Malay. J. Biochem. Mol. Biol. (2019) 22 (3)
TABLE OF CONTENTS
Nur Nadhirah Zakaria, Ahmad Fareez Ahmad Roslee, Azham Zulkharnain, Claudio Gomez-Fuentes, Mansur Abdulrasheed, Suriana Sabri, Nancy Calisto-Ulloa and Siti Aqlima Ahmad
BACTERIAL GROWTH AND DIESEL BIODEGRADATION IN THE PRESENCE OF As, Cu AND Pb BY ANTARCTIC MARINE BACTERIA
Antarctica is considered to be low-impacted by anthropogenic activities despite the rising activities occurring in the Southern Ocean. Rising human activities from within Antarctica and external sources resulted in hydrocarbon and heavy metal pollution and create more and more evidence on how much anthropogenic pollution has influenced the southern polar region. Bioremediation possibilities in these parts are very limited in terms of maximum efficiency due to its unique climatic conditions. Furthermore, heavy metals come hand in hand with hydrocarbon pollution and there is a call to obtain. In the present study, diesel degradation was inhibited the most by Pb (31.75%), As (34.35%) and lastly Cu (48.91%) in comparison to control flask (65.19%). Bacterial growth was most inhibited by Cu. Pb had little effect to the growth of bacteria in comparison to As and Cu. Growth rates were obtained by fitting the Exponential mathematical model to the data and obtaining individual growth rates and measures of good fit. ANOVA analysis of the growth obtained P values of control, As and Pb to be <0.05 while Cu gave a P>0.05.
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Syazani Darham, Claudio Gomez-Fuentes, Azham Zulkharnain, Suriana Sabri, Nancy Calisto-Ulloa, Nicolás Ramírez-Moreno and Siti Aqlima Ahmad
ISOLATION AND IDENTIFICATION OF MOLYBDENUM-REDUCING COLD-ADAPTED MARINE BACTERIA ISOLATED FROM BERNARDO O’HIGGINS RIQUELME BASE STATION, ANTARCTICA
Molybdenum (Mo) pollution is an emerging problem in some parts of the world. Traces of Mo can be found in the soil and snow even in the most remote part on earth, Antarctica. Bioremediation of Mo using microorganisms has been an up-and-coming alternative in cleaning up Mo from the environment. Mo reduction is a process that transforms sodium molybdate with an oxidation state of 5+ or 6+ to Mo-blue, a less toxic form of the compound. The objectives of this research are to screen, isolate and identify the best cold-adapted Mo-reducing bacterial strain isolated from marine water samples at Bernardo O’Higgins Riquelme Base Station, Antarctica. A total of 11 psychrotolerant strains were seen able to reduce Mo and further studied to determine their taxonomic position using phylogenetic analysis. Based on 16S rRNA identification, the strains were identified as Shewanella sp., Staphylococcus sp. and Marinomonas sp. This study suggests the potential use of the best Mo-reducing cold-adapted bacteria, strain Marinomonas sp. strain AQ5-A9, on the remediating of Mo in the Antarctic region.
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M. Ali Sarong , S. Supriyatno, Asiah M.D., Mimie Saputri, S. Safika, Asri Mursawal, Wenni Novita Sari & Agung S. Batubara
DNA HOMOLOGICAL ANALYSIS OF THE GELOINA SHELL IN REULEUNG LEUPUNG MANGROVE ECOSYSTEM, ACEH BESAR DISTRICT, ACEH PROVINCE, INDONESIA
Three species of Geloina shells were found in the Reuleung Leupung mangrove ecosystem area, Aceh Besar District, Indonesia which consists of Geloina erosa, G. expansa, and G. coaxana. The phylogenetic relationship of the three closely related species is currently not established. In this study, we analyze the DNA sequence homology of the three Geloina species and compare them with other mangrove shells species stored at NCBI GenBank. The research activities were conducted from July to September 2017, in the area of the Reuleng Leupung mangrove ecosystem where Geloina shells were obtained using destructive sampling method. The results showed that the three Geloina shells have a high similarity with Polymesoda carolina Voucher, Cyrenoida floridana Partial, Cyrenoida floridana Voucher, Glauconome rugosa Voucher, Corbicula fluminea, Glauconome viren Partial, where a very close level of phylogenetic relationship was shown at Polymesoda carolina Voucher sequence homology of 99%. We conclude that due to 99% DNA homology between G. erosa, G. expansa, and G. coaxan and Polymesoda carolina, they are of the same species.
Page 20 - 24
Ilham Abd Allah Ali Al-Saleem
Celiac Disease and Risk of Lymphoma
Celiac disease (CD) is a popular autoimmune systemic defect producing from consumption of gluten that stores a protein of barley, wheat and rye. A few numbers of individual’s celiac disease continuous atrophy of villous with a raise of mucosa intraepithelial lymphocytes (IELs) in an intestine, regardless of a rigorous gluten free diets GFD termed refractory celiac disease (RCD). Precocious detection and treatment could be related to avoid disease problems as a malignant lymphoma. The purposes of this research are to epitomize the accessible facts on the (CD) history related to lymphoma, the lymphoma kinds associated with CD , the methods that gluten excite enteropathy-associated T-cell lymphoma and celiac disease molecular grounds associated with lymphoma.
