Asian Pacific Journal of Tropical Biomedicine

Issue 2,2016 Table of Contents

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  • 1  Inhibitory actions of Pseuderanthemum palatiferum (Nees) Radlk. leaf ethanolic extract and its phytochemicals against carbohydrate-digesting enzymes
    Pawitra Pulbutr Somsak Nualkaew Sakulrat Rattanakiat Benjamart Cushnie Achida Jaruchotikamol
    2016(2):93-99. DOI: 10.1016/j.apjtb.2015.10.010
    [Abstract](17) [HTML](0) [PDF 0.00 Byte](0)
    Objective: To investigate the effects of the leaf ethanolic extract of Pseuderanthemum palatiferum (PPE) and its isolated phytochemicals, stigmasterol and sitosterol-3-O-β-Dglucopyranoside, against α-amylase and α-glucosidase enzyme activities both in vitro and in vivo. Methods: A concentration of maltose, which is a product released in α-amylase-catalyzing reaction, was used as an index of in vitro α-amylase activity. Meanwhile, in vitro α-glucosidase enzyme activity was indicated by the amount of liberated p-nitrophenol in α-glucosidase-catalyzing reaction. In vivo α-amylase and α-glucosidase enzyme activities were evaluated in the normal rats by using oral starch tolerance test and oral sucrose tolerance test, respectively. Results: PPE exerted a concentration-dependent inhibitory action against both α-amylase and α-glucosidase in vitro with the IC50 values of (11.79 ± 8.10) mg/mL and (1.00 ± 0.11) mg/mL, respectively. Stigmasterol and sitosterol-3-O-β-D-glucopyranoside also exerted an in vitro α-amylase inhibition with the IC50 values of (59.41 ± 8.22) μg/mL and (111.19 ± 9.02) μg/mL, respectively. However, these phytochemicals did not produce a concentration-dependent inhibition against in vitro α-glucosidase activity. PPE and its isolated phytochemicals significantly decreased the blood glucose levels at t = 30 min in the oral starch tolerance test. From the sucrose tolerance test, only PPE but not its isolated phytochemicals significantly caused a depletion in the blood glucose levels at t = 30 min Conclusions: These results indicate an inhibitory action against carbohydrate-digesting enzymes as the anti-diabetic mechanism of action of PPE. Nonetheless, further clinical study is required to justify its role in the treatment of diabetes.
    2  In vitro antimicrobial and larvicidal properties of wild Ricinus communis L. in Mauritius
    Sillma Rampadarath Daneshwar Puchooa
    2016(2):100-107. DOI: 10.1016/j.apjtb.2015.10.011
    [Abstract](17) [HTML](0) [PDF 0.00 Byte](0)
    Objective: To explore the phytochemical and antimicrobial activities as well as the insecticidal properties of the different sections of Ricinus communis (castor) plant in Mauritius. Methods: Qualitative and quantitative methods were used for the determination of phytochemicals in the crude leaves, pericarp, seeds, bark and root extracts obtained by using polar and non-polar solvents. The disc diffusion and micro-dilution methods were used to evaluate the antimicrobial activities of the crude solvent extracts against 13 microorganisms. The insecticidal properties of the crude extracts on larvae of Bactrocera zonata (Diptera: Tephritidae), which caused important economic losses to local fruits were also investigated. Results: All the extracts from the different parts of the plant showed antimicrobial activity against most tested microorganisms. The polar solvents' extracts of the fully mature parts of the castor plant were active against the Pseudomonas aeruginosa (ATCC 27853), Escherichia coli (E. coli) (ATCC 25922), E. coli (0145:H28 Acc. No. CP006027.1) with inhibition zones ranging from 16 mm to 19 mm and against Bacillus cereus (ATCC 11778) (B. cereus), Listeria innocua (ATCC 33090) (L. innocua). Lowest microbial inhibitory concentration was recorded for B. cereus, L. innocua, Staphylococcus aureus (ATCC 29213), E. coli (ATCC 25922) and Proteus mirabilis strain (NCTC 11938) with value of 3.2 μg/mL. The most active extract against both Gram-negative and Grampositive bacteria was extracted from the fully mature pericarps and it was the most active against E. coli (ATCC 25922), B. cereus, L. innocua and Staphylococcus aureus (ATCC 29213). In addition, the extracts obtained by using polar solvent and fully mature leaves demonstrated the strongest larvicidal activity against Bactrocera zonata (100%). Conclusions: Ricinus communis (castor) plant extracts possess larvicidal properties providing an effective eco-friendly control for fruit flies. The antimicrobial results justify the use of this plant in traditional medicine and the practice of supplementing decoctions/ concoctions with conventional antibiotics.
