Malaysian Journal of Biochemistry
& Molecular Biology
(E-ISSN: 2600-9005)
The Official Publication of the Malaysian Society for Biochemistry & Molecular Biology (MSBMB)
Indexed by SCOPUS and Malaysian Citation Index (MYCITE)
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December 2019
Malay. J. Biochem. Mol. Biol. (2019) 22 (3)
TABLE OF CONTENTS
Page 1-7
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
Abstract
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.
Page 8 - 15
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
Abstract
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.
Page 20 - 24
Ilham Abd Allah Ali Al-Saleem
Celiac Disease and Risk of Lymphoma
Abstract
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.
Page 16 - 19
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
Abstract
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 25 - 32
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
Abstract
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.
Page 38 - 42
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
Abstract
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.
Page 43 - 47
Page 48 - 54
Edmund Ui-Hang Sim and Keh-Li Yew
PUTATIVE TARGET PROTEINS OF THE RIBOSOMAL PROTEIN, RPeL27 IN NASOPHARYNGEAL CARCINOMA CELLS
Abstract
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.
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
Abstract
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 33 - 37
Harika Vemugadda, Prajna P Shetty, Monalisa Biswas, Revathi P Shenoy, Nalini K
ERYTHROCYTE ARGINASE ACTIVITY AND SERUM NITRIC OXIDE IN DIABETES MELLITUS
Abstract
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
Abstract
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.
Page 58 - 65
Yiing Jye Yap, Khuen Yen Ng, Rhun Yian Koh & Soi Moi Chye3
THE PRO-APOPTOTIC MECHANISMS OF MELATONIN IN CANCER
Abstract
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
Abstract
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.
Page 72 - 75
Umar A. A1*., Fakai, I. M & Bagudo, A. I
EXPRESSION OF RECOMBINANT HUMAN EXONUCLEASE1 (HEXO1) FROM BACTERIA
Abstract
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.
August 2019
Malay. J. Biochem. Mol. Biol. (2019) 22 (2)
TABLE OF CONTENTS
Page 1-5
Roseline Wai Kuan Yap, Mei-Hua Lin, Yoshihiro Shidoji and Wai Sum Yap
ASSOCIATION AND GENE-ENVIRONMENT INTERACTION EFFECT OF VEGFR-2 GENE POLYMORPHISM WITH MENTAL HEALTH ON METABOLIC RISK FACTORS OF CARDIOVASCULAR DISEASE IN CHINESE MALAYSIAN FEMALE ADULTS
Abstract
Female adults in Malaysia are prone to cardiovascular disease (CVD) and mental health problems. Significant associations of vascular endothelial growth factor receptor-2 (VEGFR-2) gene rs2071559 polymorphism were reported in Asian populations. This study aimed to examine the association and interaction effects of VEGFR-2 gene rs2071559 polymorphism with mental health on metabolic risk factors of CVD in Chinese Malaysian female adults. Physical measurements: body mass index (BMI), body fat percentage (BFP), blood pressure; and biomarkers: blood glucose (BG), glycated hemoglobin (HbA1c), total cholesterol, triglycerides, low-density lipoprotein cholesterol, and high-density lipoprotein cholesterol (HDL-C) were determined. Job Stress Scale (JSS), Depression, Anxiety, and Stress Scale (DASS-21), and Rhode Island Stress and Coping Inventory (RISCI) questionnaires were used to measure job stress, mental health (stress, anxiety and depression), and coping with perceived stress. A total of 81 Chinese Malaysian female adults were included. The allele frequency for rs2071559 (C; T allele) was 0.41; 0.59, and significant genetic association was obtained with HbA1c levels (p=0.034) after adjusting for potential confounders. Significant correlations were obtained for stress with BMI (r=-0.022; p=0.046), depression with BFP (r=-0.242; p=0.030); and stress coping with BG (r=0.303; p=0.006). Significant gene-environment interaction effects were obtained for rs2071559 with stress (p=0.015) and depression (p=0.038) on HDL-C levels. Significant associations and interaction effects of rs2071559 polymorphism and mental health were obtained for metabolic risk factors of CVD in Chinese Malaysian female adults. Further investigation to confirm the findings is required, including promotion of healthy mental health in prevention of CVD including metabolic risk factors.
Page 6-11
Faizan Naeem Razaliab, Adawiyah Suriza Shuibb and Nurhayati Zainal Abidin
PREDICTION OF SIGNALING PATHWAY INDUCED BY Solanum nigrum POLYSACCHARIDE FRACTION, SN-ppF3 IN ACTIVATING RAW 264.7 MACROPHAGE CELLS
Abstract
Previously, the Solanum nigrum polysaccharide fraction, SN-ppF3 was proven to have an immunomodulatory activity by classically activating RAW 264.7 murine macrophage cell line. However, the cellular pathway induced by SN-ppF3 has not to be outlined. In the present study, we predicted the possible cellular pathways induced when macrophage cells were treated with SN-ppF3. The cells were treated with SN-ppF3 for 24 hours and microscopically observed for any morphological due to the treatment. Pinocytosis analysis was carried out to revalidate SN-ppF3 capability as an immunomodulator and also to serve as cytotoxicity evaluation. To outline the signaling pathway induced the cell lysate of 24 hours SN-ppF3-treated macrophage cells were subjected to inflammation analysis through ELISA approach. After the treatment, the morphology of RAW 264.7 cells was obviously altered and pinocytosis activity was significantly increased. In response to the treatment, several phosphorylated proteins such as IκB-α, p38, and NF-κB p65 were significantly up-regulated. Our study suggested that SN-ppF3 treatment could classically activate macrophage through NF-κB pathways which closely similar to the pathway induced by LPS.
