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1.  Errata 
Riadh et al. Afr J Tradit Complement Altern Med. 2011;8(3):322–327.
DETECTION AND EXTRACTION OF ANTI-LISTERIAL COMPOUNDS FROM CALLIGONUM COMOSUM, A MEDICINAL PLANT FROM ARID REGIONS OF TUNISIA.
Riadh H, Imen F, Abdelmajid Z, Sinda F.
Should read
Hammami et al. Afr J Tradit Complement Altern Med. 2011;8(3):322–327.
DETECTION AND EXTRACTION OF ANTI-LISTERIAL COMPOUNDS FROM CALLIGONUM COMOSUM, A MEDICINAL PLANT FROM ARID REGIONS OF TUNISIA.
Hammami R, Farhat I, Zouhir A, Fedhila S
PMCID: PMC3746587
2.  Errata 
Zhou et al., Afr J Tradit Complement Altern Med. (2012) 9(3):303–314
EXTRACTS OF SALVIA MILTIORRHIZA BUNGE ON THE CYTOKINES OF RAT ENDOMETRIOSIS MODELS
Zan-Hua Zhou 1, Qing Weng 2, Jian-Hong Zhou 3*, Jue Zhou 4*
1School of Medicine, Lishui University, Lishui, 323000, Zhejiang, China
2The Third People's Hospital of Yuhang District, Hangzhou, Zhejiang, 311115, China
3Women's Hospital, School of Medicine, Zhejiang University, Hangzhou, 310006, Zhejiang, Chin, 4College of Food Science and Biotechnology Engineering, Zhejiang Gongshang University, Hangzhou, Zhejiang, 310035, China.
Email: juezhou2006@yahoo.com.cn
Acknowledgements
Dr. Jue Zhou was funded by Research Fund of Zhejiang Gongshang University (1110XJ2011023).
Should read thus
Zhou et al., Afr J Tradit Complement Altern Med. (2012) 9(3):303–314
EXTRACTS OF SALVIA MILTIORRHIZA BUNGE ON THE CYTOKINES OF RAT ENDOMETRIOSIS MODELS
Zan-Hua Zhou 1, Qing Weng 2, Jian-Hong Zhou 3*, Jue Zhou 4*
1School of Medicine, Lishui University, Lishui, 323000, Zhejiang, China
2The Third People's Hospital of Yuhang District, Hangzhou, Zhejiang, 311115, China
3Women's Hospital, School of Medicine, Zhejiang University, Hangzhou, 310006, Zhejiang, Chin, 4College of Food Science and Biotechnology Engineering, Zhejiang Gongshang University, Hangzhou, Zhejiang, 310035, China.
Email: juezhou2006@yahoo.com.cn
Acknowledgements
Dr. Jue Zhou was funded by Research Fund of Zhejiang Gongshang University (1110XJ2011023). The first two authors (Zan-Hua Zhou and Qing Weng) contributed equally to the present study
PMCID: PMC3746588  PMID: 23986924
3.  Temperament Determination for Melatonin: A Bridge from Iranian Traditional to Modern Sleep Medicine 
History acknowledged Ibn Sina, or Avicenna, the author of the highly skilled textbook of medicine “Al-Qanun Fi Al-Tibb” or “The Canon of Medicine", as one of the greatest physicians in medicine. According to this medical textbook, the explanation of the existence of a cold temperament for sleep was that during sleep hours, people tended to have a movement of the nature of the body toward the inside, which caused the body to become cold during sleep. Temperament determination for molecules, including drugs, has proved several applications. The present study tried to demonstrate that the multitasking melatonin molecule, as a sleep related hormone, had a cold temperament. The consideration of this temperament for melatonin had the potential to connect and integrate Iranian traditional medicine to current medicine, and also opened new frontiers for the physiopathology of modern sleep medicine, based on traditional medicine.
PMCID: PMC3746582  PMID: 24146459
Sleep; Melatonin; Iranian traditional medicine; Avicenna
4.  Effects of Polysaccharides from Gynostemma Pentaphyllum (Thunb.), Makino on Physical Fatigue 
Background
Gynostemma pentaphyllum (Thunb.) Makino has been reported to have a wide range of health benefits in Chinese herbal medicines. Polysaccharides from Gynostemma pentaphyllum (PGP), has been identified as one of the active ingredients responsible for its biological activities. Although many pharmacological activities of PGP have received a great deal of attention, there is limited evidence for the anti-fatigue effects of PGP. The purpose of this study was to investigate the effects of polysaccharides from PGP on physical fatigue.
Materials and method
The rats were divided into four groups, with 10 animals per group: control (C), group, low-treated (LT), group, medium-treated (MT), group, and high-treated (HT), group. The C group received distilled water, while LT, MT and HT groups were given various doses of PGP (100, 200, 400 mg/kg· d). After 30 days, forced swimming test was carried out in an acrylic plastic pool, then the exhaustive swimming time of rats and some biochemical parameters related to fatigue were measured. The data obtained showed that PGP could extend the exhaustive swimming time of the rats, as well as decrease the blood lactic acid (BLA), and blood urea nitrogen (BUN), concentrations, and increase the hemoglobin, liver glycogen and muscle glycogen concentrations.
