Asian Pacific Journal of Tropical Biomedicine

Issue 3,2014 Table of Contents

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  • 1  Isolation and structural elucidation of cytotoxic compounds from the root bark of Diospyros quercina (Baill.) endemic to Madagascar
    Fatiany Pierre Ruphin Robijaona Baholy Randrianarivo Emmanuel Raharisololalao Amelie Marie-Therese Martin Ngbolua Koto–te-Nyiwa
    2014(3):169-175. DOI: 10.1016/S2221-1691(14)60227-6
    [Abstract](40) [HTML](0) [PDF 443.34 K](154)
    Abstract:
    Objective: To isolate and characterize the cytotoxic compounds from Diospyros quercina (Baill.) G.E. Schatz & Lowry (Ebenaceae). Methods: An ethno-botanical survey was conducted in the south of Madagascar from July to August 2010. Bio-guided fractionation assay was carried out on the root bark of Diospyros quercina, using cytotoxicity bioassay on murine P388 leukemia cell lines as model. The structures of the cytotoxic compounds were elucidated by 1D and 2D NMR spectroscopy and mass spectrometry. Results: Biological experiments resulted in the isolation of three bioactive pure compounds (named TR-21, TR-22, and TR-23) which exhibited very good in vitro cytotoxic activities with the IC50 values of (0.017 5±0.0060) µg/mL, (0.089±0.005) µg/mL and (1.027±0.070) µg/mL respectively. Thus, they support the claims of traditional healers and suggest the possible correlation between the chemical composition of this plant and its wide use in Malagasy folk medicine to treat cancer. Conclusions: The ability of isolated compounds in this study to inhibit cell growth may represent a rational explanation for the use of Diospyros quercina root bark in treating cancer by Malagasy traditional healers. Further studies are, therefore, necessary to evaluate the in vivo antineoplastic activity of these cytotoxic compounds as effective anticancer drugs.
    2  Comparative micromorphological study of wild and micropropagated Dioscorea bulbifera Linn.
    Mubo A. Sonibare Adedapo A. Adeniran
    2014(3):176-183. DOI: 10.1016/S2221-1691(14)60228-8
    [Abstract](25) [HTML](0) [PDF 2.45 M](118)
    Abstract:
    Objective: To study the leaf epidermis of wild and micropropagated Dioscorea bulbifera Linn. (D. bulbifera) in order to document useful diagnostic features that may be employed for correct crude drug identification and to clear any taxonomic uncertainties in the micropropagated medicinal plant. Methods: Growth responses of micropropagated D. bulbifera were observed on Murashige Skoog medium supplemented with 6-benzylamino purine (1.0 mg/L)+α-naphthaleneacetic acid (0.2 mg/ L)+cysteine (20 mg/L) using nodal segments as explants. Leaves of the wild and micropropagated plants were studied microscopically. Results: More than 80% shoot regeneration and formation of 10%-30% whitish-brown callus were observed within 3 weeks. The highest root proliferation was obtained from Murashige Skoog medium of 6-benzylamino purine (0.05 mg/L) and α-naphthaleneacetic acid (0.01 mg/L) with mean root length of (27.00±1.25) mm and elongated single shoot of mean length (38.00±11.09) mm. Leaf epidermal features that revealed similarities between the wild and micropropagated plants included amphistomatic condition, presence of mucilage, glandular unicellular trichome with multicellular head, polygonal cells with smooth walls, stomata type and shape. Slight variations included thick cuticular wall with closed stomata in wild plant compared to thin walled opened stomata in the in vitro plant. Opening of stomata accounted for larger average stomata sizes of (7.68±0.38) µm and (6.14±0.46) µm on the adaxial and abaxial surfaces, respectively of the micropropagated plant compared to the wild. Conclusions: The diagnostic features obtained in the study could serve as a basis for proper identification for quality control for standardization of the medicinal plant.
