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Journal of Biological Sciences
eISSN: 1812-5719
pISSN: 1727-3048

Editor-in-Chief:  Mehmet Ozaslan
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Research Article
Optimization and Validation for Determination of Volatile Organic Compounds from Mediterranean Fruit Fly (Medfly) Ceratitis capitata (Diptera: Tephritidae) by Using HS-SPME-GC-FID/MS
Hasan AL-Khshemawee, Manjree Agarwal and YongLin Ren
Background and Objective: The Mediterranean fruit fly (Medfly) Ceratitis capitata (Diptera: Tephritidae), as most of the Tephritidae species, is a pest of great economic importance around the world. Volatile organic compounds (VOC) emitted by Ceratitis capitata (C. capitata) at different life stages (larvae, pupae and adults) can help us to understand the chemicals they produce when interacting. This study aimed to use GC-MS technique to determine the optimal method for accurate, rapid and cost-effective of extraction of VOC from different life stages of C. capitata. Methodology: This study used HS-SPME fibre coupled with flame ionization detection (FID) and gas chromatography with mass spectrometry (GC-MS) to determine the optimal method for accurate, rapid and cost-effective of extraction of VOC from different life stages of C. capitata. Qualitative Analysis software was used to analyse retention times and peak areas. Data were then analyzed by using SPSS. Results: Results indicated that a 4 h extraction time using 20 insects/ sample was optimal for the detection of VOC from all life stages of C. capitata. Conclusion: For saving time, 4 h as extraction time was selected. This study provide that different stage of Medfly has specific VOCs, which in turn explain the feasibility of this method as means of identifying stages of Medfly.
Research Article
Solenostemon monostachyus Modulates Inducible Nitric Oxide Synthase and mRNA Expression in Hemolytic-Induced Rats
Israel Sunmola Afolabi, Adaobi Mary-Joy Okafor, Iyanuoluwa Olubukola Osikoya, Bolaji David Akinbo, Solomon Oladapo Rotimi and Ezekiel Femi Adebiyi
Background and Objective: The 2-Butoxyethanol (2BE) rat model of human hemolytic disorders has been used widely to evaluate hemolysis and thromboembolic manifestations of different organs associated with sickle cell disease and β-thalassemia, however, studies on nitric oxide metabolism are far more scarce. This study investigated the effects of Solenostemon monostachyus leaf extract supplementation on 2BE-induced changes of inducible nitric oxide synthase (iNOS) levels and mRNA expression in female rats. Materials and Methods: Young female rats were divided into seven groups (Group A-G). All groups, except for group A, were exposed to 2BE. The treatments assignments were as follows, group C: Caffeic acid, group D: Ciklavit (herbal formulation of Cajanus cajan (L.) Mill sp.), group E-G: S. monostachyus (150, 200 and 250 mg kg–1 b.wt.), respectively. Liver and plasma iNOS levels, as well as mRNA expression of liver arginase-1 and iNOS, were assessed. Histopathological evaluation of the liver was also performed. The results were statistically analyzed for significant effects at p<0.05 using one-way analysis of variance (ANOVA). Results: Liver and plasma iNOS were significantly reduced (p<0.05) in group G, whereas liver iNOS was significantly increased (p<0.05) in group D-F compared with group B, iNOS mRNA expression was reduced, while arginase-1 mRNA expression was increased in group E-G compared with group B. Conclusion: Administration of S. monostachyus (250 mg kg–1 b.wt.) in the 2BE hemolytic rat model modulated plasma and liver iNOS levels and iNOS mRNA expression.