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Nor Nadia Fadiha Aizuddin, Izyanti Ibrahim, Rusliza Basir, Noor Embi & Hasidah Mohd Sidek
ANTI-MALARIAL AND CYTOKINE-MODULATING EFFECTS OF SODIUM TUNGSTATE IN MURINE MODELS OF CEREBRAL AND SEVERE MALARIA INVOLVE INHIBITION OF GSK3Β MEDIATED VIA ACTIVATION OF BOTH ERK AND Akt SIGNALING
Malaria remains a major global health problem, associated with high morbidity and mortality. Dysregulated production of pro-inflammatory mediators and cytokines is one of the factors that contribute to the pathogenesis of severe and cerebral malaria. Glycogen synthase kinase-3 (GSK3) which plays a pivotal role in regulating the production of pro- and anti-inflammatory cytokines is a potential target for anti-malarial therapeutics. Sodium tungstate (Na2WO4), which exhibits strong normoglycaemic effects, has been reported to cause phosphorylation of GSK3β. The present study aims to evaluate the anti-malarial and cytokine-modulating effects of Na2WO4 using Plasmodium berghei ANKA (PbA) and Plasmodium berghei NK65 (PbN), established models for cerebral and severe malaria respectively. Intraperitoneal administration of Na2WO4 into PbA- or PbN-infected mice resulted in dose-dependent chemo-suppression of parasitaemia. At 50 mg/kg body weight, Na2WO4 treatment resulted in suppression exceeding 60% and improved median survival time of infected mice (14 and 17 days for PbA- and PbN- infected animals compared to 7.5 and 12 days in non-treated control respectively). Na2WO4 treatment also significantly decreased (p>0.05) the serum levels of pro-inflammatory cytokines (TNF-α, IFN-ɤ and IL-18) whilst significantly increasing levels of anti-inflammatory cytokines (IL-10 and IL-4) in mice from both models of malarial infection. In addition, western analysis revealed that in the brain (PbA) and liver (PbN) of infected animals administered with Na2WO4, the intensities of pGSK3β (Ser9), pERK1/2 (Thr202/Tyr204), pp90RSK (Ser380) and pAkt (Ser473) were increased significantly whilst pNF-κB (Ser536) was decreased compared to controls. Taken together our results indicate that anti-malarial and cytokine-modulating effects of Na2WO4 observed involve at least in part the regulation of NF-κB through inhibition of GSK3β mediated via activation of ERK/p90RSK as well as Akt signaling. Our findings reiterate the importance of GSK3β as a molecular target for anti-malarial therapeutics.
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Anton S. Tkachenko, Galina I. Gubina-Vakulyck, Oksana I. Kauk, Anatolii I. Onishchenko, Esmira V. Shekhovtsova & Oksana A. Nakonechna
CHANGES IN S100 PROTEIN EXPRESSION IN THE CEREBELLUM OF RATS ORALLY EXPOSED TO ENERGY DRINKS
The aim of our study was to assess the rate of S100 protein expression in the cerebellum of rats orally exposed to caffeinated energy drinks (CED) during 2 and 4 weeks. Twenty WAG rats were enrolled in the experiment. They were subdivided into three groups. The animals from group 1 and group 2 (n=7 in each) were orally exposed to a caffeinated energy beverage “Black” at a dose of 12 ml per kg of body weight during two and four weeks, respectively. The control group consisted of 6 intact rats obtained drinking water instead of a caffeine-containing energy drink. Cerebellar S100 expression was evaluated immunohistochemically. Against the background of CED oral administration during two and four weeks, the density of granule cell layer becomes lower indicating neuronal loss. S100 protein was upregulated in group 1 in the entire cerebellar cortex. Thus, CED oral exposure resulted in the development of cerebellar astrogliosis. Four-week-long CED consumption resulted in S100 downregulation in the molecular and granular layers. Our findings demonstrate the damage to the cerebellum in rats exposed to CEDs with overexpression of S100, followed by a decrease in its expression.
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Ayah Rebhi Hilles, Syed Mahmood, Mohd Arifin Kaderi & Ridzwan Hashim
IDENTIFICATION OF THE BIOACTIVE COMPOUNDS OF SKIN MUCUS FROM ASIAN SWAMP EEL (Monopterus albus) USING LIQUID CHROMATOGRAPHY QUADRUPOLE-TIME-OF-FLIGHT MASS SPECTROMETRY
Asian swamp eels have been widely accepted as sources of food, especially among various Asian cultures. However, their potential values as novel sources of therapeutic agents have not been widely appreciated. Like most other tropical fishes and amphibians, the outer integumentary system of Monopterus albus is covered with mucus layers, which may act as a mechanical and biochemical barrier for their skin. The biochemical components of these mucus layers may have certain compounds that may be medically beneficial to human. The current study was interested to screen the bioactive compounds of skin mucus from the tropical Asian swamp eel (Monopterus albus) using Liquid Chromatography Quadrupole-Time-Of-Flight Mass Spectrometry (LC-QTOF-MS), for this purpose, eel skin mucus extract was used for LC-QTOF-MS analysis. The screening results for the bioactive compounds revealed different bioactive compounds which possess multiple biological properties mainly anticancer, antimicrobial, anti-inflammatory and antioxidant activities. In conclusion, the current study illustrated that eel skin mucus contain different bioactive compounds which might be consider as therapeutic-promising agents.
Page 48 - 54
Edmund Ui-Hang Sim and Keh-Li Yew
PUTATIVE TARGET PROTEINS OF THE RIBOSOMAL PROTEIN, RPeL27 IN NASOPHARYNGEAL CARCINOMA CELLS
The pathogenesis of nasopharyngeal carcinoma (NPC) is multifactorial and multigenic. Despite the identification of several NPC-associated ribosomal proteins (RPs), the roles of these factors and their interacting partners in NPC tumourigenesis are poorly understood. To date, NPC- associated RP genes are either up or down-regulated in diseased/tumour situation compared to normal condition. The ribosomal protein eL27 (RPeL27) has been known to be over-expressed at both transcript and protein levels in NPC cell lines. This hypothesis was reinforced by our study herein. More importantly, using gene knockdown (RNA interference technique) followed by 2D gel electrophoresis (2D GE) and in silico analysis; we revealed 15 proteins that are likely to interact with RPeL27 during situation of NPC tumourigenesis. These include COTL1, MAGOHB, UBE2N, NDPKA, TMED10, PSMB6, CA2, PGAM1, RPeL14, RPeS8, TPI1, PSMA2, RPeL19, GSTP1, and TPM1. Their association with RPeL27 could attribute to gene expression alteration, cell migration disruption and invasion, promotion of cancer cell survival, immune evasion, and genomic instability. Our findings provide new theoretical insights into the mechanism and involvement of RPeL27 in NPC pathogenesis. This is pertinent in understanding the molecular pathogenesis mediated by ribosomal proteins in the malignancy of the nasopharyngeal tissues.
