Asian Pacific Journal of Tropical Biomedicine

Issue 10,2013 Table of Contents

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  • 1  African eggplant (Solanum anguivi Lam.) fruit with bioactive polyphenolic compounds exerts in vitro antioxidant properties and inhibits Ca2+-induced mitochondrial swelling
    Olusola Olalekan Elekofehinti Jean Paul Kamdem Aline Augusti Bolingon Margareth Linde Athayde Seeger Rodrigo Lopes Emily Pansera Waczuk Ige Joseph Kade Isaac Gbadura Adanlawo Joao Batista Teixeira Rocha
    2013(10):757-766. DOI: 10.1016/S2221-1691(13)60152-5
    [Abstract](28) [HTML](0) [PDF 1.57 M](139)
    Objective: To evaluate the antioxidant and radical scavenging activities of Solanum anguivi fruit (SAG) and its possible effect on mitochondrial permeability transition pore as well as mitochondrial membrane potential (ΔΨm) isolated from rat liver. Methods: Antioxidant activity of SAG was assayed by using 2,2-diphenyl-1-picrylhydrazyl (DPPH), reducing power, iron chelation and ability to inhibit lipid peroxidation in both liver and brain homogenate of rats. Also, the effect of SAG on mitochondrial membrane potential and mitochondrial swelling were determined. Identification and quantification of bioactive polyphenolics was done by HPLC-DAD. Results: SAG exhibited potent and concentration dependent free radical-scavenging activity (IC50/DPPH=275.03±7.8 µg/mL). Reductive and iron chelation abilities also increase with increase in SAG concentration. SAG also inhibited peroxidation of cerebral and hepatic lipids subjected to iron oxidative assault. SAG protected against Ca2+ (110 µmol/L)-induced mitochondrial swelling and maintained the ΔΨm. HPLC analysis revealed the presence of gallic acid [(17.54±0.04) mg/ g], chlorogenic acid (21.90±0.02 mg/g), caffeic acid (16.64±0.01 mg/g), rutin [(14.71±0.03) mg/g] and quercetin [(7.39±0.05) mg/g]. Conclusions: These effects could be attributed to the bioactive polyphenolic compounds present in the extract. Our results suggest that SAG extract is a potential source of natural antioxidants that may be used not only in pharmaceutical and food industry but also in the treatment of diseases associated with oxidative stress.
    2  Bioefficacy of crude extract of Cyperus aromaticus (Family: Cyperaceae ) cultured cells, against Aedes aegypti and Aedes albopictus mosquitoes
    Fatemeh Kamiabi Zairi Jaal Chan Lai Keng
    2013(10):767-775. DOI: 10.1016/S2221-1691(13)60153-7
    [Abstract](16) [HTML](0) [PDF 886.01 K](134)
    Objective: To evaluate the growth inhibition activity of the crude extract of Cyperus aromaticus (C. aromaticus) cultured cells against the 3rd instar larvae of Aedes aegypti (Linn.) and Aedes albopictus Skuse (Ae. albopictus) under laboratory conditions, and determine the sublethal effects (EI50) of the crude extract of C. aromaticus cultured cells on some biological and morphological parameters of both Aedes mosquito species during two generations as well. Methods: The cell suspension cultures of C. aromaticus were activated from five callus lines (P4, Pa, Z1, Z6 and Ml) derived from the root explants of in vitro plantlets. The cultured cells were extracted in chloroform and used as plant material for the present study. For detection of juvenile hormone Ⅲ, the crude extracts were analyzed by HPLC. Then the crude extracts of the three C. aromaticus cultured cell lines which contained varied amounts of juvenile hormone Ⅲ [high level (P4 cell line), medium level (Z1 cell line) and low level (Ml cell line)] were tested against Aedes mosquito species. Laboratory evaluation was performed against late third instar larvae of the Vector Control Research Unit strains of Ae. aegypti and Ae. albopictus using the standard WHO method. The effects of EI50 of the C. aromaticus cultured P4 cells on fecundity, fertility, growth period, sex ratio, adult size and longevity of Aedes mosquitoes were assessed. Results: Bioassay tests presented the remarkable growth inhibition activity of the crude extracts of C. aromaticus cultured cells against the two Aedes mosquitoes. Between the two mosquito species, Ae. albopictus was more susceptible to the crude extracts with lower EI50 values. EI50 of the crude extract of C. aromaticus cultured cells (P4) increased the sterility indices in the parental generation females in both Aedes mosquito species. A significant delay in the pupal formation and adult emergence were observed in the parental generation of the both mosquito species. The sex ratio of the adult population either parental or F1 generation of the Aedes mosquito species was not significantly affected by the EI50 dosage of the crude extract of C. aromaticus cultured P4 cells. A significant decrease in the wing length of the treated adult (female and male) of Aedes aegypti as well as the treated female of Ae. albopictus were observed. Longevity of the adult female of the parental generation of both Aedes mosquitoes as well as females of F1 generation of Ae. albopictus were significantly decreased. Conclusions: The present study revealed the potential of the crude extract of C. aromaticus cultured cells in controlling vector mosquito populations in the effort to reduce the transmission of vector borne diseases.
