Open Access Short Communication

Enhanced Growth-inhibitory Effect of Microemulsified Curcumin Formulation in Human Prostate Cancer LNCaP Cells

Vaibhav Dubey, Richard Owusu-Apenten

Journal of Pharmaceutical Research International, Page 209-216
DOI: 10.9734/BJPR/2015/14475

Aim: To assess the effect of curcumin microemulsified with non-ionic surfactant surfynol 465 W or dispersed using edible oils on prostate LNCaP cancer cell viability and glutathione status.
Methodology: LNCaP cells were treated for 72-144 hr with curcumin dissolved with fish or corn oil and microemulsified using non-ionic surfactant surfynol 465 W; alternatively LNCaP cells were treated with curcumin directly dispersed in fish or corn oil (0-50 μM) for 24 -72-144 hr. Cell viability was determined using resazurin (Vision blueTM) fluorescence assay. Glutathione status was determined by monochlorobimane (MCB) assay.
Results: Treatment with 0-34 μM of microemulsified curcumin produced moderate cytotoxic effect on LNCaP cells, no 50% reduction of cell viability was observed graphically. However, when LNCaP cells were treated with curcumin dispersed with corn oil the concentration or 50% reduction of cell viability (IC50) was 12-45 μM. Similarly for cells treated with curcumin dispersed with fish oil, the IC50 was between 20-40 μM. Cytotoxic doses of curcumin dispersed with corn or fish oil increased GST status in cells by 272-656% (p =<0.01).
Conclusion: Microemulsified curcumin formulation prepared using fish or corn oil and surfynol 465 W surfactant had an inhibitory effect on viability of LNCaP cells as did direct dispersion of curcumin in fish or corn oil coupled with the ability for inducing intracellular GST status in LNCaP cells.

Open Access Original Research Article

Medicinal Plants Constituting Antimalarial Herbal Preparations in the Ghanaian Market

Samuel N. Osei-Djarbeng, Emmanuel Agyekum-Attobra, Rosemond Nkansah, Daniel Solaga, Samuel Osei-Asante, George Owusu-Dapaah

Journal of Pharmaceutical Research International, Page 153-162
DOI: 10.9734/BJPR/2015/14896

Aims: Malaria is hyper-endemic in many parts of Ghana, and a greater percentage of the population use herbal preparations to treat the disease. This study was conducted to find out the medicinal plants that constitute the components of antimalarial herbal mixtures sold in Ghana.
Place and Duration of Study: The study was conducted in Accra (the capital city of Ghana) and Kumasi (the second largest city in the country) by researchers from the Department of Pharmaceutical Sciences at Kumasi Polytechnic. It was carried out between May and July, 2012.
Methodology: The study was based on direct observation and recording of information on available antimalarial herbal products in some Pharmacies and Herbal Medicine Shops in the two cities.
Results: The study identified forty one different herbal mixtures used as antimalarials. Fifty seven plants belonging to 28 plant families, and one animal product (honey), were found to be the components of the herbal mixtures. Cryptolepis sanguinolenta and Azadirachta indica, respectively found in 29.3% and 22.0% of the preparations, were the commonest plant components.
Most of the plant components found are reported in traditional medicine or pharmacological studies to treat malaria. These antimalarial claims are indicative of indigenous knowledge in the management of malaria.
Conclusion: With the necessary product standardisation and safety investigations, some of the antimalarial herbal products may offer alternative treatment of malaria in Ghana.

Open Access Original Research Article

GC-MS Determination of Bioactive Constituents of the Methanolic Fractions of Cnidoscolus aconitifolius

Ngozi K. Achi, O. C. Ohaeri

Journal of Pharmaceutical Research International, Page 163-172
DOI: 10.9734/BJPR/2015/13893

Background: Diabetes mellitus is a major metabolic disorder affecting a huge population all over the world. Cnidoscolus species have been extensively used for the management of diabetes in folkloric medicine. The presence of diverse secondary metabolites has been reported from species of the genus Cnidoscolus. However, there has not been much information available on phytochemical components and biological activity in the leaf methanol extract of Cnidoscolus aconitifolius.
Objective: This study was designed to extract and identify some bioactive compounds in the leaf methanol fractions of C. aconitifolius which may provide insight on its pharmacological properties and its use in traditional medicine.
Place and Duration of Study: Department of Biochemistry, Michael Okpara University of Agriculture, Umudike and National Cereals Research Institute Zaria, Nigeria between June 2012 and July 2013.
Methodology: Twenty grams of the powdered sample was subjected to column chromatography over silica gel (60-120 mesh) and eluted with 100 ml each of n-hexane, petroleum ether, chloroform, methanol and respectively at the rate of 1ml/min. The eluates were concentrated and labeled as FI, F2, F3 and F4. The percentage yields of the fractions were 7.55%, 6.00%, 15.5%, and 65.00% (w/w) respectively. n-hexane and petroleum ether did not elute much of the compounds. The active methanol fraction of C. aconitifolius extract (F4) which showed the highest hypoglycaemic effect as identified by Oral Glucose Tolerance Test (OGTT) in rats was taken for Gas Chromatography- Mass Spectrometry (GC-MS) analysis for separation of the bioactive components. GC-MS analysis was performed using a GC-MS (Model: QP2010 PLUS SHIMADZU, JAPAN) comprising a AOC-20i auto-sampler and gas-chromatograph interfaced to a mass spectrometer.
Results: The bioactive organic components of the GC-MS analysis provided peaks of six different phytochemical compounds, with their retention time (RT) and peak area (PA) in addition to minor constituents. The major compounds are dodecanoic acid-1, 2, 3- propanetriyl ester ( RT:25.74, PA: 51.18%), cyclotetradecane (RT: 23.39, PA:15.59%), eicosanoic acid (RT:20.61, PA:18.47%); octadecanoic acid (RT: 16.82, PA:1.21%), 4-nitrosophenyl-beta-phenyl propionate (RT: 11.53, PA: 4.38%), benzene acetic acid, phenyl malonic acid and 3-oxo-4-phenylbutyronitrile (RT:10.34, PA: 9.17%). The presence of these compounds in the plant extract may at least be responsible for one of the pharmacological properties of C. aconitifolius and thus recommended as plant of phyto-pharmaceutical importance

