Asian Pacific Journal of Tropical Biomedicine

591–598

Authors: Luciane Corbellini Rufatto, Denis Amilton dos Santos, Flavio Marinho, João Antonio Pegas Henriques, Mariana Roesch Ely, Sidnei Moura

Propolis has been used worldwide for years in folk medicine and currently marketed by the pharmaceutical industry. In Brazil, propolis was classified into 13 groups based on their organoleptics and physicochemical characteristics. The 13th type named red propolis has been an important source of investigation since late 90s. Their property comes from the countless compounds, including terpenes, pterocarpans, prenylated benzophenones and especially the flavonoids. This last compound class has been indicated as the responsible for its potent pharmacological actions, highlighting the antimicrobial, antiinflammatory, antioxidant, healing and antiproliferative activities. The red propolis can also be found in other countries, especially Cuba, which has similar features as the Brazilian. Therefore, with the compilation of 80 papers, this review aims to provide a key reference for researchers interested in natural products and discovery of new active compounds, such as from propolis.

603–613

Authors: Prashant Y. Mali*, Shital S. Panchal

The intention of current review is to make available up-to-date information on morphology, ecological biodiversity, medicinal uses, phytochemistry and pharmacological activities on different parts of Euphorbia tirucalli (E. tirucalli). This plant has a number of medicinal uses. Latex of E. tirucalli is vesicant and rubefacient which is used for rheumatism, warts, cough, asthma, ear-ache, tooth-ache and neuralgia. It acts as a purgative in small doses while in big doses it is bitter irritant and emetic. Milky juice is alexiteric, carminative and purgative. It is useful in whooping cough, gonorrhea, asthma, leprosy, dropsy, dyspepsia, enlargement of spleen, colic, jaundice and stone in bladder. The fresh milky juice is good alternative in syphilis and a good application in neuralgia. A decoction of branches is used in gastralgia and colic. Bark is used in treatment of fractures. Poultices prepared from the stem are useful to repair the broken bones. Boiled root liquid acts as an emetic in cases of snake-bite and for infertility in women. The wood is used for rafters, toys and veneering purposes. It is also useful against leprosy and foot paralysis subsequent to childbirth. E. tirucalli is reported to have euphol, b-sitosterol, euphorbol hexacosonate, cycloeuphordenol, cyclotirucanenol, tirucalicine, tri-methyl ellagic acid, gallic acids, terpenic alcohol, isoeuphorol, taraxasterol, tirucallol, euphorone, euphorcinol, euphorbins, 12-deoxy-4 b-hydroxyphorbol-13-phenyl acetate-20- acetate, 12, 20-dideoxyphorbol-13-isobutyrate, glut-5-en-3- b-ol, 3,3 0-di-O-methylellagic acid, euphorbin-A (polyphenol), tirucallin-A (7) (tannin), tirucallin-B (11), euphorbin-F (14) (dimers), cycloartenol, 24-methylenecycloartenol, ingenol triacetate, 12-deoxy-4 bhydroxyphorbol-13-phenyl acetate-20-acetate, taraxerone, euphorginol, taraxerol, campesterol, stigmasterol, palmitic acid, linoleic acid, b-amyrin, etc. active phytoconstituents. E. tirucalli have possessed activity in human-lymphocytes, analgesic, anthelmintics, antiarthritic, antibacterial/antifungal/antimicrobial, anti-HIV, anti-inflammatory, antioxidant, antiviral, biodiesel production, CNS depressant/neuropathic pain, cytotoxicity/ anticancer, genotoxic/mutagenic, hepatoprotective, insect repellants, immunomodulatory, larvicidal, molluscicidal/ovicidal/piscicidal, myelopoiesis, proteolytic/chitinolytics pharmacological activities. There is a need to isolate dynamic constituents, their biological trial, molecular mechanisms, experimental protection and legalization of therapeutic uses of E. tirucalli. The collected information will be helpful to locate up study protocol for recent drugs and Ayurvedic formulation expansion in curative and treat a variety of ailments. 1. Introducti

619–623

Authors: Rooh Ullah, Shafiq Ahmed Tariq*, Naeem Khan, Nawaz Sharif, Zia Ud Din, Khalid Mansoor

Objective: To evaluate the antihyperglycemic potential of Tamarix aphylla (T. aphylla) leaves in STZ–NIC induced diabetes in Wister Albino rats. Methods: Acute toxicity study was done to check the toxicity of T. aphylla (L. Karst) methanol extract. T. aphylla leaves extract was administered intraperitoneally (100 mg, 250 mg and 400 mg/kg body weight per day) to diabetic Wister rats for 21 days. The various parameters were studied including fasting blood glucose levels, haemoglobin and glycosylated haemoglobin. Results: The treatment groups with the extract at three dose levels expressively abridged the intensities of blood glucose and Glycosylated Haemoglobin. The earlier detected reduced level of plasma haemoglobin of the diabetic rats was raised to near normalcy with treatment of extract. Conclusions: The results of the current study confirm that the leaves extract of T. aphylla are nontoxic and have antidiabetic nature.

629–632

Authors: Malek Besbes Hlila, Kaouther Majouli, Hichem Ben Jannet, Mahjoub Aouni, Maha Mastouri, Boulbaba Selmi

Objective: To examine the potential antimicrobial activity of Euphorbia paralias L. (Euphorbiaeae) leaves and stems extracts. Methods: The antimicrobial activity was tested against six microbial strains: Escherichia coli ATCC 8739, Bacillus subtilis ATCC 6633, Salmonella enterica CIP 8039, Staphylococcus aureus ATCC 6538, Pseudomonas aeruginosa ATCC 9027 and Candida albicans ATCC 90028 by two different methods, the disk method and the dilution method. Results: Our results showed the important antimicrobial activity of the chloroform extract of the stems towards the majority of the strains by using both methods. Bacillus subtilis was the most sensitive strain (MIC = MBC = 15 mg/mL). Conclusion: Thus, some extracts of Euphorbia paralias can be used in the treatment of infectious diseases caused by microbes.

640–646

Authors: Lydia Enyonam Kuatsienu, Charles Ansah, Michael Buenor Adinortey

Objective: To evaluate the toxic potential of Launaea taraxacifolia leaf extract (LTE) in rats within 14 d of oral administration and also assess the potential of LTE in protecting against kidney injury induced by gentamicin using rat model. Methods: The protective ability of LTE was done after sub-acute toxicity evaluation has been carried out. Acute Kidney Injury (AKI) was induced by gentamicin at a dose of 160 mg/kg intraperitoneal i.p. Parameters and indicators considered include mortality, clinical signs, body and organ weights, haematological and clinical chemistry parameters. Gross examination and histopathological assessment was also done on selected internal organs. Results: There were no treatment-related deaths or changes in clinical signs, haematological and clinical chemistry indices during sub-acute toxicity studies with the exception of creatinine levels. This was confirmed by micrographs obtained from histopathological analysis. Co-administration of LTE with 160 mg/kg of gentamicin (i.p) markedly decreased the levels of urea and creatinine when compared to negative control group. Histological studies of kidney tissues showed an insignificant change in tubular epithelium in LTE plus gentamicin treated group compared to LTE treated only. Conclusions: Data obtained show that ethanolic leaf extract of Launaea taraxacifolia is non-toxic within a 14 d administration at a maximum dose of 1 000 mg/kg bwt and also possesses the ability to protect against gentamicin-induced kidney damage in rats at a dose of 300 mg/kg bwt.