Result
The data obtained showed that different doses of PGP could extend the exhaustive swimming time of the rats, as well as decrease the BLA and BUN concentrations, and increase the hemoglobin, liver glycogen and muscle glycogen concentrations, which suggests that PGP had significant anti-fatigue effects on rats.
Conclusion
PGP may be of use as a potential anti-fatigue agent, but there is a need for further research on long-term use in order to show its positive effects on physical fatigue.
PMCID: PMC4202428  PMID: 25371572
polysaccharides from Gynostemma pentaphyllum (Thunb.) Makino; physical fatigue; forced swimming test; rats
5.  Virulence Genotype and Phylogenetic Groups in Relation to Chinese Herb Resistance Among Escherichia Coli from Patients with Acute Pyelonephritis 
Background
Clinical isolates of herb-resistant uropathogenic E. coli were isolated. It was possible that the virulence genotypes and phylogenetic background of E. coli differed between Chinese herb-resistant E. coli and -susceptible isolates. For this purpose, the prevalence of virulence factors (VFs) and phylogenetic background, with regard to Chinese herb resistance, among E. coli strains causing acute pyelonephritis from China were investigated.
Materials and Methods
E. coli isolates from patients with acute pyelonephritis were used in this study. Standard disc diffusion methodology was used to test the susceptibility of Chinese herbal concoction against E. coli strains. Multiplex PCR amplifications employed three markers (chuA, yjaA, and TSPE4.C2) to classify E. coli isolates into one of four phylogenetic groups (group A, B1, B2, or D). The isolates were also tested for 14 virulence-associated traits (VFs) of uropathogenic E. coli.
Results
A total of 115 E. coli strains were isolated. 79 (68.7%) were susceptible and 36 (31.3%) were resistant to the herbal concoction. 20.9% of the isolates encoded three or more of VFs for which they were screened, with 13.9% in susceptible isolates and 36.1% in resistant isolates. The key VFs (fyuA and/or iutA siderophores) present in >80% of isolates. The papA and papC adhesins were detected in the majority of resistant isolates (72.2% and 63.9% respectively). 78.5% of susceptible isolates belong to phylogenetic groups A, while 83.3% of resistant isolates belong to group B2.
Conclusion
PapA and papC are significant VFs with an essential role in contributing to Chinese herb-resistance. Chinese herb-resistance is associated with a shift towards more virulent strains and B2 phylogenetic group.
PMCID: PMC4202444  PMID: 25371588
Escherichia coli; Virulence factors; Phylogenetic group; Chinese herb-resistance
6.  Chinese Herb-Resistance and Adherence to Human Uroepithelial Cells of Uropathogenic Escherichia Coli 
Background
In order to define the virulence factors between Chinese herb-resistant uropathogenic E. coli and susceptible strains, the UPEC isolates were classified into two groups according to its resistance to Chinese herbs.
Materials and Methods
The susceptibility profile of strains was determined by disk diffusion method. PCR systems were used to detect genes encoding papC, Aer, hly and cnf1. Isolated human urothelial cells were incubated in vitro and investigated with light microscope immunohistochemistry. Adhesion of E. coli to urothelial cells was studied in vitro.
Results
The results showed that, among the 105 UPEC isolates, 18 were resistant to the herbal concoction. Cnf1 and papC occurred in ≥66.7%, of herb-resistant isolates, while, hly and Aer occurred in 22.2% and 27.8% of strains respectively. Only one gene (Cnf1) occurred in >40%, of Herb-susceptible isolates. Other genes were also found in susceptible isolates: papC (20.7%), hly (11.5%), and Aer (6.9%). Light microscopy and immunochemical investigations demonstrated the normal pelvic transitional epithelial cells cultured. The adherence of strains in both groups increased in 30 min., and reached its peak at 60, (Susceptible E. coli) or 120 min., (Resistant E. coli). The adhesion of the susceptible bacteria to human uroepithelial cells was significantly lower compared with that of the resistant E. coli (p<0.05).
Conclusion
These findings revealed that, Chinese herb-resistant uropathogenic E. coli isolates that are hemolytic, and have Aer, papC, hly, Cnf1 genes are more able to be uropathogenic and adherent.
PMCID: PMC3957250  PMID: 24653562
Escherichia coli; Adherence; Virulence genes; Human uroepithelial cells; Chinese herb-resistance
7.  Effects of Ixeris Chinensis (Thunb.) Nakai Boiling Water Extract on Hepatitis B Viral Activity and Hepatocellular Carcinoma 
Background
Hepatitis B virus (HBV) infection and hepatocellular carcinoma are major diseases that affect the Taiwanese population. Therefore, the development of an alternative herbal medicine that can effectively treat these diseases is a research target. In this study, we tested Ixeris Chinensis (Thunb.) Nakai boiling water extract (ICTN BWE) in vitro and analysed its effects on the HBV and liver cancer.