Research Article
Bioactivity of Fungi Trichoderma reesei Associated with Sponges Stylissa flabelliformis Collected from National Park West Bali, Indonesia
Erna Prawita Setyowati, SUT Pratiwi, Triana Hertiani and Oka Samara
Objective: In this study, antimicrobial and cytotoxic properties of ethyl acetate extract of fermented Trichoderma reesei (T. reesei )fungi associated with the sponge Stylissa flabelliformis (S. flabelliformis) was tested. Materials and Methods: Fermentation of T. reesei was carried out in the saboroud dextrosa saline broth. The fermented filtrate was extracted using ethyl acetate. The ethyl acetate extract was tested for its antimicrobial and cytotoxic activity. Thin-layer chromatography and gas chromatography mass spectrometry were performed to determine the metabolite classes of compounds responsible for antimicrobial and cytotoxic activity. Results: Ethyl acetate extract of T. reesei was active against the bacteria Staphylococcus aureus ATCC 29213, Eschericia coli ATCC 25922 and Candida albicans ATCC 10231. The ethyl acetate extract was also active against cancer cells T47D and Raji cells, with IC50 values of 270 and 470 μg mL–1, respectively. Conclusion: An ethyl acetate extract of Trichoderma reesei fermentations showed antimicrobial and cytotoxic activity. The compounds contained in the extract of T. reesei are terpenoids or phenyl propane derivatives.
Research Article
Structure and Molecular Dynamic Regulation of FKBP35 from Plasmodium knowlesi by Structural Homology Modeling and Electron Microscopy
Jovi Silvester, Herman Umbau Anak Lindang, Lee Ping Chin, Lau Tiek Ying and Cahyo Budiman
Background and Objective: The 35 kDa FK506-binding proteins (FKBP35) of plasmodium parasite is a member of peptidyl proly cis-trans isomerase consisting of FK506-binding domain (FKBD) and tetratricopeptide repeat domain (TPRD). A comprehensive understanding of structure and function relationship of this protein is needed as a platform for development of novel antimalarial drugs with no resistance effect. However, structural study of full-length FKBP35 is hampered by some issues on molecular size and dynamic due to its structural flexibility. Therefore, this study aimed to analyze full-length structure of FKBP35 from Plasmodium knowlesi (Pk FKBP35) and determine its dynamics using structural homology modeling and a transmission electron microscope (TEM). Methodology: Structural homology modeling was constructed using SWISS-MODEL and further validated using RAMPAGE, Global model quality estimation (GMQE), QMEAN statistical parameters and VERIFY3D. For electron microscopy analysis, purified Pk FKBP35 was placed on EM grid, negatively stained using 2% uranyl acetate and recorded under TEM. The image was then processed using ImageJ to classify the molecular shape of Pk FKBP35 based on circularity index. Flexibility analysis was conducted under PredyFlexy web server. Result: The 3D model of Pk FKBP35 was successfully built based on the template of 1p5q. The structure consists of 6 β-sheet and 10 α-helix secondary structures that dominates FKBD and TPRD, respectively, with high similarity to the domains of its homologous from P. falciparum and P. vivax. Negatively stained electron micrograph showed that Pk FKBP35 assumes in three conformations of elongated, hook and circular shapes, with preference conformation being circular shapes (72%). Meanwhile, flexibility prediction demonstrated that FKBD region is more flexible than TPRD. Conclusion: Three conformations concluded that Pk FKBP35 is a dynamic protein due to its flexibility properties. This dynamic might be important for acquiring the substrates. FKBD was found to modulate the flexibility of Pk FKBP35, probably due to the functional role of this catalytic domain and structurally dominated by β-sheet structure, which is more flexible than α-helix structure. Further, 3D model of Pk FKBP35 also suggested that the linker between the domains might involve in the structural dynamic.