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Harika Vemugadda, Prajna P Shetty, Monalisa Biswas, Revathi P Shenoy, Nalini K
ERYTHROCYTE ARGINASE ACTIVITY AND SERUM NITRIC OXIDE IN DIABETES MELLITUS
The study is to estimate and correlate the erythrocyte arginase activity and serum nitric oxide levels in normal, prediabetes and type 2 diabetes mellitus. This is a case control study with total 124 samples which were grouped as normal, pre-diabetes and diabetes based on HbA1C values. Blood samples were collected from Clinical Biochemistry laboratory, Kasturba Hospital, Manipal, after the completion of HbA1C analysis. Erythrocyte Arginase activity is estimated by measuring the ornithine formed by Chinard reaction and Arginase activity is expressed as ornithine released per minute per gram hemoglobin under assay conditions. Nitric oxide is estimated by reducing the serum nitrate to nitrite by using Griess reaction method. The increase in arginase activity was seen in both prediabetes and diabetes compared to normals. Compared to normal group, there was significant decrease in nitric oxide level in pre diabetes (P=0.013) however decrease is not significant in type 2 diabetes. Significant positive correlation between the arginase and nitric oxide levels is seen in normals and type 2 diabetes whereas negative correlation in prediabetes. Increase in arginase activity is indirectly affecting the nitric oxide levels and causing the macrovascular (atherosclerosis, hypertension, gangrene of foot, diabetic neuropathy) and microvascular (diabetic nephropathy, diabetic retinopathy) complications in type 2 diabetic patients.
Page 55 - 57
Norshidah Harun, Khairul Anwar Azmi, Fazrin Adhwa Shaifulbahri & Gouri Kumar Dash
ANTIBACTERIAL SUSCEPTIBILITY TEST OF Channa striatus FILLETS AND MUCUS AGAINST Staphylococcus aureus AND Pseudomonas aeruginosa
Channa striatus is an indigenous fresh water carnivorous air breathing fish species and widely distributed in Malaysia. This white boneless meaty and tender taste edible fish is both a popular food of choice and a natural remedy in traditional medicine due to its pharmacological activities such antimicrobial, anti-inflammatory, cell proliferation and many more. Due to these salutary values of this natural product, Channa striatus often being studied in countless times in order to determine valuable information which researcher may gain. Therefore, the determination of effect exerted from both extracts particular in antibacterial activity was conducted to provide antibacterial effectivity for future formulation development. In this study, we followed the method of Wei et al. (2015) for the collection of Channa striatus mucus. Whilst for the fillets, the extraction method of Morachis-Valdez et al. (2017) and Susakate, S. et al. (2019) were referred. As both products were strongly claimed to have beneficial towards wound healing, the antibacterial investigation was carried out against two important pathogens causing wound infection. Comparing the two, Channa striatus has shown a better inhibition zone against Pseudomonas aeruginosa compared to Staphylococcus aureus. As conclusion, the potential values of Channa striatus fillets and mucus extracts need to be studied in greater detail covers the aspects of raw materials, extraction standardization to the formulation development in order to create a new natural sources product that potentially could act as alternative agent to fight bacterial infection and heal wound much quicker than natural healing process.
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Yiing Jye Yap, Khuen Yen Ng, Rhun Yian Koh & Soi Moi Chye3
THE PRO-APOPTOTIC MECHANISMS OF MELATONIN IN CANCER
Cancer is a leading cause of morbidity and mortality worldwide. Various therapies including chemotherapy, radiation therapy, surgery and hormone therapy have been used in the past decades to treat cancers. However, most treatments are associated with unwanted side effects; therefore a better anticancer approach that has less severe side effects and better efficacy is needed. Melatonin is an endogenous indolamine hormone that is mainly produced and secreted by pineal gland. It has oncostatic properties and induces apoptosis in cancer cells. This review describes the pro-apoptotic mechanisms of melatonin on a variety of tumor cells.
Page 66 - 71
Talal Sabhan Salih, Muhammad Abdul-Ghani Muhammad and Mohammed Abdul-Razaq Ibraheem
MOLECULAR PHYLOGENETIC ANALYSIS USING 16S RIBOSOMAL RNA AND MULTIPLE HOUSEKEEPING GENES OF A BIOACTIVE Streptomyces sp. ISOLATED FROM MOSUL, IRAQ
The genus of Streptomyces is the most well known candidate producer of antibiotics and many pharmaceutically important secondary metabolites. Multilocus sequence analysis (MLSA) has shown to be a promising method for streptomycetes taxonomy. In this study, a new bioactive Streptomyces sp. designated TMM19 was isolated from Mosul city, Iraq. The isolate grown strong on all International Streptomyces Project (ISP) media and exhibited different colours of aerial mycelia. S. sp. TMM19 strain displayed bioactivity against Staphylococcus aureus (ATCC 43300), Enterococcus faecalis (ATCC 51299) and Bacillus subtilis (ATCC 23857) pathogens. The results of phylogenetic analyses showed that S. sp. TMM19 was most related to Streptomyces sp. ZFG47 (accession no. CP030073.1) and S. actuosus (accession no. CP029788.1) based on the 16S rRNA genes tree, and to S. sp. Z022 (accession no. CP033073.1) and S. puniciscabiei (accession no. CP017248.1) based on MLSA of three protein-coding genes (atpD, recA and rpoB). The merged 16S rRNA and the three housekeeping genes phylogenetic tree showed that S. sp. TMM19 was again most related to sp. ZFG47 and S. actuosus, this might be explained by the highly conserved of 16S rRNA gene among Streptomyces strains. The 16S rRNA gene of Streptomyces sp. TMM19 strain was deposited in the GenBank database (NCBI) under the accession no. MN062265.1.