    3  Vasorelaxant activity of extracts obtained from Apium graveolens: Possible source for vasorelaxant molecules isolation with potential antihypertensive effect
    Vergara-Galicia Jorge Jimenez-Ramirez Luis ángel Tun-Suarez Adrián Aguirre-Crespo Francisco Salazar-Gómez Anuar Estrada-Soto Samuel Sierra-Ovando ángel Hernandez-Nu?ez Emmanuel
    2013(10):776-779. DOI: 10.1016/S2221-1691(13)60154-9
    [Abstract](20) [HTML](0) [PDF 324.98 K](134)
    Objective: To investigate the vasorelaxant effect of organic extracts from Apium graveolens (A. graveolens) which is a part of a group of plants subjected to pharmacological and phytochemical study with the purpose of offering it as an ideal source for obtaining lead compounds for designing new therapeutic agents with potential vasorelaxant and antihypertensive effects. Methods: An ex vivo method was employed to assess the vasorelaxant activity. This consisted of using rat aortic rings with and without endothelium precontracted with norepinephrine. Results: All extracts caused concentration-dependent relaxation in precontracted aortic rings with and without endothelium; the most active extracts were Dichloromethane and Ethyl Acetate extracts from A. graveolens. These results suggested that secondary metabolites responsible for the vasorelaxant activity belong to a group of compounds of medium polarity. Also, our evidence showed that effect induced by dichloromethane and ethyl acetate extracts from A. graveolens is mediated probably by calcium antagonism. Conclusions: A. graveolens represents an ideal source for obtaining lead compounds for designing new therapeutic agents with potential vasorelaxant and antihypertensive effects.
    4  Antiplasmodial, cytotoxic activities and characterization of a new naturally occurring quinone methide pentacyclic triterpenoid derivative isolated from Salacia leptoclada Tul. (Celastraceae) originated from Madagascar
    Fatiany Pierre Ruphin Robijaona Baholy Randrianarivo Emmanuel Raharisololalao Amelie Marie-Therese Martin Ngbolua Koto-te-Nyiwa
    2013(10):780-784. DOI: 10.1016/S2221-1691(13)60155-0
    [Abstract](8) [HTML](0) [PDF 735.29 K](130)
    Objective: To validate scientifically the traditional use of Salacia leptoclada Tul. (Celastraceae) (S. leptoclada) and to isolate and elucidate the structure of the biologically active compound. Methods: Bioassay-guided fractionation of the acetonic extract of the stem barks of S. leptoclada was carried out by a combination of chromatography technique and biological experiments in viro using Plasmodium falciparum and P388 leukemia cell lines as models. The structure of the biologically active pure compound was elucidated by 1D and 2D NMR spectroscopy and mass spectrometry. Results: Biological screening of S. leptoclada extracts resulted in the isolation of a pentacyclic triterpenic quinone methide. The pure compound exhibited both in vitro a cytotoxic effect on murine P388 leukemia cells with IC50 value of (0.041±0.020) µg/mL and an antiplasmodial activity against the chloroquine-resistant strain FC29 of Plasmodium falciparum with an IC50 value of (0.052±0.030) µg/mL. Despite this interesting anti-malarial property of the lead compound, the therapeutic index was weak (0.788). In the best of our knowledge, the quinone methide pentacyclic triterpenoid derivative compound is reported for the first time in S. leptoclada. Conclusions: The results suggest that furthers studies involving antineoplastic activity is needed for the development of this lead compound as anticancer drug.