654–659

Authors: Akintayo Ogundajo, Benjamin Okeleye, Anofi Omotayo Ashafa

Objectives: To investigate antimicrobial and cytotoxic potentials as well as chemical constituents of extracts from Macaranga barteri (M. barteri). Methods: Antimicrobial activity was carried out using micro-dilution, cell culture and GC–MS methods were employed to determine the cytotoxicity and chemical constituents of the extracts respectively. Results: Marked activity was observed in methanol (ME) fraction [MIC50: (0.097 7– 6.250 0) mg/mL] compared to hexane and ethyl acetate fractions. Aeromonas hydrophila (environmental strain) and Shigella sonnei (ATCC 29930) were the most susceptible pathogens to ME and ciprofloxacin (Cl) at MIC50 value of 0.097 7 and < 0.019 5 mg/mL respectively. Cryptococcus neoformans (ATCC 66031) was susceptible to ME at 0.195 3 mg/mL compared to fluconazole at 10.000 0 mg/mL. Decreased viability of the Vero cells was observed at the concentrations of 0.1–1.0 mg/mL. The lethal dose (LC50) of hexane, ethyl acetate and methanol fractions were recorded at (0.30 ± 0.07), (0.52 ± 0.05) and (0.22 ± 0.04) mg/mL, respectively. Some of the compounds identified from ME were caryophyllene (25.21%), neophytadiene (11.90%), a-humulene (7.67%), phytol (4.40%), ethyl ester hexadecanoic acid (4.04%) and nerolidol (2.83%) which were known to have various antimicrobial activities. Conclusions: Methanol fraction of M. barteri is a potent and safe antimicrobial and antifungal alternative which can be useful in the search for new antimicrobial drugs. The study also confirmed the orthodox usage of M. barteri in combating infectious diseases.

666–669

Authors: Nunthawan Nowwarote, Thanaphum Osathanon

Objective: To investigate a dysregulation of Notch signaling in oral lichen planus (OLP) using public available microarray dataset. Methods: A mRNA expression profiling dataset from Gene Expression Omnibus was downloaded. Differential gene expression between OLP and normal oral epithelium was examined using NetworkAnalyst. The dysregulated genes related to Notch signaling were identified. Results: Thirteen genes in Notch signaling pathway were significantly differential expressed between OLP and normal epithelium. OLP samples significantly increased the mRNA levels of HEYL, APH1B, CNTN1 and PSEN2. Whilst, ITCH, HES1, TLE2, DLK2, DTX2, NOTCH3, JAG2, RFNG, and SPEN were downregulated in OLP groups. Conclusions: Notch signaling was dysregulated and may participate in pathophysiologic process in OLP.

505–512

Authors: Vagish Kumar Laxman Shanbhag

Curcumin is a widely researched natural product and is known to possess anticarcinogenic properties. Chronic lymphocytic leukemia is a type of leukemia that principally affects patients with age higher than 60 years. Since the toxicity of conventional drugs exceeds the benefits of treating this leukemia type, patients are treated only in the advanced symptomatic stages. The current article reviews curcumin, its general actions and targets in cancer, and specifically that of it in chronic lymphocytic leukemia.

524–532

Authors: Oluwaseyi Tunrayo Taiwo, Sammy Olufemi Sam-Wobo, Olufunmilayo Ajoke Idowu, Adewale Oladele Talabi, Adewale Matthew Taiwo

Objective: This study compared the prevalence of intestinal helminths in Water, Sanitation and Hygiene (WASH) intervention and non-intervention communities in Abeokuta, Nigeria. Methods: Stool samples were collected from 225 respondents in a study carried out between July and November, 2014. Stool samples were examined for presence of helminths using Formol-Ether concentration method. Data collected from stool samples were analyzed using SPSS for Windows (version 16). Results: Results showed that, at the intervention community, 88 out of 113 respondents were infected with at least one helminth infection while at non-intervention community, 80 out of 112 respondents were infected. This result revealed overall helminth prevalence of 78% at Mawuko and 71% at Isolu. In both intervention (Mawuko) and non-intervention (Isolu) communities, hookworm was the most prevalent helminth observed (21% and 18%, respectively) followed by Ascaris lumbricoides (13% and 13%, respectively), Taenia sp. (4% and 3%, respectively) and Trichostrongylus sp. (1% and 3%, respectively). Cases of single infections of Trichuris trichiura, Strongyloides stercoralis and Hymenolepsis nana were observed only at the non-intervention community. However, S. stercoralis occurred in the multiple infections observed at the intervention community. Significantly more (p < 0.05) cases of infections were observed in male (46%) at the intervention community than female (31%) while in non-intervention community females (39%) were significantly (p < 0.05) more infected than their male counterparts (32%). Conclusions: This study concludes that the Community Led-Total Sanitation intervention programme, which was executed in Mawuko was not effective as expected.

538–543

Authors: Nathalie Heil, Karent Bravo, Andres Montoya´ , Sara Robledo, Edison Osorio

Objective: This study was designed to investigate the wound healing activity of aqueous extracts of Ullucus tuberosus (U. tuberosus) using in vitro models. Methods: Lyophilized pulp and acetone extracts of U. tuberosus were produced using ultrasound extraction. The capacity for collagenase activation was evaluated using fluorescence detection of the enzymatic activity. Then, the influence of U. tuberosus extracts on cell proliferation, cell migration and synthesis of the extracellular matrix (ECM) proteins, metalloproteinase (MMP-1) and pro-collagen was analyzed using human dermal fibroblasts in culture. Results: An increase in collagenase activity of 12% supports the utility of U. tuberosus as an agent for scar treatment. In addition, the extracts showed an increase in the proliferation and migration of human dermal fibroblasts and the production of pro-collagen and MMP-1 after treatment with U. tuberosus extracts. The increase in proliferation, migration and pro-collagen levels positively influenced the regeneration of scarless tissue during the proliferation phase, whereas the increase in MMP-1 may have favored the wound healing process during the remodeling and cellular differentiation phases. Conclusion: The results of this study show for first time that U. tuberosus is a promising candidate to support scarless tissue regeneration.

549–554

Authors: Ali Jelowdar, Abdollah Rafiei, Mohammad Reza Abbaspour, Iran Rashidi, Mahmood Rahdar

Objective: To evaluate the efficacy of combined ABZ and PZQ and their solid lipid nanoparticles in chemoprophylaxis of cystic echinococcosis (CE). Methods: ABZ and PZQ loaded solid lipid nanoparticles (SLNs) were prepared by high shear homogenization and microemulsion congealing techniques with some minor modification. Nanoparticles average size, polydispersity index (PDI), and particle size distribution were determined by scanning electron microscopy (SEM) and photon correlation spectroscopy. Forty females BALB/c were experimentally infected by protoscoleces (PSC) and randomly divided into four equal groups of 10 mice. After the end of the 3 months treatment period and 2 months rest, mice were sacrificed and the peritoneal cavity was opened for removal, counting, measuring, and histological analysis of hydatid cyst. Results: The results indicated that ABZ and PZQ chemoprophylaxis treatment reduced the wet weight and size of developed cysts 77.3% and 79%, respectively. The corresponding result for the ABZ and PZQ loaded SLNs was 83% and 85%, respectively. Conclusions: This study for the first time demonstrated that ABZ and PZQ loaded SLNs is superior to free ABZ and PZQ for the chemoprophylaxis of CE in mice.

