to their diverse pharmacological properties, including antioxidants and hepatoprotective activities. The protective effects of aqueous extract of Persea americana (AEPA) against carbon tetrachloride (CCl₄)-induced hepatotoxicity in male albino rats was investigated.

Materials and Methods

Liver damage was induced in rats by administering a 1:1 (v/v) mixture of CCl₄ and olive oil [3 ml/kg, subcutaneously (sc)] after pre-treatment for 7 days with AEPA. Hepatoprotective effects of AEPA was evaluated by estimating the activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) and levels of total bilirubin (TBL). The effects of AEPA on biomarkers of oxidative damage (lipid peroxidation) and antioxidant enzymes namely, catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx) and glutathione S-transferase (GST) were measured in liver post mitochondrial fraction.

Results

AEPA and Reducdyn® showed significant (p<0.05) hepatoprotective activity by decreasing the activities of ALT, AST, ALP and reducing the levels of TBL. The activities of antioxidant enzymes, levels of malondialdehyde and protein carbonyls were also decreased dose-dependently in the AEPA-treated rats. Pre-treatment with AEPA also decreased the serum levels of glutathione significantly.

Conclusion

These data revealed that AEPA possesses significant hepatoprotective effects against CCl₄-induced toxicity attributable to its constituent phytochemicals. The mechanism of hepatoprotection seems to be through modulation of antioxidant enzyme system.     

The efficacy of the crude root bark extracts of Erythrina abyssinica on rifampicin resistant Mycobacterium tuberculosis

Introduction

Tuberculosis (TB) is one of the leading causes of morbidity and mortality with a global mortality rate at two million deaths per year while one third of the world''s population is infected with the TB bacillus.

Ojective

To determine the efficacy of the crude extracts of Erythrina abyssinica root bark on rifampicin-resistant TB.

Methods

Crude extracts of root bark of Erythrina abyssinica, were screened against three strains of Mycobacterium tuberculosis including rifampicin-resistant TMC-331. Susceptibility tests used the disc diffusion method and were done on solid Middle brook 7H10, while the Minimum Inhibitory concentration (MICs) and minimum bactericidal concentration (MBCs) were determined by the Microtitre plate method using Middle brook 7H9 broth.

Results

The total crude methanol extract showed activity against all the three strains of mycobacterium used, at 50mg/ml and diameters of zones of inhibition of up to 26 mm. Erythrina abyssinica total crude methanol extract showed the highest activity on the pan sensitive strain H37Rv (0.39±0.0 mg/ml) and the rifampicin resistant strain (TMC-331) (2.35±1.11 mg/ ml) and was also active on Mycobacterium avium (0.39±0.mg/ml. The values for isoniazid were 0.25µg/ml and 9.38µg/ml for H37Rv and TMC-331 respectively, while for rifampicin the MIC value was 0.25µg/ml for H37Rv but it was not active on TMC-331. Acute toxicity test gave an LD50 of 776.2mg/kg body weight while the phytochemical analysis showed the presence of alkaloids, tannins and flavones.

Conclusion

The conclusion from the study was that Erythrina abbyssinica has antimycobacterial activity and reasonable safety that merits further research.     

Evaluation of the Antioxidant Activity of Root Extract of Pepper Fruit (Dennetia Tripetala), and It's Potential for the Inhibition of Lipid Peroxidation

Background

The antioxidant properties of ethanolic root extract of pepper fruit (Donnetia tripetala), and its effect on lipid peroxidation of some fresh beef tissues during frozen storage were investigated.

Materials and Methods

The antioxidant parameters were assessed using standard methods, while malondialdehyde levels of different fresh beef tissue sections treated with the extract prior to freezing, were estimated in a colorimetric reaction with thiobarbituric acid.

Results

The H₂O₂-scavenging ability of the extract was similar to that of ascorbic acid, with a maximum scavenging power of 55.61 ±4.98%, and an IC₅₀ value of 86µg/ml. The extract exhibited a concentration-dependent ferric ion-reducing power, although this was significantly lower relative to that of the ascorbic acid (p < 0.05). The total phenolic content was 212.5 ± 0.002 mg/g, while the nitric oxide-scavenging ability was 64.33 ± 0.2% after 150 min. The capacity of the extract to inhibit lipid peroxidation in frozen heart muscle slices was significantly higher than that of vitamin C (p < 0 .05), but comparable to vitamins C and E in frozen testes and kidney slices.

