Asian Pacific Journal of Tropical Biomedicine

Issue 5,2015 Table of Contents

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  • 1  Ebola virus disease: past, present and future
    Harish Rajak Deepak Kumar Jain Avineesh Singh Ajay Kumar Sharma Anshuman Dixit
    [Abstract](14) [HTML](0) [PDF 419.13 K](87)
    Ebola virus disease is one of the most deadly ailments known to mankind due to its high mortality rate (up to 90%) accompanying with the disease. Ebola haemorrhagic fever (EHF) is an infectious disease of animal that can be transmitted to both human and non-human primates. The first epidemic of EHF occurred in 1976 in the Democratic Republic of the Congo. The incubation period of ebola is less than 21 days. Ebola virus infections are depicted by immune suppression and a systemic inflammatory response that leads to damage of the vascular, coagulation and immune systems, causing multi-organ failure and shock. Five genetically distinct members of the Filoviridae family responsible for EHF are as follows: Zaire ebolavirus, Sudan ebolavirus, Côte d’Ivoire ebolavirus, Bundibugyo ebolavirus and Reston ebolavirus. The ongoing 2014 West Africa ebola epidemic has been considered as the most serious panic in the medical field with respect to both the number of human cases and death toll. The natural host for ebola virus is unknown, thus it is not possible to carry out programs to regulate or abolish virus from transmission to people. The ebola virus infection provides little chance to develop acquired immunity causing rapid progression of the disease. It is pertinent to mention that at present, there is no antiviral therapy or vaccine that is helpful against ebola virus infection in humans. The impediment of EHF necessitates much better understanding of the epidemiology of the disease, particularly the role of wildlife, as well as bats, in the spread of ebola virus to humans.
    2  The past, current and future trends in DNA vaccine immunisations
    Sidgi Syed Anwer Abdo Hasson Juma Khalifa Zayid Al-Busaidi Talal Abdulmalek Sallam
    [Abstract](21) [HTML](0) [PDF 587.66 K](80)
    This review focuses on DNA vaccines, denoting the last two decades since the early substantiation of preclinical protection was published in Science in 1993 by Ulmer et al. In spite of being safely administered and easily engineered and manufactured DNA vaccine, it holds the future prospects of immunization by inducing potent cellular immune responses against infectious and non-infectious diseases. It is well documented that injection of DNA plasmid encoding a desired gene of interest can result in the subsequent expression of its products and lead to the induction of an immune response within a host. This is pertinent to prophylactic and therapeutic vaccination approach when the peculiar gene produces a protective epitope from a pathogen. The recent studies demonstrated by a number of research centers showed that these immune responses evoke protective immunity against several infectious diseases and cancers, which provides adequate support for the use of this approach. We attempt in this review to provide an informative and unbiased overview of the general principles and concept of DNA vaccines technology with a summary of a novel approach to the DNA vaccine, present investigations that describe the mechanism(s) of protective immunity provoked by DNA immunization and to highlight the advantages and disadvantages of DNA immunisation.
    3  Statistical issues in randomised controlled trials: a narrative synthesis
    Bolaji Emmanuel Egbewale
    [Abstract](8) [HTML](0) [PDF 173.69 K](83)
    Randomised controlled trials (RCTs) are gold standard in the evaluation of treatment efficacy in medical investigations, only if well designed and implemented. Till date, distorted views and misapplications of statistical procedures involved in RCTs are still in practice. Hence, clarification of concepts and acceptable practices related to certain statistical issues involved in the design, conduct and reporting of randomised controlled trials is needed. This narrative synthesis aimed at providing succinct but clear information on the concepts and practices of selected statistical issues in RCTs to inform correct applications. The use of tests of significance is no longer acceptable as means to compare baseline similarity between treatment groups and in determining which covariate(s) should be included in the model for adjustment. Distribution of baseline attributes simply presented in tabular form is however, rather preferred. Regarding covariate selection, such approach that makes use of information on the degree of correlation between the covariate(s) and the outcome variable is more in tandem with statistical principle(s) than that based on tests of significance. Stratification and minimisation are not alternatives to covariate adjusted analysis; in fact they establish the need for one. Intention-to-treat is the preferred approach for the evaluation of primary outcome measures and researchers have responsibility to report whether or not the procedure was followed. A major use of results from subgroup analysis is to generate hypothesis for future clinical trials. Since RCTs are gold standard in the comparison of medical interventions, researchers cannot afford the practices of distorted allocation or statistical procedures in this all important experimental design method.