    3  The effects of glycyrrhizic acid and glabridin in the regulation of CXCL5 inflammation gene on acceleration of wound healing
    Hong Yung Yip Melissa Su Wei Poh Yoke Yin Chia
    2016(2):108-113. DOI: 10.1016/j.apjtb.2015.10.009
    [Abstract](32) [HTML](0) [PDF 0.00 Byte](0)
    Objective: To evaluate the anti-inflammatory property of both glycyrrhizic acid (GA) and glabridin in reducing inflammation to accelerate wound regeneration on 3T3-L1 and NIH-3T3 fibroblast cell lines. Methods: Cell proliferation and viability assay (MTT assay), scratch wound healing assays, and quantitative real-time PCR were conducted to investigate the effects on cell proliferation, cell migration, and expression of CXC chemokine ligand 5 inflammation gene respectively. Results: Results showed that at a low concentration of 1 × 10−8 mol/L, glabridin down regulated cell proliferation in NIH-3T3 significantly, suggesting its involvement in ERK1/2 signaling pathway. GA and glabridin significantly accelerated cell migration through wound healing in both 3T3-L1 and NIH-3T3 and significantly down regulated the expression of CXC chemokine ligand 5 in 3T3-L1 at concentration 1 × 10−8 mol/L, indicating the possible involvement of nuclear factor-ĸB and cyclooxygenase 2 transcriptions modulation. Conclusions: Both GA and glabridin can serve as potential treatment for chronic in- flammatory disease, and glabridin as an oncogenic inhibitor due to its anti-proliferative property.
    4  Neurotropic effects of aspartame, stevia and sucralose on memory retention and on the histology of the hippocampus of the ICR mice (Mus musculus)
    Lejan Miguel Alabastro Villareal Rachelle Anne Montes Cruz Michael Bagui Ples Rodel Jonathan Santos Vitor II
    2016(2):114-118. DOI: 10.1016/j.apjtb.2015.11.001
    [Abstract](6) [HTML](0) [PDF 0.00 Byte](0)
    Objective: To identify the effects of the consumption of non-nutritive sweeteners on memory retention and on the histology of the hippocampus. Methods: In this study, 20 mice were used to determine if there is an effect of consuming the maximum allowable dose of the non-nutritive sweeteners on the memory retention and on the histology of the hippocampus. The mice were distributed into four groups and the treatments were given via oral gavage: Group 1 (water), Group 2 (aspartame: 1 000 mg/kg), Group 3 (stevia: 1 000 mg/kg) and Group 4 (sucralose: 16 000 mg/kg). Treatments were administered to the different experimental groups for 32 days, after which memory retention was tested using the two-day water maze protocol. After the tests, the mice were sacrificed and the brain was analyzed histologically for neurotrophic effects. Results: Based on the results of the two-day water maze protocol, there were no differences between the non-nutritive sweeteners and the control group. However, stevia showed high cellular apoptosis followed by aspartame, sucralose and control group. Conclusions: There was no significant effect on the memory of the mice. It showed histologically however, that stevia had a significant neurotropic effect compared to the other sweeteners.