Materials and Methods
We used a human liver cancer cell line (Hep3B, a cell line that continuously secretes HBV particles into a medium) as an experimental model for the screening of various ICTN BWE concentrations and their effects on the HBV in vitro.
Results
Our results showed that 75 µg/mL ICTN BWE downregulated the relative expression of the hepatitis B virus surface antigens (HBsAg) to 77.1%. Using the human liver cancer cell lines HuH-7 and HepG2, and 3-(4,5-dimethylthiazol-zyl)-2,5-diphenyl tetrazolium bromide (MTT) and tumour clonogenic assays, we then showed that ICTN BWE inhibits hepatocellular carcinoma growth.
Conclusion
Fluorescent microscopy of DAPI(4′,6-Diamidino-2-phenylindole)-stained nuclei and DNA fragmentation assays confirmed the inhibitory effects of ICTN BWE on liver tumour cell growth through induction of apoptosis.
PMCID: PMC3957264  PMID: 24653576
herbal medicine; Ixeris Chinensis (Thunb.) Nakai; antihepatocellular carcinoma; apoptosis; antihepatitis B virus
9.  Chinese Herbal Medicine: A Safe Alternative Therapy for Urinary Tract infection in Patients with Renal Insufficiency 
We have used reduced doses of Chinese herbs for estimation of urinary tract infections (UTIs) patients with stable impairment of renal function. A total of 33 adult female patients with moderately impaired renal function and symptomatic UTIs were included in this study. Urine cultures were carried out. Patients were monitored clinically and with various laboratory tests. Chinese herbal concoction divided by milligrams of creatinine per 100 ml were orally administrated for ten days. Three patients were excluded from final analysis. Most of the patients responded symptomatically to treatment. Chinese herbs eradicated the primary pathogen in 68.7% of the patients at the day 10 of treatment. Two patients relapsed (one had abbreviated courses of therapy) 6 to 8 days posttreatment. Organisms which recurred included Escherichia coli and Pseudomonas aeruginosa. Bacterial reinfections occurred 5 to 8 days posttreatment in four patients. Adverse reactions observed among the 30 patients were rare. Nausea (6.67%) and mild elevation of hepatic enzymes (3.33%) were probably drug related. Nausea disappeared when the therapy ended. Elevated hepatic enzymes resumed at the 2-week follow-up. Two patients demonstrated slight increases in serum creatinine on day 10 of treatment. One patient had a 12.5% elevation over baseline and the other had a 13.0% elevation. Serum creatinine values had improved in these two patients at 4-week follow-up. Chinese herbal medicine was effective and safe in the treatment of UTIs with renal insufficiency.
PMCID: PMC3746634  PMID: 23983345
Chinese herbs; Urinary tract infection; Renal insufficiency; Dose adjustment
10.  Prevalence and Acceptability of Male Circumcision in South Africa 
Background
The objectives of the current national study were to determine the rates of self-reported circumcision among South African men and, more importantly, evaluate the acceptability of male circumcision in South Africa by uncircumcised adult men and all adult women.
Materials and Methods
The study based on a population-based survey included a nationally representative subgroup of 6654 men aged 15 years and older who where included in the analysis on male circumcision prevalence, and a subgroup of 6796 women aged 15 to 49 years who were included in the analysis on male circumcision acceptance.
Results
An overall prevalence of self-reported male circumcision of 42.8% was found. Among the Black African population group the prevalence of male circumcision was 48.2%, 32.1% were traditionally and 13.4% were medically circumcised. Among males not circumcised 45.7% of 15–24 years olds indicated that they would consider being circumcised compared to 28.3% among 25–49 years olds. In multivariate analysis among non-circumcised men Black African and Coloured population groups and having heard of the HIV protective effect of male circumcision were significant predictors for male circumcision acceptability, and among women with a non-circumcised sexual partner, Black African and Coloured population groups and higher education were predictors for male circumcision acceptability.
Conclusion
The study found high rates and high acceptability of male circumcision. Findings associated with the acceptability of male circumcision can be used to increase awareness of the benefits of male circumcision for HIV prevention.
PMCID: PMC4202407  PMID: 25392591
Male circumcision; prevalence; acceptability; national population-based survey; South Africa
11.  Gongronema Latifolium Delays Gastric Emptying of Semi-Solid Meals in Diabetic Dogs 
The aim of the study was to investigate sonographically the effect of Gongronema latifolium on gastric emptying of semi-solid meals in diabetic dogs. Twenty-five alloxan-induced diabetic dogs were randomly allotted into five groups of five dogs each in a randomised placebo-controlled study. These are placebo, prokinetic dose, low dose, moderate dose and high dose groups. The placebo group served as the control. The low, moderate and high dose groups ingested methanolic leaf extract of G. latifolium at 100 mg/kg, 250 mg/kg, 500 mg/kg respectively, while the prokinetic group ingested 0.5 mg/kg of metoclopramide. After a 12-hour fast, each group ingested its treatment capsules 30 minutes before the administration of test meal. Measurements of gastric emptying and blood glucose levels were obtained from each dog 30 minutes before and immediately after the ingestion of a test meal, every 15 minutes for another 4 hours and then every 30 minutes for further 2 hours. Gastric emptying of the moderate and high dose groups were 227.8 ± 9.9 min and 261.3 ± 19.3 min respectively and significantly (p < 0.0001) slower than the placebo control group of 143.0 ±17.8 min. The gastric emptying of the low dose group (169.8 ± 3.8) and control group did not differ significantly (p > 0.05). A strong inverse relationship between gastric emptying and the incremental blood glucose levels was noted in the diabetic dogs after the ingestion of Gongronema latifolium (r = −0.90; p < 0.0001). Gonogronema latifolium delayed gastric emptying in diabetic dogs.