    4  The three newest emerging influenza H10N8, H5N6 and H6N1: comparison of its morbidity and mortality
    Somsri Wiwanitkit Viroj Wiwanitkit
    [Abstract](5) [HTML](0) [PDF 133.77 K](71)
    5  Preliminary investigation on the prevalence of malaria and HIV co-infection in Mae Sot District, Tak Province of Thailand
    Siwalee Rattanapunya Wanna Chaijaroenkul Jiraporn Kuesap Ronnatrai Ruengweerayut Kesara Na-Bangchang
    [Abstract](9) [HTML](0) [PDF 175.15 K](81)
    Objective: To preliminarily investigate the prevalence of HIV co-infection in patients with malaria in Mae Sot District, Tak Province of Thailand. Methods: The study was a retrospective study on blood samples collected from a total of 256 patients with malaria (all species and severity) who attended Mae Tao clinic for migrant workers, Tak Province during 2005-2007 (148 samples) and 2010-2012 (108 samples). Malaria diagnosis was performed based on microscopic examination of patients' blood smears. Chemiluminescent microparticle immunoassay and gel particle passive agglutination were employed for the detection of HIV antigen in patients' plasma. Results: Plasmodium falciparum (P. falciparum) and Plasmodium vivax (P. vivax) are the two predominant malaria species with the ratio of about 1: 1 to 1.5:1. Most of the P. falciparum cases were presented with acute uncomplicated signs and symptoms with highest parasitemia of 1 045 000 asexual parasites/μL bloods. The prevalence of malaria and HIV co-infection during 2005-2007 was 1.35% (2/148 cases, 1 each for P. falciparum and P. vivax co-infection), but was increased to 2.78% (3/108 cases, 2 and 1 for P. falciparum and P. vivax co-infection, respectively) during 2010-2012. Conclusions: The increasing trend of prevalence of malaria and HIV co-infection in Mae Sot, Tak province was of a great concern on either pharmacodynamics or pharmacokinetics aspect. The study in a larger numbers of malaria patients in different endemic areas throughout the country with different time periods is underway.
    6  Reno-protective effect of garlic extract against immobilization stress induced changes in rats
    Syed Kashif Zaidi Shakeel Ahmed Ansari Ghulam Md. Ashraf Mohammad Alam Jafri Shams Tabrez Naheed Banu
    [Abstract](19) [HTML](0) [PDF 592.38 K](77)
    Objective: To examine immobilization stress-induced antioxidant defense alterations in rat kidney and the antioxidant effects of aqueous garlic extract in pre and post stress extract treatments. Methods: Albino rats were treated with aqueous extract of garlic both before and 6 h of immobilization stress. Pro-oxidant eminence of rat kidney was assessed by determining the levels of glutathione, thiobarbituric acid reactive substances, aspartate aminotransferase, alanine aminotransferase, glucose, uric acid, alkaline phosphatase and antioxidant enzymes activities. Results: In response to 6 h of immobilization stress, a significant rise in the level of kidney enzymes was recorded. However, antioxidant enzyme activities showed a sharp decline. Conclusions: The extract treatment before and after the stress reverted the activities of above mentioned enzymes towards their control values. Hence, garlic extract can be given as nutritional supplement for scavenging the free radicals generated in rat kidney.