Page 12-16
Abdulmoneim M. A.Kannah, Abdul Sattar J. Z. Al-Hayani and Mahmoud I. Al-Jubouri
THE BIOLOGICAL ABILITY OF Chara sp. ALGAE TO ABSORB NICKEL FROM THE ENVIRONMENT
Abstract
Chlorophyta algae (Chara Sp.) were selected and dried. The effective groups of algae responsible for adsorption using infrared (COOH, C = O and CH) were identified and negative groups such as (OH), (P = O), (CO3), (CH2), (Cl) and (Br) on the surface of the algae, and (2) g and treated with a series of different concentrations (0.5,10,20,40,100) mg L-1 of nickel ion where the coefficients were incubated at a temperature of (25 ° C). After the incubation period, the equilibrium stabilizers were extracted and the nickel was estimated and mathematically described according to the Equations (single and two surfaces Langmuir, Freundlich, Temkin, Dubinin and Polani) respectively for nickel. It was found that the values of the coefficient of selection were significant for all equations, which gives a clear indication of the possibility of using any of these six equations, but the equation of the two surface Langmuir is the most efficient in the mathematical description of adsorption, In addition, this equation showed a very large correlation between the actual adsorption values and the calculated values of this equation because they have the least standard error (SE) (0.0002) and the highest coefficient of determination (R2) (1.00), Which earns the advantage in mathematical description on the rest of the equations. The value of the maximum adsorption capacity (Xm) on the first and second surface of algae (1.60) and (7.47) mg. Kg -1, while the binding capacity (K) on the first and second surfaces (75.23) and (41.81) L.mg-1, respectively.
Page 17-20
Ahmed AbdulJabbar Suleiman, HananYassin Muhsin, Rafid A. Abdulkareem and Farah Amer Abed
EFFICIENCY OF PRIMER DESIGN TOOLS IN EVALUATION OF TWO MOLECULAR METHODS TO DETECT TWO SINGLE NUCLEOTIDE POLYMORPHISMS RELATED WITH ATHEROSCLEROSIS
Abstract
Atherosclerosis is one of the most important coronary artery disease(CAD) caused by lipid accumulation, hypertension, smoking ,and many other factors such as environmental and genetic factors, its recorded that genetic variations in rs10757278, rs1333049correlated with CAD.In present study,100 blood samples were collected(50CAD patients and 50 appeared to be healthy controls), who referred to Ibn-Albytar general hospital/in Bagdad city for heart disease from February to March 2019.Genotyping for two SNPs rs10757278, rs1333049 were done by Allele specific PCR and Tetra ARMS technique.The results revealed that, Tetra ARMS technique is more accurate to detect the mutant and normal cases than allele specific PCR. In conclusion, Tetra ARMS technique was suggested to be more specific, sensitive and accurate method used as a molecular biology tool to detect genetic polymorphisms.
Page 21-25
Hala Mouayed Radif, Mustafa Nadhim Owaid and Rusol Mohammed Al-Bahrani
EVALUATION INFLUENCE OF EXTRACT OF Terfezia claveryi DESERT TRUFFLE, PEZIZACEAE, AGAINST Streptococcus pneumoniae, IN VIVO
Abstract
This study aims to use extract of desert truffle Terfezia claveryi for treatment the infected mice by Streptococcus pneumoniae. The bacterium S. pneumoniae isolated from patients infected with Pneumonia. Sputum diagnosis of bacteria depended on serological, biochemical and hemolysis tests to identify S. pneumoniae. Aqueous extract of T. claveryi was used as an antibacterial agent in vivo and studied all the changes in the liver and kidney. The changes in kidney function tests, lipid profile and the liver Enzymes also have studied after taking place the infection by this bacterium and compared with the treatment by aqueous extract of T. claveryi. Using extract of this truffle was amazing in the cases of Blood Urea, Serum Creatinin, cholesterol, triglyceride, HDL, LDL, GOT, GPT, and ALP which returned significantly (p<0.05) to the normal state like in the control. The infected liver and kidney tissues by the bacterium exhibited degeneration, necrosis and hemorrhage while they returned to normal state after the treatment by aqueous extract of T. claveryi. Hence, the introduction of T. claveryi in the pharmacological field is remarkable, especially in the treating of S. pneumoniae infections.
Page 26-36
Ifeanacho Mercy O., Ikewuchi Catherine C. and Ikewuchi Jude C.
NUTRIENT AND BIOACTIVE PHYTOCHEMICAL COMPOSITIONS OF Cnidoscolus aconitifolius
Abstract
Cnidoscolus aconitifolius is consumed as a vegetable in many parts of Southern Nigeria. Presently, there is no information regarding the allicins alkaloids, carotenoids, glycosides, phytosterols, saponins and terpenoids profiles of the leaves and stems of this vegetable. The proximate, minerals, vitamins, amino acid, carotenoids, saponins, glycosides, phytosterols, terpenoids, alkaloids and allicins profiles of the leaves and stems of Cnidoscolus aconitifolius were determined using standard methods. The leaves and stems had high fibre (14.0 - 15.2 g/100g), carbohydrate (40.2 - 41.2 g/100g) and protein (22.2 - 24.5 g/100g). These were equivalent to about 56.0-66.9% daily value for fibre, 44.4-49.0% daily value protein and 13.4-15.1% daily value carbohydrate. They had high contents of selenium, copper, manganese, iron, cobalt, magnesium (leaves only) and vitamins E, B9, C and K (stems only), carotenoids, saponins and glycosides; and moderate phytosterols and terpenoids. Their proteins were rich in essential amino acids (42.7-45.6%). Ten known carorenoids were detected in the stems and leaves, consisting mainly of carotene (43.7-46.1%), lutein (20.8-22.5%) and neo-xanthin (10.92-12.99%). Sapogenin (62.99-64.56%) and saponine (24.5-28. 9%) were the most abundant of the eleven saponins detected. Of the twelve known glycosides were detected, artemetin (65.8-67.6%) and digitoxin (24.8-27.7%) were the most abundant. Seven phytosterols were detected, and consisted mainly of sitosterol (63.6-71.3%), stigmasterol (10.9-13.6%) and 5-avenasterol (10.6-11.4%). Beta-amyrin (52.2-66.3%), alpha-amyrin (18.5-31.6%) and lupeol (14.8-15.9%) were the most abundant of the five phytosterols detected. This result indicates that the leaves and stems of Cnidoscolus aconitifolius are good sources of nutrients and bioactive phytochenicals that can support human health and nutrition. This rich profile makes them potential sources of nutraceuticals.