PMCID: PMC3847424  PMID: 24311844
Diabetes; Gastric emptying; Gongronema latifolium; Sonography; Postprandial blood glucose; Antral area
12.  Effect of Tao-Hong-Si-Wu-Tang, A Traditional Chinese Herbal Medicine Formula, on Physical Fatigue in Mice 
Tao-Hong-Si-Wu-Tang (THSWT) is a famous traditional Chinese herbal medicine formula, which has traditionally been used in China for about one thousand years. The present study investigated the effect of THSWT on physical fatigue. 32 male mice were randomly divided into 4 groups with 8 in each group. All were administered orally and daily for 28 days. Group I received isotonic saline solution as control; Group II, III and IV obtained 5, 10 and 20ml/ kg body weight of THSWT solutions, respectively. After 28 days, the anti-physical fatigue effect of THSWT was evaluated by using a forced swimming test, along with the determination of blood lactic acid, blood urea nitrogen (BUN), liver glycogen and muscle glycogen contents. The data showed that THSWT could extend exhaustive swimming time of mice, as well as decrease the BLA and BUN contents and increase the liver glycogen and muscle glycogen contents. The results support that THSWT had anti-physical fatigue effect.
PMCID: PMC3746359  PMID: 24082327
Tao-Hong-Si-Wu-Tang; physical fatigue; forced swimming test; mice
13.  Effect of Purificatory Measures Through Cow's Urine and Milk on Strychnine and Brucine Content of Kupeelu (Strychnos Nuxvomica Linn.) Seeds 
Strychnos nux vomica Linn.(Loganaceae) commonly known as Nux vomica (Kupeelu), is a poisonous plant and its seeds are used widely in Ayurvedic system of medicine since time immemorial. Ayurveda advocates that nux vomica seeds are to be administered in therapeutics only after going through certain purificatory measures (Shodhana). There are more than six media: cow's urine (Go mutra), cow's milk (Go dugdha), cow's ghee (Go ghrita), Kanji (thin gruel), castor oil (Eranda taila) and fresh ginger juice (Ardraka swarasa) etc., which have been reported in different classical texts of Ayurveda for proper processing of nux vomica seeds. In this study, an attempt has been made to purify the seeds by using three different methods as described in ancient treatise by using cow's urine and cow's milk as media alone and together. This study revealed that all the methods studied reduced the toxicity of strychnine and brucine contents in comparison to the raw seeds as determined by HPTLC. Out of these three methods maximum reduction in strychnine and brucine contents was found when the seeds were purified by keeping them in cow's urine for seven days followed by boiling in cow's milk for three hrs.
PMCID: PMC3746529  PMID: 23983327
Kupeelu; Strychnos nuxvomica; Shodhana; strychnine; Ayurveda; brucine; Cow's milk; Cow's urine
14.  The Anti-Oxidant Effects of Ginger and Cinnamon on Spermatogenesis Dys-function of Diabetes Rats 
Background
Diabetes rats have been linked to reproductive dysfunction and plant medicine has been shown to be effective in its treatment. Antioxidants have distinctive effects on spermatogenesis, sperm biology and oxidative stress, and changes in anti-oxidant capacity are considered to be involved in the pathogenesis of chronic diabetes mellitus. Ginger and cinnamon are strong anti-oxidants and have been shown to reduce oxidative stress in the long-term treatment of streptozotocin (STZ)-induced diabetes in animal models. The present study examined the influence of combined ginger and cinnamon on spermatogenesis in STZ-induced diabetes in male Wistar rats.
Materials and Methods
Animals (n = 80) were allocated randomly into eight groups, 10 each: Group 1: Control rats given only 5cc Normal saline (0.9% NaCl) daily;Group2: rats received ginger (100mg/kg/rat) daily; Group 3: rats received cinnamon (75mg/kg) daily; Group 4: rats received ginger and cinnamon, (100mg/kg/rat ginger and 75mg/kg cinnamon) daily; Group 5: Diabetic control rats received only normal saline. Group 6: Diabetic rats received 100mg/kg/day ginger; Group 7: Diabetic rats received 75mg /kg/ day cinnamon; Group 8: Diabetic rats received ginger and cinnamon (100mg/kg/day and 75mg/kg /day). Diabetes was induced with 55 mg/kg, single intra-peritoneal injection of STZ in all groups. At the end of the experiment (56th day), blood samples were taken for determination of testosterone, LH,FSH, total anti-oxidant capacity, and levels of malondialdehyde, SOD, Catalase and GPX. All rats were euthanized, testes were dissected out and spermatozoa were collected from the epididymis for analysis.