Research Article
Antioxidant and Biochemical Evaluation of Thaumatococcus daniellii Seeds in Rat
Shalom Nwodo Chinedu, Franklyn Nonso Iheagwam, Chisom Juliet Anichebem, Gbemisola Beatrice Ogunnaike and Opeyemi Christianah Emiloju
Background and Objective: Despite numerous reports of medicinal uses of Thaumatococcus daniellii (T. daniellii) plant, there remains a dearth of information on the in vivo effect of the seed. In this study, the antioxidant and biochemical effects of T. daniellii seeds in the liver and kidney of male wister rats were assessed. Materials and Methods: Seeds were macerated with ethanol and filtrate was concentrated to yield an ethanolic crude extract. Rats were orally dosed with vitamin C, 500, 1000 and 1500 mg kg–1 of the extracts for 14 days. Antioxidant and biochemical parameters were evaluated. Liver histology was examined. Data were subjected to one-way analysis of variance and expressed as Mean±SEM. Results: It was observed that T. daniellii seed reduced the amount of total body weight gain but did not have any effect on organ weight (p<0.05). Treatment also resulted in significant (p<0.05) increase in hepatic SOD and GSH. It was only at the highest dose that renal GSH increased significantly (p<0.05). These antioxidant effects were associated with maintaining bilirubin concentration and reducing AST activity in the liver (p<0.05). Histopathological observations were in correlation with the biochemical results showing that there was no pathologic abnormality. Conclusion: The results of this study indicate that T. daniellii seed is a source of natural antioxidant and may be exploited for the treatment of kidney and liver diseases.
Research Article
Hepatoprotective Activity of Pineapple (Ananas comosus) Juice on Isoniazid-induced Rats
Novi Yantih, Yahdiana Harahap, Wahono Sumaryono, Rianto Setiabudy and Lestari Rahayu
Background and Objective: Pineapple fruit (Ananas comosus) can inhibit the activity of cytochrome 2E1 (CYP2E1). Isoniazid (INH) is one of the most important antituberculosis drugs and it undergoes hydrolysis in the liver via an enzymatic reaction with CYP2E1, resulting in the formation of hepatotoxic compounds. Extracts of ethanol and water from pineapple fruit can decrease alanine transaminase (ALT) and aspartate transaminase (AST) levels in rats, the increased ALT and AST levels are directly proportional to the damage of liver function. The aim of this study was to evaluate the hepatoprotective activity of pineapple juice in INH-induced rats. Materials and Methods: Rats were divided into four groups. The normal group (Group 1) was treated with water, the negative group (Group 2) was induced with INH, the positive group (Group 3) was treated with silymarin and the test group (Group 4) was treated with pineapple juice. The treatments for all groups were administered orally for 8 weeks. Rat blood was collected at 0, 2, 4 and 8 weeks after the start of treatment. Levels of ALT and AST were determined using the photometric method and the rat livers were taken for histopathological testing during week 8. Data were analyzed using the one-way analysis of variance test with a 95% confidence interval. Results: Based on the data analysis, pineapple juice exhibited hepatoprotective activity, as it decreased the ALT and AST levels in the rats after 4 weeks of treatment. Conclusion: Pineapple juice, protected the rats’ livers by inhibiting the central venous diameter widening, although the data analysis showed that the liver function in these rats was not as good as that in the positive controls.
Research Article
CD14 Expression and Microbial Infection in Bladder Tumours
Hind Mizhir Mousa, Amna Nassief Jassem and Munther Jaffer Hussain
Background and Objective: CD14 is a molecule involved in non-specific immunity that plays important roles in immune cell activation and signaling in response to bacterial cell wall derived components. The current study aimed to determine the diversity of bacterial infection in urinary bladder tumours and studying the immunoexpression of CD14 in bladder tissues associated with bacterial infection. Materials and Methods: In total, 65 patients with bladder tumours with urine and paraffin embedded tissue samples were enrolled in the study, among these patients, 50 had bladder carcinoma (BC) and 15 had benign tumours (BT). Specific culture media and biochemical tests were used to isolate and identify bacterial species and immunohistochemistry (IHC) was used to detect CD14 tissue expression in both groups. Chi-square test was used to compare the difference between percentages of this study by using SAS. Results: The results showed that 37 out of 50 BC patients were positive for gram-negative bacteria, 3 out of 50 patients were positive for gram-positive bacteria and 10 out of 50 patients were negative growth. All BT patients showed presence of gram-negative bacteria. IHC staining showed that CD14+ was significantly over expressed in BC patients compared with BT patients (82% vs. 46.7%, p< 0.01). CD14 expression was also high in gram-negative bacteria infected bladder tissues in patients with both BC and BT and its expression was significantly correlated with this type of bacterial infection (p<0.01). Conclusion: A significant association between CD14 expression and bacterial infected tissues was found and concluded that CD14 activation driven by microbial inflammation probably contributes to bladder cancer.