Page 37-40
Md Monjurul Ahasan and Sultana Umma Habiba
RESPONSE OF TREHALOSE AS A CARBON SOURCE ON ORGANOGENESIS OF PROTOCORM-LIKE BODIES (PLBS) IN Dendrobium kingianum CULTURED IN VITRO
Abstract
Plant growth and development are largely dependent on the availability of carbohydrates as well as to serve as energy and carbon source in vitro culture. The aim of these studies was to know the response of trehalose on organogenesis of protocorm-like bodies (PLBs) in and selection of satisfactory application procedure in Murashige and Skoog (MS) medium for getting better response in this aspect. First experiment results revealed that there were no significant differences among treatments. In case of second experiment, maximum number of PLBs (19.4) was produced in the medium where PLBs were treated in 5 g/L trehalose for 2 days and then transferred to the sucrose (20 g/L) medium and were kept for five weeks when compared with the control (10.1). Third experiment showed PLBs formation rate was 100% at blue and red LED light treatments whereas shoot formation was highest (3.8) at red LED light.
Page 41-45
Azantee Yazmie Abdul Wahab,Muhammad Lokman Md Isa, Haiyuni Mohd Yassim, Yukmil Amru Salleh, Afzan Mat Yusof, Redzuan Nul Hakim Abdul Razak, Nik Fakhuruddin Abdul Razak, Hussin Muhammad and Muhammad Alif Mazlan
IDENTIFICATION OF HYALURIC ACID SYNTHESIS 2 (HAS2) AND GREMLIN 1 (GREM1) GENE EXPRESSIONS IN HUMAN CUMULUS CELLS AS A BIOMARKER FOR OOCYTE QUALITY
Abstract
Nowadays, the criteria for oocyte selection is based on morphological criteria however, it needs further improvement to select the best embryo quality. The gene expression in the cumulus cell plays important role in signaling for follicular development as well as for oocyte quality. The aim of this study is to investigate the present of HAS2 and GREM1 gene expression in the cumulus cells that can become a useful marker for oocyte quality. A preliminary study was performed on cumulus cells derived from four different patients that undergo assisted reproductive technique treatment. Cumulus cells were isolated from 4 patients and the expression of HAS2 and GREM1 was analyzed by using reverse transcriptase polymerase chain reaction (rt- PCR). The results shown that HAS2 and GREM1 were expressed in grade 3 oocytes, whereas, the genes were absent in grade 4 oocytes. This showed that the expression influenced the oocyte quality. Hence, the measurement of HAS2 and GREM1 expressions in cumulus cells would reliably useful tool for selecting competence oocytes with greater chances to be fertilized in assisted reproductive technique.
Page 46-51
Gayathiri Verasoundarapandian, Syazani Darham and Siti Aqlima Ahmad
TOXICITY OF MOLYBDENUM AND MICROBIAL APPLICATION IN MOLYBDENUM REDUCTION FOR BIOREMEDIATION: A MINI REVIEW
Abstract
Molybdenum (Mo) is one the most widely used trace element in many industries and exhibit an important role in humans, animals and plant. Large quantities of hazardous Mo waste released by anthropogenic activities from industrialisation and advancement of technology tremendously increase the burden on the aquatic and soil environments. High accumulation or prolonged exposure of heavy metal such as Mo can cause deleterious health effects on ruminants and aquatic biota as its low toxicity towards human. However, the toxicity of Mo has been reported in spermatogenesis and embryo of mice and fish respectively. Its pollution from several hundreds to thousands of part per million has been documented in water and soils worldwide. Increased level of Mo can pollute the river and bring severe damage to the ecosystem. Bioremediation of heavy metals by microbes in removing the pollutants became more crucial in addressing worldwide environmental pollutions. The mini review summarises the applications of microorganisms in Mo reduction that would be beneficial to future studies on environmental reduction of Mo.
Page 52-59
Nur Nadhirah Zakaria, Zakaria Man, Azham Zulkharnain and Siti Aqlima Ahmad
PSYCHROTOLERANT BIOSURFACTANT-PRODUCING BACTERIA FOR HYDROCARBON DEGRADATION: A MINI REVIEW
Abstract
Biosurfactants are a structurally diverse group of surface-active substances synthesised by microorganisms. All biosurfactants have tremendous potential ranging from medicine to environmental applications especially in hydrocarbon remediation. Petroleum pollution is a major issue in both cold and temperate climate countries. These hydrocarbon pollutants have low solubility and high solid-water distribution ratios, thus limiting the interaction between microbial cells. Petroleum pollution is a major issue in both cold and temperate climate countries. In Antarctica, due to the recalcitrant nature of hydrocarbon components coupled with the region’s extremely weather conditions, there were difficulties faced by bioremediation approaches. However, using biosurfactant in hydrocarbon bioremediation increases the bioavailability of hydrocarbon, thus expediting bioremediation. Few studies have reported on psychrotolerant bacterial species that are able to degrade hydrocarbon and produce biosurfactants. This review focuses on psychrotolerant bacteria with the potential to synthesise biosurfactants and degrade hydrocarbons.
Page 60-63
Haziq Ab. Wahid, Noor Syamila Othman, Azmir Ahmad, Siti Aesah @ Naznin Muhammad, Kahairi Abdullah, Wan Ishlah Leman, Muhammad Izani Mohd. Shiyuti, Luqman Rosla and Mohd. Arifin Kaderi
EXPRESSION OF MICRORNA-101 IN FORMALIN-FIXED PARAFFIN- EMBEDDED SAMPLES OF NASOPHARYNGEAL CARCINOMA
Abstract
Nasopharyngeal carcinoma (NPC) is among the five most common malignancies in Malaysia. Most NPC patients are diagnosed at late stages of the disease which complicates the clinical management of the patients. Identification of new reliable biomarker is crucial to improve early diagnosis of NPC and increase the survival rate of patients. Recent study found that microRNAs (miRNAs), particularly miR-101, were involved in the tumorigenesis of head and neck cancer where NPC samples were included in the study. This study was conducted to observe the expression of miR-101 in NPC tumour tissues and compare its consistency with previous study as a step towards finding the new biomarker for NPC. The biopsy samples were obtained from hospitals and verified histologically using hematoxylin and eosin method for tissue classification. Total RNA was extracted from NPC tissues and normal nasopharyngeal epithelium tissues. The expression of miR-101 in NPC was quantified using quantitative polymerase chain reaction (qPCR) method. The differential expression of miR-101 in NPC as compared to normal nasopharyngeal epithelium tissues was analysed using 2-ΔΔCT calculation. The significance of the differential expression was analysed using SPSS software. Five samples have been verified as NPC and three samples were normal nasopharyngeal epithelium. The differential expression calculation found that miR-101 was downregulated in NPC as compared to normal nasopharyngeal epithelium tissues, which consistent with previous study. However, the differential expression was not significant. Therefore, our finding provides a preliminary result towards embarkment of a larger and comprehensive study.