    5  Identification of an anticancer compound against HT-29 cells from Phellinus linteus grown on germinated brown rice
    Tae-Il Jeon Chang-Hwa Jung Jeong-Yong Cho Dong Ki Park Jae-Hak Moon
    2013(10):785-789. DOI: 10.1016/S2221-1691(13)60156-2
    [Abstract](22) [HTML](0) [PDF 889.05 K](123)
    Objective: To isolate and identify the anticancer compound against proliferation of human colon cancer cells from ethyl acetate (EtOAc) extract of Phellinus linteus grown on germinated brown rice (PB). Methods: EtOAc extract of PB was partitioned with n-hexane, EtOAc, and water-saturated n-butanol. Anticancer compound of n-hexane layer was isolated and identified by HPLC and NMR, respectively. Cytotoxicity against HT-29 cells was tested by SRB assay. Results: The n-hexane layer obtained after solvent fractionation of PB EtOAc extracts showed a potent anticancer activity against the HT-29 cell line. Atractylenolide I, a eudesmane-type sesquiterpene lactone, a major anticancer substance of PB, was isolated from the n-hexane layer by silica gel column chromatography and preparative-HPLC. This structure was elucidated by one- and two-dimensional NMR spectroscopic data. Atractylenolide I has not been reported in mushrooms or rice as of yet. The isolated compound dose-dependently inhibited the growth of HT-29 human colon cancer cells. Conclusions: Atractylenolide I might contribute to the anticancer effect of PB.
    6  Production of microbial medium from defatted brebra (Milletia ferruginea) seed flour to substitute commercial peptone agar
    Berhanu Andualem Amare Gessesse
    2013(10):790-797. DOI: 10.1016/S2221-1691(13)60157-4
    [Abstract](17) [HTML](0) [PDF 1.03 M](164)
    Objective: To investigate and optimize microbial media that substitute peptone agar using brebra seed defatted flour. Methods: Defatted process, inoculums preparation, evaluation of bacterial growth, preparation of cooked and hydrolyzed media and growth turbidity of tested bacteria were determined. Results: Two percent defatted flour was found to be suitable concentration for the growth of pathogenic bacteria: Escherichia coli (ATCC 25922) (E. coli), Pseudomonas aeruginosa (ATCC 27853), Salmonella (NCTC 8385) and Shigella flexneri (ATCC 12022) (S. flexneri), while 3% defatted flour was suitable for Staphylococcus aureus (ATCC 25923) (S. aureus). E. coli (93±1) and S. flexneri (524±1) colony count were significantly (P≤0.05) greater in defatted flour without supplement than in supplemented medium. E. coli [(3.72×109±2) CFU/mL], S. aureus [(7.4×109±2) CFU/mL], S. flexneri [(4.03×109±2) CFU/mL] and Salmonella [(2.37×109±1) CFU/mL] in non-hydrolyzed sample were statistically (P≤0.05) greater than hydrolyzed one and commercial peptone agar. Colony count of Salmonella [(4.55×109±3) CFU/mL], S. flexneri [(5.40×109±3) CFU/mL] and Lyesria moncytogenes (ATCC 19116) [(5.4×109±3) CFU/mL] on raw defatted flour agar was significantly (P≤0.05) greater than cooked defatted flour and commercial peptone agar. Biomass of E. coli, S. aureus, Salmonella and Enterococcus faecalis in non-hydrolyzed defatted flour is highly increased over hydrolyzed defatted flour and commercial peptone broth. Conclusions: The defatted flour agar was found to be better microbial media or comparable with peptone agar. The substances in it can serve as sources of carbon, nitrogen, vitamins and minerals that are essential to support the growth of microorganisms without any supplements. Currently, all supplements of peptone agar are very expensive in the market.