Conclusion

These results suggest that the root extract of D. tripetala is rich in antioxidants which can be applied to meat preservation during refrigerated storage.     

Bioactivity-Guided Isolation of Anticancer Agents from Bauhinia Kockiana Korth

Background

Flowers of Bauhinia kockiana were investigated for their anticancer properties.

Methods

Gallic acid (1), and methyl gallate (2), were isolated via bioassay-directed isolation, and they exhibited anticancer properties towards several cancer cell lines, examined using MTT cell viability assay. Pyrogallol (3) was examined against the same cancer cell lines to deduce the bioactive functional group of the phenolic compounds.

Results

The results showed that the phenolic compounds could exhibit moderate to weak cytotoxicity towards certain cell lines (GI₅₀ 30 – 86 µM), but were inactive towards DU145 prostate cancer cell (GI₅₀ > 100 µM).

Conclusion

It was observed that pyrogallol moiety was one of the essential functional structures of the phenolic compounds in exhibiting anticancer activity. Also, the carboxyl group of compound 1 was also important in anticancer activity. Examination of the PC-3 cells treated with compound 1 using fluorescence microscopy showed that PC-3 cells were killed by apoptosis.     

Antimicrobial Activity of Methanolic Extracts of Sambucus Ebulus and Urtica Dioica Against Clinical Isolates of Methicillin Resistant Staphylococcus Aureus

Background

Increase in the emergence of drug -resistant pathogens led to the development of natural antimicrobials. In this study the antimicrobial effect of methanolic extracts of Sambucus ebulus and Urtica dioica on 16 skin and wound infections isolates of methicillin resistant S. aureus have been studied.

Material and Methods

Solvent extraction procedure was done using soxhlet apparatus for extracting antimicrobial agents from freeze dried plants. Antibacterial activity was measured using agar well diffusion method.

Results

The MIC of Sambucus ebulus and Urtica dioica extracts against the standard strain of S. aureus ATCC 6538 were determined using the micro dilution method at 15 mg and 20 mg respectively. All the test bacteria were found sensitive to the Sambucus ebulus extract and only one isolate was resistant to Urtica dioica extract.

Conclusion

Extracts of Sambucus ebulus and Urtica dioica possess antibacterial potency against MRSA isolates and may be used as a natural antiseptics and antimicrobial agents in medicine.     

Hypoglycaemic activity of commelina africana and ageratum conyzoides in relation to their mineral composition

Background

Many plants with antidiabetic properties probably act in part through their content of fibre, vitamins, bioactive or mineral content

Objectives

This study investigated the mineral, proximate, phytochemical compositions and hypoglycaemic effect of Commelina africana and Ageratum conyzoides extracts in diabetic rats, and the likely relationship between this property and the mineral, proximate and phytochemical compositions of the plants.

Methods

The plants were subjected to mineral, proximate composition and phytochemical analysis. Attempt was made to see (if any) the relationship between the hypoglycaemic effect and the mineral, proximate compositions and phytochemistry of the plants. Alloxan-induced diabetic animals were administered 500mg/kg body weight aqueous extracts of the plants and glibenclamide as the reference hypoglycaemic agent.

Results

Aqueous extract of Ageratum conyzoides reduced fasting blood glucose of experimental animals by 39.1% while Commelina africana reduced the same by 78.0%. Alkaloids, cardenolides, saponins, and tannins were detected in both plants. Anthraquinones was absent in C. africana but a trace of it was detected in A. conyzoides. The hypoglycaemic effect of Commelina africana was comparable with the reference hypoglycaemic agent. Ageratum conyzoides showed comparably weaker hypoglycaemic effect than exhibited by reference hypoglycaemic agent. Comparatively, Commelina africana had higher mineral concentrations (except Na) than Ageratum conyzoides.

Conclusions

Plants'' extracts minerals (magnesium, potassium and iron) and bioactive components (alkaloids and cardenolides) seemingly enhanced their hypoglycaemic effect. Furthermore, these minerals, alkaloids and cardenolides could be helpful in ameliorating complications of diabetes like hypertension and cardiovascular disease.     