Open Access Original Research Article

Comparison of Three Nutriceutical Food Supplements for the Treatment of Infertility

Frank Comhaire

Journal of Pharmaceutical Research International, Page 173-180
DOI: 10.9734/BJPR/2015/15105

Aims: Approximately one out of every 8 couples is confronted with a problem of infertility. The diagnostic and therapeutic management, taking into account the WHO recommendations, may be complemented by nutriceutical food supplementation. The present study compares the original nutriceutical (Qualisperm®) with two recently marketed products (Proxeed plus®, and PROfertil®).
Study Design: The ingredients and their quantities of the three nutriceuticals were listed and compared, and their effectiveness on semen quality was assessed. The probability of conception per month was calculated, as were the numbers needed to treat, and the time to pregnancy, whenever available.
Results: All three nutriceuticals improve semen quality. The pregnancy rate per month of Proxeed plus was 3.7%, it was 5% for PROfertile, and 11% when Qualisperm was combined with WHO-recommended treatment of the male partner. The numbers needed to treat were 5.0 (CI:3.58-8.18) for Proxeed plus, 9.4 (CI: 4.46-94.9) for PROfertil, and 2.8 (CI: 1.7-8.7) for Qualisperm added to WHO-recommended treatment. The combination of Proxeed plus with the non-steroidal anti-inflammatory agent Cinnoxicam generated a number needed to treat of 2.9 (CI: 2.32-3.98). The time to pregnancy was reduced to half when combining WHO-recommended treatment with Qualisperm, as compared to either WHO recommended treatment alone, or treatment with Proxeed plus with Cinnoxicam added. In a placebo-controlled double-blind randomised trial of in vitro fertilisation the number needed to treat of Qualisperm, given together with a specific oil to both partners, was 4.
Conclusion: Since confounding factors may have influenced the findings of the present comparison, the results should be interpreted with care. It is concluded that all three nutriceuticals may improve fertility, but that the efficiency of Qualisperm is best documented.

Open Access Original Research Article

Histomorphological Evaluation of Reproductive Organs Following Piper betel (Linn.) Leaf Stalk Extract Administration in Male Albino Rats

V. Vengaiah, A. Govardhan Naik, C. Changamma

Journal of Pharmaceutical Research International, Page 181-191
DOI: 10.9734/BJPR/2015/14247

Aim: To evaluate the histological effects of Piper betel leaf stalk extract on reproductive organs and the liver in male rats
Study Design: Aqueous P. betel leaf stalk extract was administered to rats at the dose of 50 mg/Kg/day by oral gavage for 15 days. Twenty four hours after the last dose, the animals were autopsied. Testis, epididymis, seminal vesicles, prostate and the liver were isolated and fixed immediately in Bouin’s fluid for 24 hours. The tissues were dehydrated in various grades of alcohol, cleaned in xylene and after embedding in paraffin, blocks were prepared. Paraffin sections of 5µ thickness were cut with rotary microtome and processed for staining.
Place and Duration of Study: Place of study was S.V. University, Department of Zoology, Tirupati, India and experiment lasted for 6 months.
Results: P. betel leaf stalk extract administration caused seminiferous tubular derangement and sloughing of germ cells in testes. The histological changes were more pronounced in caput epididymis, reduction in lumen size due to creeping in of mucosal folds and lowering of secretion. Hepatic cell proliferation and Kupffer cell activation were also observed in liver of extract treated rats.
Conclusion: Piper betel leaf stalk extract is deleterious to histology or architecture of reproductive organs, which may affect spermatogenic function of the testis.