    3  15-Lipoxygenase inhibition of Commelina benghalensis, Tradescantia fluminensis, Tradescantia zebrina
    Cean Socorro M. Alaba Christine L. Chichioco-Hernandez
    2014(3):184-188. DOI: 10.1016/S2221-1691(14)60229-X
    [Abstract](24) [HTML](0) [PDF 308.71 K](247)
    Abstract:
    Objective: To evaluate the 15-lipoxygenase inhibitory activity of the methanol leaf extracts of Commelina benghalensis, Tradescantia fluminensis (T. fluminensis) and Tradescantia zebrina. Method: The inhibitory activity was evaluated using a spectrophotometric assay by observing the increase in absorbance at 234 nm due to the formation of the product 13- hydroperoxyoctadecadienoic acid. The extracts were also tested for the presence of terpenoids, saponins, tannins, flavonoids, steroids, phenolic compounds, alkaloids and cardiac glycosides. Results: All the extracts inhibited the action of 15-lipoxygenase at a concentration of 0.2 µg/mL. T. fluminensis and Tradescantia zebrina exhibited higher than 50% inhibition with T. fluminensis at 87.2%. T. fluminensis was partitioned with ethyl acetate and hexane and their IC50 values were determined at 8.72 µg/mL and 98.04 µg/mL, respectively. Conclusions: T. fluminensis is a potentially good source of 15-lipoxygenase inhibitors.
    4  Phytochemical constituents and antibacterial activity of some green leafy vegetables
    Ramesa Shafi Bhat Sooad Al-Daihan
    2014(3):189-193. DOI: 10.1016/S2221-1691(14)60230-6
    [Abstract](12) [HTML](0) [PDF 297.51 K](162)
    Abstract:
    Objective: To investigate the antibacterial activity and photochemicals of five green leafy vegetables against a panel of five bacteria strains. Methods: Disc diffusion method was used to determine the antibacterial activity, while kanamycin was used as a reference antibiotic. The phytochemical screening of the extracts was performed using standard methods. Results: All methanol extracts were found active against all the test bacterial strains. Overall maximum extracts shows antibacterial activity which range from 6 to 15 mm. Proteins and carbohydrates was found in all the green leaves, whereas alkaloid, steroids, saponins, flavonoids, tannins were found in most of the test samples. Conclusions: The obtain result suggests that green leafy vegetables have moderate antibacterial activity and contain various pharmacologically active compounds and thus provide the scientific basis for the traditional uses of the studied vegetables in the treatment of bacterial infections.
    5  Acute and subacute oral toxicity study on the flavonoid rich fraction of Monodora tenuifolia seed in albino rats
    Raphael Chukwuma Ekeanyanwu Obioma Uzoma Njoku
    2014(3):194-202. DOI: 10.1016/S2221-1691(14)60231-8
    [Abstract](21) [HTML](0) [PDF 1.30 M](129)
    Abstract:
    Objective: To investigate the effects of the flavonoid rich fraction of Monodora tenuifolia seed on the haematology, histopathology and liver profile of Wistar albino rats. Methods: Toxicity study was investigated on the flavonoid rich fraction of Monodora tenuifolia in rats administered different concentrations orally for 28 d using standard laboratory procedures. Results: The LD50 of the flavonoid rich fraction was found to be above 5 000 mg/kg body weight in mice observed for 48 h. After the Day 14, biochemical markers of liver injury such as serum alanine aminotransferase, and aspartate aminotransferase decreased significantly (P<0.05 at doses of 100 and 200 mg/kg body weight and P<0.01 at 400 mg/kg) while serum alkaline phosphatase increased non-significantly (P>0.05). There was non-significant (P>0.05) effect observed across the groups in the levels of serum total protein, albumin, globulin, urea and creatinine. The result of histological examination showed various degrees of peribiliary hepatitis after the Day 14 which fizzled out after the Day 28. Conclusions: The result therefore suggests that the seed extract is potentially safe.