    5  Sulforaphene and sulforaphane in commonly consumed cruciferous plants contributed to antiproliferation in HCT116 colon cancer cells
    Piman Pocasap Natthida Weerapreeyakul
    2016(2):119-124. DOI: 10.1016/j.apjtb.2015.11.003
    [Abstract](20) [HTML](0) [PDF 0.00 Byte](0)
    Objective: To analyze two isothiocyanates (sulforaphene and sulforaphane) and their antiproliferative effect of 11 indigenous cruciferous vegetables. Methods: Phytoconstituents identification was conducted by high performance liquid chromatography and gas chromatography-mass spectrometer techniques. The antiproliferation was evaluated in colon cancer cell line HCT116 by MTT assay. Results: Isothiocyanate identification by high performance liquid chromatography showed that broccoli, cabbage, “Khi-Hood” (Raphanus sativus L. var. caudatus Alef) and Chinese radish contained isothiocyanates sulforaphane. Sulforaphene and sulforaphane in broccoli, cabbage and “Khi-Hood” were characterized by the gas chromatography-mass spectrometer analysis. Antiproliferation screening by MTT assay found that the potent plants which possessed IC50 below 50 μg/mL were cabbage and “Khi-Hood”, while the others had low antiproliferation with IC50 higher than 50 μg/mL. Difference in antiproliferation was probably due to difference existed phytochemical constituents in each plant. “Khi-Hood” possessed the highest antiproliferation against HCT116 with the lowest IC50 at (9.42 ± 0.46) μg/mL. The IC50 of chemotherapeutic drug (mitomycin C) was (19.12 ± 1.00) μg/mL, while both melphalan and 5-fluorouracil possessed the IC50 value higher than 50 μg/mL. Conclusions: Commonly consumed cruciferous vegetables exerted varied antiproliferation and isothiocyanate contents. High isothiocyanate content in “Khi-Hood” was contributed to high antiproliferation. Among 11 plants studied, “Khi-Hood” could be an alternative chemopreventive diet.
    6  Nutritional quality and safety aspects of wild vegetables consume in Bangladesh
    Miah Mohammed Abdus Satter Mohammed Murtaza Reza Linkon Khan Syeda Absha Jabin Nusrat Abedin Mohammed Faridul Islam Badhan Shaha
    2016(2):125-131. DOI: 10.1016/j.apjtb.2015.11.004
    [Abstract](20) [HTML](0) [PDF 0.00 Byte](0)
    Objective: To evaluate the nutritional composition, including major minerals, essential trace elements and toxic heavy metals of five different wild vegetables Dhekishak (Dryopteris filix-mas), Helencha (Enhydra fluctuans), Kalmishak (Ipomoea aquatica), Patshak (Corchorus capsularis) and Shapla stem (Nymphaea stellata) and their safety aspects. Methods: Proximate parameters moisture, ash, fat, fiber, protein, carbohydrate and energy; major minerals Na, K, Ca and Mg; trace elements Fe, Zn and Cu; and toxic heavy metals Pb, Cd, Cr, Ni and Hg were evaluated in the selected wild vegetables using the standard food analysis techniques. Results: The results from nutritional analysis showed that all the wild vegetables used in this study had a low content of crude fat and high content of moisture, ash, crude protein, crude fiber, carbohydrate and energy having the recommended dietary allowances. The vegetables were also rich in major minerals Na, K, Ca and Mg, sufficient in essential trace elements Fe, Cu and Zn while the heavy metals Pb, Cr and Ni were detected higher in amount in all the vegetables except Patshak than the limits recommended by Food and Agriculture Organization/World Health Organization. The heavy metals Cd and Hg were not detected in any vegetable. Conclusions: The outcome of this study suggests that the wild vegetables have very good nutritional potential to meet the recommended dietary allowances, but special awareness should be taken for public health concern about the high level of Pb, Cr and Ni which exceed the Food and Agriculture Organization/World Health Organization recommended limits for the metals in vegetables.
    7  Bioprospecting optimal phenology for bioactive molecules in native golden yellow Pleurotus citrinopileatus Singer
    Godfrey Nattoh Erastus Gatebe Fredrick Musieba Julius Mathara
    2016(2):132-142. DOI: 10.1016/j.apjtb.2015.10.012
    [Abstract](19) [HTML](0) [PDF 0.00 Byte](0)
    Objective: To bioprospect optimal phenological phases as source of novel molecules from native golden yellow Pleurotus citrinopileatus across four phenologies in both aqueous and ethanol extracts, and identify novel molecules responsible for these activities. Methods: Standard qualitative assay, Folin–Ciocalteu assay; aluminium chloride spectrophotometric, 2, 2-diphenyl-1-picrylhydrazyl, 2, 2’-azinobis (3-ethylbenzothiazoline-6- suslfonic acid, ferricyanide reducing antioxidant power were used to determine total flavonoid, polyphenols, radical scavenging, and reducing power. Spectrophotometric methods were used for lycopene, β-carotene, and total carotenoids, while liquid chromatography quadrupole time of flight mass spectrometry was used for identification and comparative quantitation of polyphenols and flavonoids across the four phenological states. ChemSpider™ database was used for the identification of compounds based on their empirical formula, accurate mass and literature review of previously reported compounds in mushroom. Results: Primordial phases exhibited higher contents of secondary metabolites than mature basidiocarps. Polyphenols content differed across physiological phases with primordials exhibiting significant high contents (P < 0.05) [(13.803 ± 0.797) mg gallic acid equivalent/g dry weight]. Distribution of total flavonoids was significantly different (P < 0.05) across physiological states and ranged from (3.311 ± 0.730) to (14.824 ± 0.890) mg quercetin equivalent g dry weight. Ten polyphenol acids and seven flavonoids compounds identified varied across these phases with primordials exhibiting relatively high peak areas. Total antioxidant activities showed a positive correlation with total polyphenols (r = 0.969; P < 0.05) and total flavonoids (r = 0.960; P < 0.05) across these phenologies. Conclusions: These findings provide evidence that primordials of golden yellow mushroom as opposed to their fruiting bodies are potent sources of bioactive health molecules.
