Asian Pacific Journal of Tropical Biomedicine

Issue 10,2012 Table of Contents

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  • 1  Making darkness visible: breaking the silence on HIV travel restrictions
    Alcides Troncoso
    [Abstract](8) [HTML](0) [PDF 202.94 K](61)
    2  Studies on bioprospecting potential of a gastropod mollusc Cantharus tranquebaricus (Gmelin, 1791)
    G Sarumathi M Arumugam S Kumaresan T Balasubramanian
    [Abstract](32) [HTML](0) [PDF 317.01 K](69)
    Objective: To study the biological activities of the tissue extract of Cantharus tranquebaricus (C. tranquebaricus). Methods: Crude extract of gastropod was tested for inhibition of bacterial growth. Antibacterial assay was carried out by disc diffusion method and the activity was measured accordingly based on the inhibition zone around the disc impregnated with gastropod extract. Molecular weight of the extract was determined by using SDS-PAGE. Plasma coagulation, Fibrin plate assay and substrate SDS-PAGE were used to determine the effect of sample on plasma coagulation, fibrin (ogen) olytic and proteolytic activity. Results: The maximum inhibition zone (10 mm) was observed against Vibrio cholera (V. cholera) and minimum inhibition zone (2 mm) was noticed against Proteus mirablis (P. mirablis). The molecular weight was determined as 47-106 kDa. The tissue extract shows proteolytic activity above 48 kDa. SDS-PAGE analysis of fibrinogen after incubation with the tissue extract showed fibrinogenolytic activity. In plasma coagulation assay C. tranquebaricus tissue extract showed procoagulant property and it coagulated chicken plasma within 150 s, while control took 5 min to clot. The 9 HU hemolytic units were found against chicken blood and also exhibit high level of brine shrimp lethality. Conclusions: This study suggests that C. tranquebaricus could be used as potential source for isolating bioactive compounds, since it is explored first time and found with promising results.
    3  The effect of polymorphism in gene of insulin-like growth factor-I on the serum periparturient concentration in Holstein dairy cows
    A Mirzaei H Sharifiyazdi MR Ahmadi T Ararooti A Rowshan Ghasrodashti A Kadivar
    [Abstract](18) [HTML](0) [PDF 296.83 K](67)
    Objective: To investigate the relationship between polymorphism within the 5'-untranslated region (5'-UTR) of IGF-I gene and its periparturient concentration in Iranian Holstein dairy cows. Methods: Blood samples (5 mL, n = 37) were collected by caudal venipuncture from each animal into sample tubes containing the EDTA and DNA was extracted from blood. In order to measure IGF-I concentration the collection of blood samples (n = 111) was also done at 14 d before calving (prepartum), 25 and 45 d postpartum. Results: We found evidence for a significant effect of C to T mutation in position 512 of IGF-I gene on its serum concentration in dairy cows in Iran. Cows with CC genotype had significantly higher concentration (Mean±SD) of IGF-I at 14 d prepartum (91.8±18.1) μg/L compared to those with TT genotype (73.3±14.4) μg/L (P=0.04). A significant trend (quadratic) was found for IGF-I concentration, as higher in CC cows compared to ones with TT genotype, during the 14 d before calving to 45 d postpartum (P=0.01). Conclusions: We concluded that C/T transition in the promoter region of IGF-I gene can influence the serum concentration of IGF-I in periparturient dairy cows.