Open Access Original Research Article

New Cembranoid Diterpenes from Sarcophyton trocheliophorum

Khaled A. Shaaban, Mohamed A. Ghani, Mohamed Shaaban

Journal of Pharmaceutical Research International, Page 192-201
DOI: 10.9734/BJPR/2015/14757

Aims: To isolate the unpolar bioactive diterpenes in pure forms from the soft coral Sarcophyton trocheliophorum, collected from Red Sea at the Egyptian coasts, and to identify their structures using diverse spectroscopic means (NMR [1D&2D] and MS). Additionally, to study the antimicrobial and cytotoxicity of the isolated compounds compared with the main crude extract of the soft coral.
Study Design: Collection of the soft coral, Sarcophyton trocheliophorum from Red Sea at coast of Hurgada, east Egypt, its homogenization, maceration in suitable organic solvent for extraction of the desired bioactive compounds and their purification by a series of different chromatographic techniques, identification of the compounds by mass spectrometry and NMR (1D&2D) spectroscopy, and then determination of the antimicrobial and cytotoxicity for them compared with their original crude extract.
Place and Duration of Study: Institute of Organic and Biomolecular Chemistry, University of Göttingen, Germany; and Chemistry of Natural Compounds Department, National Research Centre, Egypt, between August, 2008 and December, 2010.
Methodology: The soft coral Sarcophyton trocheliophorum was homogenized in a blender, macerated with chloroform-methanol, and the chloroform layer was concentrated in vacuo; while aqueous methanol solution was re-extracted with n-butanol, and the latter was concentrated to dryness. The resultant whole crude extract was fractionated using several subsequent column chromatographies on silica gel, and the fast fractions containing the unpolar components were purified by different column chromatographies (silica gel, Sephadex column and preparative TLC) to deliver the desired diterpenes in pure form. Structures of the afforded diterpenes were identified by mass spectrometry (ESI and HRESI-MS), NMR analysis (1H,13C NMR, and 2D NMR) and by comparison with reference data. The antimicrobial activity was determined by disc diffusion assay, while the cytotoxicity was determined using brine shrimp assay.
Results: Two new unpolar diterpenes, cis-cembrene C (1) and cis-cembrenene C (2) were isolated along with (+)-sarcophine (3), (+)-sarcophytoxide (4) and (-)-sarcophytoxide (5), from the chloroform-soluble extract of soft coral Sarcophyton trocheliophorum, collected from Red Sea. Four additional unpolar sesquiterpenes; ß-elemene (6), caryophyllene (7), alloaromadendrene (8) and 1-methyl-4-(5-methyl-1-methylene-hex-4-enyl)-cyclohexene; bisabolene (9) were identifed by GC-MS analysis. Structures of the new diterpenes 1-2 were identified by spectroscopic data (1H, 13C, 1H-1H COSY, HMQC, HMBC, HREI-MS and HRESI-MS) interpretation and comparison with related structures. The antimicrobial and cytotoxic activities of compounds 1-5 were evaluated in comparison with the original soft coral crude extract.
Conclusion: The soft coral Sarcophyton trocheliophorum is a prolific source for production of numerous bioactive compounds with diverse structures and biological activities which can be exploited for their commercial production.

Open Access Original Research Article

In vitro Antimicrobial Activity of Commercially Available Melaleuca alternifolia (Tea Tree) Oil on Some Selected Clinical Pathogens

Ephraim Ogbaini-Emovon, Helmut Schuster, Jarret Waze, Kalu Eziyi Iche

Journal of Pharmaceutical Research International, Page 202-208
DOI: 10.9734/BJPR/2015/14544

Aim: To evaluate the In vitro antimicrobial activity of Melaleuca alternifolia oil (tea tree oil), (TTO) on selected clinical isolates of fungi and bacteria, including some multidrug resistant strains commonly associated with nosocomial infection.
Study Design: Laboratory experimental study.
Place and Duration of Study: Public Health England (PHE), Microbiology laboratory, Southampton University Hospital, England, May 2014.
Methodology: Three different concentrations (1%, 5% and 10%) of commercially available TTO were prepared (%v/v). Using an improvised disc diffusion antibiotic susceptibility testing method, the activity of TTO was tested against pure reference strains of Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, clinical isolates of Candida albicans, Pseudomonas aeruginosa, Streptococcus pyogenes, Bacteroides fragilis, methicillin- resistant Staphylococcus aureus (MRSA), extended spectrum beta lactamase Escherichia coli (ESBL) vancomycin-resistant Enterococcus faecium (VRE), and Carbapenemase-producing Klebsiella pneumoniae (CRKP). The zones of inhibition were measure for each organism at the various concentrations of TTO.
Results: All the organisms tested showed susceptibility to TTO at concentration range of 5% - 10%, except Pseudomonas aeruginosa. Staphylococcus aureus ATCC 25923, MRSA, and VRE did not show any susceptibility at TTO concentration of 1%. The zones of inhibition for the susceptible organisms ranged between 10mm-19mm at 5% concentration and 15 mm- 36 mm at 10% concentration of TTO.
Conclusion: TTO has a broad spectrum of In vitro activity against clinical pathogens. The essential oil or its active ingredient could be of potential benefit in the treatment of fungal, common bacterial and multidrug-resistant bacteria usually associated with hospital-acquired infections as well as mixed bacterial infections involving anaerobic bacteria.