    8  Effects of variations in culture media and hormonal treatments upon callus induction potential in endosperm explant of Barringtonia racemosa L.
    Nurul Izzati Osman Norrizah Jaafar Sidik Asmah Awal
    2016(2):143-147. DOI: 10.1016/j.apjtb.2015.10.007
    [Abstract](28) [HTML](0) [PDF 0.00 Byte](0)
    Objective: To induce callus from the medicinally valuable species, Barringtonia racemosa L. (B. racemosa) whereby the formation of callus is essential for micropropagation studies and in vitro plant secondary metabolites production. Methods: The callus induction potential in B. racemosa was assessed from endosperm explant cultured on different culture media and plant hormonal treatments. Lloyd and McCown's woody plant medium and Murashige and Skoog's medium were used in the study as culture media. On the other hand, various concentrations and combinations of 2,4-dichlorophenoxyacetic acid (1.0–2.0 mg/L) and kinetin (0.5–2.5 mg/L) had been incorporated in the culture media to exert the effects of auxin and cytokinin on callus induction. Results: From the present study, it was found that the profuse [(1.681 ± 0.770) g fresh weight, (0.239 ± 0.239) g dry weight] and friable callus formation was optimally produced with desirable morphology and considerable percentage of callus induction (56.70%) in endosperm explants cultured on 1.0 mg/L 2,4-dichlorophenoxyacetic acid and 1.5 mg/L kinetin in Murashige and Skoog's medium. Conclusions: A reliable protocol for inducing callus formation of profuse and friable morphology in endosperm explant of B. racemosa had therefore been successfully established.
    9  Evaluation of entomopathogenic Bacillus sphaericus isolated from Lombok beach area against mosquito larvae
    Bambang Fajar Suryadi Bagyo Yanuwiadi Tri Ardyati Suharjono Suharjono
    2016(2):148-154. DOI: 10.1016/j.apjtb.2015.10.013
    [Abstract](20) [HTML](0) [PDF 0.00 Byte](0)
    Objective: To isolate, characterize and evaluate toxicity of Bacillus sphaericus (B. sphaericus) from beach area of Lombok Island. Methods: Soil was collected from determined locations and suspended in sterile physiological saline water. After heat shock was applied, suspension was spread on NYSM agar medium. Colonies grown were then observed and isolated. Colony, cell morphology, and biochemical/physiological characteristics were tested and compared to B. sphaericus 2362 as standard. Initial toxicity testing was done against three species of mosquito larvae (Culex quinquefasciatus, Anopheles aconitus and Aedes aegypti) and isolates that showed more than 50% larvae killing will be assayed to obtain LC50 and LC90 values within 48 h. PCR technique were conducted to obtain 16s rDNA amplicon for sequencing and to detect toxin-expressing genes (using multiplex PCR). Results: Twenty isolates of B. sphaericus have been collected from 20 determined locations and their characteristics were in agreement with standard B. sphaericus characteristics. Bioassay testing showed that four isolates (namely isolate MNT, SLG, TJL2 and PLG) were mildly toxic against all larvae. The rests were either low toxic or non-toxic at all. Phylogenetic analysis showed that all four isolates were clustered with other known mildly and highly toxic strains. The multiplex PCR result showed four toxic isolates owned 1–2 bands from Bin toxin genes and three bands from Mtx toxin genes, whereas 16 isolates with low to non-toxic characteristics showed only three bands from Mtx toxin genes. Conclusions: Four toxic isolates of B. sphaericus were isolated from beach area of Lombok Island. They showed mild toxicity against larvae of three mosquito species.