    4  Characterization of cytotoxic compound from marine sediment derived actinomycete Streptomyces avidinii strain SU4
    Sudha S Masilamani Selvam M
    [Abstract](30) [HTML](0) [PDF 310.28 K](65)
    Objective: To investigate the cytotoxic activity of actinomycete isolated from marine sediment. Methods: In the present study the DNA was isolated and the ITS region of 16s rRNA was amplified by polymerase chain reaction, using two universal bacterial primers, 1492R (5′- GGTTACCTTGTTAC GACTT-3′) and Eubac27F (5′-AGAGTTTGATCCTGGCTC AG-3′). The amplified products were purified using TIANgel mini purification kit, ligated to MD18-T simple vector (TaKaRa), and transformed into competent cells of Escherichia coli DH5α. 16S rRNA gene fragment was sequenced using forward primer M13F (-47) and reverse primer M13R (-48). Blast search sequence similarity was found against the existing non-redundant nucleotide sequence database thus, identified as Streptomyces sp SU, Streptomyces rubralavandulae strain SU1, Streptomyces cacaoi strain SU2, Streptomyces cavourensis strain SU3, Streptomyces avidinii strain SU4, Streptomyces globisporus strain SU5, Streptomyces variabilis strain SU6, Streptomyces coelicolor strain SU 7. Among the eight identified isolates, one actinomycete Streptomyces avidinii strain SU4 was selected for further study. Results: Crude extract of the actinomycete isolate exhibited IC50 in 64.5 μg against Hep-2 cell line, 250 μg in VERO cell line. This value is very close to the criteria of cytotoxicity activity for the crude extracts, as established by the American National Cancer Institute (NCI) is in IC50 < 30 μg /mL. The GC MS analysis showed that the active principle might be 1,2-benzenedicarboxylic acid, bis(2-methylpropyl) ester (12.17%), isooctyl phthalate (15.29%) with the retention time 15.642 and 21.612, respectively. Conclusions: This study clearly proves that the marine sediment derived actinomycetes with bioactive metabolites can be expected to provide high quality biological material for high throughout biochemical and anticancer screening programs. These results help us to conclude that the potential of using metabolic engineering and post genomic approaches to isolate more bioactive compounds and make their possible commercial application is not far off.
    5  In vitro inhibition activity of polyphenol-rich extracts from Syzygium aromaticum (L.) Merr. & Perry (Clove) buds against carbohydrate hydrolyzing enzymes linked to type 2 diabetes and Fe2+-induced lipid peroxidation in rat pancreas
    Stephen Adeniyi Adefegha Ganiyu Oboh
    [Abstract](29) [HTML](0) [PDF 319.04 K](72)
    Objective: To investigate and compare the inhibitory properties of free and bound phenolic extracts of clove bud against carbohydrate hydrolyzing enzymes (alpha-amylase & alpha-glucosidase) and Fe2+ -induced lipid peroxidation in rat pancreas in vitro. Methods: The free phenolics were extracted with 80% (v/v) acetone, while bound phenolics were extracted from the alkaline and acid hydrolyzed residue with ethyl acetate. Then, the interaction of the extracts with alpha-amylase and alpha-glucosidase was subsequently assessed. Thereafter, the total phenolic contents and antioxidant activities of the extracts were determined. Results: The result revealed that both extracts inhibited alpha-amylase and alpha-glucosidase in a dose-dependent manner. However, the alpha-glucosidase inhibitory activity of the extracts were significantly (P<0.05) higher than their alpha-amylase inhibitory activity. The free phenolics (31.67 mg/g) and flavonoid (17.28 mg/g) contents were significantly (P<0.05) higher than bound phenolic (23.52 mg/ g) and flavonoid (13.70 mg/g) contents. Both extracts also exhibited high antioxidant activities as typified by their high reducing power, 1,1 diphenyl-2- picrylhydrazyl (DPPH) and 2, 2-azinobis- 3-ethylbenzo-thiazoline-6-sulfonate (ABTS) radical scavenging abilities, as well as inhibition of Fe2+ -induced lipid peroxidation in rat pancreas in vitro. Conclusions: This study provides a biochemical rationale by which clove elicits therapeutic effect on type 2 diabetes.
    6  Pharmacognostical, phytochemical and pharmacological evaluation for the antipyretic effect of the seeds of Saraca asoca Roxb.