Results
Sperm numbers, percentages of sperm viability and motility, and total serum testosterone increased in ginger and cinnamon and combined ginger and cinnamon treated diabetic rats compared with control groups. Serum testosterone, LH and FSH were higher compared to control group and also serum anti-oxidants (TAC, SOD, GPX and catalase) all were increased at the end of treatment. Combined ginger and cinnamon showed more intense increase in all parameters compare to ginger and cinnamon alone. Most of the results were significant (P<0.05).
Conclusion
We concluded that combined ginger and cinnamon have significant beneficial effects on the sperm viability, motility, and serum total testosterone, LH,FSH and serum anti-oxidants' level and could be effective for maintaining healthy sperm parameters and male reproductive function in diabetics.
PMCID: PMC4202389  PMID: 25392573
Ginger; Cinnamon; Streptozotocin; Spermatogenesis; rat
15.  RAPD and SSR Based Genetic Diversity Analysis of Elite-2 Set of Synthetic Hexaploid Wheats 
Background
Synthetic hexaploid wheats are artificially reconstituted hexaploid wheats that possess high genetic variation which could be utilized for the development of new improved wheat varieties. One such group of synthetic wheats is called the Elite-II set of synthetic wheats that are derived from crossing durum wheat with different Aegilops tauschii wheats.
Materials and Methods
In the current study genetic diversity was investigated among 18 Elite-II synthetic hexaploid wheat lines at DNA level. Two types of molecular markers i.e. RAPD and SSR were used for this purpose.
Results
Both types of markers proved useful in estimating the overall genetic diversity among these lines. Based on RAPD data range of genetic distances in these lines was from 0 to 100 percent. Seven D-genome specific SSRs were also used to get further estimation of the genetic diversity contributed by Aegilops tauschii parent. On the basis of results obtained it is inferred that the Aegilops tauschi accessions used in the production of these synthetic lines were genetically different and they contributed to the enhancement of genetic variation in the synthetic lines. These results could be helpful for future genome mapping programs.
Conclusion
The overall extensive genitive diversity indicates that these lines are good candidates for development of improved wheat varieties by crossing with cultivated wheat varieties.
PMCID: PMC4202390  PMID: 25392574
Elite-II; molecular markers; RAPD; SSR; synthetic hexaploid wheat
16.  Anti-Oxidant Effects of Pomegranate Juice on Saccharomyces Cerevisiae Cell Growth 
Background
Pomegranate juice has a number of positive effects on both human and animal subjects.
Material and methods
Four groups were used in this study. i: Control group, ii: H2O2 group, iii: Pomegranate juice (PJ) group and iv: PJ + H2O2 group. Following the sterilization method for pomegranate juice (10%) and H2O2 (6% v/v), Saccharomyces cerevisiae cultures were added and the cultivation incubated at 35°C for 72 hours. Fatty acids and vitamin concentrations were measured using HPLC and GC and the total protein bands profile were determined by SDS-PAGE.
Results
According to our results statistically significant differences have been determined among the study groups in terms of fatty acids and vitamin (p<0,05). Fatty acid synthesis, vitamin control and cell density increased in groups to which PJ was given in comparison with the control group (p<0,05). Pomegranate juice increased vitamins, fatty acids and total protein expression in Saccharomyces cerevisiae in comparison with the control.
Conclusion
Pomegranate juice has a positive effect on fatty acid, vitamin and protein synthesis by Saccharomyces cerevisiae. Accordingly, we believe that it has significantly decreased oxidative damage thereby making a positive impact on yeast development.
PMCID: PMC4202391  PMID: 25392575
Pomegranate juice; SDS-PAGE; fatty acid; vitamin
17.  Chemical Composition Profiling and Antifungal Activity of the Essential Oil and Plant Extracts of Mesembryanthemum Edule (L.) Bolus Leaves 
Background
Essential oil from Mesembryanthemum edule leaves have been used by the Eastern Cape traditional healers for the treatment of respiratory tract infections, tuberculosis, dysentery, diabetic mellitus, laryngitis and vaginal infections. The investigation of bioactive compounds in the essential oil of this plant could help to verify the efficacy of the plant in the management or treatment of these illnesses.
Materials and methods
Various concentrations of the hydro-distilled essential oil, ranging from 0.005–5 mg/ml, were tested against some fungal strains, using the micro-dilution method. Minimum inhibitory activity was compared with four other different crude extracts of hexane, acetone, ethanol and aqueous samples from the same plant. The chemical composition of the essential oil, hexane, acetone and ethanol extracts was determined using GC-MS.