    7  In-vitro antimicrobial effectiveness of herbal-based mouthrinses against oral microorganisms
    Ju Ying Teh Rabiah Rawi Siti Suraiya Md Noor Haslina Taib Suharni Mohamad
    [Abstract](37) [HTML](0) [PDF 779.66 K](79)
    Objective: To evaluate the in vitro antimicrobial effectiveness of commercial herbal-based mouthrinses against oral microorganisms. Methods: A total of three mouthrinses (OX, Pesona and Watsons) were tested for their antimicrobial activity against six oral organisms, Streptococcus mutans (S. mutans), Streptococcus sobrinus (S. sobrinus), Lactobacillus salivarius (L. salivarius), Staphylococcus aureus (S. aureus), Pseudomonas aeruginosa (P. aeruginosa) and Candida albicans (C. albicans) by standard agar-disk diffusion assay. Oradex mouthrinse containing 0.12% chlorhexidine gluconate and sterile distilled water was served as positive and negative controls, respectively. Results: All mouthrinse formulations were effective in inhibiting the growth of S. mutans, S. sobrinus, L. salivarius and C. albicans. Among the tested mouthrinses, Pesona was the only effective mouthrinse against S. aureus and P. aeruginosa, similar to Oradex mouthrinse. Pesona mouthrinse formulation appears to be as effective as Oradex mouthrinse formulation to kill S. aureus and P. aeruginosa. Statistical analysis showed no significant difference among the tested formulations regarding their antimicrobial activities (P > 0.05). Conclusions: Pesona was not the only herbal mouthrinse effective in inhibiting the growth of S. mutans, S. sobrinus, L. salivarius and C. albicans in vitro. All tested formulations were effective against those strains. Our findings may serve as a guide for selecting a kind of herbal mouthrinses as well as providing information to the dental professionals about the efficacy of these products.
    8  Anticancer activity of Aloe vera and Calligonum comosum extracts separetely on hepatocellular carcinoma cells
    Maram Shalabi Kh. Khilo Mahmoud M. Zakaria Mahmoud G. Elsebaei Walied Abdo Walaa Awadin
    [Abstract](32) [HTML](0) [PDF 7.41 M](76)
    Objective: To investigate the in vitro anticancer effect of Aloe vera (A. vera) and Calligonum comosum (C. comosum) extracts against hepatocellular carcinoma (HepG2) cells. Methods: HepG2 cells were tested against different doses of A. vera and C. comosum. Viability of the cells was assessed by MTT assay. Evaluation of apoptosis and DNA damage in HepG2 cells were performed using annexin V apoptosis detection kit. The expression of p53 and anti-apoptotic (Bcl-2) were tested by real time-PCR and flow cytometer analyser. Hematoxylin and eosin stained sections from untreated and treated HepG2 cells were observed using light microscopy. Results: The IC50 values of A. vera and C. comosum extracts were (10.45 ± 0.31) and (9.60 ± 0.01) μg/mL respectively. The extracts separately increased cytotoxicity against HepG2 cells in a time and dose dependent manners. Also, it apparently induced apoptosis through increase P53 and decrease Bcl-2 genes expressions. Conclusions: The results indicated that the extracts could have anti-hepatocarcinogenic effect, at least in part, through modulation of apoptosis.