    Sasmal S Majumdar S Gupta M Mukherjee A Mukherjee PK
    [Abstract](13) [HTML](0) [PDF 278.93 K](67)
    Objective: To conduct a systemic evaluation of the medicinal value of seeds which include macroscopic and microscopic characterization, physiochemical evaluation, preliminary phytochemical screening and experimental antipyretic activity. Methods: Saraca asoca seed was studied for pharmacognostical, phytochemical and other recommended methods for standardizations. Also, the acetone extract of the seeds was evaluated for acute toxicity study and antipyretic activity using Brewer’s yeast induced pyrexia in Wistar rats at oral doses of 300 mg/kg and 500 mg/kg. Results: After phytochemical screening, the acetone extract showed the presence of saponin, tannins and flavonoids which inhibit pyrexia. The therapeutic efficacy achieved at both the dose levels of the research drug and standard drug aspirin (100 mg/kg) showed significant (P<0.01) antipyretic activity when compared to the control group. The highly significant antipyretic effect exhibited at the dose of 500 mg/kg was also found to be sustainable in nature. Conclusions: The antipyretic effect of the acetone extract showed significant results in rats at the dose of 500 mg/kg after following the standard pharmacognostical and phytochemical methods.
    7  In-vitro antimicrobial activity of marine actinobacteria against multidrug resistance Staphylococcus aureus
    Sathish Kumar SR Kokati Venkata Bhaskara Rao
    [Abstract](10) [HTML](0) [PDF 300.89 K](71)
    Objective: To investigate the antibacterial activity of marine actinobacteria against multidrug resistance Staphylococcus aureus (MDRSA). Methods: Fifty one actinobacterial strains were isolated from salt pans soil, costal area in Kothapattanam, Ongole, Andhra Pradesh. Primary screening was done using cross-streak method against MDRSA. The bioactive compounds are extracted from efficient actinobacteria using solvent extraction. The antimicrobial activity of crude and solvent extracts was performed using Kirby-Bauer method. MIC for ethyl acetate extract was determined by modified agar well diffusion method. The potent actinobacteria are identified using Nonomura key, Shirling and Gottlieb 1966 with Bergey's manual of determinative bacteriology. Results: Among the fifty one isolates screened for antibacterial activity, SRB25 were found efficient against MDRSA. The ethyl acetate extracts showed high inhibition against test organism. MIC test was performed with the ethyl acetate extract against MDRSA and found to be 1 000 μg/mL. The isolated actinobacteria are identified as Streptomyces sp with the help of Nonomura key. Conclusions: The current investigation reveals that the marine actinobacteria from salt pan environment can be able to produce new drug molecules against drug resistant microorganisms.
    8  Studies on the seasonal variations in the proximate composition of ascidians from the Palk Bay, Southeast coast of India
    Ananthan G Karthikeyan MM Selva Prabhu A Raghunathan C
    [Abstract](25) [HTML](0) [PDF 309.14 K](65)
    Objective: To investigate the seasonal fluctuations of the proximate composition of the ascidians muscle. Methods: The moisture content was estimated by drying 1 g of fresh tissue at a constant temperature at 105 ℃ for 24 h.The loss of weight was taken as moisture content. The total protein was estimated using the Biuret method. The total carbohydrate in dried sample was estimated spectrophotometrically following the phenol- sulphuric acid method. The lipid in the dried sample tissue was gravimetrically estimated following the chloroform-methanol mixture method. Ash content was determined gravimetrically by incinerating 1 g dried sample in muffle furnace at about 550 ℃ for 6 h and results are expressed in percentage. Results: It was found very difficult to compare the monthly variations, as all the ten species, exhibited wide fluctuations in their proximate compositions. For the sake of convenience, average seasonal values were calculated by summing the monthly values. Conclusions: The proximate composition of the 10 commonly available ascidians showed high nutritive value and hence these groups especially solitary ascidians can be recommended for human consumption in terms of pickles, soup, curry and others after ensuring the safety of consumers.