Page 64-68
Atsadaporn Pakdeelun and Jantaporn Thongekkaew
SCREENING AND OPTIMIZATION OF β-GLUCOSIDASE PRODUCTION BY Candida sp. JK9/1
Abstract
A total of 73 yeast strains were screened for an extracellular β-glucosidase producing ability by using substrate p-nitrophenyl-β-D-glucoside (pNPG). Among them strain JK9/1 showed the maximum β-glucosidase activity which was further identified as Candida sp. strain JK9/1 based on ITS1-5.8S-ITS2 and D1/D2 domains of the subunit (26S) ribosomal DNA. Optimum conditions for β-glucosidase production were determined using different biophysical and biochemical parameters. The maximum -glucosidase production was obtained in the optimum medium containing 1.5% (w/v) of xylose as a carbon source and 0.02 % (w/v) KNO3 as additional inorganic nitrogen, with an initial medium pH of 4.0 and at 25oC for 6 days of cultivation. Under optimal conditions, Candida sp. strain JK9/1 produced the enzyme of 7.21 ± 0.19 U.mL-1, which was 4.2 times higher than the amount before optimization (1.71±0.03 U.mL-1). This can contribute its β-glucosidase producing ability for a great potential to be used in the biotechnological application.
Page 69-73
Laith Sofian Younis, Ali Aziz Abid and Saad Tawfiq Rasheed
EFFECT OF G(129)R POLYMORPHISM IN GROWTH DIFFERENTIATION FACTOR 9 GENE ON AWASSI EWES THAT BREED OUT OF SEASON
Abstract
Growth differentiation factor 9 (GDF9) is a member of the TGF β superfamily that plays a critical role in ovarian follicular development and ovulation rate. The recent study was performed to identify the linkage between GDF9 mutation (G(129)R) and seasonality in awassi sheep. Thirty mature non-pregnant ewes with were used in this study between September/ 2018 to January/2019. Fifteen ewes were lambed at September and Novamber/2018, which considered Seasonal group, and the Non-Seasonal group which 15 ewes lambed in mid-December and January/2019. Two primers were utilized to amplify exon I of GDF9 gene by polymerase chain reaction (PCR) after DNA extraction from blood specimens. Polymorphisms were revealed via sequencing and compared with the sequencing of the ovine GDF9 gene in NCBI data bases. The results revealed one single nucleotide polymorphism (SNP) G(129)R in chromosome 5 of GDF9 gene (exon I) when compared with Sequence ID: FJ429111.1, without resulted in an amino acid. Two genotypes (GG and GA) were observed with higher significant differences (P<0.01) between genotypic frequencies of G(129)R locus. The results showed that mutant heterozygote genotype (GA) recorded highly significant increased (P<0.001) in non-seasonal ewes (60.00) as compared with wild homozygote (GG) genotype (40.00). In contrast, genotype frequency (GG) was recorded higher significantly increased (P<0.001) in seasonal ewes (63.00) when compared with (GA) genotype (37.00). As a conclusion, exon I polymorphisms of GDF9 gene in Awassi breed have an expected effect on the Seasonality and the mutant genotype find majorly in out of Season awsassi ewes.
APRIL 2019
Malay. J. Biochem. Mol. Biol. (2019) 22 (1)
TABLE OF CONTENTS
Page 1-6
Tengku Athirrah Tengku Mazuki, Mohd Yunus Shukor and Siti Aqlima Ahmad
BIOREMEDIATION OF PHENOL IN ANTARCTIC: A MINI REVIEW
Abstract
Aromatic compounds are widely distributed in nature and considered among the most prevalent pollutants in the environment. In recent years, the natural supply of phenolic substances has been greatly increased due to the release of industrial by-products into the environment. One of pollutants associated with the products and waste of chemical and petroleum industries is phenol, a compound that is highly toxic to most living organisms. Pollutions involving phenolic compounds have been documented even in extreme environments including the Antarctic. The application of indigenous microbes capable of degrading phenol obtained from phenol-polluted environments in hopes to remediate phenol has been proven effective even though phenol is harmful to most microbes. The degradation of phenol in cold polluted environments requires the use of microorganisms that can function at low temperatures and withstand the toxicity of phenol. This paper describes various sources of phenol, microorganisms involved in the biodegradation including aerobe and anaerobe, toxicity of the phenol towards living organism as well as the pollutions caused by the phenolic compounds.
Page 7-10
AbdulMushin M. Shami
EFFECT OF PEPTIDES EXTRACTED FROM Lactobacillus casei AGAINST PATHOGENIC
BACTERIA WITH ANTIOXIDANTS
Abstract
This study aims to determine the antibacterial activity of peptides from Lactobacillus casei against selected pathogenic bacteria, such as methicillin-resistant Staphylococcus aureus (MRSA), supported by evidence of antioxidant activities. Well diffusion, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) assays were used to test antibacterial activity against methicillin-resistant Staphylococcus aureus. DPPH (2, 2-diphenyl- 1- picrylhydrazyl) and superoxide dismutase (SOD) assays were used to evaluate antioxidant activity. HPLC and gel filtration were used for purification of the peptides. Scanning electron microscope was applied to investigate the mode of attachment of the peptides on target microbial membranes. Lactobacillus casei BL 23 were isolated and identified from different raw milk sources. Acidic methanolic extraction method demonstrated antibacterial properties against MRSA, S. aureus, Escherichia coli, Bacillus cereus, and Pseudomonas aeruginosa, along with antioxidant properties for the extracts. The infrared spectroscopic analysis of peptide indicated the presence of O-H, C-C, C=N, and C-H. The mode of action of these substances showed morphological changes in the cells’ walls and membranes, which disrupts the cytoplasmic content of the tested bacteria. It was concluded that peptides extracted from Lactobacillus casei as possesses significant antibacterial and antioxidant activities. The results suggest that these peptides can be a new source for developing nutraceuticals or pharmaceuticals with an antibacterial and antioxidant activities.