    7  Anti-inflammatory and antioxidant activities of ethanolic extract of aerial parts of Vernonia patula (Dryand.) Merr.
    Arpona Hira Shubhra Kanti Dey Md. Sariful Islam Howlader Arif Ahmed Hemayet Hossain Ismet Ara Jahan
    2013(10):798-805. DOI: 10.1016/S2221-1691(13)60158-6
    [Abstract](19) [HTML](0) [PDF 799.47 K](125)
    Objective: To investigate the inflammatory and antioxidant activities of ethanolic extract of aerial part of Vernonia patula (Dryand.) Merr (EAV). Methods: The anti-inflammatory activity of EAV was studied using carrageenan and histamine- induced rat paw edema test at different doses (100, 200 and 400 mg/kg body weight). DPPH free radical scavenging, nitric oxide scavenging, reducing power and Fe2+ ion chelating ability were used for determining antioxidant activities. Results: The EAV, at the dose of 400 mg/kg, showed a significant anti-inflammatory activity (P<0.01) both in the carrageenan and histamine-induced oedema test models in rats, showing 62.86% and 64.42% reduction in the paw volume comparable to that produced by the standard drug indomethacin (67.26% and 66.01%) at 5 h respectively. In DPPH free radical scavenging test, IC50 value for EAV was found fairly significant 36.59 µg/mL when compared to the IC50 value of the reference standards ascorbic acid 8.97 µg/mL. The IC50 values of the extract and ascorbic acid were 47.72 and 12.39 µg/mL, respectively in nitric oxide scavenging assay. The IC50 value of the EAV (33.59 µg/mL) as percentage of Fe2+ ion chelating ability was also found significant compared to that of EDTA (9.16 µg/mL). The maximum absorbance for reducing power assay was found to be 1.928 at 100 µg/mL when compared to 2.449 for standard ascorbic acid. The total phenolic content was 198.81 mg/g of gallic acid equivalent. Acute toxicity test showed that the plant might be safe for pharmacological uses up to a dose level of 3 200 mg/kg of body weight in rats. Conclusions: Therefore, the obtained results suggest the acute anti-inflammatory and antioxidant activities of the EAV and thus provide the scientific basis for the traditional uses of this plant part as a remedy for inflammations.
    8  Production of asiaticoside from centella (Centella asiatica L. Urban) cells in bioreactor
    Nguyen Hoang Loc Nguyen Thi Duy Nhat
    2013(10):806-810. DOI: 10.1016/S2221-1691(13)60159-8
    [Abstract](16) [HTML](0) [PDF 1.12 M](130)
    Objective: To investigate the effects of some culture conditions on production of asiaticoside from centella (Centella asiatica L. Urban) cells cultured in 5-L bioreactor. Methods: The centell cell suspension culture was conducted in 5-L bioreactor to investigate the growth and asiaticoside accumulation under various conditions. Asiaticoside content was determined by HPLC analysis. Results: The results showed that the cell growth and asiaticoside accumulation peaked after 24 d of culture at an agitation speed of 150 r/min and aeration rate of 2.5 L/min. The cell biomass reached a maximum value of 302.45 g fresh weight (31.45 g dry weight) and growth index of 3.03 with inoculum size of 100 g. However, asiaticoside content was the highest (60.08 mg/g dry weight) when culture was initiated with an inoculum size of 50 g. Conclusions: The present study found the suitable conditions for growth of centella cells and their asiaticoside production in bioreactor.
    9  In vitro treatments of Echinococcus granulosus with fungal chitosan, as a novel biomolecule
    Bahman Rahimi-Esboei Mahdi Fakhar Aroona Chabra Mahboobeh Hosseini
    2013(10):811-815. DOI: 10.1016/S2221-1691(13)60160-4
    [Abstract](16) [HTML](0) [PDF 1.04 M](157)
    Objective: To determined the antiparasitic activity of the isolated chitosan from Penicillium viridicatum, Penicillium aurantiogriseum and commercial chitosan against protoscolicidal of hydatid cysts were determined. Methods: After isolating chitosan from fungal cell walls, four concentrations (50, 100, 200, 400 μg/ mL) of each type of prepared chitosan and commercial chitosan were used for 10, 30, 60, and 180 min, respectively. Results: Among different type of chitosan, commercial chitosan with the highest degree of deacetylation showed high scolicidal activity in vitro. Fungal chitosan could be recommended, as good as commercial chitosan, for hydatic cysts control. Conclusions: It seems to be a good alternative to synthetic and chemical scolicidal.