    9  Antibacterial activity of sequentially extracted organic solvent extracts of fruits, flowers and leaves of Lawsonia inermis L. from Jaffna
    E Christy Jeyaseelan S Jenothiny MK Pathmanathan JP Jeyadevan
    [Abstract](25) [HTML](0) [PDF 252.00 K](67)
    Objective: To reveal the antibacterial activity of sequentially extracted different cold organic solvent extracts of fruits, flowers and leaves of Lawsonia inermis (L. against) some pathogenic bacteria. Methods: Powders of fruits, flowers and leaves of L. inermis were continuously extracted with dichloromethane (DCM), ethyl acetate and ethanol at ambient temperature. The dried extracts were prepared into different concentrations and tested for antibacterial activity by agar well diffusion method, and also the extracts were tested to determine the available phytochemicals. Results: Except DCM extract of flower all other test extracts revealed inhibitory effect on all tested bacteria and their inhibitory effect differed significantly effect was showed by ethyl acetate extract of flower against (P<0.05). The highest inhibitory Staphylococcus aureus (S. aureus) and Pseudomonas aeruginosa (P. aeruginosa), and ethyl acetate extract of fruit on Escherichia coli (E. coli) and Bacillus subtilis (B. subtilis). The ethyl acetate and ethanol extracts of flower, fruit and leaf expressed inhibition even at 1 mg/ 100 μl against all test bacteria. Among the tested phytochemicals flavonoids were detected in all test extracts except DCM extract of flower. Conclusions: The study demonstrated that the ethyl acetate and ethanol extracts of fruit and flower of L. inermis are potentially better source of antibacterial agents compared to leaf extracts of respective solvents.
    10  Characterization and phylogenetic analysis of novel polyene type antimicrobial metabolite producing actinomycetes from marine sediments: Bay of Bengal India
    Valan Arasu M Asha KRT Duraipandiyan V Ignacimuthu S Agastian P
    [Abstract](10) [HTML](0) [PDF 317.51 K](71)
    Objective: To isolate and indentify the promising antimicrobial metabolite producing Streptomyces strains from marine sediment samples from Andrapradesh coast of India. Methods: Antagonistic actinomycetes were isolated by starch casein agar medium and modified nutrient agar medium with 1% glucose used as a base for primary screening. Significant antimicrobial metabolite producing strains were selected and identified by using biochemical and 16S rDNA level. Minimum inhibitory concentrations of the organic extracts were done by using broth micro dilution method. Results: Among the 210 actinomycetes, 64.3% exhibited activity against Gram positive bacteria, 48.5 % showed activity towards Gram negative bacteria, 38.8% exhibited both Gram positive and negative bacteria and 80.85 % isolates revealed significant antifungal activity. However, five isolates AP-5, AP-18, AP-41 and AP-70 showed significant antimicrobial activity. The analysis of cell wall hydrolysates showed the presence of LL-diaminopimelic acid and glycine in all the isolates. Sequencing analysis indicated that the isolates shared 98.5%-99.8% sequence identity to the 16S rDNA gene sequences of the Streptomyces taxons. The antimicrobial substances were extracted using hexane and ethyl acetate from spent medium in which strains were cultivated at 30℃for five days. The antimicrobial activity was assessed using broth micro dilution technique. Each of the culture extracts from these five strains showed a typical polyene-like property. The lowest minimum inhibitory concentrations of ethyl acetate extracts against Escherichia coli and Curvularia lunata were 67.5 and 125.0 μg/mL, respectively. Conclusions: It can be concluded that hexane and ethyl acetate soluble extracellular products of novel isolates are effective against pathogenic bacteria and fungi.
    11  Toxicity profile of ethanolic extract of Azadirachta indica stem bark in male Wistar rats
    Anofi Omotayo Tom Ashafa Latifat Olubukola Orekoya Musa Toyin Yakubu
    [Abstract](26) [HTML](0) [PDF 337.85 K](72)
    Objective: To investigate the toxic implications of ethanolic stem bark extract of Azadirachta indica (A. indica) at 50, 100, 200 and 300 mg/kg body weight in Wistar rats. Methods: Fifty male rats of Wistar strains were randomly grouped into five (A-E) of ten animals each. Animals in Group A (control) were orally administered 1 mL of distilled water on daily basis for 21 days while those in Groups B-E received same volume of the extract corresponding to 50, 100, 200 and 300 mg/kg body weight. Results: The extract did not significantly (P>0.05) alter the levels of albumin, total protein, red blood cells and factors relating to it whereas the white blood cell, platelets, serum triacylglycerol and high-density lipoprotein cholesterol decreased significantly (P<0.05). In contrast, the final body weights, absolute weights of the liver, kidney, lungs and heart as well as their organ-body weight ratios, serum globulins, total and conjugated bilirubin, serum cholesterol, low-density lipoprotein cholesterol and computed atherogenic index increased significantly. The spleen-body weight ratio, alkaline phosphatase, alanine and aspartate transaminases, sodium, potassium, calcium, feed and water intake were altered at specific doses. Conclusions: Overall, the alterations in the biochemical parameters of toxicity have consequential effects on the normal functioning of the organs of the animals. Therefore, the ethanolic extract of A. indica stem bark at the doses of 50, 100, 200 and 300 mg/kg body weight may not be completely safe as an oral remedy and should be taken with caution if absolutely necessary.