    6  Chromatographic finger print analysis of anti-inflammatory active extract fractions of aerial parts of Tribulus terrestris by HPTLC technique
    Mona Salih Mohammed Mohamed Fahad Alajmi Perwez Alam Hassan Subki Khalid Abelkhalig Muddathir Mahmoud Wadah Jamal Ahmed
    2014(3):203-208. DOI: 10.1016/S2221-1691(14)60232-X
    [Abstract](21) [HTML](0) [PDF 649.09 K](124)
    Abstract:
    Objective: To develop HPTLC fingerprint profile of anti-inflammatory active extract fractions of Tribulus terrestris (family Zygophyllaceae). Methods: The anti-inflammatory activity was tested for the methanol and its fractions (chloroform, ethyl acetate, n-butanol and aqueous) and chloroform extract of Tribulus terrestris (aerial parts) by injecting different groups of rats (6 each) with carrageenan in hind paw and measuring the edema volume before and 1, 2 and 3 h after carrageenan injection. Control group received saline i.p. The extracts treatment was injected i.p. in doses of 200 mg/kg 1 h before carrageenan administration. Indomethacin (30 mg/kg) was used as standard. HPTLC studies were carried out using CAMAG HPTLC system equipped with Linomat Ⅳ applicator, TLC scanner 3, Reprostar 3, CAMAG ADC 2 and WIN CATS-4 software for the active fractions of chloroform fraction of methanol extract. Results: The methanol extract showed good antiedematous effect with percentage of inhibition more than 72%, indicating its ability to inhibit the inflammatory mediators. The methanol extract was re-dissolved in 100 mL of distilled water and fractionated with chloroform, ethyl acetate and n-butanol. The four fractions (chloroform, ethyl acetate, n-butanol and aqueous) were subjected to anti-inflammatory activity. Chloroform fraction showed good anti-inflammatory activity at dose of 200 mg/kg. Chloroform fraction was then subjected to normal phase silica gel column chromatography and eluted with petroleum ether-chloroform, chloroform-ethyl acetate mixtures of increasing polarity which produced 15 fractions (F1-F15). Only fractions F1, F2, F4, F5, F7, F9, F11 and F14 were found to be active, hence these were analyzed with HPTLC to develop their finger print profile. These fractions showed different spots with different Rf values. Conclusions: The different chloroform fractions F1, F2, F4, F5, F7, F9, F11 and F14 revealed 4, 7, 7, 8, 9, 7, 7 and 6 major spots, respectively. The results obtained in this experiment strongly support and validate the traditional uses of this Sudanese medicinal plant.
    7  Survey on cattle ticks in Nur, north of Iran
    Ashkan Ghasemi Moghaddam Mozafar Razavi Seyed Mehrshad Rasouli Sajad Hosseinzade Mohammad Mehdi Darvishi Arash Rakhshanpour Mohammad Taghi Rahimi
    2014(3):209-212. DOI: 10.1016/S2221-1691(14)60233-1
    [Abstract](15) [HTML](0) [PDF 998.99 K](122)
    Abstract:
    Objective: To survey the prevalence of cattle ticks in Nur County and prepare a list of tick fauna in this district. Methods: This investigation was carried out on 150 head of cattle ticks of rural areas of Nur city which is located in Mazandaran province during spring and summer seasons of 2011. The collected ticks were identified using light microscope and available systematic keys. Results: A total number of 1 563 ticks were isolated from examined cattle and their genus and numbers including: Ixodes ricinus 51% (111 male and 691 female) and Boophilus annulatus 49% (83 male and 678 female), respectively. Conclusions: Results of the current investigation indicate the presence of two species of acarine ectoparasites which have potential health risk Ixodes ricinus and Boophilus annulatus. More studies are required to increase our data concerning ticks and other ectopreasites of ruminants in other areas of Mazandaran province and should be noted to their ability in transmission of infectious agents.