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Umar A. A1*., Fakai, I. M & Bagudo, A. I
EXPRESSION OF RECOMBINANT HUMAN EXONUCLEASE1 (HEXO1) FROM BACTERIA
Human exonuclease1 (hExo1) directly involves in many events of DNA processing such as replication, mismatch repair (MMR), and double strand break repair (DSBR), and can also act as a sensor to lesion and inducer of apoptosis in the event of DNA lesion. Human Exo1 protein was functionally expressed in E. coli strain BL21-CodonPlus (DE3)-RIL. Expression of hEXo1 in E. coli under the transcriptional regulation of the T7 promoter yielded a soluble cytosolic monomeric protein with an apparent molecular mass of 110 kDa, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Recombinant hExo1 was purified to near homogeneity using a two-step purification procedure that involves affinity chromatography using heparin column and size-exclusion chromatography. A yield of about 100 μg of the hExo1 protein could be purified from 500 mL of E. coli strain BL21-CodonPlus (DE3)-RIL cells. The purified protein was analysed by LC-ESI-tandem MS on a Q-TOF2 mass spectrometer fitted with a nanoflow ESI (electrospray ionization) source and confirmed to be hExo1.
Page 62 - 68
Malay. J. Biochem. Mol. Biol. (2020) 23 (1)
TABLE OF CONTENTS
Tengku Athirrah Tengku-Mazuki, Azham Zulkharnain, Kavilasni Subramaniam, Peter Convey, Claudio Gomez-Fuentes and Siti Aqlima Ahmad
EFFECTS OF ZINC (Zn) AND CHROMIUM (Cr) ON THE PHENOL-DEGRADING BACTERIA GROWTH KINETICS
Heavy metals occur naturally within the earth crust; however, anthropogenic activities can artificially introduce these elements into the environment. Despite being the foremost isolated continent, Antarctica is not free from human contamination. Heavy metals are well-known to be the powerful inhibitors of xenobiotics biodegradation activities. A microbial growth model was presented for bacteria cell growth in the biodegradation of phenol containing heavy metals such as zinc (Zn) and chromium (Cr) ions. The Gompertz model was used to estimate three main growth parameters namely lag phase (λ), maximum growth rate (μmaz), and maximum cell number at the stationary phase (Nmax). Bacterial growth for both heavy metals was shown to be properly fit towards the curve with a high value of R2 and low square root of the variance of residuals (RSME) value. The effect of heavy metals at 1.0 ppm showed that Cr has a considerable effect on bacteria consortium, inhibiting the degradation of phenol, while Zn has no effect, removing 100% of phenol. The predicted biokinetic from this model suggests the suitability of the bacteria consortium to be used in phenol removal.
Page 5 - 11
Mohamad Khairi Zainol, Sasi Kala Surianarayanan, Mohd Aidil Adhha Abdullah, Hasmadi Mamat and Zamzahaila Mohd Zin
EFFECT OF HYDROLYSIS TIME ON ANTIOXIDANT AND ANTIMICROBIAL PROPERTIES OF JACK BEAN (Canavalia ensiformis) PROTEIN HYDROLYSATE
Jack Bean or Kacang Koro (Canavalia ensiformis) is one of the under-exploited tropical dry beans. This study was aimed to determine the effect of hydrolysis time on physicochemical properties, antioxidative and antimicrobial activity of Jack Bean protein hydrolysate (JBPH). The physicochemical properties of JBPH were evaluated based on protein content, WHC, OHC, degree of hydrolysis, foam stability and foaming capacity. The antioxidative activity of JBPH was measured using DPPH, hydroxyl radical scavenging, superoxide radical scavenging and FRAP. Well diffusion method was used to study antimicrobial activity of JBPH. The highest protein content (33.16±0.03%) obtained in JBPH that hydrolysed for 150 min. The degree of hydrolysis was showed for JBPH highest at 120 min (51.79±0.28%). The size of the microstructure of JBPH analysed using SEM were decrease with hydrolysis time. FTIR analysis confirmed that JBPH comprised of three major components (Region I, II and III). Water holding capacity of JBPH was the highest for the sample hydrolysed for 60 min (63.87±0.72%) while oil holding capacity depicts the highest by it at 180 min (57.17±1.19%). Foaming capacity and foam stability decreased with hydrolysis time. JBPH produced at 120 min hydrolysis time showed the highest inhibition toward DPPH (42.44%) and hydroxyl radicals (20.01%). FRAP and superoxide radical scavenging, JBPH at 90 min showed the highest inhibition (91.15±0.05 µM and 64.33%). JBPH also showed antimicrobial properties by inhibits the growth of P. aeruginosa. The best hydrolysis time to produce JBPH with the highest physicochemical properties was found at 120 min.