Results
GC/MS analysis of the essential oil resulted in the identification of 28 compounds, representing 99.99% of the total oil. Phytoconstituents of hexane, acetone and ethanol extracts yielded a total peak chromatogram of fifty nine compounds. A total amount of 10.6% and 36.61% of the constituents were obtained as monoterpenes and oxygenated monoterpenes. Sesquiterpene hydrocarbons (3.58%) were relatively low compared to the oxygenated sesquiterpenes (9.28%), while the major concentrated diterpenes and oxygenated diterpenes were 1.43% and 19.24 %, respectively and phytol 12.41%. Total amount of fatty acids and their methyl esters content, present in the oil extract, were found to be 19.25 %. Antifungal activity of the oil extract and four solvent extracts were tested against five pathogenic fungal strains. The oil extract showed antifungal activity against Candida albican, Candida krusei, Candida rugosa, Candida glabrata and Cryptococcus neoformans with MIC ranges of 0.02 0.31 mg/ml. Hexane extract was active against the five fungal strains with MICs ranging between 0.02–1.25 mg/ml. Acetone extracts were active against C. krusei only at 0.04mg/ml. No appreciable antifungal activity was found in either ethanol or water extracts when compared with commercial antibiotics.
Conclusion
The profile of chemical constituents found in M. edule essential oil and its antifungal properties support the use of M. edule by traditional healers as well as in the pharmaceutical and food industries as a natural antibiotic and food preservative.
PMCID: PMC4202392  PMID: 25392576
Mesembryanthemum edule; Essential oil; GC/MS; Antifungal activity; Opportunistic fungi
18.  Biocompatibility of Bio Based Calcium Carbonate Nanocrystals Aragonite Polymorph on NIH 3T3 Fibroblast Cell Line 
Background
Currently, there has been extensive research interest for inorganic nanocrystals such as calcium phosphate, iron oxide, silicone, carbon nanotube and layered double hydroxide as a drug delivery system especially in cancer therapy. However, toxicological screening of such particles is paramount importance before use as delivery carrier. In this study we examine the biocompatibility of CaCO3 nanocrystal on NIH 3T3 cell line.
Material and Methods
Transmission and field emission scanning electron microscopy (TEM and FESEM) were used for the characterisation of CaCO3 nanocrystals. Cytotoxicity and genotoxic effect of calcium carbonate nanocrystals in cultured mouse embryonic fibroblast NIH 3T3 cell line using various bioassays including MTT, and Neutral red/Trypan blue double-staining assays. LDH, BrdU and reactive oxygen species were used for toxicity analysis. Cellular morphology was examined by scanning electron microscopy (SEM) and confocal fluorescence microscope.
Results
The outcome of the analyses revealed a clear rod-shaped aragonite polymorph of calcium carbonate nanocrystal. The analysed cytotoxic and genotoxicity of CaCO3 nanocrystal on NIH 3T3 cells using different bioassays revealed no significance differences as compared to control. A slight decrease in cell viability was noticed when the cells were exposed to higher concentrations of 200 to 400 µg/ml, while increase in ROS generation and LDH released at 200 and 400 µg/ml was observed.
Conclusions
The study has shown that CaCO3 nanocrystal is biocompatible and non toxic to NIH 3T3 fibroblast cells. The analysed results offer a promising potential of CaCO3 nanocrystal for the development of intracellular drugs, genes and other macromolecule delivery systems.
PMCID: PMC4202393  PMID: 25392577
Biocompatibility; Calcium carbonate; nanocrystals; drugs and Cockle shells
19.  Ultrastructure and Elemental Analysis of Hypoxis Hemerocallidea: A Multipurpose Medicinal Plant 
Background
Herbal medicine is a popular means of medical management in some parts of the world especially in Africa. Hypoxis hemerocallidea Fisch.C.A.Mey. & Avé-Lall, also known as African potato of the Hypoxidaecae family, is one of the medicinal plants that have enjoyed long usage as an herbal medicine in South Africa. In this study, the morphology and elemental constituents of H. hemerocallidea leaf was investigated to correlate the functional role of the ultrastructure in the production of therapeutic compounds.
Materials and Methods
Fresh leaves of H. hemerocallidea were prepared for analysis using standard methods. The ultrastructure and crystal deposits of the plant were assessed using scanning electron microscopy (SEM), and energy dispersive x-ray (EDX).
Results
It was observed that the leaves were characterised by multicelullar glandular and non glandular trichomes which are sparsely distributed over the entire surfaces. The glandular trichomes (GTs) in H. hemerocallidea leaf have boulbous heads which are probably filled with secretions, while the non glandular trichomes were long, fibrous and sparse. EDX-SEM of Hypoxis hemerocallidea leaf revealed that carbon, oxygen, nitrogen and silicon are the major components of the deposits, while other elements such as iron, sulphur, sodium, calcium, magnesium, potassium, manganese, iodine, chromium and iodine were present in small but variable amounts.
Conclusion
The presence of these elements which are crucial to maintaining good health, in addition to other bioactive constituents might be accountable for the multipurpose therapeutic uses of Hypoxis hemerocallidea in the treatment of cancers, HIV/AIDS related diseases, urinary tract infections, cardiovascular disorders, diabetes and other chronic ailments of humans.