563–568

Authors: Muawiya Musa Abarshi, Edward Oyedeji Dantala, Sanusi Bello Mada

Objective: To investigate the level of heavy metals (Cu, Zn, Fe Mn Ni, Pb and Cd) in muscles, gills and liver tissue of Croaker fish from oil spilled rivers of Bonny and Finima, Niger Delta region, Nigeria. Methods: Twenty four Croacker fishes (Genyonemus lineatus) weighing between 250 and 260 g and 24–26 cm length were collected from each sampling sites (Bonny and finima rivers). The frozen fishes were thawed and dried at 105
C until they reach a constant weight. The dried samples were homogenized and digested with 10 ml tri-acid mixture (HNO3: HClO4:H2SO4), and then the digested samples were diluted to 100 ml with deionized water. Heavy metals (Cu, Ni, Zn, Pb, Mn, Iron Fe, and Cd) concentrations were determined by atomic absorption spectrophotometer. Results: Our results revealed that concentrations and distribution patterns of all heavy metals studied varied significantly (P < 0.05) amongst the fish tissues analyzed and sampling sites. Moreover, liver tissue of fish caught from Finima creek accumulated the highest concentrations of Cu (52.64 ± 3.01 mg/g dry wt), Zn (166.50 ± 6.45 mg/g dry wt) and Fe (801.50 ± 14.15 mg/g dry wt) in comparison to the liver of fish caught from Bonny river in which the levels of Cu, Zn and Fe were (45.00 ± 2.79), (49.90 ± 2.91) and (216 ± 6.11 mg/g dry wt), respectively. In addition, Mn, Ni, Pb and Cd exhibited their highest concentrations in gills from both locations. As expected, muscle tissue contained the least concentrations of all metals investigated from both sampling sites. Furthermore, all the metals investigated exhibited highest concentration in fish collected from Finima creek. These abnormal high level heavy metals accumulation observed in this location could be linked to the frequent crude oil spills as well as industrial activity around the area which might get discharged into Finima creek. In general, the mean concentrations of some toxic heavy metals investigated exceed the recommended maximum permissible limits set by the Joint FAO/WHO committee. However, some metals are within the acceptable limits. Conclusions: In conclusion, our data showed the abnormal higher concentration of these metals and this might be toxic to the fish and other aquatic organisms directly or by extension to humans that frequently consumed such contaminated fishes.

577–584

Authors: Samuel Odeyemi, Anthony Afolayan, Graeme Bradley

Objective: The bulb of Albuca bracteata Jacq., and Albuca setosa commonly used in Eastern Cape Province of South Africa for the treatment of several types of disease conditions including diabetes, was investigated for their phytochemical contents and antioxidant activities. Methods: The antioxidant activity was determined by evaluating the effect of various solvent extracts (acetone, methanol, ethanol and water) on 2,2 – diphenyl-2- picrylhydrazyl (DPPH) scavenging activity, 2,2 0 – azino-bis (3-ethylbenzthiazoline-6- sulfonic acid diammonium salt (ABTS), ferric reducing power, hydrogen peroxide and nitric oxide scavenging activity while the various extracts were also analysed for phytochemical their contents. Results: The results obtained indicate that polyphenols, proanthocyanidins, saponins and alkaloids are present in higher concentrations. The composition of phenols (117 mg/g), flavonols (26.28 mg/g), proanthocyanidins (84.85 mg/g) and flavonoids (5.36 mg/g) were significantly higher in the acetone extract while saponin and alkaloids contents were higher in the aqueous extract. Antioxidant studies of the extracts for nitric oxide, hydrogen peroxide, ABTS and DPPH showed high antioxidant potential in a concentration-dependent manner that was not significantly different from Vitamin C, BHT and Rutin used as standard references. The methanol extract showed higher ferric reducing potential compared to the aqueous extract but was significantly different from Vitamin C, BHT and Rutin. Conclusion: This implies that the bulb of Albuca bracteata may serve as a natural source of antioxidants, which together with the numerous phytochemicals present, could account for its folkloric use as a medicinal plant.

587–590

Authors: Nurul Suhaiza Hassanudin, Zubaidah Abdul Wahab, Khalid Ibrahim, Fadzilah Mohd Nor

Infections caused by Strongyloides stercoralis (S. stercoralis) in human are generally asymptomatic, however in immunocompromised individual, hyperinfection may develop with dissemination of larvae to extra-intestinal organs. The diagnosis could be easily missed due to asymptomatic presentation and insufficient exposure towards the infection itself, which may lead to low index of suspicion as a consequence. In this report, a case of a Malaysian male with underlying diabetes mellitus, hypertension, cerebrovascular accident, bullous pemphigus and syndrome of inappropriate antidiuretic hormone secretion who initially complained of generalized body weakness and poor appetite without any history suggestive of sepsis is presented. However, he developed septicemic shock later, and S. stercoralis larvae was incidentally found in the tracheal aspirate that was sent to look for acid fast bacilli. Regardless of aggressive resuscitation, the patient succumbed due to pulmonary hemorrhage and acute respiratory distress syndrome. It was revealed that the current case has alarmed us via incidental finding of S. stercoralis larvae in the tracheal aspirate, indicating that the importance of the disease should be emphasized in certain parts of the world and population respectively.

385–388

Authors: Afzan Mat Yusof, Najat Hashim, Muhammad Lokman Md Isa

Objective: To identify the prevalence of Cryptosporidium from goats in three types of farm management systems in Terengganu, Malaysia and to determine the Cryptosporidium species infecting goats by using 18S rRNA. Methods: A total of 478 fecal samples were randomly collected from goats in three farms; 199 samples were collected from intensive farm, 179 samples from semi-intensive farm and 100 samples from extensive farm. The samples were processed by using formolether concentration technique and stained by using modified Ziehl–Neelsen. Positive samples were performed by using nested PCR analysis by using 18S rRNA. Results: Out of 478 goats, 207 (43.3%) were found to be infected with Cryptosporidium. Goats reared under the intensive farm management system reported the highest prevalence of infection (49.7%), followed by intensive farm management system (41%) and the lowest prevalence was reported in the goats reared under semi-intensive management system (37.4%). Conclusions: The identified species found in goat was Cryptosporidium parvum. Future study on the zoonotic transmission of Cryptosporidium parvum in goats needs to be done in order to find the source of transmission of this parasite.

397–400

Authors: Andi Nur Fitriani Abubakar, Suminar Setiati Achmadi, Irma Herawati Suparto

Objective: To determine the structure of triterpenoid isolated from avocado seeds and the cytotoxic effect on MCF-7 and HepG2 cells. Methods: The powder sample was macerated with ethanol, followed with separation of the extract by column chromatography. The target compound was monitored on thin layer chromatography plate and reagent Lieberman–Buchard. The isolated compound was characterized by spectral analysis, mainly ultraviolet, infrared, and liquid chromatographymass spectroscopy and their spectroscopic data with those reported in literature were compared. In vitro cytotoxic activity was investigated against Vero, MCF-7, and HepG2 cell lines using MTT assay. Results: A triterpenoid compound was isolated from ethanol extract. The extracts, fraction (F3), and the isolated compound showed a significant cytotoxic activity against all investigated cell lines. MTT assay showed that the triterpenoid isolate inhibited cell proliferation of MCF-7 and HepG2 cell line with the IC50 values of 62 mg/mL and 12 mg/ mL, respectively, and was safe to normal cells. Conclusions: The results of the present study reveal that triterpenoid from avocado seeds have the potential for further development as anticancer agents.