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Mohd Sahaid Kalil and Ahmad Razi Othman
THE EFFECT OF Rhodobacter sphaeroides CULTIVATION TECHNIQUES FOR HYDROGEN GAS PRODUCTION
Rhodobacter sphaeroides NCIMB 8523 is a facultative anaerobic bacterium that can generate hydrogen (H2) under aerobic and anaerobic conditions. To compare hydrogen production by R. sphaeroides, three parameters were chosen in this report. First, hydrogen gas production was compared under aerobic and anaerobic conditions. Second, under the different culture chamber (headspace) condition and finally, the effect of different culture volume towards hydrogen production were evaluated. Gas chromatography with thermal conductivity detector (GC-TCD) analysis was used to measure total H2 production, approximately 9.09 mL and 3.57 mL of H2 for anaerobic and aerobic respectively. Different headspace was tested under anaerobic condition and H2 production was measured. Hydrogen gas production increased proportionally with the increment of the headspace area percentage. Hydrogen production for this test was at 4.18 mL, 6.51 mL, 7.65 mL and 10.74 mL for 50%, 30%, 20% and 0% of headspace area respectively. Different culture volume (250 mL and 500mL) was chosen in H2 production under anaerobic condition. Results showed that higher H2 production achieved with 250 mL culture volume with H2 volume at 10.44 mL and 7.58 mL for 250 mL and 500 mL respectively. Obtained results showed that R. sphaeroides NCIMB 8523 produce higher H2 gas under anaerobic conditions at 0% of headspace with optimum culture volume at 250 mL.
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Wai Meng Lau, Menaga Subramaniam and Yang Mooi Lim
MASLINIC ACID ENHANCES IMMUNOGENICITY IN A TIME-DEPENDENT MANNER IN RAJI CELLS
The survival mechanism of tumourigenic cells has evolved to escape the vigilant scrutiny of immune responses. As such, immediate immune recognition of virus-infected cells is necessary to eradicate and delay or prevent the onset of tumour development. Although maslinic acid has been long identified as a prospective chemopreventive agent, its role as an immunogenicity enhancer is not reported. In Raji cells, maslinic acid significantly induced the expression of various genes in the antigen processing and presentation pathway over 48 hours. This observation suggests a new perspective on the mode of action of maslinic acid in targeting the immunogenicity of Raji cells, thereby preventing the onset of tumorigenesis.
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Khadijah Nabilah Mohd Zahri, Azham Zulkharnain, Salihu Ibrahim, Claudio Gomez-Fuentes, Suriana Sabri, Nancy Calisto-Ulloa and Siti Aqlima Ahmad
KINETIC ANALYSIS ON THE EFFECTS OF LEAD (Pb) AND SILVER (Ag) ON WASTE CANOLA OIL (WCO) BIODEGRADATION BY SELECTED ANTARCTIC MICROBIAL CONSORTIUM
Canola oil is used in most of Antarctic research station base and the possibility of discharging the waste canola oil (WCO) through the pipe in the kitchen is high, which leads to environmental pollution. Consortium bacteria form the Antarctic was isolated in degrading the WCO and tested regarding the degradation of oil with the presence of heavy metals. In this study, lead (Pb) and silver (Ag) were used to determine the behaviour of the bacteria consortium to degrade the WCO. The presence of lead allowed the degradation of oil to increased 48% to 56% while the availability of silver prevented the bacterial to grow and degrade the contaminants. Many types of data are best analysed by the fitting curve. The bacterial growth was fitted using both and linear and nonlinear regression curve where the exponential growth equation was used in a nonlinear curve. Bacterial growth with lead shown to be properly fit towards the curve with a high value of R2 and low-value RMSE. In addition, there was no significant difference between linear and exponential regression curves for both conditions of the bacteria with heavy metals, lead and silver.
Page 24 - 29
Lubna Abdallah, Marah Hantash, Amany Shayeb, Maryan Jaber, Sereen Jaber, Abdullah Dweikat, Hameed Hamdan and Osama Hidmi
IN VITRO EFFECT OF CAMEL WHEY ON COPPER INACTIVATED GLUTATHIONE-S-TRANSFERASES
The main aim of the present study was to determine the in vitro effect of some heavy metals in general and copper in particular on the activity of sheep liver glutathione-s-transferases (GSTs). Besides that, the in vitro effect of camel whey on copper inactivated GSTs was also studied in this research. The effect of heavy metals zinc, nickel, cobalt, manganese, copper and camel whey on GSTs activity was studied by spectrophotometric method using 1-chloro-2,4-dinitro-benzene (CDNB) as substrate. Results showed that copper ion had the most potent inhibitory effect on GSTs activity when compared to other studied heavy metals as it directly reduced the enzyme residual activity to reach 29.3% at 2 mM concentration. In addition to that, the further increase in incubation time between heavy metals and GSTs lead to an increase in inhibitory behavior of all heavy metals under study. Furthermore, the role of camel whey on copper inactivated GSTs was clearly showed that it can reduced the inhibitory effect of copper on enzyme activity prior to its addition. The acquired residual activity after direct whey addition increased from 0.76% to 16.48% at 2 mM copper concentration. Moreover, the pre-incubation of GSTs with camel whey for a certain period caused better effect than its direct addition. Hence, the residual activity of GSTs increased from approximately zero to 38.5% at 2 mM. In conclusion, camel whey play an important role in the activation of copper inactivated GSTs.