PMCID: PMC4202394  PMID: 25392578
mineral elements; Hypoxis hemerocallidea; multipurpose medicinal plant; scanning electron microscope; trichomes; ultrastructure; EDX-MS analysis
20.  Laxative Activities of Cassia Sieberiana and Senna Obtusifolia 
Background
The root and stem bark of Cassia sieberiana DC. (Caesalpiniaceae) and the root of Senna obtusifolia (Linn) Irwin and Barneby (Caesalpiniaceae), used for constipation in Nigeria, were assayed for laxative properties in male albino rats using the official senna leaf (Senna alexandrina Mill. family Caesalpiniaceae) as the reference standard. This is with a view to finding alternative laxative drug to official senna which is presently being imported into Nigeria from the United Kingdom.
Materials and Methods
The mean percentage of wet faeces in rats, an indication of laxative activity, were obtained using established methods. The laxative activity was established at 500 mg/kg after the infusion of the drug was orally administered on male albino rats following established methods while a set of data was analyzed at 95 % confidence level.
Results
At 500 mg/kg, Senna obtusifolia root gave about 45 % wet faeces while Cassia sieberiana root gave about 40 % wet faeces while at the highest dose of 700 mg/kg, they produced 60 % and 38 % wet faeces, respectively. At these two doses, the official Senna gave 50.6 % and 66 % wet faeces, respectively. Thus, S. obtusifolia and C. sieberiana roots exhibited 89 % and 80 % of the potency of S. alexandrina (the official drug), respectively. The analysis of variance revealed a significant statistical difference in the levels of wet faeces produced by rats dosed with C. sieberiana root.
Conclusion
The results have shown that the roots of the two species could be developed as mild laxative drugs for children and pregnant women for whom the official senna will be contraindicated.
PMCID: PMC4202395  PMID: 25392579
Cassia sieberiana; Senna obtusifolia; Gastrointestinal; Laxative
21.  Phytotherapeutic Activity of Euphorbia Cyparissias Extracts on Ixodidae (Acari) Female Ticks 
Background
Given its numerous biologically active components, Euphorbiaceae has been found to be a large plant family and polyvalent with quite interesting therapeutic activity that can be studied.
Materials and Methods
The ixodicidal activity of Euphorbia cyparissias extracts was studied in vitro and in vivo. Tested concentrations were 10, 5, 2, 1, 0.5 and 0.25%.
Results
For the in vitro study, conducted on field-collected female specimens of Dermacentor marginatus and Haemaphysalis punctata, the efficacy results showed that the ticks died after exposure in the case of 10, 5, and 2% tincture concentrations. The effects appeared after 30 minutes and became more visible 120 minutes after each exposure. The statistical differences regarding the used concentrations were found to be: F = 6.51, df = 5, P < 0.001. The in vivo study of the efficacy of E. cyparissias concentrations was performed on 35 naturally infested sheep and on 30 bovines parasitized with Ixodes ricinus, sprayed with tincture and glycerinate dilutions (bovines) on days 0 and 7. The results revealed detrimental effects on the survivability of female ticks, the most prominent being the reduction of their movement capacity. In sheep in vivo efficiency observed within 24 hrs varied, between 1 and 23% for D. marginatus and between 7 and 27% for H. punctata and respectively between 2 and 53% after 24 hrs, for I. ricinus, comparable effects being also found 72 hrs after the second administration of Euphorbia extracts.
Conclusion
Extracts from E. cyparissias may be used, with results, as an ecologic alternative tick control management method, being a cheap solution, with a sizeable role in reducing the use of synthetic and/or other harming and resistance source ixodicidal conditionings.
PMCID: PMC4202396  PMID: 25392580
bio-control; Euphorbia; Acari: Ixodidae; ruminants
22.  The Inhibitory Activity of the Extracts of Popular Medicinal Herbs on CYP1A2, 2C9, 2C19 and 3A4 and the Implications for Herb-Drug Interaction 
Background
Studies have suggested an increasing practice of concurrent herb-drug consumption. One of the major clinical risks of such concomitant herb-drug use is pharmacokinetic herb-drug interaction (HDI). This is brought about by the ability of phytochemicals to inhibit or induce the activity of metabolic enzymes. The aim of this study was to investigate the potential of the crude aqueous extracts of three popular medicinal herbs used in South Africa to inhibit major cytochrome P450 (CYP) enzymes.
Materials and Methods
The extracts of Bowiea volubilis, Spirostachys africana and Tulbaghia violacea were incubated with human liver microsomes (HLM) to monitor the phenacetin O-deethylation, diclofenac 4′-hydroxylation, S-mephenytoin 4′-hydroxylation and testosterone 6β-hydroxylation as respective probe reactions for CYP1A2, CYP2C9, CYP2C19 and CYP3A4. The inhibitory activity, where observed, was profiled against the extract concentration.
Results
Extracts of Bowiea volubilis inhibited the metabolic activity of CYP1A2 and CYP3A4 with IC50 values of 92.3 ± 5.5 µg/mL and 8.1 ± 0.6 µg/mL respectively. Similar observation with Spirostachys africana showed inhibitory activity against CYP1A2 and CYP3A4 with respective IC50 values of 14.3 ± 0.6 µg/mL and 47.4 ± 2.4 µg/mL. Tulbaghia violacea demonstrated relatively weak inhibitory activity against CYP1A2 (767.4 ± 10.8 µg/mL) and CYP2C9 (921 ± 15.3 µg/mL).