406–410

Authors: Edy Setiti Wida Utami, Sucipto Hariyanto, Yosephine Sri Wulan Manuhara

Objective: To study asymbiotic seed germination and mass propagation of Dendrobium lasianthera which is one of the endangered medicinal orchids using seeds. Methods: The 14 weeks old hand pollinated seeds were sown on Vacin and Went (VW) solid medium supplemented with various concentrations of peptone (1, 2, 3 g/L) and without peptone which was used as control treatment. At the 4, 8, and 12 weeks after the seeds were sown, seed germination and shoot formation were investigated. To evaluate the role of organic nutrient additives on subsequent shoot development and root formation, particular shoots with about 1 cm length contains 1–2 leaves obtained from the seeds germination was cultured on VW medium additives with different of organic nutrient: 15% coconut water, 2 g/L peptone, 150 g/L banana homogenate, and without organic nutrient was used as control. After 16 weeks of culture, the plantlet height, number of leaves, number of roots, leaf length and root length were recorded. Results: The supplementation of 2 g/L peptone in VW medium was proven to be suitable concentration for seed germination (100%) and shoot formation with 84.0% the protocorm development to phase 5 (shoot). VW medium containing 15% coconut water was effectively improved the shoot development, with well developed roots and leaves compared to the other treatment and 95% of acclimatized plantlets survived. Conclusions: This protocol is an efficient way for the in vitro mass propagation of this Dendrobium lasianthera.

416–420

Authors: Kanchanat Ternchoocheep, Damrassamon Surangkul, Sukhgij Ysothonsreekul

Objective: To investigate the effect of asiatic acid (AA) on the differentiated human neuroblastoma SH-SY5Y cells cytotoxic-induced by cholesterol. Methods: Human neuroblastoma SH-SY5Y cells were either exposed to different concentrations of AA or treated with different doses of cholesterol to reveal their responding viability by MTT assay. The selective 1 mmol/L concentration of AA was then used to test for either the protective or the recovery effects on the cells treated with 250 mmol/L concentration of cholesterol. Results: AA has a propensity to directly increase the viability of differentiated human neuroblastoma SH-SY5Y cells. Cholesterol has significant cytotoxic effect on those cells in a concentration-dependent manner. AA has the ability to slightly recover the viability of the differentiated culture cytotoxic-induced by cholesterol but could not protect those cells from cytotoxic-induced by cholesterol. Conclusions: High concentrations of cholesterol were observed to be harmful to the neurons and AA had a slight effect of reducing neuronal death caused by cholesterol.

428–431

Authors: Wimonrat GunTang, Nathamon Kamonvoradej, Chitchanok Chomchat, Sangrawee Suriyakan2, Sangob Sanit3, Jintana Wongwigkarn, Nophawan Bunchu, Damrongpan Thongwat, Supaporn Lamlertthon

Objective: To investigate the prevalence of Candida spp. and the virulence factors of Candida albicans (C. albicans) isolated from external surfaces of blow flies collected from Mae Sot, Tak Province, Thailand. Methods: The blow flies were collected by sterile sweep nets from three areas in Mae Sot. Yeast isolation was first performed on Sabouraud dextrose agar (SDA) supplemented with chloramphenicol and cycloheximide. The yeast isolates were then identified by using chromogenic agar, a yeast identification test kit, a germ tube formation test and a carbohydrate utilization test. The b-hemolysis was determined on 7% sheep blood agar, while phospholipase activity was measured on SDA agar supplemented with 10% egg yolk suspension. Antifungal susceptibility testing was determined by broth micro-dilution testing against ketoconazole and amphotericin B. Results: The prevalence rate of Candida spp. on the external surfaces of the blow flies was 78.1%. All C. albicans isolated from the blow fly demonstrated b-hemolysin and potent phospholipase activities and 47.1% of C. albicans were resistant to ketoconazole with MIC values 128 mg/mL. Conclusions: The results indicate that blow flies could play an essential role in the transmission of potentially pathogenic and antifungal resistant C. albicans into the environment. Further investigation on other virulence factors and genetic relatedness among isolates from the blow flies, the environment and clinical specimens is required to confirm this role.

437–442

Authors: Guntur Berlian, Olivia Mayasari Tandrasasmita, Raymond Rubianto Tjandrawinata

Objective: To study the regulation of trombinol on thrombopoietin, an essential regulator of thrombocyte production. Methods: Effect of trombinol on thrombopoietin regulation was evaluated at the mRNA and protein levels in human hepatoma HepG2 cells. The mRNA expressions were revealed by PCR and real-time PCR, while the protein expressions were analyzed using western blotting and human ELISA kit. Statistical differences between the test were determined by student's t-test with P < 0.05 was considered statistically significant. Results: Trombinol significantly increased the expression of thrombopoietin at the level of mRNA and protein secretion in HepG2 cell lines. Trombinol with the concentration of 15 mg/mL, positively induces 2.5-fold of thrombopoietin expression. Up-regulation of GABP, a transcription factor of thrombopoietin, is suggested to be involved in cellular regulatory mechanisms of trombinol. Here, our result shows convincing evidence that trombinol affects the thrombopoietin productions in vitro. This molecular explanation of thrombopoietin's stimulating function is in line with the traditional use of Psidium guajava for treatment of diseases involving thrombocytopenia. Conclusions: Thrombopoietin stimulating function of trombinol could be potentially considered as one of alternative treatment for thrombocytopenia-related cases, including post chemotherapy shock, dengue fever and liver failure.

450–454

Authors: Yatnita Parama Cita, Achmad Suhermanto, Ocky Karna Radjasa, Pratiwi Sudharmono

Objective: To explore secondary metabolite of bacteria-associated Xestospongia testudinaria from Tanjung Kasuari, Sorong, Papua. Methods: The antimicrobial activities of extracts against two Gram-positive bacteria (Staphylococcus aureus) and three Gram-negative bacteria (Pseudomonas aeruginosa, Eschericia coli and Salmonella typhi) were determined by disk diffusion dilution method. Results: The test showed that of 15 isolates of symbiont bacteria, 6 isolates were successfully isolated and coded, namely, Xp 4.1, Xp 4.2, Xp 4.3, Xp 4.4, Xp 4.5 and Xp 4.6. Of the six bacterial isolates, isolated Xp 4.2 was found to have more powerful antibacterial activity than any other isolates of symbiont bacteria. Antibacterial activity assay for the n-hexane soluble fractions, ethyl-acetate soluble fractions, and n-buthanol soluble fractions revealed more powerful anti-bacterial activity than any other soluble fractions. Phytochemical screening showed alkaloid and steroid/triterpenoid, while identification for isolate of Xp 4.2 bacterial showed bacteria. Conclusions: Metabolites of bacterial associated with marine sponge Xestospongia testudinaria promise to be developed into antibacterial agents.