    12  Evaluation of in-vitro antibacterial activity and anti-inflammatory activity for different extracts of Rauvolfia tetraphylla L. root bark
    B. Ganga Rao P. Umamaheswara Rao E. Sambasiva Rao T. Mallikarjuna Rao V. S. Praneeth. D
    [Abstract](35) [HTML](0) [PDF 293.54 K](60)
    Objective: To assess the in-vitro antibacterial activity and anti-inflammatory activity of orally administered different extracts (Hydro-alcoholic, methanolic, ethyl acetate and hexane) of Rauvolfia tetraphylla (R. tetraphylla) root bark in Carrageenan induced acute inflammation in rats. Methods: In-vitro antibacterial activity was evaluated for extracts against four Gram positive and four Gram negative bacteria by using cylinder plate assay. Hydro-alcoholic extract (70% v/v ethanol) at 200, 400 and 800 mg/kg doses and methanolic, ethyl acetate and hexane extracts at doses 100, 200 and 400 mg/kg were tested for anti-inflammatory activity in Carrageenan induced rat paw oedema model and paw thickness was measured every one hour up to 6 hrs. Results: All extracts of R. tetraphylla root bark showed good zone of inhibition against tested bacterial strains. In Carrageenan induced inflammation model, hydro-alcoholic and methanolic extract of R. tetraphylla root bark at three different doses produced significant (P<0.001) reduction when compared to vehicle treated control group and hexane, ethyl acetate extracts. Conclusions: In the present study extracts of R. tetraphylla root bark shows good in-vitro antibacterial activity and in-vivo anti-inflammatory activity in rats.
    13  In vitro antimicrobial effects of grape seed extract on peri-implantitis microflora in craniofacial implants
    Binit Shrestha M.L. Srithavaj Theerathavaj Sroisiri Thaweboon Boonyanit Thaweboon
    [Abstract](9) [HTML](0) [PDF 238.97 K](63)
    Objective: To determine the antimicrobial effects of grape seed on peri-implantitis microflora. Methods: The grape seed extract was tested against peri-implantitis microflora most commonly found in craniofacial implants including reference strains of Staphylococcus aureus (S. aureus), Escherichia coli (E. coli), Candida albicans (C. albicans) and clinical strains of S. aureus, Klebsiella pneumonia (K. pneumonia) and Candida parapsilosis (C. parapsilosis) by disk diffusion test. Minimum inhibitory concentrations (MIC) and minimum cidal concentrations (MCC) were determined using modified agar dilution millpore method. The extract was further combined with polyethylene glycol and propylene glycol, and was tested for antimicrobial effects. Results: Grape seed extract showed positive inhibitory effects with S. aureus at MIC of 0.625 mg/mL and MCC of 1.25 mg/mL respectively. However the extracts showed minimal or no reactivity against strains of E. coli, K. pneumonia, C. parapsilosis and C. albicans. The use of grape seed extract in combination with polyethylene glycol and propylene glycol also showed dose dependent inhibitory effect on S. aureus. Conclusions: The results of the study showed that grape seed has potential antimicrobial effects which can be further studied and developed to be used in the treatment of infected skin-abutment interface of craniofacial implants.
    14  Cytotoxicity of methanol extracts of Elaeis guineensis on MCF-7 and Vero cell lines
    Soundararajan Vijayarathna Sreenivasan Sasidharan
    [Abstract](30) [HTML](0) [PDF 288.91 K](73)
    Objective: To investigate the cytotoxic effect of Elaeis guineensis methanol extract on MCF-7 and Vero cell. Methods: In vitro cytotoxicity was evaluated in by MTT assay. Cell morphological changes were observed by using light microscope. Results: The MTT assay indicated that methanol extract of the plant exhibited significant cytotoxic effects on MCF-7. Morphological alteration of the cell lines after exposure with Elaeis guineensis extract were observed under phase contrast microscope in the dose dependent manner. Conclusions: The results suggest the probable use of the Elaeis guineensis methanol extract in preparing recipes for cancer-related ailments. Further studies on isolation of metabolites and their in vivo cytotoxicity are under investigation.