    9  Green synthesis, antimicrobial and cytotoxic effects of silver nanoparticles mediated by Eucalyptus camaldulensis leaf extract
    Afrah Eltayeb Mohammed
    [Abstract](25) [HTML](0) [PDF 384.69 K](77)
    Objective: To investigate the environmental-friendly extracellular biosynthetic technique for the production of the silver nanoparticles (AgNPs) by using leaf extract of Eucalyptus camaldulensis (E. camaldulensis). Methods: The NP were characterized by colour changes and the UV-visible spectroscopy. The cytotoxic effects of prepared AgNPs was detected against four types of pathogenic bacteria, including two Gram-negative bacteria (Pseudomonas aeruginosa and Escherichia coli) and two Gram-positive bacteria (Staphylococcus aureus and Bacillus subtilis) by using agar well diffusion method. Results: A peak absorption value between 400-450 nm for the extract and the colour change to dark brown were corresponding to the plasmon absorbance of AgNPs. On the other hand, aqueous extract of E. camaldulensis leaves could be effective against tested microorganisms which showed inhibition zones of 9.0-14.0 mm. Furthermore, biologically synthesized AgNPs had higher ability to suppress the growth of the tested microorganisms (12.0-19.0 mm). Conclusions: Our findings indicated that extracellular synthesis of AgNPs mediated by E. camaldulensis leaf extract had an efficient bactericidal activity against the bacterial species tested. The exact mechanism of the extracellular biosynthesis of metal NP was not well understood. Further studies are needed to highlight the biosynthesis process of AgNPs and also to characterize the toxicity effect of these particles.
    10  Weight losing, antihyperlipidemic and cardioprotective effects of the alkaloid fraction of Hunteria umbellata seed extract on normal and triton-induced hyperlipidemic rats
    Adejuwon Adewale Adeneye Peter Anthony Crooks
    [Abstract](8) [HTML](0) [PDF 1.50 M](74)
    Objective: To investigate the weight losing, antihyperlipidemic and cardioprotective effects of the alkaloid fraction of Hunteria umbellata (H. umbellata) seed. Methods: Adult female Wistar rats (weight range: 120-150 g) were randomly divided into 4 and 5 treatment groups in the normal and triton-induced hyperlipidemic models, respectively. and were daily treated for 14 d before they were humanely sacrificed under inhaled diethyl ether anesthesia. About 5 mL of whole blood was obtained by cardiac puncture from each treated rat, from which serum for lipids assay was subsequently separated. Tissue samples of livers of treated rats were harvested and processed for histopathological analysis. Results: Repeated daily oral treatments of normal rats with 25 and 50 mg/kg/day of alkaloid fraction of H. umbellata resulted in significant (P<0.05 and P<0.001) and dose-dependent weight loss, and decreases in the serum triglyceride, total cholesterol and low density lipoprotein cholesterol, while significantly (P<0.001) increased the serum levels of high density lipoprotein cholesterol fraction. Similarly, oral pre-treatments with 25 and 50 mg/ kg/day of alkaloid fraction of H. umbellata for 14 d before induction of hyperlipidemia with triton WR-1339 significantly (P<0.01, P<0.001) and dose-dependently attenuated increases in the average body weights, serum levels of triglyceride, total cholesterol and low density lipoprotein cholesterol while also significantly (P<0.01, P<0.001) and dose-dependently attenuated significant (P<0.001) decrease in the serum high-density lipoproteincholesterol levels when compared to the untreated control values. However, the results obtained for 50 mg/kg of alkaloid fraction of H. umbellata in both normal and triton WR-1339-induced hyperlipidemic rats were comparable to that recorded for 20 mg/kg of simvastatin. Similarly, oral pretreatments with 25 and 50 mg/kg/day of alkaloid fraction of H. umbellata significantly improved the histological lesions of fatty hepatic degeneration induced by triton WR-1339 treatment. Conclusions: Overall, results of this study showed that repeated oral treatments with 25 and 50 mg/kg/day of alkaloid fraction of H. umbellata elicited weight losing, antihyperlipidemic and cardioprotective effects in triton WR-1339 induced hyperlipidemic rats that were mediated via de novo cholesterol biosynthesis inhibition.