460–465

Authors: Enayatollah Seydi, Shabnam Babaei, Amir Fakhri, Jalal Pourahmad

Objective: To explore the cytotoxicity effects of Caspian cobra (Naja oxiana or N. oxiana) venom on hepatocytes and mitochondria obtained from the liver of HCC rats. Methods: In this study, HCC was induced by diethylnitrosamine (DEN), as an initiator, and 2-acetylaminofluorene (2-AAF), as a promoter. Rat liver hepatocytes and mitochondria for evaluation of the selective cytotoxic effect of N. oxiana venom were isolated and mitochondria and cellular parameters related to apoptosis signaling were then determined. Results: Our results showed a raise in mitochondrial reactive oxygen species (ROS) level, swelling in mitochondria, mitochondrial membrane potential (Djm) collapse and release of cytochrome c after exposure of mitochondria only isolated from the HCC group with the crude venom of the N. oxiana (12.5, 25, and 50 mg/mL). This crude venom also induced caspase-3 activation (P < 0.001) in the hepatocytes obtained only from the HCC rat liver. Conclusions: Based on the over all results, we suggested that N. oxiana may be considered as a promising complementary therapeutic agent for the treatment of HCC.

473–477

Authors: Rajiv Ravi, Zary Shariman Yahaya

Objective: To investigate the prevalence, phylogenetics and DNA barcoding of Zeylanicobdella arugamensis (Z. arugamensis) from crimson snapper (Lutjanus erythropterus), Jerejak Island, Penang, Malaysia. Methods: Experiment was conducted with 200 fish specimens of cultured Lutjanus erythropterus from Jerejak Island, Penang, Peninsular Malaysia. The water temperature and length for each fish were measured prior to parasites examination. Next, the morphological identification of parasites was performed. Genomic DNA from parasites was extracted for further molecular analysis. After PCR amplification, phylogenetic tree was constructed. The lowest Bayesian information criterion scores showed that the most compatible model is Tajima and Nei. Finally, data sets of cytochrome oxidase subunit I gene sequence and trace file have been submitted to Barcode of Life Data System. Results: The prevalence rate of Z. arugamensis was recorded to be 11.5%, and the intensity was 1.48. The low intensity was due to the water temperature recorded in this study (32.9–33.2
C). All the individuals of Z. arugamensis recorded in this study showed a close relationship with species that were recorded in NCBI database (Z. arugamensis DQ414344, Aestabdella leiostomi DQ414305, Pterobdella amara DQ414334 and Cystobranchus meyeri DQ414315) but less relationship with Aestabdella abditovesiculata DQ414300. Finally, the DNA sequences submitted to Barcode of Life Data System in accordance to species have already obtained Barcode Index Number as BOLD: ACM3477. Conclusions: This study has provided an overview of sequence divergence at cytochrome oxidase subunit I gene, DNA barcodes and parasite prevalence of Z. arugamensis.

483–491

Authors: I. Made Artika, Chairin Nisa Ma'roef

Emerging viruses are viruses whose occurrence has risen within the past twenty years, or whose presence is likely to increase in the near future. Diseases caused by emerging viruses are a major threat to global public health. In spite of greater awareness of safety and containment procedures, the handling of pathogenic viruses remains a likely source of infection, and mortality, among laboratory workers. There is a steady increase in both the number of laboratories and scientist handling emerging viruses for diagnostics and research. The potential for harm associated to work with these infectious agents can be minimized through the application of sound biosafety concepts and practices. The main factors to the prevention of laboratory-acquired infection are well-trained personnel who are knowledgable and biohazard aware, who are perceptive of the various ways of transmission, and who are professional in safe laboratory practice management. In addition, we should emphasize that appropriate facilities, practices and procedures are to be used by the laboratory workers for the handling of emerging viruses in a safe and secure manner. This review is aimed at providing researchers and laboratory personnel with basic biosafety principles to protect themselves from exposure to emerging viruses while working in the laboratory. This paper focuses on what emerging viruses are, why emerging viruses can cause laboratory-acquired infection, how to assess the risk of working with emerging viruses, and how laboratory-acquired infection can be prevented. Control measures used in the laboratory designed as such that they protect workers from emerging viruses and safeguard the public through the safe disposal of infectious wastes are also addressed.

493–504

Authors: Khaled Mohamed Mohamed Koriem

ultiple sclerosis (MS) is the most famous autoimmune disease attacking the central nervous system. It attacks people from age 20–50 years old and the females' attacks double than males' attacks. MS is an autoimmune disease affecting principally the central nervous system that causes nerve sheath demyelination, followed by axon damage and paralysis. MS symptoms include muscle weakness, weak reflexes, muscle spasm, diffi- culties in movement and unbalance. Many factors may be responsible for MS: microorganism, virus, smoking, stress, environmental toxins, contaminated diet and gout. MS is widely spread in the population in North Europe and this is related to lack of vitamin D due to decrease of sunlight exposure. MS biomarkers include nitric oxide, interleukin-6, nitric oxide synthase, fetuin-A and osteopontin. MS is not a genetic disease (not transferred from parents into next generations) but MS appears when leukocyte antigen system-related genes are changed in human chromosome 6. The physiology of MS patients is controlled by numbers of biological processes such as activation of immuneinflammatory, oxidative and nitrosative stress pathways. MS includes two main steps: (1) myelin sheath destruction and formation of lesions and, (2) inflammation. Four types of MS can be distinguished: relapsing-remitting, primary progressive, secondary progressive and progressive relapsing. Nine treatments have been accepted for relapsingremitting MS type: interferon b-1a, interferon b-1b, mitoxantrone, natalizumab, glatiramer acetate, fingolimod, dimethyl fumarate, teriflunomide, and alemtuzumab. However, the only treatment used is mitoxantrone for progressive MS with many side effects. Complementary treatments are also used in MS treatments such as vitamin D, Yoga, medicinal plants, oxygen therapy, acupuncture and reflexology.

275–279

Authors: Sylvie Linsuke1*,#, Liliane Mpabanzi, Sabin Nundu, Faustin Mukunda, Pascal Lutumba, Katja Polman

Objective: To improve knowledge and practice of health staff as well as the availability of material resources for diagnosis and management of schistosomiasis in two endemic provinces of DRC (Kinshasa and Bas-Congo). Methods: Structured interviews were performed using questionnaires with staff from 35 healthcare facilities in 9 health zones (HZ) of Kinshasa and 2 HZ in Bas-Congo. Results: Schistosomiasis was reported to be present in all the included HZ. Health staff knew the most important symptoms of schistosomiasis, but advanced symptoms were more accurately reported in Bas-Congo. Knowledge of symptoms related to schistosomiasis such as anemia (P = 0.0115) and pollakiuria (P = 0.0260) was statistically different in both two provinces. Kato-Katz technique and urine filtration were unavailable in both provinces. Parasitological diagnosis was mostly performed using the direct smear method. PZQ was available in 70% of the health facilities, all situated in Bas-Congo. Diagnosis and treatment mostly relied on symptoms and cost more in urban area than in rural. Conclusions: Though knowledge on schistosomiasis among health staff appears sufficient, substantial efforts still must be made to improve the availability of diagnostic tools and treatment in the health facilities in DRC.