Conclusion
The results suggest the potential for HDI between the herbs and the substrates of the affected enzymes, if sufficient in vivo concentration is attained.
PMCID: PMC4202397  PMID: 25392581
Cytochrome P450; drug metabolism; enzyme inhibition; herb-drug interaction; liver microsomes
23.  Cytotoxicity of Selected Medicinal Plants Used in Mt. Frere District, South Africa 
Background
In South African traditional medicine, some are plants known to combat pediatric diseases and are commonly used by traditional healers. The aim was to evaluate cytotoxicity effects of plants.
Materials and methods
The ground plant material was exhaustively extracted using methanol, acetone and water separately for 72 hrs. These organic solvents were removed from filtrates using a rotavapour. Stock solutions were prepared at 40 mg/ml Dimethyl sulfoxide (DMSO) and test solutions were transferred into vials and 10 brine shrimps introduced in each. The number of dead shrimps was counted to ascertain toxicity. Ten A. salina nauplii (larva) were transferred into each sample vial and filtered brine solution was added to make 5 ml. The nauplii were counted macroscopically in the stem of the pipette against a lighted background. A drop of dry yeast suspension was added as food to each vial. Probit analysis was used to determine the concentration at which lethality to brine shrimp represents 50 % (LC50).
Results
All the tested extracts showed that the concentration is directly proportional to death of brine shrimps. Fifty percent lethality (LC50) of the tested crude extract ranged between 4.1 and 4.6 µg/ml with methanol extract of B. abyssinica being the lowest and T. acutiloba the highest.
Conclusion
This study revealed that 100% of plant crude extracts screened for activity against Artemisia salina larvae showed strong cytotoxicity below 10 µg/ml and plant species with LC50 values < 1000 µg/ml may not make good paediatric remedies due to their inherent toxicity.
PMCID: PMC4202398  PMID: 25392582
Medicinal plants; traditional pediatrics; cytotoxicity
24.  Chemopreventive Activity of Methanol Extract of Melastoma Malabathricum Leaves in DMBA-Induced Mouse Skin Carcinogenesis 
Background
Melastoma malabathricum L. Smith (family Melastomaceae) is a shrub that has been used by the Malay practitioners of traditional medicine to treat various types of ailments. The present study aimed to determine the chemopreventive activity of methanol extract of M. malabathricum leaves (MEMM) using the standard 7,12-dimethylbenz(α)anthracene (DMBA)/croton oil-induced mouse skin carcinogenesis model.
Materials and Methods
In the initiation phase, the mice received a single dose of 100µl/100 µg DMBA (group I–V) or 100µl acetone (group VI) topically on the dorsal shaved skin area followed by the promotion phase involving treatment with the respective test solutions (100 µl of acetone, 10 mg/kg curcumin or MEMM (30, 100 and 300mg/kg)) for 30 min followed by the topical application of tumour promoter (100µl croton oil). Tumors were examined weekly and the experiment lasted for 15 weeks.
Results
MEMM and curcumin significantly (p<0.05) reduced the tumour burden, tumour incidence and tumour volume, which were further supported by the histopathological findings.
Conclusion
MEMM demonstrated chemoprevention possibly via its antioxidant and anti-inflammatory activities, and the action of flavonoids like quercitrin.
PMCID: PMC4202399  PMID: 25392583
Melastomaceae; skin cancer; anti-carcinogenic activity
25.  Foliar Micro-Morphology of Gasteria Bicolor Haw. (Asphodelaceae) from South Africa 
Background
The succulent genus, Gasteria, which comprises 16 species, is endemic to South Africa and has its main centre of distribution in the Savanna Region of the Eastern Cape. Whereas G. bicolor has been investigated phyto-chemically and pharmacologically, not much data concerning the anatomical and micro-morphological features can be found in literature.
Materials and Methods
This study was undertaken, using light and scanning electron microscopy to obtain information on the micro-morphological features of this important medicinal plant to facilitate its identification and authentication. The elemental composition of the leaf was determined by energy dispersive X-ray spectroscopy (EDXS).
Results
The epidermal cells are either hexagonal or pentagonal in form, and are compactly arranged with undulate anti-clinal cell walls. The epidermal cell width was approximately 50 µm. Stomata apertures are elliptical and the upper epidermis of the leaf has paracytic stomata which are slightly raised above the epidermal surface with 4 to 5 subsidiary cells surrounding each stoma. Based on the EDXS microanalysis, the mineral crystals present at the level of the mesophyll of G. bicolor were probably mixtures of calcium oxalate, calcium sulphate and silica.
Conclusion
The co-occurrence of aluminum suggests the potential role of the crystals in detoxification of aluminum and heavy metals, as reported previously.
PMCID: PMC4202400  PMID: 25392584
Foliar micro-morphology; Gasteria bicolor; light microscopy; scanning electron microscopy

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