Page 11-16
Aloysius Duran Corebima, Mariana Rengkuan, Harissudin Masrur, Jahidin & Mohamad Nur Ibrahim
ANALYSIS OF MUTATIONS IN THE SUB - UNIT II CYT. OXIDASE GENE (COX2) OF Tarsius tarsier FROM BUTON ISLAND INDONESIA
Abstract
Tarsius spreading over Sulawesi mainland and on the surrounding islands including Buton Island is still known as an interesting animal to be studied, especially related to gene mutation. The objective of this descriptive explorative research is to analyze mutations at partial sequence of subunit II (COX2) cyt. oxidase gene of Tarsius tarsier form Buton. The sequencing of PCR product produced a base sequence of 560 nt. The gene sequencing results were then aligned with the genes of Tarsius dentatus and Carlito syrichta taken from Genebank (C. syrichta is assumed as the Sulawesian tarsier ancestor). It can be seen that the total number of the invariable sites and variable sites were 432 and 128 respectively, but the total number of mutation was 130. Subtitution mutations were found in 128 sites; the total numbers of transition and transversion mutations were 89 and 16 respectively without calculating the double and single subtitution at the same site of transition. Most of subtitution mutations in COX2 gene sequences occur in the third base of each codon.
Page 17-21
Maizatul Farhain Ismail, Nurul 'Azyyati Sabri, Saiful Nizam Tajuddin, Lee Chin Mei, Siti Hatijah Mortan, Ab. Rahim Mohd-Hairul
ISOLATION AND IDENTIFICATION OF DENITRIFYING BACTERIA FROM INDIGENOUS MICROORGANISMS
Abstract
The objective of this study was to isolate and identify novel potential denitrifying bacteria from two different Kuantan areas which are Jubli Perak Agricultural Park, Kuantan (3o50'49.6"N 103o18'06.1"E) and Felda Lepar Hilir, Kuantan (3o40'41.4"N 103o03'24.7"E). Indigenous Microorganisms (IMOs) available in the locations were cultivated by fermentation of steamed rice in the location together with brown sugar. Serial dilutions of the fermented medium were spread on Jensen’s agar and incubated at 30 oC for eight days. A total of six colonies were subjected to various biochemical analysis including Gram staining, catalase, methyl red, carbohydrate fermentation and nitrate reduction tests. All bacterial isolates were subjected to genomic DNA extraction and PCR amplification of 16S rRNA genes using 27F and 1942R primers. All the amplified product of 16S rRNA genes from the bacterial isolates were purified and sent for sequencing. BLASTn and phylogenetic analysis based on 16S rRNA gene sequences shown all the isolates belong to Bacillus spp. and clustered into two main clusters.
Page 22-26
Aminu Sahalu Bello, Nasir Abdullahi, Hadiza Abdullahi and Abdullahi A. Imam
MOLECULAR MARKERS OF ACT RESISTANCE AMONG MALARIA PAEDIATRIC PATIENTS ATTENDING PUBLIC HEALTH HOSPITAL IN KANO STATE-NIGERIA
Abstract
The spread of artemisinin resistant P. falciparum compromises the therapeutic efficacy of artemisinin combination therapies (ACT) and is considered the greatest threat to current global initiatives to control and eliminate malaria. The relationship of artemisinin resistance to treatment failure has been unclear. This is particularly relevant in Kano North-west Nigeria, where artemisinin combination therapies (ACT) are recommended for P. falciparum. Fifty paediatrics patients were recruited in which 60% of them were males, the mean age of participants is 8.7years, minimum and maximum age were 3 and 14years, while 40% of them were female, within the age of 11 to 14years. The propeller domain gene of K13, a molecular marker of artemisinin resistance, was sequenced successfully in 33 out of 50 P. falciparum isolates collected from the recruited participants residing in Kano munincipal, Fagge, Dala, Nassarawa ,Gwale, Kumbotso and Tarauni local govt areas of Kano state. K13 propeller domain mutations were found in 30.3% (10/33) of the samples isolated. A total of 16 unique mutations were detected, of which twelve were non-synonymous while four were synonymous. Two blood samples isolates obtained from patients from Kano munincipal and Fagge local government area of Kano state consist of more than one single nucleotide polymorphism. The 12 different amino acids positions affected include: Leu440Phe, Tyr441Leu, Phe439Val, Gly670Ala, Met671Lys, Asn672Asp, Leu673Term, Tyr682Phe, Ser694Ille, Gln701Leu and Trip726Leu.These findings suggest that K13-propeller mutations are present in artemisinin- sensitive parasites as well as reduced in-vitro susceptibility to the ACT which provides baseline prevalence of K13-propeller mutations in Kano-Northwest Nigeria.
Page 27-31
Citrawati Dyah Kencono Wungu, Mochamad Amin, S. Eriaty N. Ruslan, Priyo Budi Purwono, Ulfa Kholili, Gwenny Ichsan Prabowo, Ummi Maimunah , Poernomo Boedi Setiawan, Maria Inge Lusida, Soetjipto, Retno Handajani
HEPATITIS B VIRUS X GENE MUTATION WITH PREDOMINANCE A1762T+G1764A DOUBLE MUTATION ON CHRONIC LIVER DISEASE PATIENTS IN SURABAYA, INDONESIA
Abstract
Background: HBV infection is a major problem worldwide, especially in developing countries including Indonesia. Mutations in the HBV gene X are commonly found in patients with CLD, especially in cirrhosis and hepatocellular carcinoma. Mutations in the X gene can cause loss of stability, increased transactivation function, and decreased anti-apoptotic ability of HBx protein.
Aim: The aim of this study was to detect HBV X gene mutations in CLD patients in Surabaya.
Methods: This was a cross sectional research taking samples at Dr. Soetomo General Hospital, Surabaya, Indonesia. This study used nested PCR by targeting HBV X gene. Samples showing positive HBV DNA PCR results were followed by sequencing and X gene mutation analysis by comparing sequencing results with reference strains.
Results: In this study, 30 samples of CLD patients with positive HBsAg in Dr. Soetomo Surabaya were obtained. From the results of the multiple alignments, 12/30 samples (40%) had mutations on HBV X region which overlapped with Core Promoter region. There were 3 types of substitution mutations on HBV X gene (C1632T, T1753A/C/G, A1762T, and G1764A) with the dominant mutation types were A1762T and G1764A mutations, in which both mutations were found together as double mutation.