288–293

Authors: Zulaikha Zahran, Nik Mohd Izham Mohamed Nor, Hamady Dieng, Tomomitsu Satho, Abdul Hafiz Ab Majid

Objective: To test the effectiveness of conidial spore formulations [Aspergillus tubingensis (A. tubingensis) and Trichoderma harzianum (T. harzianum)] against tropical bed bugs, Cimex hemipterus. Methods: Spore formulations were made from two fungal strains, T. harzianum and A. tubingensis. The bed bugs were exposed to the conidial spores placed soaked onto a fabric cloth for 1 h and the mortality counts were recorded daily until 14 days. Results: Mean survival times based on Kaplan–Meier survival analysis showed no significant differences between all the concentrations in both the fungal isolates: T. harzianum and A. tubingensis. However, the evaluation of both the isolates in terms of virulence resulted in low lethal hours in all the concentrations except for the high concentration of A. tubingensis (LT50 = 44.629 h) at the conidial exposure of 1 × 106 spores/ mL. Rapid mortality of the bed bugs was observed from Day 6 to Day 12, ranging from 13% to 90% in all three concentrations of A. tubingensis. With reference to the T. harzianum exposure, the concentration of 1 × 104 spores/mL displayed a gradual increase in the percentage mortality of 90 on Day 14. Conclusions: Approaches to the bed bugs treatment should be explored in-depth using a natural biological agent like fungus especially A. tubingensis to reduce this pest population, in order to replace chemical methods.

300–305

Authors: Emanuel Dani Ramdani, Ujiatmi Dwi Marlupi, James Sinambela, Raymond Rubianto Tjandrawinata

Objective: To identify and isolate the chemical compounds of Phaleria macrocarpa (P. macrocarpa) fruit ethanolic extract. Methods: Dried fruit of P. macrocarpa was extracted with 90% ethanol and partitioned between n-hexane/H2O and ethyl acetate/H2O. The organic layer was fractionated by various stationary phase and identified by using combined data of 1D [(proton nuclear magnetic resonance (NMR), carbon-13 NMR)], 2D-NMR (heteronuclear multiplequantum correlation and heteronuclear multiple-bond correlation), and mass spectrum. Results: Purification of n-hexane and ethyl acetate fractions from ethanolic extract of P. macrocarpa fruit resulted in isolation of nine compounds. Conclusions: A new compound was isolated and identified as glyceryl pentacosanoate. Also, two xanthones, which are 1,7-dihydroxy-3,6-dimethoxyxanthone and 1,6,7- trihydroxy-3-methoxyxanthone, are firstly reported to be isolated from P. macrocarpa.

314–320

Authors: Mohammad Bagher Rezvani, Mohammad Niakan, Mohammad Kamalinejad, Fateme Sadat Ahmadi, Faeze Hamze

Objective: To investigate the effect of Iranian honey, cinnamon and their combination against Streptococcus mutans bacteria. Methods: Nine experimental solutions were examined in this study, including two types of honey (pasteurized and sterilized), two types of cinnamon extract (dissolved in distilled water or dimethyl sulfoxide) and five different mixtures of cinnamon in honey (prepared by admixing 1%–5% w/w of cinnamon extract into 99%–95% w/w of honey, respectively). Meanwhile, each of mentioned agent was considered as the first solution while it was diluted into seven serially two-fold dilutions (from 1:2 to 1:128 v/v). Therefore, eight different concentrations of each agent were tested. The antibacterial tests were performed through blood agar well diffusion method, and the minimum inhibitory concentration (MIC) was determined. Ultimately, the data were subjected to statistical analysis incorporating Two-way ANOVA and Bonferroni post hoc tests (a = 0.01). Results: The highest zone of inhibition was recorded for the mixtures of honey and cinnamon while all the subgroups containing 95%–99% v/v of honey were in the same range (P < 0.01). The MIC for both honey solutions were obtained as 500 mg/mL whereas it was 50 mg/mL for both cinnamon solutions. Moreover, the MIC related to all honey/cinnamon mixtures were 200 mg/mL. Conclusions: A profound synergistic effect of honey and cinnamon was observed against Streptococcus mutans while there was no significant difference among extracts containing 99%–95% v/v of honey admixing with 1%–5% v/v of cinnamon, respectively.

332–338

Authors: Apinun Limmongkon1*, Prachya Janhom1, Ampika Amthong1, Muenfan Kawpanuk1, Patcharaporn Nopprang1, Jirarat Poohadsuan1, Thapakorn Somboon1, Suwaree Saijeen2, Damratsamon Surangkul1, Metawee Srikummool1, Tantip Boonsong1

Objective: To investigate the change in total phenolic compounds, antioxidant activity, and resveratrol content of five different germinated peanut cultivars. Methods: The germinated sprouts of five peanut cultivars (Kalasin1, Kalasin2, Konkaen, Konkaen4, and Tainan9) were extracted with 80% ethanol and collected as crude extract. The antioxidant capacities were determined with 2,2-diphenyl-1-picrylhydrazyl and ferric ion reducing antioxidant power method. The total phenolic compound was measured using the Folin–Ciocalteau assay. The qualification and quantification of resveratrol was performed by high performance liquid chromatography method. Results: Among the five cultivars, a three-day germination of Kalasin1 showed the highest phenolic content [(40.67 ± 2.62) mg gallic acid/g dry weight], expressed the highest 2,2-diphenyl-1-picrylhydrazyl antioxidant value [(80.51 ± 1.47) mmol/L Trolox/g dry weight], and ferric ion reducing antioxidant power antioxidant value [(171.33 ± 8.59) mmol/L ascorbic acid/g dry weight]. However, the high performance liquid chromatography result of Kalasin2 significantly increased to the highest resveratrol content of (6.44 ± 1.26) mg/g dry weight on the second day of germination. Conclusions: The variation of phytochemical content in the peanut sprout is due to the effect of the peanut cultivar and the germination period.

347–351

Authors: Rokhaya Fall, Saliou Ngom, Dienaba Sall, Mbacke Semb ´ ene ` 3, Abdoulaye Samb

Objective: To isolate and identify the compounds in the essential oils from the leaves of Callistemon viminalis (D.R.) and Melaleuca leucadendron (Linn.) collected in Dakar, Senegal. Methods: The essential oils from the leaves of these two myrtaceaes were extracted by steam distillation and analyzed by gas chromatograph and gas chromatography–mass spectrometer. Results: A total of 34 constituents were identified in the oil of Callistemon viminalis and the major compounds were 1.8-cineole (58.12%), limonene (9.72%), a-terpineol (9.56%), geranial (6.02%), d-elemene (3.53%), myrcene (2.96%) and a-pinene (2.49%). For the essential oil of Melaleuca leucadendron, 43 constituents were identified, and 1.8-cineole (28.87%), epiglobulol (23.06%), a-pinene (12.22%), limonene (11.65%) and a-terpineol (7.06%) were major compounds. Conclusions: Considering properties of the identified major compounds, essential oils of both studied myrtaceae could be used in the medicine field including the food, pharmaceutical and cosmetic industry.

358–366

Authors: Rigo Baluyot Villacorta, Kristine Faith Javier Roque, Giovanni Alarkon Tapang, Sonia Donaldo Jacinto

Objective: To examine three plant extracts [Lumnitzera racemosa (Combretaceae) (L. racemosa), Albizia procera (Fabaceae) (A. procera) and Cananga odorata (Annonaceae)] for their potential as source of photosensitizers in photodynamic therapy. Methods: Human mammary adenocarcinoma (MCF-7) cells were treated with the plant extracts, which were irradiated with 5.53 mW and 0.553 mW broadband light. Cell viability was assessed using MTT assay and induction of apoptosis was determined using terminal deoxynucleotidyl transferase-dUTP nick end labeling assay. Results: The crude ethanolic extracts, independently, were nontoxic against cancer and non-cancer cells but when irradiated with 5.53 mW broadband light, L. racemosa and A. procera extracts were cytotoxic against MCF-7 with IC50 of 11.63 mg/mL and 10.73 mg/mL, respectively. With 0.553 mW broadband light, the IC50 values were higher at 17.14 mg/mL and 19.59 mg/mL, respectively. Photoactivated L. racemosa and A. procera extracts were found to be more cytotoxic against MCF-7 than the non-cancer cell line, human dermal fibroblast-neonatal. Moreover, the cytotoxicity of the extracts was mediated by apoptosis. Conclusions: Two of the plant extracts used, L. racemosa and A. procera were toxic and induced apoptosis to mammary cell adenocarcinoma, MCF-7 when photoactivated. These extracts were also more toxic to human cancer than non-cancer cell lines.