Expectorant and Antitussive Effect of Hedera helix and Rhizoma coptidis Extracts Mixture

Purpose

This study aims to investigate the additive effect of the Hedera helix (HH) and Rhizoma coptidis (RC) extracts mixture on antitussive and expectorant activities in animals.

Materials and Methods

The expectorant assay was performed with phenol red secretion in mice trachea. Mice or guinea pigs were randomly divided into groups of 8 each, including negative and positive control groups. After gastric administration of the test extracts in mice, 2.5% phenol red solution (0.2 mL) was intraperitoneally injected. Trachea was dissected and optical density of tracheal secretion was measured. After gastric administration of the test extracts in guinea pigs, the antitussive activities were assessed using a citric acid-induced cough measurement.

Results

The extracts of HH and RC significantly increased tracheal secretion and inhibited cough. The mixture of HH and RC extracts in a 1:1 concentration at a dose of 200 mg/kg showed a more potent effect on phenol red secretion (25.25±3.14) and cough inhibition (61.25±5.36) than the individual use of each extracts [phenol red secretion; HH 13.39±4.22 (p=0.000), RC 20.78±2.50 (p=0.010), cough inhibition; HH 9.89±4.14 (p=0.010), RC 30.25±7.69 (p=0.000)]. A 3:1 ratio mixture of HH to RC demonstrated an optimal expectorant effect (p<0.001), and this mixture showed expectorant and antitussive effects in a dose-dependent manner.

Conclusion

This study provides evidence for antitussive and expectorant effect of a 3:1 mixture of HH and RC, which may be a useful therapeutic option for respiratory diseases.     

Antibacterial agents from the leaves of Crinum purpurascens herb (Amaryllidaceae)

Background

Typhoid fevers and urogenital infections continue to be serious health problems in developing countries. In our search for therapeutic agents from natural sources with potential for the treatment of typhoid fevers and urogenital infections, extract and compounds were obtained from Crinum purpurascens and tested.

Methods

Two alkaloids (4,5-ethano-9,10-methylenedioxy-7-phenanthridone or hippadine (1) and 4,5-ethano-9-hydroxy-10-methoxy-7-phenanthridone or pratorimine (2)) and one steroid (â-D-glucopyranoside of sitosterol (3)) were isolated from the CH₂Cl₂/MeOH (1:1) leaf extract of Crinum purpurascens and screened for antibacterial activity using both agar diffusion and broth dilution techniques.

Results

For the CH₂Cl₂/MeOH extract, the MIC values obtained were 3 mg/ml (against P. aeruginosa), 4 mg/ml (against E. coli, K. pneumoniae and S. aureus) and 6 mg/ml (against S. typhi and S. paratyphi B), whereas the MBC values varied between 7 and 12 mg/ml. For compound 1, the MIC values varied between 200 and 250 µg/ml, whereas the MBC value was 300 µg/ml against all the bacteria strains used. Compound 2 did not show any antimicrobial activity against these bacteria strains. For compound 3, the MIC values varied between 250 and 300 µg/ml, whereas the MBC values were 300 µg/ml (against S. typhi and S. paratyphi B) and > 300 µg/ml (against the other bacteria strains).

Conclusion

These data suggest that C. purpurascens leaf extract contains antibacterial agents which could be used in the treatment of typhoid fevers and urogenital infections.     

In-vitro antimicrobial activity of selected honeys on clinical isolates of Helicobacter pylori

Background

Helicobacter pylori is a gram-negative bacterium incriminated in gastroduodenal ulcers, and mucosa-associated lymphoid tissue lymphoma imposing a major burden on health care systems worldwide. Honeys have been shown to have in vitro activity against microaorganisms and suitable for use in ulcers, infected wounds and burns.

Objective

The study was aimed at evaluating the antimicrobial potential of honeys (Manuka−, Capillano®, Eco- and Mountain) at different concentrations (10%v/v, 20%v/v, 50%v/v and 75%v/v) against clinical isolates of H. pylori.

Methods

H. pylori was isolated from gastric biopsies of patients with gastroduodenal pathologies following standard microbiological procedures. Antimicrobial susceptibility of the isolates to different honey varieties was determined by the disk diffusion assay. Also, the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the most potent honey was determined by the agar dilution method. Data were analysed using the Fisher exact test and statistical significance considered at p<0.05.