Page 30 - 36
Mohammed, Z.M, Haruna, Z.K, Abdullahi, Z.I, Hussein, S.A, Kurfi, B.G, Habila, J.D, Abdullahi, M.H, Aliyu, A and Ibrahim, A
ESTIMATION OF ANTIOXIDANT PROPERTIES OF Combretum micranthum METHANOL LEAF EXTRACT AS AN INDEX FOR NEUROPROTECTION AGAINST LEAD INDUCED TOXICITY IN MICE
Combretum micranthum is a medicinal plant used for the treatment of various diseases in the North-Eastern part of Nigeria with little or no scientific basis. It is thus the aim of this research to validate antioxidant capacity of Combretum micranthum (CM) methanol leaf extracts as an index for its neuroprotective potential against lead induce toxicity in the brain of mice. Thirty six (36) Swiss Albino mice (19 - 22g body weight) were used for the study, randomized equally into six groups of six animals each and treated for the period of 14days: Normal Control (Distilled water), Negative control (40mg/kg lead acetate), Group III (lead acetate + 100 mg/kg b.w. extract), Group IV (lead acetate + 50 mg/kg b.w. extract), Group V (lead acetate + 25 mg/kg b.w. extract) and Positive control (lead acetate + 0.5 mg/kg b.w diazepam). The study assayed for the neuroprotective potential of Combretum micranthum by following in vitro tests (DPPH, Metal Chelating, and Reducing Power assays) and in-vivo tests (levels of glutathione (GSH), catalase (CAT) and superoxide dismutase (SOD)). The study indicated a significant neuroprotective effect of the Combretum micranthum methanol extract in terms of its IC50 values (DPPH: 0.467 mg/ml, Metal Chelating: 0.290 mg/ml, and Reducing Power: 0.004 mg/ml) in the in vitro tests and also indicated a significant increase of GSH, CAT and SOD levels in the brain of mice administered doses of the extract, as compared to those mice treated with 40 mg/kg lead acetate alone, which showed decreased level in the antioxidants (GSH: 23.34±1.43µG/ML, CAT: 8.65± 0.77U/mg and SOD: 10.93±0.30 U/mg) in the in vivo tests. Also, the extract was found, using GC-MS, to contain propargylamine, guanidine and acetic acid, compounds known to possess antioxidant functions. Thus, findings of the present study suggested Combretum micranthum to be a potential plant in preventing oxidative stress and brain damage that may be induced by lead administration.
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Muhjah Falah Hassan, Rabab Zahir Al-Yasiry and Rana A. Ghaleb
POSSIBLE PREDISPOSING AND PREDICTABLE FACTORS FOR THE DEVELOPMENT OF OVARIAN HYPER STIMULATION SYNDROME (OHSS) IN WOMEN WITH POLYCYCTIC OVARY SYNDROME (PCOS) WHOM UNDERGONE INTRACYTOPLASMIC SPERM INJECTION (ICSI)
Ovarian hyperstimulation syndrome (OHSS) is a serious, iatrogenic problem which complicates ovulation induction/controlled ovarian stimulation (COS) during any assisted cycle. It usually occurs due to hCG which administered following exogenous gonadotropin stimulation for triggering final oocyte maturation. Women with polycystic ovary syndrome (PCOS) are at high risk but not all PCOS women develop OHSS. It can be effectively prevented and treated when diagnosed early. The study aims to know the possible factors which predispose the women with PCOS to develop OHSS and what are the predictors of its' development during COS/ICSI cycle to be avoided if possible, in the future by careful pre-cycle evaluation and intra-cycle monitoring of the predisposed women. The study included 53 females who seek ICSI treatment for infertility due to PCOS. They were followed up retrospectively and divided according to the development of OHSS into two groups. Group I (n=10): females who developed OHSS and group II (n=43): females who did not develop OHSS following COS/ICSI program. Assessment of demographic data, body mass index(BMI), cycle day 2 hormonal profile, TVUS for endometrial thickness(ET), the total dose of gonadotropins, duration of stimulation cycles, type of oocyte maturation trigger, estrogen and ET at the day of trigger and a total number of retrieved oocytes was done to know possible predisposing and predictable causes for developing OHSS. The study showed women who developed OHSS were insignificantly younger with a higher BMI than those who did not, p-value= 0.24 and 0.08 respectively. Cycle day 2 estrogen, LH, prolactin and ET were higher, total dose and duration of the stimulation cycle were less, most of them were triggered by GnRH agonist, having a higher level of estrogen and thicker ET at the day of trigger and produced higher number of oocytes. Conclusion: Despite being a significant, proved risk factor for OHSS, not all women with polycystic ovaries develop OHSS during COS/ICSI. Young age group women, those with a high BMI and who had high cycle day 2 E2, LH and Prolactin are more liable to develop OHSS. Women with PCOS who produced a high number of oocytes and had high serum E2 level on the day of oocyte maturation trigger are more likely to develop OHSS.
Page 41 - 46
Rashad Ayad Al-Heety
ASSOCIATION OF CIRCULATING MICRO-RNAs WITH CLINICAL FEATURES IN IRAQI PATIENTS WITH GRAVES DISEASE
MicroRNAs (miRNAs) play an important role in the development and functions of the immune system and involved in the pathogenesis of autoimmune diseases mainly by regulating gene expression. Circulating miRNAs can be used as a diagnostic and therapeutic target for a variety of autoimmune diseases. This study aims to assess the existence of any possible association between the expression of circulating miR-146a-5p, miR-142-3p, and let-7b with clinical features of Graves disease (GD). Forty patients with GD and forty healthy controls were involved in this study. Patients were divided into groups based on the presence or absence of clinical features (goiter and/or orbitopathy). The expression of circulating miR-146a-5p, miR-142-3p, and let-7b was determined by two steps Reverse Transcription Polymerase Chain Reaction (RT-PCR) technique. Results obtained show that there is a significant elevation (p<0.01) in the expression of miR-146a-5p, miR-142-3p and let-7b in serum of patients compared with the control group, while there is non-significant overexpression (p>0.05) in patients with goiter and patients with orbitopathy compared to patients without goiter and orbitopathy. It was found that miR-146a-5p and miR-142-3p were positively correlated with the level of thyroid-stimulating hormone receptor antibody (TSHR-Ab), while let-7b is negatively correlated. In conclusion, the up-regulation of miR-146a-5p, miR-142-3p and let-7b in patients and the positive correlation with TSHR-Abs indicating that circulating miRNAs could be used as biomarkers and targets for treatment, however, there is no association with its clinical features.