    10  Anti-hyperglycemic effects of aqueous Lenzites betulina extracts from the Philippines on the blood glucose levels of the ICR mice (Mus musculus)
    Faizal Rajeeb Mangudadatu Hussin Rodel Jonathan Santos Vitor II Julie Ann Oraa Joaquin Melody Mendoza Clerigo Anamy Ma. Caterial Paano
    2016(2):155-158. DOI: 10.1016/j.apjtb.2015.04.013
    [Abstract](21) [HTML](0) [PDF 0.00 Byte](0)
    Objective: To examine the anti-hyperglycemic effects of aqueous Lenzites betulina (L. betulina) extracts on normoglycemic glucose-loaded mice. Methods: Different doses of aqueous extract from L. betulina were administered to 45 ICR mice (Mus musculus) to determine whether there was an effect of L. betulina extracts on the blood glucose level of the ICR mice. Aqueous extracts of L. betulina were orally gavaged to mice using oral glucose tolerance test. A total of five groups were used to determine the effect of the fungi on blood glucose of the mice. Group A (positive control) was given 16.7 μg/kg glimepiride; Group B (negative control) was given distilled water; Group C (low dosage) was given 200 mg/kg aqueous extract; Group D (mid dosage) was given 400 mg/kg aqueous extract and Group E (high dosage) was given 800 mg/kg aqueous extract. Baseline blood glucose value was firstly acquired before induction of hyperglycemia through D-glucose, after which another check on blood glucose was made after 0.5 h. Immediately, after the acquisition of hyperglycemic blood glucose level, the individual administration of treatments were done. After that, three blood collections were done spanning 3 h with 1 h interval. Results: The low dose (200 mg/kg) and the mid dose (400 mg/kg) of L. betulina extracts were significantly different (P < 0.05) from their respective baseline values throughout the whole experiment with the latter surpassing its baseline value during the 3rd hour. On the other hand, the high dose (800 mg/kg) during the 1st hour after administration was not significantly different (P > 0.05) from its corresponding baseline value, acting faster than the positive control (glimepiride), which only became significantly different (P < 0.05) at the 2nd hour. Conclusions: Aqueous L. betulina extract is able to produce hypoglycemic effects on the mice with all doses, which are able to normalize blood glucose levels at varying times.
    11  Evaluation of imatinib mesylate (Gleevec) on KAI1/CD82 gene expression in breast cancer MCF-7 cells using quantitative real-time PCR
    Seyed Ataollah Sadat Shandiz Marjan Khosravani Sepideh Mohammadi Hassan Noorbazargan Amir Mirzaie Davoud Nouri Inanlou Mojgan Dalirsaber Jalali Hamidreza Jouzaghka Fahimeh Baghbani-Arani Behta Keshavarz-Pakseresht
    2016(2):159-163. DOI: 10.1016/j.apjtb.2015.10.006
    [Abstract](6) [HTML](0) [PDF 0.00 Byte](0)
    Objective: To evaluate the effect of imatinib mesylate on cell viability, anti cancer effect through modulation of KAI1/CD82 gene expression in breast cancer MCF-7 cell line. Methods: The effects of imatinib mesylate on cell viability in MCF-7 cell line were assessed using MTT assay and IC50 value was determined. GAPDH and KAI1/CD82 were selected as reference and target genes, respectively. Quantitative real time PCR technique was applied for investigation of KAI1/CD82 gene expression in human breast cancer MCF-7 cells. Subsequently, the quantity of KAI1 compared to GAPDH gene expressions were analyzed using the formula; 2−ΔΔCt. Results: Imatinib was showed to have a dose-dependent inhibitory effect on the viability of MCF-7 cells. CD82/GAPDH gene expression ratios were 1.322 ± 0.030 (P > 0.05), 2.052 ± 0.200 (P < 0.05), 2.151 ± 0.270 (P < 0.05) for 10, 20 and 40 mmol/L of imatinib concentrations. Conclusions: Based on the present data, imatinib mesylate might modulate metastasis by up-regulating KAI1/CD82 gene expression in human breast MCF-7 cancer cell line.

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