    8  Long-term spatial memory and morphological changes in hippocampus of Wistar rats exposed to smoke from Carica papaya leaves
    Aboyeji Lukuman Oyewole Bamidele Victor Owoyele
    2014(3):213-218. DOI: 10.1016/S2221-1691(14)60234-3
    [Abstract](8) [HTML](0) [PDF 1.51 M](116)
    Abstract:
    Objective: To investigate the effects of smoking of dried leaves of Carica papaya (pawpaw) based on ethnopharmacological information which indicated that smoking of papaya leaves could influence motor performance and learning. Methods: Twenty-four rats were used for the study, and were grouped into four groups. Groups 1 served as the control (not exposed to papaya leaves smoke), while Groups 2, 3 and 4 were exposed to smoke from 6.25 g, 12.50 g and 18.75 g of dry pawpaw leaves respectively in a smoking chamber twice daily for 21 d with each exposure lasting for 3 min. Lastly, hippocampus was harvested in each group for histological study. Results: The results showed that there were significant (P<0.05) increases in mean of recall latencies of long-term spatial memory in the animal administered the high dose while the other groups had significantly (P<0.05) lower frequencies. Histological investigation showed signs of mild neural degeneration in high dose group and hypochromic appearance of the Nissl substance in all treated groups. Conclusions: In conclusion, the findings from this study has demonstrated that smoking of papaya leaves has the ability to maintain an intact long-term spatial memory at all doses but retrieving such memory is faster with the low and medium dosages.
    9  Anti-oxidative and anti-inflammatory effects of Tagetes minuta essential oil in activated macrophages
    Parastoo Karimian Gholamreza Kavoosi Zahra Amirghofran
    2014(3):219-227. DOI: 10.1016/S2221-1691(14)60235-5
    [Abstract](16) [HTML](0) [PDF 993.70 K](121)
    Abstract:
    Objective: To investigate antioxidant and anti-inflammatory effects of Tagetes minuta (T. minuta) essential oil. Methods: In the present study T. minuta essential oil was obtained from leaves of T. minuta via hydro-distillation and then was analyzed by gas chromatography-mass spectrometry. The antioxidant capacity of T. minuta essential oil was examined by measuring reactive oxygen, reactive nitrogen species and hydrogen peroxide scavenging. The anti-inflammatory activity of T. minuta essential oil was determined through measuring NADH oxidase, inducible nitric oxide synthase and TNF-α mRNA expression in lipopolysacharide-stimulated murine macrophages using realtime PCR. Results: Gas chromatography-mass spectrometry analysis indicated that the main components in the T. minuta essential oil were dihydrotagetone (33.86%), E-ocimene (19.92%), tagetone (16.15%), cis-β-ocimene (7.94%), Z-ocimene (5.27%), limonene (3.1%) and epoxyocimene (2.03%). The T. minuta essential oil had the ability to scavenge all reactive oxygen/reactive nitrogen species radicals with IC50 12-15 μg/mL, which indicated a potent radical scavenging activity. In addition, T. minuta essential oil significantly reduced NADH oxidase, inducible nitric oxide synthaseand TNF-α mRNA expression in the cells at concentrations of 50 μg/mL, indicating a capacity of this product to potentially modulate/diminish immune responses. Conclusions: T. minuta essential oil has radical scavenging and anti-inflammatory activities and could potentially be used as a safe effective source of natural anti-oxidants in therapy against oxidative damage and stress associated with some inflammatory conditions.
    10  Hesperidin as a preventive resistance agent in MCF-7 breast cancer cells line resistance to doxorubicin
    Rifki Febriansah Dyaningtyas Dewi P.P. Sarmoko Nunuk Aries Nurulita Edy Meiyanto Agung Endro Nugroho
    2014(3):228-233. DOI: 10.1016/S2221-1691(14)60236-7
    [Abstract](22) [HTML](0) [PDF 1.05 M](144)
    Abstract:
    Objective: To evaluate of hesperidin to overcome resistance of doxorubicin in MCF-7 resistant doxorubicin cells (MCF-7/Dox) in cytotoxicity apoptosis and P-glycoprotein (Pgp) expression in combination with doxorubicin. Methods: The cytotoxic properties, 50% inhibition concentration (IC50) and its combination with doxorubicin in MCF-7 cell lines resistant to doxorubicin (MCF-7/Dox) cells were determined using MTT assay. Apoptosis induction was examined by double staining assay using ethidium bromide-acridine orange. Immunocytochemistry assay was performed to determine the level and localization of Pgp. Results: Single treatment of hesperidin showed cytotoxic activity on MCF-7/Dox cells with IC50 value of 11 µmol/L. Thus, combination treatment from hesperidin and doxorubicin showed addictive and antagonist effect (CI>1.0). Hesperidin did not increase the apoptotic induction, but decreased the Pgp expressions level when combined with doxorubicin in low concentration. Conclusions: Hesperidin has cytotoxic effect on MCF-7/Dox cells with IC50 of 11 µmol/ L. Hesperidin did not increased the apoptotic induction combined with doxorubicin. Cochemotherapy application of doxorubicin and hesperidin on MCF-7/Dox cells showed synergism effect through inhibition of Pgp expression.