Conclusion: X gene mutations were found in 40% CLD patients in Surabaya with the dominant mutation was in the form of double mutation A1762T and G1764A in 30% CLD patients in this study. The mutation was found mostly in advance stage of CLD.
Page 32-36
Anatolii I. Onishchenko, Anton S. Tkachenko, Oksana A. Nakonechna, Yevgen M. Korniyenko, Igor M. Ryshchenko, Alexander V.Tsygankov and Yevgen O. Posokhov
A STUDY OF CELL MEMBRANES IN NASAL EPITHELIAL CELLS FROM PATIENTS WITH CHRONIC RHINOSINUSITIS WITH NASAL POLYPS BY MEANS OF A FLUORESCENT PROBE
Abstract
Aim. To assess the state of membranes in nasal epithelial cells obtained from the patients with chronic rhinosinusitis with nasal polyps (CRSwNP) with the help of the fluorescent probe 2-(2'-ОН-phenyl)-5-phenyl-1,3-oxazole.
Methods. The state of membrane phospholipid bilayer in suspensions of nasal epithelial cells isolated from ten patients with CRSwNP was evaluated using the fluorescent probe 2-(2'-ОН-phenyl)-5-phenyl-1,3-oxazole that reacts on the physico-chemical properties of its microenvironment. Changes in fluorescence spectra were determined using a Thermo Scientific Lumina fluorescence spectrometer (Thermo Fisher Scientific) 1 hour after the addition of the probe to nasal epithelial cell suspensions.
Results. CRSwNP was found to be associated with a higher rate of nasal epithelial cell membrane hydration in the region of phospholipid glycerol moiety, carbonyl groups and aliphatic chains of fatty acids attached to the carbonyl groups.
Conclusion. Our findings suggest that CRSwNP is accompanied by the elevated hydration rate of the most polar region, namely polar heads of phospholipids of nasal epithelial cell membranes.
Page 37-43
Deborah Anna Van Oosterhout, Nor Amira Jasmi, Sarah Abdullah Alhaddad, Thur Sina Alkesah and Shariff Halim
Musa accuminata ETHONOLIC EXTRACT IN ALLEVIATING MORPHINE INDUCED DEPENDENCE IN RATS
Abstract
Background and Aim: Morphine is a highly addictive narcotic drug which is used for the treatment of moderate to severe pain. Prolonged use of morphine induces side effects which include addiction followed by a dependent and withdrawal effect. Increasing oxidative stress is proposed as one of the mechanisms that lead to opioid dependency and withdrawal effects. This current study was therefore carried out to determine the effects of Musa Accuminata ethanol leaf extract on the level of Malondialdehyde and GSH in the Ventral Tegmental Area (VTA) region of morphine addicted rats.
Materials and method: Fifty Sprague-Dawley rats were divided into 5 groups (n= 10) and were administered intraperitoneally with increasing dose of morphine sulphate twice daily (2.5mg/kg) (4 groups), normal saline (negative group). Treatment groups were given three doses of Musa Accuminata leaf extract ethanol extract tested (250,500 and 1000mg/kg) via oral gavage for 30 days. Toxicity test was done with the administration of 5000mg/kg of Musa accuminata and its toxicity symptoms were observed for 7 days. Withdrawal symptoms were observed on days one, seven, fourteen, twenty-one and thirty on days of treatment. The rats were sacrificed and its VTA regions were harvested using the Elisa Test kit.
Results: Results showed a significant reduction in withdrawal symptoms (p<0.05) for the treated groups after 30 days of treatment, as compared to the untreated groups. Toxicity results showed no symptoms of toxicity. In parallel to the withdrawal behaviour, level of MDA(p<0.05) also decreased significantly in treatment groups as compared to the untreated groups. In addition, the level of GSH increased significantly (p<0.05) in the brain of rats treated Musa Accuminata.
Conclusion: Musa Accuminata leaf extract has showed evidence that it canreduce the symptoms of withdrawal by alleviating the oxidative stress in VTA region.
Page 44-52
Diajeng Sekar Adisuri and Adeline Chia Yoke Yin
HUMAN PAPILLOMAVIRUS: RECENT ADVANCEMENTS IN STUDY MODELS AND PIPELINE PROPHYLACTICS AND THERAPEUTICS FOR ONCOGENIC HPV
Abstract
The incidence of sexually transmitted infections (STIs) worldwide has increased at an unacceptably alarming rate despite the advances in screening methodologies, diagnosis, and treatment. HPV infection has become a global concern, infecting both men and women without any discrimination between the genders. Caused by Human Papillomavirus, a double-stranded DNA virus with an epithelial tissue tropism, according to the Centre of Disease Control (CDC) in 2018, it is thought to have infected around 80 million people—about one in four— in the United States. In general, HPV is thought to be responsible for more than 90% of both anal and cervical cancers, approximately 70% of vaginal and vulvar cancers, and more than 60% of penile cancers. Although prophylactic vaccines are available (Gardasil, Gardasil 9, Cervarix), none of these vaccines exert any therapeutic effects on existing HPV infections. In this review, the advances made in the attempts of replicating the lifecycle of HPV virus both in vitro and in vivo will be laid out. The different aspects of existing prophylactic vaccines as well as ones that are being developed will also be addressed. Further, this review will peek into the pipeline drugs that have undergone clinical trials focusing on treating existing HPV infections that are being developed as well as the current trends/strategies of combating this silent but malicious infection.