Results

All the four honey varieties exhibited antibacterial activity. The strongest inhibitory activity (82.22%) was demonstrated by Mountain honey at 75%v/v, followed by Capillano® and Manuka− honeys (75.56%), and Eco-honey (73.36%) at the same concentration. However, no statistically significant difference (p>0.05) was noted between the honeys at different concentrations. The MIC and MBC concentrations of Mountain honey were in the range 0.117 – 0.938ì/mL and 0.366 – 2.965ìg/mL respectively. The antimicrobial potential of these honeys at different concentrations were highly comparable to clarithromycin, the positive control.

Conclusion

These honeys may contain compounds with therapeutic potential against our local isolates of H. pylori.     

Development of a Quantitative Sandwich Enzyme-Linked Immunosorbent Assay for Detecting the MPT64 Antigen of Mycobacterium tuberculosis

Purpose

Tuberculosis (TB) is a major infectious disease and is responsible for two million deaths annually. For the identification and quantitation of Mycobacterium tuberculosis (M. tuberculosis), a causative agent of TB, a sandwich enzyme-linked immunosorbent assay (ELISA) against the MPT64 protein of M. tuberculosis, an antigen marker of the M. tuberculosis complex, was developed.

Materials and Methods

The MPT64 protein was expressed, and anti-MPT64 monoclonal antibodies were prepared. A sandwich ELISA was established using recombinant MPT64 protein and anti-MPT64 monoclonal antibodies. The sandwich MPT64 ELISA was evaluated using reference and clinical mycobacterial strains.

Results

The sandwich MPT64 ELISA detected MPT64 protein from 2.1 ng/mL to 250 ng/mL (equivalent to 1.7×10⁴ CFU/mL and 2.0×10⁶ CFU/mL). All 389 clinical M. tuberculosis isolates tested positive in the sandwich MPT64 ELISA (sensitivity, 100%), and the assay showed no cross reactivity to any tested nontuberculous mycobacterial strain (specificity, 100%).

Conclusion

The sandwich MPT64 ELISA is a highly sensitive and quantitative test for MPT64 protein, which can identify M. tuberculosis.     

Cytotoxicity,Phytochemical Analysis and Antioxidant Activity of Crude Extracts from Rhizomes of Elephantorrhiza Elephantina and Pentanisia Prunelloides

Background

Elephantorrhiza elephantina (Ee) and Pentanisia prunelloides (Pp) are two medicinal plants which are widely used to remedy various ailments including diarrhoea, dysentery, inflammation, fever, rheumatism, heartburn, tuberculosis, haemorrhoids, skin diseases, perforated peptic ulcers and sore joints in southern Africa (South Africa, Swaziland, Botswana and Zimbabwe). The following study was conducted to explore the in vitro cytotoxicity, antioxidant properties and phytochemical profile of the two medicinal plants.

Materials and Methods

The cytotoxicity of the aqueous and methanol extracts and fractions of both species was studied using the brine shrimp lethality tests (BST) for the first time.

Results

The results demonstrated that the lethality (LC₅₀) for crude extracts for both plants ranged between 1.8 and 5.8 ppm and was relatively greater than that for the methanol, ethyl acetate and chloroform fractions of the extracts which ranged between 2.1 ppm and 27 ppm. This suggested that crude extracts were more potent than their respective fractions, further explaining that the different fractions of phytochemicals in these plant species work jointly (in synergy) to exert their therapeutic efficacy. Both aqueous and methanol extracts of the two medicinal plants demonstrated a high degree of antioxidant capacity against the DPPH radical with the Duh and Yen inhibition percentage ranging between 4.5% and 72%. Phytochemical studies of the rhizome extracts showed that the major compounds present include flavonoids, tannins, anthocyanidins, anthraquinones, triterpenoids (oleanolic acid), the steroidal saponin Diosgenin, the sugars, rhamnose, glucuronic acid, Arabinose and hexoses.

Conclusion

This is the first report of the detection and isolation of diosgenin and oleanolic acid from the rhizome extracts of Ee and Pp. All structures were determined using spectroscopic/spectrometric techniques (1H NMR and 13C and LC-ESI-MS) and by comparison with literature data.