    15  Morpho-anatomical and physicochemical studies of Fumaria indica (Hausskn.) Pugsley
    Prakash Chandra Gupta Ch V Rao
    [Abstract](32) [HTML](0) [PDF 342.45 K](73)
    Objective: To study morpho-anatomical characters and physicochemical analysis of Fumaria indica (F. indica) (Hausskn.) Pugsley, (Fumariaceae), an important medicinal plant used extensively for treating a variety of ailments in various system of indigenous medicine. Methods: Evaluation of the different parts of the plant was carried out to determine the morpho-anatomical, physicochemical, phytochemical and HPTLC fingerprinting profile of F. indica and other WHO recommended methods were performed for standardization. Results: Morpho-anatomical studies showed compound and pinnatifid leaf, 4 to 6 cm in length, linear and oblong in shape and anomocytic arrangement of stomata, thin walled parenchymatous cells, scattered, sclerenchymatous, capped vascular bundles and radiating medullary rays. Physicochemical studies showed foreign matter 0.2%, loss on drying 6.8%, total ash 16.77%, alcohol and water soluble extractives 8.92% and 20.26%, respectively, sugar 17.75%, starch 22.97% and tannins 2.37%. Phytochemical evaluation revealed the presence of carbohydrate, alkaloids, flavonoids, saponins, tannins and sterol. Thin layer chromatography was carried out with different solvents and the best solvent system was chloroform and methanol in 80:20 ratio and revealed 12 spots with different Rf value under UV light 366λ. Conclusions: The results of the study can serve as a valuable source of information and provide suitable standards for identification of this plant material for future investigations and applications.
    16  Barbaloin: A concise report of its pharmacological and analytical aspects
    DK Patel K Patel V Tahilyani
    [Abstract](12) [HTML](0) [PDF 265.00 K](76)
    Barbaloin is C-glucoside of aloe emodin anthrone which is found in the plant name as Aloe vera is a perennial succulent (Liliaceal), also called the healing plant. Barbaloin have variety of pharmacological activity such as strong inhibitory effect on histamine release, anti-inflammatory, cathartic, antiviral, antimicrobial, anticancer, antioxidant activity and alternative for pharmaceutical or cosmetic applications. The peak amount of barbaloin was reached about 3 h after oral administration. Concentration of barbaloin in Aloe vera leaves was shown to depend on the leaf part, age, and position of the leaf. Young leaves contain more barbaloin compared to old one. Various researches have been done on barbaloin but still the relationship between the barbaloin and its overall effect has not been clarified. A more specific perceptive of the pharmacological activities of barbaloin is required to develop for pharmaceutical purpose. Many attempts have been made regarding its isolation, biological activity to examine their effects, and clarify their functional mechanism. This review gives a brief idea about its uses, ethnomedicinally and commercially important analytical techniques and their pharmacological activities.
    17  Primary tuberculosis of the thyroid gland: a case report
    Sant Parkash Kataria Parul Tanwar Sneh Singh Sanjay Kumar
    [Abstract](33) [HTML](0) [PDF 318.74 K](68)
    Tuberculosis of the thyroid gland is an uncommon disease and primary involvement of thyroid is even more rare. It is a rare disease even in countries in which tuberculosis is endemic. The diagnosis is often difficult as the clinical presentation has no distinct characteristics. Clinical course of the disease may resemble toxic goiter or acute thyroiditis or may follow a subacute or chronic growth pattern without specific symptomatology. Histologically presence of necrotizing epithelioid cell granulomas along with langhans type giant cells are the hallmark of thyroid tuberculosis. Demonstration of acid fast bacilli by ZN staining confirms the diagnosis, but this stain is frequently negative in tissue sections.

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