    11  Prevalence and antimicrobial resistance of Salmonella spp. in raw retail frozen imported freshwater fish to Eastern Province of Saudi Arabia
    Nasreldin Elhadi
    2014(3):234-238. DOI: 10.1016/S2221-1691(14)60237-9
    [Abstract](21) [HTML](0) [PDF 286.78 K](139)
    Abstract:
    Objective: To determine the proportion of imported frozen fish contaminated with Salmonella among retail food stores and supermarkets in the Eastern Province of Saudi Arabia. Methods: A total of 223 frozen freshwater fish purchased from different supermarkets and grocery stores were analyzed for the presence of foodborne pathogen Salmonella. The isolation of Salmonella was determined and confirmed by using the methods of US Food and Drug Administration’s Bacteriological Analytical Manual, CHROMagar Salmonella plus, biochemical tests and API 20E strips. Antimicrobial susceptibilities of Salmonella isolates were determined by the disk diffusion method on Muller-Hinton agar, as described by Kirby-Bauer, in accordance with the guidelines of the Clinical and Laboratory Standards Institute. Results: Out of the total 223 fish samples (20 of catfish, 18 of carfu, 20 of mirgal, 25 of milkfish, 35 of mackerel, 75 of tilapia, and 30 of rohu), 89 (39.9%) were tested positive for Salmonella. The prevalence of positive samples were reported for the freshwater fish of pangas (60.0%, n=12), carfu (27.7%, n=5), mirgal (35.0%, n=7), milkfish (52.0%, n=13), mackerel (31.4 %, n=11), tilapia imported from Thailand (64.0%, n=16), tilapia imported from India (28.0%, n=14), rohu imported from Thailand (26.6%, n=4) and rohu imported from Myanmar (46.6%, n=7). A total of 140 isolates of Salmonella spp. were yielded from at least seven different types of frozen freshwater fish imported from 5 different countries and were tested for their susceptibility to 16 selected antimicrobial agents. The highest antibiotic resistance was observed to tetracycline (90.71%) followed by ampicillin (70%) and amoxicillin-clavulanic acid (45%). Conclusions: The obtained results of this study shows that these raw retail imported frozen freshwater fish are contaminated with potentially pathogenic Salmonella spp. And the study recommend and suggest that there is a need for adequate consumer measures.