    11  Evaluation of antidiabetic activity of plants used in Western Sudan
    Maha Abdulrahman Alamin Ahmed Ibrahim Yagi Sakina Mohamed Yagi
    [Abstract](28) [HTML](0) [PDF 1.28 M](76)
    Objective: To investigate the traditional antidiabetic uses of some indigenous Sudanese plants on streptozotocin-induced diabetes rats. Methods: Diabetic rats were treated with a 400 mg/kg dose of aqueous extracts of five plant species orally for 2 h (acute) or 14 days (chronic). In acute model blood glucose levels were monitored at specific intervals. In the chronic model blood samples were collected from overnight fasted diabetic rats on day 15 to estimate blood glucose level. And the body weight, serum lipid profile and activities of liver and kidney enzymes were measured. Histopathological observations of liver sections were also studied. Results: In the case of acute treatment, aqueous extracts of Tinospora bakis (T. bakis), Nauclea latifolia (N. latifolia) and Randia nilotica (R. nilotica) at 400 mg/kg significantly lowered (P < 0.05) blood glucose levels in diabetic rats whereas, chronic treatment of diabetic rats with 400 mg/kg of T. bakis, N. latifolia, R. nilotica and Mitragyna inremis proved to have significant (P < 0.05) antihyperglycemic effect and have the capacity to correct the metabolic disturbances associated with diabetes. Histopathological studies showed that the aqueous extracts of these four plants reinforced the healing of liver. However, Striga hermonthica aqueous extract did not exert any antihyperglycemic effect to diabetic rats. Conclusions: This study demonstrated that T. bakis, N. latifolia, R. nilotica and Mitragyna inremis have therapeutic value in diabetes and related complications and thus supporting the traditional uses of these plants in Sudanese traditional medicine.
    12  Susceptibility of Anopheles maculipennis to different classes of insecticides in West Azarbaijan Province, Northwestern Iran
    Ali Reza Chavshin Farrokh Dabiri Hassan Vatandoost Mulood Mohammadi Bavani
    [Abstract](5) [HTML](0) [PDF 312.95 K](75)
    Objective: To determine the susceptibility status of Anopheles maculipennis (An. maculipennis) against the major insecticides used in the health sectors in West Azarbaijan Province, Northwestern Iran. Methods: Unfed 3-5 days old adult females of An. maculipennis were collected across the West Azarbaijan Province and were subjected to evaluation of their susceptibility following World Health Organization recommended protocol against six insecticides (permethrin, deltamethrin, propoxur, bendiocarb, malathion and dieldrin) belonging to four different classes. Results: In this study, 916 specimens of An. maculipennis were examined against the insecticides which indicated that An. maculipennis was tolerant to permethrin, deltamethrin and dielderin, but displayed resistance against propoxur, bendiocarb and malathion. Conclusions: The pattern of resistance in An. maculipennis could be attributed to the agricultural landscapes, agricultural pesticides used and the exposure of the mosquitoes to insecticides. Logical cooperation is needed between the agriculture and health sectors to ensure the judicious use of pesticides in each sector and the management of probable resistance.
    13  Limited genetic diversity among Plasmodium falciparium isolates using nested PCR in Jazan Area, Saudi Arabia
    Omima M. Eida Amany M. Eida
    [Abstract](32) [HTML](0) [PDF 1.11 M](80)
    Objective: To establish molecular characterization of Plasmodium falciparum field isolates in Jazan Area of Saudi Arabia measured with highly polymorphic genetic marker, i.e. the merozoite surface protein 2 (MSP 2). Methods: Blood samples were collected from 128 clinically suspected patients attending both Jazan and Sabia hospitals, Kingdom of Saudi Arabia. Both hospitals reflected urban and rural settings respectively. Analysis of central polymorphic region of MSP 2 (3D7 and FC27 allelic families) was performed using nested PCR for malaria patients. Results: For MSP 2 allelic families of Plasmodium falciparum, 16 cases (53.3%) carried FC27 type and 14 cases (46.7%) carried 3D7 type, whereas no malaria cases harbored both allelic types. The present study showed that in urban area, 80% of FC27 fragments were 500 bp while in rural area it was 45.5% (P = 0.08). The FC27 400 bp allele was more prevalent in patients from rural than those from the urban area (P = 0.08). The most prevalent infecting 3D7 allele was the 3D7 300 bp in both areas. In the present study, there were no multiple infections. Conclusions: The limited genetic diversity which was observed in Jazan (considered as an endemic area) may be attributed to the small sample size or sustained malaria control program.