373–377

Authors: Bora Korkut1*, Duygu Korkut2, Funda Yanikoglu1, Dilek Tagtekin

Objective: To determine quantitatively the amount of demineralization and the ability of commercially available products and an experimental cream to inhibit or reverse orthodontic related demineralization. Methods: A total of 20 patients who were 25–35 years old and having orthodontic treatment for 6–8 months were chosen. Caries risk assessments were done for each patient and ones with “moderate risk” were included. Patients with fixed orthodontic appliances were divided into 4 groups (5 patients each) including one control and 3 study groups. All patients used same toothpaste 2 times a day during the 3 weeks study period. Additional to the toothpaste first study group used MI Paste Plus (GC, Tokyo, Japan), second study group used Remin Pro (Voco, Cuxhaven, Germany) and third group used an experimental remineralizing cream per day for 3 weeks. Maxillary central and lateral incisors of each patient were examined by FluoreCam (Daraza Therametric Technologies, USA) device. The examinations were performed at baseline and at the end of 1st, 2nd and 3rd weeks. Results: According to the FluoreCam measurements the control group showed significant amount of demineralization at the end of 3 weeks, moreover the amount of demineralization has gradually increased in time. At the end of the study all 3 study groups showed significant amount of remineralization and the amount of remineralization for all the 3 study groups has gradually increased in time. However the amount of remineralization for 3rd study group was lesser than the 1st and 2nd study groups. The remineralization amounts for the 1st and 2nd study groups were determined to be identical. Conclusions: This study demonstrated that demineralization is measurable around orthodontic brackets and the demineralization can be completely inhibited and/or reversed by the use of commercially available remineralization products.

181–182

Authors: Somsri Wiwanitkit, Viroj Wiwanitkit

Malaria is an important tropical mosquito borne infection. It is still the present global public health issue. The management of malaria requires antimalarial drugs. The resistance to antimalarial drugs is a very big problem. The genetic variant is proposed to be an important factor affecting susceptibility to antimalarial drug. Here, the authors studied the change in molecular weight due to important pfatp6 and pfmdr1 polymorphisms and further implied the interrelationship with susceptibility to antimalarial drug. The greatest change can be seen in case of G639D (of pfatp6 polymorphism) while the least change can be seen in the case of N1042D (of pfmdr1 polymorphism). The results from some studies imply that there must be other factors that affect the susceptibility to antimalarial drugs. Those factors might be protein conformation factors, epigenetic factors or environmental factors. Further studies on these aspects should be carried out. It is concluded for possible role of epigenetic phenomenon.

188-192

Authors: Nadine Alvarez, Daymí Serpa, Ramlah Kadir, Yanely Tirado, Reinier Borrero, Sonsire Fernandez, Rub
en Cabrera, Yolanda Valdes, Caridad Zayas, Reinaldo Acevedo, Luis Izquierdo, María Elena Sarmiento, Mohd-Nor Norazmi, Jos
e Luis P
erez, Armando Acosta

Objective: To characterize the immunogenicity and the induction of cross-reactive responses against Mycobacterium tuberculosis (M. tuberculosis) of a proteoliposome (PL) from Mycobacterium bovis Bacillus Calmette–Gu
erin (BCG) with and without alum hydroxide (AL) as adjuvant (PLBCG-AL and PLBCG, respectively) in BALB/c mice. Methods: BALB/c mice were inoculated with phosphate buffer solution, BCG, PLBCG and PLBCG-AL. The humoral immunogenicity was determined by ELISA [immunoglobulin G (IgG), IgG1 and IgG2a] and the cellular immunogenicity was evaluated in vivo by delayed type hypersensitivity. The humoral cross-reactive response against M. tuberculosis was determined by Western blot. Results: Sera from animals immunized with PLBCG-AL and PLBCG showed significant increase in specific total IgG and IgG1 antibodies and the presence of cross-reactive antibodies against M. tuberculosis antigens, which were more intense with the use of alum as adjuvant. Mice immunized with PLBCG and PLBCG-AL also showed a specific cellular response in vivo. Conclusions: The cellular and humoral immunogenicity of PLBCG and the capacity to induce cross-reactive responses against M. tuberculosis is in agreement with the protective capacity previously demonstrated by this vaccine candidate and supports the continuation of its evaluation in further stages.

198–207

Authors: Sidgi Syed Anwer Abdo Hasson

Objective: To explore whether a DNA immunization approach targeting the major haemorrhage molecule of a prothrombin activator-like metalloproteinase from Echis ocellatus (E. ocellatus) venom could be conceived to inspire antibodies with more prominent specificity and equal adequacy to current conventional antivenoms systems. Methods: The isolated DNA EoMP-6 was used as the template for PCR amplification using the EoDC-2-specific forward and reverse primers. A PCR product of approximately 700 bp was obtained and cloned into pSecTag-B expression vector where anti-EoDC-2 antibodies were generated and analysed for their efficacy to neutralise local haemorrhage in vitro and in vivo. Results: Our results suggest that the generated anti-EoDC-2 showed a remarkable efficacy by (a) interfering with the interaction of the recombinant disintegrin “EoDC-2” isolated from the E. ocellatus as well as other viper species to the a2b1-integrins on platelets; (b) complete inhibition of the catalytic site of the metalloproteinase molecules in vitro using an adaptation antibody zymography assay. Furthermore, it has a polyspecific potential and constitutively expressed significant inhibition by cross-reaction and neutralised venom-induced local haemorrhage exerted by different viper species in vivo. The potential characteristic of EoDC- 2 against one part (the non-catalytic domain) as opposed to the whole molecule to neutralise its haemorrhagic activity is of crucial importance as it represents a novel approach with greater immunological specificity and fewer hazards, if any, than conventional systems of antivenom production, by exposure large animals that usually being used for the current antivenom production to a less injurious than expression of the whole molecule containing the catalytic metalloprotease domain. Hence, we report for the first time that our preliminary results hold a promising future for antivenom development. Conclusions: Antibodies generated against the E. ocellatus venom prothrombin activatorlike metalloprotease and disintegrin-cysteine-rich domains modulated and inhibited the catalytic activity both in vitro and in vivo of venom metalloproteinase disintegrin cysteine rich molecules. Thus, generating of venom specific-toxin antibodies by DNA immunization offer a more rational treatment of snake envenoming than conventional antivenom.