Page 53-59
Farouk R Melek, Dalia O Saleh, Amina M Medhat, Abdel Razik H Farrag, Neveen S Ghaly and Sara M Baraka
ANTIDIABETIC AND ANTIOXIDANT ACTIVITIES OF Phoenix dactylifera L. SEED EXTRACT IN STREPTOZOTOCIN-INDUCED DIABETIC RATS
Abstract
Phoenix dactylifera seeds are popularly used in Egypt for treating diabetes mellitus (DM). The antidiabetic activity of aqueous methanolic seed extracts of two cultivars of P. dactylifera namely Sammany (PDSE) and Hayany (PDHE) was investigated in streptozotocin (STZ)- induced DM in male albino rats. The PDSE and PDHE at doses 70, 140 and 280 mg/kg b.w. and gliclazide (10 mg/kg b.w.) were given separately for 14 days after a single administration of STZ (52.5 mg/kg, i.p.). The dose of 280 mg/kg b.w. of PDSE and PDHE showed a marked reduction in serum glucose level by about 55.4% and 56% of STZ-induced group, respectively. Also, PDSE and PDHE succeeded in decreasing serum total cholesterol, triglycerides, α-amylase and malondialdehyde levels with a significant value (P<0.05) in a dose dependent manner, while the serum levels of C-peptide, total antioxidant capacity and reduced glutathione were significantly elevated. From the histopathological studies, PDSE and PDHE showed a beneficial effect against diabetic complications that appeared in hepatic lobules and pancreatic islets. Phytochemical investigation on the two studied cultivars resulted in identifying 24 phenolic compounds. Conclusion: the results obtained in this study support the popular use of P. dactylifera seeds in the treatment of DM.
Page 60-66
Alyssa Danielle Fonseka, Hong Hao Chan, Chooi Ling Lim, Soi Moi Chye, Anna Pick Kiong Ling and Rhun Yian Koh
TARGETING AUTOPHAGY IN ALZHEIMER’S DISEASE: AN OVERVIEW
Abstract
Autophagy is a self-degradation process that acts as a housekeeper to remove aggregated and misfolded proteins, intracellular pathogens, as well as damaged organelles with the aid of lysosomes. Autophagy is essential for the survival and homeostasis of post-mitotic cells such as neurons, as these cells lack of the capability to dilute toxicants or damaged organelles by cell division. Lately, increasing attention has been focused on the role of autophagy in the degradation of misfolded proteins and neuronal cell death in Alzheimer’s disease (AD). In this review, current evidence on the role of autophagy in AD and therapeutic strategies targeting autophagy are discussed.
Page 67-73
Fatemeh Alian, Mohsen Nabi-Afjadi , Saeed Habibollahi, Masoud Negahdary and Abbasali Palizban
ASSOCIATION BETWEEN THE RS7903146 (C/T) POLYMORPHISM OF THE TCF7L2 AND SERUM CONCENTRATION OF COPPER IN TYPE 2 DIABETES PATIENTS
Abstract
Type 2 diabetes (T2D) is a multifactorial illness, while genetic and environmental factors are involved in the progression of the disease. The aim of this study was to evaluate the serum level of copper in diabetic and healthy patients and the association of it with rs7903146 (C/T) polymorphism of the TCF7L2. In this study, 132 samples were considered and evaluated. The DNA was extracted from all whole blood samples. Subsequently, genotyping was performed. Then, genotype and allele frequencies were evaluated in both healthy and patient groups. In addition, the serum copper level was measured. Data analysis was performed using SPSS software. The found data showed that the mean of the serum copper level was significantly increased, in diabetic patients with TT genotype (385.56 ±30.38), in comparison with healthy subjects (135.37 ±19.18) (p=0). Moreover, a significant increase in the mean of serum copper level in diabetic patients with genotype CT (387.19 ±34.94), compared to healthy subjects (124.73 ±10.31) (p=0), as well as a significant increase in the mean of serum copper level in diabetic individuals with CC genotype (429.29 ±44.36), compared to healthy subjects (141.78 ±12.35) were observed (p=0). Finally results showed that the SNP, rs7903146 is a risk factor for T2D, in specific T allele in both homozygous and heterozygous states. Findings of this study also showed that the serum copper level in diabetic patients was higher than the healthy ones. However, there was no significant difference in the serum copper level in diabetic patients with CC and TT genotype.
Page 74-78
Muhammad Farhan, Hamna Rafiq, Hira Rafi, Maira Saleem and Arshi Saba
DETERMINATION OF METABOLIC AND PSYCHOLOGICAL CONSEQUENCES OF QUERCETIN REPEATED ADMINISTRATION IN MALE RATS: AGE DEPENDENT STUDY
Abstract
Dietary Nutrients are the determinant of healthy life, high levels of lipids are the contributing factors of metabolic syndromes and high levels of lipid may affect the psychological factors i.e depressive symptoms which appears in late adulthood, but the composition of modern diet with bid proportion of fats and poor quality of nutrition in developing countries has made its higher risk for young people as well. Nutraceuticals are supplements containing therapeutic effects for several diseases. Flavonoids are secondary metabolites of plants primarily possess antioxidant property and various biochemical and physiological effects. The antioxidative property of flavonoids makes them focus of researchers for last decades. Quercetin is a flavonoid of subgroup flavonol, reported to alleviate stress induced psychological interferences. The study aimed to observe the effects of quercetinin different age group as well as in different environmental conditions. Study included twelve young (4-6 week) and twelve old rats (16-20) which were divided in to two groupwater and quercetin. Rats were treated with quercetin (100 mg/kg) for two weeks.Food intake in grams, body weight change in grams and behavioral tests which include cage crossing/ 10 minutes in home cage activity test and square crossed/ 5 minutes in open field arena test were conducted every week of administration. Possible anxiolytic effects of quercetin were monitored by using light dark transition box, a known anxiolytic model. After two weeks animals decapitated and blood collected for biochemical estimations. The study concluded that quercetin produced effects in young as well as old rats.
Page 79-86
Kavilasni Subramaniam, Tengku Athirrah Tengku Mazuki, Mohd Yunus Shukor and Siti Aqlima Ahmad
ISOLATION AND OPTIMISATION OF PHENOL DEGRADATION BY ANTARCTIC ISOLATE USING ONE FACTOR AT TIME
Abstract
Phenol has been classified as a priority pollutant by the United States Environmental Protection Agency (US EPA) due to its high toxicity. The increased level of phenol concentration in the Antarctic environment causes a significant risk to the aquatic and terrestrial lives there due to its persistence, biomagnification and accumulation in the food chain. The biodegradation of phenol in Antarctica is considered highly challenging owing to its harsh and extremely cold climate. Therefore, actions of bioremediation are crucial to overcome this problem. To date, little data is available regarding the biodegradation of phenol by aboriginal Antarctic bacteria, and reports on the utilisation of phenol as sole carbon source by strains isolated from Antarctic soil are scarce. In the present study, bacteria isolated from Antarctic soil and identified as Arthrobacter sp. strain AQ5-15 based on 16S rRNA sequence was screened and optimised through conventional method for efficient phenol degradation. Based on preliminary screening, AQ5-15 strain was capable of completely degrading 0.5 g/L phenol within 108 h at 10°C. A study on the effects of significant factors including nitrogen source and concentration, salinity, pH and temperature was carried out to optimise the conditions for phenol degradation. Finding revealed that this strain is a psychrotolerant with optimum temperature at 20°C and prefer neutral or near-neutral condition for phenol degradation. The basic knowledge obtained from this study will provide the benefits on custody of the Antarctic environment especially in removing phenol and its derivatives at low temperature.