    10  In vitro activity of natural honey alone and in combination with curcuma starch against Rhodotorula mucilaginosa in correlation with bioactive compounds and diastase activity
    Moussa Ahmed Noureddine Djebli Saad Aissat Baghdad Khiati Abdelmalek Meslem Salima Bacha
    2013(10):816-821. DOI: 10.1016/S2221-1691(13)60161-6
    [Abstract](26) [HTML](0) [PDF 781.33 K](123)
    Objective: To evaluate the in vitro activity and synergism of the combinations of natural honey and curcuma starch against Rhodotorula mucilaginosa in correlation with total phenolic, flavonoid contents, and diastase activity. Methods: The Folin-Ciocalteu test was used to determine the total polyphenols content and the flavonoid content was analyzed using by the aluminum chloride method. The antifungal activity of the natural honey, determined by an agar well diffusion assay and agar incorporation method. Results: Total phenolic content varied from (63.93±0.11) to (95.36±6.08) mg GAE/100 g honey as gallic acid equivalent. Total flavonoids content varied from (5.41±0.04) to (9.94±0.54) mg CE/100 g. Diastase activity values were between (7.3±2.8) and (26±2.8). The zone inhibition diameter for the six honey samples without starch ranged between 6 and 20 mm. When starch was mixed with honey and then added to well, a zone inhibition increase diameter 7 and 21 mm. The percentage increase was noticed with each variety and it ranged between 5% and 62.5%. The minimal inhibitory concentrations for the six varieties of honey without starch against Rhodotorula mucilaginosa ranged between 28% and 36% (v/v). When starch was incubated with honey and then added to media, a minimal inhibitory concentration drop has been noticed with each variety. It ranged between 6.66 % and 20% (w/v). No significant correlation was established between diastase activity and bioactive compounds. Conclusions: The mixture of curcuma starch and honey could lead to the development of new combination antibiotics against Rhodotorula infections
    11  Seroprevalence of avian influenza (H9N2) in broiler chickens in Northwest of Iran
    Abolfazl Ghaniei Manoochehr Allymehr Ali Moradschendi
    2013(10):822-824. DOI: 10.1016/S2221-1691(13)60162-8
    [Abstract](30) [HTML](0) [PDF 669.76 K](150)
    Objective: To demonstrate seroprevalence of avian invluenza (H9N2) subtybe in broiler chickens in Northwest of Iran. Materials: A total of 310 blood samples were collected from 25 broiler flocks in slaughterhouses of West Azarbayjan, Iran. Serum samples were subjected to haemagglutination inhibition test. Results: The test showed 40.6% of positive serums. Mean antibody titer of avian influenza virus differed between geographical locations in this survey. Conclusions: High prevalence of avian influenza virus antibodies in serum of birds emphasize that avian influenza has an important role in respiratory complexes in broiler chickens in this region, and probably throughout Iran. Biosecurity measures, monitoring and surveillance programs, and to some degree vaccination are effective tools to prevent introduction of H9N2 infection and its economic losses.