214–221

Authors: Fateheya Mohamed Metwally, Hend Mohamed Rashad, Hanaa Hamdy Ahmed, Asmaa Ahmed Mahmoud, Ehab Ragaa Abdol Raouf, Aboelfetoh Mohamed Abdalla

Objective: To elucidate the molecular mechanisms of the potent anti-obesity effect of Moringa oleifera Lam. (M. oleifera) ethanolic extract and to clarify the link between these mechanisms and the associated metabolic and vascular risks in the experimental model of visceral obesity. Methods: M. oleifera ethanolic extract was orally administered at 600 mg/kg body weight in obese female rats daily for 12 weeks. At the end of treatment, body weight was determined, and the atherogenic index, coronary artery index, glucose level, insulin resistance status, liver and kidney functions were assessed. Also, the mRNA of leptin, adiponectin and resistin in visceral adipose tissue was determined by quantitative real time-PCR. Results: The results showed that M. oleifera extract down-regulated mRNA expression of leptin and resistin, while it up-regulated adiponectin gene expression in obese rats relative to untreated obese control counterparts. This amelioration of genes expression was paralleled by a reduction in body weight and improvement of the atherogenic index and coronary artery index, as well as glucose level and insulin resistance value without adverse effects on liver or kidney functions, versus the untreated obese control ones. Conclusions: It is reasonable to assume that the anti-obesity, anti-atherogenic and antidiabetic properties of M. oleifera are mechanistically achieved via working directly on the adipokines of the visceral adipose tissue. Therefore, M. oleifera may be a good therapeutic candidate for the symptoms of metabolic syndrome.

227–233

Authors: Ghodsieh Bagherzade, Maryam Manzari Tavakoli, Mohmmad Hasan Namaei

Objective: To synthesis silver nanoparticles (AgNPs) by using extract of saffron (Crocus sativus L.) wastages and to test their antibacterial activity against six bacteria. Methods: In this paper, the synthesis of AgNPs using aqueous extract of saffron wastage as a green method without any chemical stabilizer and reducer is demonstrated. The synthesized AgNPs were determined by UV–vis spectrum, high resolution transmission electron microscopy, X-ray diffraction, and Fourier transmission infrared spectroscopy analysis. Results: UV–vis spectrum showed a peak at 450 nm due to excitation of surface plasmon vibrations. Fourier transmission infrared spectroscopy showed that nanoparticles were capped with plant secondary metabolites. X-ray diffraction analysis also demonstrated that the size range of the synthesized nanoparticles was 12–20 nm. Transmission electron microscope image illustrated AgNPs with spherical shape and an average size of 15 nm. The result of antibacterial activities showed that the biosynthesized AgNPs had an inhibiting activity against Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumonia, Shigella flexneri and Bacillus subtilis. Conclusions: The biosynthesized AgNPs showed significant antibacterial effect against Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumonia, Shigella flexneri and Bacillus subtilis, so, it can be used in biomedical applications.

240–244

Authors: Swastika Oktavia, Nastiti Wijayanti, Bambang Retnoaji

Objective: To examine the anti-angiogenic potential of Artocarpus heterophyllus (A. heterophyllus) seed extract in chicken chorioallantoic membrane (CAM). Methods: This study used chicken CAM ex ovo culture to examine the potential antiangiogenic activity of A. heterophyllus seed methanolic extract. Basic fibroblast growth factor was used to induce the ectopic formation of blood vessels on CAM treated with extract. Blood vessel number was assessed by macroscopic and microscopic observation, and compared and analyzed for all treatments and controls. Results: Macroscopic observation revealed that a dose of 35 mg/mL of methanolic extract of A. heterophyllus seeds could inhibit basic fibroblast growth factor-induced angiogenesis by 61% in chicken CAM ex ovo culture. This concurred with microscopic observations on the histological structure of blood vessels, which indicated that extract treatment repressed the formation of new blood vessels. Conclusions: This is the first study to report the anti-angiogenic effect of methanolic extract derived from A. heterophyllus seeds and its potential as a candidate for future anticancer therapy.

249–252

Authors: Atheer Awad Mehde, Faridah Yusof, Wesen Adel Mehdi, Raha Ahmed Raus, Layla Othman Farhan3, Jwan Abdulmohsin Zainulabdeen, Zaima Azira Zainal Abidin, Hamid Ghazali, Azlina Abd Rahman

Objective: To prove probable relations between serum E3 SUMO-protein ligase NSE2 (NSMCE2) concentration, peroxynitrite related to oxidative stress in nephrolithiasis patients. Methods: A total of 60 patients with nephrolithiasis and 50 healthy volunteers were involved in this study. Colorimetric method was used to detect blood urea, creatinine, uric acid, protein, albumin, total antioxidant status, total oxidant status, peroxynitrite, nitric oxide and oxidative stress index. Glutathione, NSMCE2 and superoxide dismutase were measured by ELISA. Results: A significant increase in level of peroxynitrite, total oxidant status, NSMCE2 and oxidative stress index in patients was observed, while total antioxidant status and glutathione were significantly decreased. Conclusions: The study concluded that serum NSMCE2 significantly correlated with peroxynitrite and oxidative stress in patients with nephrolithiasis.

257–261

Authors: Andriy Synytsya, Jutamart Monkai, Roman Bleha, Anna Macurkova, Tomas Ruml, Juhee Ahn, Ekachai Chukeatirote

Objective: To evaluate the in vitro antimicrobial property of three different partitioned extracts (petroleum ether, ethanol and water) prepared from some fungal mycelia. Methods: Seven fungal mycelia were prepared, initially extracted with acidified ethanol (0.2 mol/L HCl in 80% ethanol), yielding the raw crude extracts. The obtained extracts were then further partitioned with petroleum ether (F1), ethanol (F2) and water (F3). All the fractions were tested for antimicrobial activity using the disc diffusion assay. Results: Our data showed that all the fractions could inhibit the testing bacteria. However, the inhibitory activity was found to be dependent on (i) the fungal strains used; (ii) the solvent extracted; and (iii) the testing bacteria assayed. In general, the ethanolic extracts (F2) derived from all fungi displayed highest inhibitory activity against the testing bacteria except for Chaetomium sp. Conclusions: The findings of the present study concluded that the extracts prepared from the fungal mycelia had the bioactive compounds with antibacterial property. This study is a pioneering work and further study should be carried out for development of the new drug leads.

265–274

Authors: Khaled Mohamed Mohamed Koriem

Infectious diseases resemble a great threat to the human health according to World Health Organization where about 17% of all deaths (z9.2 million deaths) in 2013 recorded are related to infectious diseases. The pathogenic microorganisms such as bacteria, viruses, fungi and parasites are the principle causes of infectious diseases. Ebola, AIDS, dengue, hepatitis, malaria, tuberculosis and schistosomiasis are among 216 infectious diseases found where the immunity represents the first line defense in infection. Lipidomic includes examination of different biological lipids in the biological cell. The lipidomic research covers all aspects of individual lipid molecule including its structure, function, connection with other cell constituents such as protein, lipid, and metabolite in both health and disease conditions. Details of cell biology obtained from different pathogens (viruses, bacteria, and parasites) provide a great data on molecular structure of hostpathogen relation and consequently on infection process. The lipids here play a very important role in many processes involved in host-pathogen relations. The role of lipid in host-pathogen link includes many processes in (1) structural host constituents, (2) host recognition, (3) intracellular transferring, and (4) energy and resource homeostasis during pathogen duplication. There are many lipid phosphatases, kinases, and lipases molecules that greatly involved in these processes and controlling pathogen expression and infection progress. The cell lipid metabolism depends on an adequate energy stores that push the infection to be accelerated and disease symptoms to be appeared. Consequently, future lipidomics studies are the basic for detecting the lipid role in host-pathogen relations which help in therapy advances and biomarkers development.