Research Article
Herbal Medicine Use by People in Jordan: Exploring Believes and Knowledge of Herbalists and Their Customers
Reem A. Issa and Iman A. Basheti
Background and Objective: Many customers visit the local herbal medicine stores to seek treatments for their acute and chronic health conditions. No previous study has investigated customer and herbalist believes and knowledge with regards to the safe and efficient use of herbal medicines in Jordan. This study aims to explore herbal medicine use amongst the public in Jordan and to identify the most frequent herbal medicine used for the treatment of varied health conditions. Secondary aim includes assessing believes and knowledge of herbalists and customers on the use of herbal medicine. Methodology: A questionnaire based survey was conducted in order to explore believes and knowledge of herbalists and customers towards the safe and efficient use of herbal medicines in Amman, Jordan. Data were analyzed by SPSS. Results: The majority of herbalists (64%) believed that use of herbal medicine by customers with chronic conditions can help in their treatment or in reducing the doses of their conventional medicines. Herbalists (69%) tend to always refer their customers to their medical doctor/pharmacist when using herbal medicine and conventional medicine simultaneously. Herbalist's believes in herbal medicine use was not affected by their length of study experience, geographic location or their level of education. To treat their chronic conditions 58.5% of the customers were using herbal medicine. A high proportion (65.1 and 74.5%) of customers reported to have high believes in herbal medicine efficacy and safety over conventional medicine. Herbalists chose work experience (50.6%), family and friends, published papers, books and the internet to be their sources in obtaining their knowledge on herbal medicine use. Customers chose the herbalist to be their first choice as a source of knowledge on herbal medicine. Conclusion: There was a high believe in herbal medicine use by both herbalists and customers in Jordan. Customers obtain their knowledge primarily from the herbalists and herbalists obtain their knowledge primarily from their work experience. An urgent need for the concerned regulatory bodies to provide reliable source of herbal medicinal education pertaining to herbal medication use safety and efficacy is called for.
Short Communication
Development of Internal PCR Control (IPC) for Human Mitochondrial DNA Typing Kit
Ishar Seri Mirianti, Abdullah Nur Azeelah and Zainuddin Zafarina
Background and Objective: Amplification of target sequences has become a necessary technique in molecular research and has been used for various applications such as forensic investigation, genetic tracking, disease diagnosis and much more. The objective of this study was to develop an internal PCR control (IPC) for mitochondrial DNA (mtDNA) typing using allele specific PCR (asPCR) method. Materials and Methods: A total of 5 non-competitive IPC with 300 bp in length were synthesized by incorporating the mtDNA target sequence in the fragment. Both target sequences and IPCs shared similar forward primers. However, reverse primers were designed for each IPC in order to obtain the desired fragment length. Each developed IPC consist of 3 to 6 selected mt DNA SNPs that was used as control to determine the presence of variant in the target sequence, without the need of DNA sequencing. A total of 20 mtDNA SNPs from both coding and control regions were selected for asPCR (16 148, 3552, 16 355, 195, 1872, 1709, 16 108, 16 335, 16 274, 8440, 13 626, 16 291, 9080, 3705, 4491, 146, 1719, 16 093, 3027 and 7684). Results: The IPCs for all selected mtDNA SNPs in asPCR were successfully developed and amplified. The successful amplified IPCs were observed in either wild type lane or variant type lane. Conclusion: Development and incorporation of IPC is important and necessary as these fragments act as control to monitor the performance and amplification results of the asPCR.

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