    14  Chemical profiling of Curcuma aeruginosa Roxb. rhizome using different techniques of solvent extraction
    Sanimah Simoh Alizah Zainal
    [Abstract](32) [HTML](0) [PDF 208.71 K](80)
    Objective: To investigate the possible phytochemical constituents of Curcuma aeruginosa Roxb. (C. aeruginosa) rhizome using two different techniques of direct solvent extraction. Methods: Two different techniques of direct solvent extractions, i.e. methyl tert-butyl ether (MTBE) extraction and two-phase methanol/chloroform (M/C) system, were used in this study. The analysis of the phytochemical constituents in MTBE and M/C extracts was performed using gas chromatography-mass spectrometry/mass spectrometry. The mass spectra of the compounds was matched with the NIST 08 mass spectral library. Results: The present study revealed that the extraction using two-phase M/C have resulted in higher metabolite coverage compared to the extraction with MTBE. Direct solvent extraction using MTBE revealed the presence of 27 compounds; whereas, M/C allowed the extraction of 18 and 36 compounds in polar (methanol) and nonpolar (chloroform) fractions respectively. The major compounds detected in the MTBE extract that based on the peak area percentage were methenolone (16.64%), cycloisolongifolene, 8,9-dehydro-9-formyl- (15.93%), labd-13- en-15-oic acid,8,12-epoxy-12-hydroxy-γ-lactone (10.77%), propiolic acid, 3-(1-hydroxy)-2 isopropyl-1,5-methylcyclohexyl) (7.84%), 4-oxo-β-isodamascol (5.17%), velleral (3.11%) and Z-α-farnesene (2.00%). The most prevailing major compounds identified in the polar fraction of the M/C extraction were α-D glucopyranoside, 1,3,4,6 tetrakis-O-(TMS) (trimethylsilyl)-β- D-fructofuranosyl 2,3,4,6-tetrakis-O-(TMS)- (38.08%), d-glucose, 2,3,4,5,6-pentakis-O-(TMS)-, O-methyloxime (14.61%), D-fructose, 1,3,4,5,6-pentakis-O-(TMS)-, O-methyloxime (5.28%), isocitric acid (TMS) (3.06%), oxalic acid, bis (TMS) ester (2.96%), hexadecanoic acid, TMS ester (2.16%), citric acid, ethyl ester, tri-TMS (1.91%) and butanedioic acid, [(TMS) oxy]-, bis (TMS) ester (1.14%); whereas in the nonpolar extract, among the major compounds detected were cycloisolongifolene, 8, 9-dehydro -9-formyl (15.70%), propiolic acid, 3-(1-hydroxy-2- isopropyl-5-methylcyclohexyl) (11.09%), stearic acid, TMS ester (2.78%), hexadecanoic acid, TMS ester (2.33%), oleic acid, TMS ester (1.62%), curzerene (1.56%); Z-α-farnesene (1.52%), germacrone (1.41%) and β-elemene (1.33%). Conclusions: It was evident from the results that C. aeruginosa rhizome extracted using two different techniques of solvent extractions (MTBE and M/C) contained various chemical classes of compounds including terpenoids, sterols, organic acids, fatty acids and sugars. Different methods of extraction have led to different compounds extraction for C. aeruginosa rhizome. The results also indicated that the plant was a source of phytochemical importance.
    15  Medley of infections-a diagnostic challenge
    Raghavendra Bhat Parul Kodan Meenakshi A Shetty
    [Abstract](22) [HTML](0) [PDF 142.67 K](78)
    We present a rare case of multiple infections coexisting together. This is one of the rarest cases of four infections which coexisted together in our patient. It is an alarming for the physicians to be aware of such infections as early prompt diagnosis can be lifesaving.

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