Page 47 - 51
Riri Andri Muzasti and Dewi Fuji Lestari
DIAGNOSTIC AND PROGNOSTIC VALUE OF INTERLEUKIN-6 FOR PRURITUS IN PATIENTS WITH END STATE RENAL DISEASE UNDERGOING REGULAR HEMODIALYSIS
Chronic kidney disease (CKD) is a national health problem because the incidence tends to increase, has a poor prognosis, and requires high costs. Pruritus is estimated to occur in 20% - 90% of patients with end-stage renal disease (ESRD) undergoing dialysis. Hemodialysis (HD) patients tend to develop inflammation, which involves several proinflammatory cytokines, such as Interleukin-6 (IL-6). Previous studies have shown that IL-6 is a central regulator of the inflammatory process and is found in dermal nerve fibers. We conducted a cross-sectional analytic study to determine the cut-off value of IL-6 levels as a marker of pruritus in ESRD patients undergoing HD. Patients were asked to fill out a Pauli-Magnus questionnaire, and we checked their serum IL-6, calcium, phosphate, and parathyroid hormone levels. Out of the 100 patients who met the inclusion and exclusion criteria, we found that age, HD frequency, and IL-6 levels were significantly associated with pruritus (p = 0.003; p = 0.021 and p = 0.001). The cut-off value of IL-6 as a predictor of pruritus in ESRD patients undergoing regular HD was 101.55 mg/ml, with a sensitivity of 98.1% and specificity of 95.8%. From this study, we conclude that IL-6 levels >101.55 mg/ml was a powerful predictor of pruritus in ESRD patients undergoing regular HD.
Page 52 - 56
Aida Atiqah Mohamad Noor, Catrenar De Silva, Baskaran Gunasekaran, Shafinaz Abd Gani, Murni Marlina Abd Karim and Siti Aqlima Ahmad
IDENTIFICATION OF MARINE BACTERIA ISOLATED FROM MARINE SOIL SEDIMENTS AND THEIR ABILITY TO BIOSYNTHESISE AgNPs EXTRACELLULARLY
Over the past few years, nanoparticles synthesis is one of the most active research in the nanotechnology field. The synthesis can be done chemically, physically and biologically. However, some researchers prefer to synthesise it biologically or also known as biosynthesis or ‘green synthesis’ because it is believed to be safer, environmentally friendly and cost-effective. In this work, we report the extracellular synthesis of 20 isolated marine bacteria from marine soil sediment which were identified and evaluated to synthesise silver nanoparticles (AgNPs). This was done by the addition of silver nitrate (AgNO3) solution with the cell-free supernatant of the isolated marine bacteria at room temperature. The marine bacteria were identified using 16S rRNA identification and neighbour-joining phylogenetic tree were constructed. Identification results showed that the isolated bacteria consist of 19 Serratia sp. and 1 Providencia sp. The biosynthesised AgNPs colloids were evaluated using morphological and optical analysis. AgNPs were observed for colour change and determined using ultraviolet-visible (UV-Vis) spectrophotometer. The existence of surface plasmon resonance peak at 400 to 450 nm is evidence of AgNPs formation.
Page 57 - 61
Dhiey A. Al-Aameri and Bareq N. Al-Nuaimi
MUTATIONS IN ERGOSTEROL 11 GENE OF FLUCONAZOL RESISTANT Candida albicans ISOLATED FROM DIFFERENT CLINICAL SAMPLES
Fluconazole was used to test the susceptibility of Candida albicans isolated from different clinical samples, and to detect mutations in ERG11 gene, and their relationship to fluconazole resistance. Forty-eight isolates of Candida albicans were tested for susceptibility using the disc diffusion method (M-44). ERG11 genes of six isolates were amplified (four resistant, two susceptible) and sequenced. The sequenced genes were analyzed to detect the mutations. Out of 48 isolates of Candida albicans, 4 (8%) were resistant to fluconazole. Sixteen-point mutations were detected included 13 silent mutations, and three missense mutations. The mutations of A945C (E266D) and G1609A (V488I) were found only in susceptible Candida albicans isolates, while the mutation of G1456A (V437I) was detected only in resistant Candida albicans isolates. Candida albicans had a high susceptibility against fluconazole. The amino acid substitutions of E266D and V488I have no role in fluconazole resistance, while the substitution of V437I may have a role in developing resistance against fluconazole. Multiple point mutations in ERG11 gene may develop resistance to fluconazole.
Page 62 - 68
Aisyah Mohd Ismail and Farida Zuraina Mohd Yusof
EFFICIENCY OF COMBINED EFFECTS OF LACTIC ACID BACTERIA (Lactobacillus sp.) AND HIGH OXYGEN ATMOSPHERE ON REDUCING THE POSTHARVEST DECAY OF STRAWBERRY FRUITS
Strawberry is one of the most delicate and perishable fruits which have a short postharvest life span estimated to be less than 5 days. It is prone to rapid dehydration, physiological disorders, bruising, mechanical injuries and infections caused by several pathogens which rapidly reduce the quality of the ripe fruit. The traditional application of chemical fungicides to control the postharvest diseases of strawberry fruits has several disadvantages, such as the adverse effects of pesticide residues on humans and favours the increase of pathogen-resistant strains. The present work used Lactobacillus casei as the bio-control agent along with the action of high oxygen atmosphere packaging to reduce the postharvest decay of strawberry fruits. Moreover, Lactic Acid Bacteria (LAB) is generally recognized as safe (GRAS) and usually matches all recommendations for food products. The dual culture assay results showed that the Lactobacillus casei did not inhibit the growth of fungal pathogen as no inhibition zone produced, but it slowed down their growth compared with the controls. Meanwhile, the in-vivo screening proven that no symptoms of infection seen on the fruits until the fifth day of storage for the strawberries which were treated with the biological control bacteria and highest concentration (80%) of modified oxygen. Hence, our results support the potential of Lactobacillus casei as a bio-control agent integrated with a high oxygen atmosphere against the postharvest rot of strawberry fruits.