    12  Prevalence and antimicrobial resistance of Salmonella spp. in raw retail frozen imported freshwater fish to Eastern Province of Saudi Arabia
    Nazaire A?zoun Rock A?kpon Martin Akogbéto
    2014(3):239-243. DOI: 10.1016/S2221-1691(14)60238-0
    [Abstract](18) [HTML](0) [PDF 475.52 K](126)
    Abstract:
    Objective: To determine the proportion of imported frozen fish contaminated with Salmonella among retail food stores and supermarkets in the Eastern Province of Saudi Arabia. Methods: A total of 223 frozen freshwater fish purchased from different supermarkets and grocery stores were analyzed for the presence of foodborne pathogen Salmonella. The isolation of Salmonella was determined and confirmed by using the methods of US Food and Drug Administration’s Bacteriological Analytical Manual, CHROMagar Salmonella plus, biochemical tests and API 20E strips. Antimicrobial susceptibilities of Salmonella isolates were determined by the disk diffusion method on Muller-Hinton agar, as described by Kirby-Bauer, in accordance with the guidelines of the Clinical and Laboratory Standards Institute. Results: Out of the total 223 fish samples (20 of catfish, 18 of carfu, 20 of mirgal, 25 of milkfish, 35 of mackerel, 75 of tilapia, and 30 of rohu), 89 (39.9%) were tested positive for Salmonella. The prevalence of positive samples were reported for the freshwater fish of pangas (60.0%, n=12), carfu (27.7%, n=5), mirgal (35.0%, n=7), milkfish (52.0%, n=13), mackerel (31.4 %, n=11), tilapia imported from Thailand (64.0%, n=16), tilapia imported from India (28.0%, n=14), rohu imported from Thailand (26.6%, n=4) and rohu imported from Myanmar (46.6%, n=7). A total of 140 isolates of Salmonella spp. were yielded from at least seven different types of frozen freshwater fish imported from 5 different countries and were tested for their susceptibility to 16 selected antimicrobial agents. The highest antibiotic resistance was observed to tetracycline (90.71%) followed by ampicillin (70%) and amoxicillin-clavulanic acid (45%). Conclusions: The obtained results of this study shows that these raw retail imported frozen freshwater fish are contaminated with potentially pathogenic Salmonella spp. And the study recommend and suggest that there is a need for adequate consumer measures.
    13  Katayama syndrome in patients with schistosomiasis
    Shailendra Kapoor
    2014(3):244-244. DOI: 10.1016/S2221-1691(14)60239-2
    [Abstract](6) [HTML](0) [PDF 201.20 K](114)
    Abstract:
    14  An update on Ayurvedic herb Convolvulus pluricaulis Choisy
    Parul Agarwa Bhawna Sharma Amreen Fatima Sanjay Kumar Jain
    2014(3):245-252. DOI: 10.1016/S2221-1691(14)60240-9
    [Abstract](26) [HTML](0) [PDF 583.80 K](123)
    Abstract:
    Convolvulus pluricaulis Choisy (C. pluricaulis) is a perennial herb that seems like morning glory. All parts of the herb are known to possess therapeutic benefits. The plant is used locally in Indian and Chinese medicine to cure various diseases. It is used in Ayurvedic formulation for chronic cough, sleeplessness, epilepsy, hallucinations, anxiety etc. Based on the comprehensive review of plant profile, pharmacognosy, phytochemistry, pharmacological and toxicological data on the C. pluricaulis, there will be more opportunities for the future research and development on the herb C. pluricaulis. Information on the C. pluricaulis was collected via electronic search (using Pub Med, SciFinder, Google Scholar and Web of Science) and library search for articles published in peer-reviewed journals. Furthermore, information also was obtained from some local books on ethnopharmacology. This paper covers the literature, primarily pharmacological, from 1985 to the end of 2012. The C. pluricaulis is an important indigenous medicine, which has a long medicinal application for liver disease, epileptic disease, microbial disease, cytotoxic and viral diseases, central nervous system (CNS) disease in Ayurvedic medicine, traditional Chinese medicine and other indigenous medical systems. The isolated metabolites and crude extract have exhibited a wide of in vitro and in vivo pharmacological effect, including CNS depression, anxiolytic, tranquillizing, antidepressant, antistress, neurodegenerative, antiamnesic, antioxidant, hypolipidemic, immunomodulatory, analgesic, antifungal, antibacterial, antidiabetic, antiulcer, anticatatonic, and cardiovascular activity. A chemical study of this plant was then initiated, which led to the isolation of carbohydrats, proteins, alkaloids, fatty acids, steroids, coumarins, flavanoids, and glycosides as active chemicals that bring about its biological effects. A series of pharmacognostical studies of this plant show that it is a herb, its stem and leaves are hairy, more over it has two types of stomata, anisocytic and paracytic. A herb, C. pluricaulis has emerged as a good source of the traditional medicine for the treatment of liver disease, epileptic disease, microbial disease, cytotoxic and viral diseases, and CNS disease. Pharmacological results have validated the use of this species in traditional medicine. All the parts of the herb are known to possess therapeutic benefits. Expansion of research materials would provide more opportunities for the discovery of new bioactive principles from C. pluricaulis.

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