    12  First detection of Leishmania infantum DNA in wild caught Phlebotomus papatasi in endemic focus of cutaneous leishmaniasis, South of Iran
    Rassi Yavar Karami Hadi Abai Mohammad Reza M Mohebali Bakshi Hasan Oshaghi Mohammad Ali Rafizadeh Sina Bagherpoor Hagigi Habib Hosseini Abodolrahim Gholami Manuchehr
    2013(10):825-829. DOI: 10.1016/S2221-1691(13)60163-X
    [Abstract](29) [HTML](0) [PDF 819.76 K](128)
    Objective: To identify the vectors and reservoirs of cutaneous leishmaniasis in the endemic focus of Farashband, Fars Province, South of Iran. Methods: Sticky papers and Sherman trap were used for collection of sand flies and rodents, respectively. Polymerase chain reaction (PCR) of kDNA, ITS1-rDNA were used for identification of Leishmania parasite in sand flies as well as rodents. Results: Totally 2 010 sand flies were collected and the species of Phlebotomus papatasi Scopoli was the common specimen in outdoors and indoors places. PCR technique was employed on 130 females of Phlebotomus papatasi. One of them (0.76%) was positive to parasite Leishmania major (L. major) and one specimen (0.76%) was positive to Leishmania infantum. Microscopic investigation on blood smear of the animal reservoirs for amastigote parasites revealed 16 (44%) infected Tatera indica. Infection of them to L. major was confirmed by PCR against kDNA loci of the parasite. Conclusions: The results indicated that Phlebotomus papatasi was the dominant species circulating two species of parasites including L. major and Leishmania infantum among human and reservoirs. Furthermore, Tatera indica is the only main host reservoir for maintenance of the parasite source in the area.
    13  Mucormycosis in a diabetic ketoacidosis patient
    G.?Sree Vijayabala Rajeshwari G Annigeri Ramachandran Sudarshan
    2013(10):830-833. DOI: 10.1016/S2221-1691(13)60164-1
    [Abstract](19) [HTML](0) [PDF 1.54 M](115)
    Oral cavity is considered to be a kaleidoscope for body’s general health. Many systemic conditions do present with diverse oral manifestations. Mucormycosis involving the oral cavity is one such entity that presents as necrosis of bone in immunocompromised patients. Mucormycosis is an opportunistic fungal infection that mainly affects the patients with uncontrolled diabetes mellitus. Hereby, we report a case of mucormycosis involving the palate in a patient with diabetic ketoacidosis.
    14  Review on pharmacological and toxicologyical effects of oleum azadirachti oil
    Khaled MM Koriem
    2013(10):834-840. DOI: 10.1016/S2221-1691(13)60165-3
    [Abstract](21) [HTML](0) [PDF 814.99 K](133)
    Oleum azadirachti consists of the oil obtained from dried seeds of Azadirachta indica A. Juss. (family: Meliaceae). Local names of Azadirachta indica A. Juss. are Abodua, aforo-oyinbo, anwe egyane, arista, azad dirakht, azadarakht, azedarach and bead tree. Indigenous to India, and widely distributed in South and South-East Asia and cultivated in Africa, the South Pacific Islands, South and Central America and Australia, and in southern Florida and California, United States of America, it is a straight-boled deciduous tree, which is 6-25 m high. Bark is dark- brown, externally fissured with a buff inner surface and fibrous fracture. Leaves alternately arranged, pinnately compound and up to 40 cm long, and composed of 8-18 short-petiolate narrow-ovate, pointed and curved toothed leaflets, 3-10 cm long and 1-4 cm wide arranged in alternate pairs. The major constituents are oxidized tetranortriterpenes including azadirachtin (azadirachtin A), azadiriadione, epoxyazadiradione, azadirone, nimbidin, nimbin, deacetylnimbin, salannin, gedunin, mahmoodin, 17-hydroxydiradione and related derivatives. It is of various medicinal uses, such as a contraceptive for intravaginal use, a mosquito repellent, and treatment of vaginal infections, treatment of gastric ulcers, cardiovascular disease, malaria, rheumatism and skin disorders, external applications for treatment of septic wounds,ulcers and boils, treatment of allergic skin reactions, asthma, bruises, colic, conjunctivitis, dysmenorrhoea, fever, gout, headache, itching due to varicella, kidney stones, leukorrhoea, psoriasis, scabies, sprains and muscular pain, and wounds. It is also used as an emmenagogue, tonic, stomatic and vermicide. In conclusion, the plant oil had antifertility, antihyperglycaemic, anti-inflammatory, antimicrobial, antiviral, antiulcer, estrogenic, immune, contraceptive, antibacterial, insect repellent, and skin treatment effects.

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