Page 69 - 76
Aliaksandr Chirkin, Mershed S. Altani, Nadezhda Stepanova and Anna Chirkina
CHARACTERIZATION OF BIOCHEMICAL HEALTH MARKERS DURING ATHLETIC ACTIVITY IN PUBERTY
Over the past half-century, there has been a steady decrease in the age at which athletes begin to achieve rapid success in competitive sports. Aged-related changes occur in metabolism and its regulation during puberty. To fully understand age-related performance in competitive sports, researchers must study gender-dependent metabolic characteristics of athletes during puberty. The aim of this work was a comparative analysis of metabolic markers in athletes during puberty. The study population included 430 female adolescents (54 - control group, 376 - athletes) and 900 male adolescents (144 - control group, 756 - athletes). Blood serum was collected from the study population, and 24 biochemical markers and 7 coefficients for metabolic characteristics were assessed. The article presents data on constant and variable biochemical biomarkers of blood serum of adolescents of both sexes at puberty. Three categories of biochemical health markers were assessed. To assess the course of puberty without regular physical exertion and during sports, the following biochemical markers should be determined: the first group (independent of gender)- two markers (atherogenic index and Glucose / HDL cholesterol coefficient); the second group (during puberty in males) is the glucose level; the third group (during puberty in females) - 6 markers (the content of triglycerides, calcium, potassium, the value of TIBC, CPK activity and CPK / ALP coefficient). Conclusions: Changes in these markers can indicate negative effects on the most important biochemical and physiological processes in puberty.
Page 77 - 85
Angelo Mark P. Walag and Romeo M. Del Rosario
TOTAL FLAVONOIDS CONTENT, TOTAL PHENOLICS CONTENT, AND ANTIOXIDANT ACTIVITIES OF Acanthaster planci AND Linckia laevigata COLLECTED FROM CARMEN, AGUSAN DEL NORTE, PHILIPPINES
Marine invertebrates are known to contain metabolites with unprecedented diversity in terms of their molecular structures and bioactivities. The majority of the invertebrate phyla have been accounted for but only several studies on antioxidant activities have been made on Philippine sea stars. The main aim of this research is to determine the total flavonoids content, total phenolics content, and antioxidant activities of ethyl acetate and methanolic extract of Linckia laevigata and Acanthaster planci. Standard methods in plants were utilized to determine TFC and TPC while TEAC, DPPH, and FRAP assays were used to evaluate antioxidant activities. No flavonoids were detected in both species for the two solvents utilized while total phenolics were detected in both species and solvents utilized. Total phenolics were noted to be higher in methanol compared to ethyl acetate extract. A. planci also consistently recorded higher TPC for both solvents compared to L. laevigata. Consistently in DPPH and TEAC, A. planci had higher antioxidant capacity compared to L. laevigata for both solvents utilized. Overall, lower antioxidant capacities were observed for all three assays which could be due to the method of sample preparation, especially in the drying of samples which could have affected the natural antioxidants present in the body parts of the sea stars. It is recommended that conservation measures be implemented to protect these species which are a potential source of novel antioxidant compounds.
Page 86 - 89
Baskaran, A. and Karthikeyan, V.
ANALYSIS OF PHYLOGENY AND EVOLUTIONARY DIVERGENCE OF matK and rbcL SEQUENCES OF Barleria longiflora L.f
DNA barcodes are short parts of DNA used to identify an unknown specimen of a species. The matK and rbcL gene is widely used in systematic studies to resolve divergences at many taxonomic levels. Barleria longiflora L.f belongs to the family Acanthaceae which is endemic in distribution. The collected specimens were subjected to sequence analysis of matK and rbcL gene and the same was deposited in Gene Bank and an accession number has been assigned for the same (Accession No: KR861702.1 and KR861703.1). The data were further analyzed for the construction of neighbor joining tree and to infer the evolutionary divergence among the maximum identical sequences retrieved from NCBI Gene Bank through BLAST search. The results of the search revealed that the matK and rbcL sequence of B. longiflora has not been sequenced earlier from any part of the world. The matK and rbcL sequence of B. longiflora may be used for the identification of this species reported from any part of the world through BLAST analysis if the identical sequences are submitted to Gene Bank in future.
Page 90 - 98
Soon Tsuey Ning, Adeline Chia Yoke Yin, Yap Wei Hsum and Tang Yin-Quan
ANIMAL VENOM-DERIVED ANTIMICROBIAL PEPTIDES: NOVEL AND IMPROVED WEAPON FOR CANCER TREATMENT
Cancer is the second leading cause of human death worldwide. Conventional standard anticancer therapies such as chemo-, hormonal-therapies and radiation, are always accompanied with undesired severe side effects and toxicities due to their low specificity. Small peptides are being identified as potential anticancer agents as they could specifically target cancer cells without disrupting normal tissues, thus enable them to be a better alternative agent for the prevention and management of cancer. The increased expression of anionic phospholipids on cancer cell surface renders them more susceptible towards cationic antimicrobial peptides (AMPs). Knowing animal venoms could impair multiple hallmarks of cancer, AMPs isolated from animal venoms might be a new strategy for cancer treatment. In this review, we present the AMPs identified from animal venoms and discuss their multiple mechanism of action against cancer cells.