Page 87-92
Mardhiah Zainal Abidin, Nor Liana Aqilah Mohd Yatim, Nuralina Najwa Hamsah Zollapi, Mohd Badrin Hanizam Abdul Rahim, Shafinaz Abd Gani, Muhajir Hamid and Mohd Ezuan Khayat
OPTIMIZATION OF ULTRASONIC-ASSISTED EXTRACTION OF PHENOLIC COMPOUND FROM GOLDEN CHICKEN FERN (Cibotium barometz) RHIZOME VIA RESPONSE SURFACE METHODOLOGY
Abstract
There are a lot of medical potentials from Cibotium Barometz that can be exploited due to its secondary metabolites, specifically the phenolic compounds. Therefore, numerous studies have been employed to study the optimization of phenolic compounds extraction from other medical beneficial plants. However, until today there are no definite experiment has been conducted to study the optimization of phenolic compounds extraction of C. Barometz. Hence, this study was designed to systemically optimize the extraction process of phenolic compounds from C. Barometz by using response surface methodology (RSM). The variables were evaluated by using three-factor Box-Behnken experimental design. The three process variables were; ethanol concentration (20-100%), extraction time (10-60 min) and solid-to-liquid ratio (1:20 - 1:100; g: mL) while the independent variable is the total phenolic content (TPC). The optimum extraction condition obtained from RSM are 38.99% ethanol concentration, 47.51 min extraction time, and 1:59.68 (g: mL) ratio under ultrasonic assisted extraction (UAE). Net antioxidant activity was determined by scavenging activity of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, where the lowest IC50 obtained was from ethanol extract via RSM approach (IC50 value 817.87±23.75 μg/mL) which values lie within the range of standard error of the standard (IC50 value 242.53±22.76 μg/mL). The results show that the extraction of C. Barometz can be systemically optimized by using the variables obtained from the RSM method.
Page 93-96
Usunobun Usunomena, Ugbeni C. Osezele and Agu K. Chukwunonso
PROTECTIVE EFFECT OF Celosia argentea AQUEOUS LEAF EXTRACT ON LIVER FUNCTION IN CARBON-TETRACHLORIDE (CCl4)-INDUCED HEPATOTOXICITY
Abstract
Carbon tetrachloride (CCl4) is a solvent used in preparation of many industrial organic compounds whose exposure through inhalation and ingestion is found with high concentration in liver, muscles, fat tissue, kidney and blood. The aim of this study was to examine the protective effect of Celosia argentea aqueous leaf extract in Carbon tetrachloride (CCl4)- induced hepatotoxicity. Wistar male albino rats received either 10ml/kg CCl4 (30% v/v in olive oil) intraperitoneally or 200mg/kg Celosia argentea leaf and 100mg/kg Vitamin C prior to CCl4 administration. Hepatotoxicity was assessed by recording changes in liver function enzymes, liver synthetic molecules and histopathological approaches. Abnormal levels of liver function enzymes such as aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), gamma-glutamyltransferase (γ-GT) and liver synthetic molecules (Bilirubin, total protein, albumin and globulin) in blood were observed in rats administered CCl4. However, treatment with 200mg/kg Celosia argentea leaf or 100mg/kg Vitamin C significantly (p<0.05) decreased the levels of ALT, AST, ALP, γ-GT, Bilirubin and also significantly increased total protein and albumin. Histopathological analysis showed that CCl4 caused significant structural liver damages including inflammation, severe parenchymal changes, necrosis, hemorrhage and congestion which were significantly returned towards normalcy with either the Celosia argentea leaf or the Vitamin C. The study showed that Celosia argentea leaf ameliorates CCl4-induced hepatotoxicity which can be attributed to the phyto-agents in the leaf.
Page 97-105
A. S. Mukhamedzhanova, Z. Alikulov, G. A. Shalakhmetova, A. N. Antipov, K. S. Isayeva and G. T. Kazhybayeva
ACTIVATION AND DETERMINATION OF MOLYBDENUM OF XANTHINE OXIDASE FROM ANIMAL MILK
Abstract
Xanthine oxidase (XO) in goat, camel and horse milk is in a molybdenum-free form, which means that it is inactive. The enzymatic activity is realized through exogenous molybdate (Mo) and thiols during milk heat-treatment at 85 °C for 10 minutes. Mo atoms bind to sulfhydryl (- SH) groups of molybdenum cofactor (Moco) in the active center of XO. However, during the enzyme heat-treatment, the SH-group is oxidized with oxygen. Therefore, different antioxidants were used for the protection of SH-groups of the cofactor. XO nitrate reductase (NR) and nitrite reductase (NiR) activity is determined only by the amount of nitrite: for NR, nitrite is the product of a catalytic reaction, and for NiR, nitrite is a substrate. Sulfanilamide and N-(1- Naphthyl)-Ethylenediamine are used to determine the amount of nitrite. We have discovered that in the presence of phosphate and such antioxidants as dithiothreitol, ascorbate and dithionite, exogenous Mo was discolored with sulfanilamide to blue (maximum absorption at 695 nm). When using cysteine or glutathione as antioxidants, the molybdate did not discolor with sulfanilamide, i.e. they do not interfere with the determination of nitrite. The use of dithiothreitol, phosphate and sulfanilamide led to the presenting of exogenous molybdate in the molecule of XO micelles of cream fat globules, thereby the enzyme was activated and a method for determining molybdenum in biological materials was found by the example of animal milk.
Page 106-109
Tang Ting Fang, Siti Hatijah Mortan, Lee Chin Mei, Noor Suhana Adzahar and Ab. Rahim Mohd-Hairul
ISOLATION AND IDENTIFICATION OF LACTIC ACID BACTERIA FROM SELECTED LOCAL MEDICINAL PLANTS
Abstract
This study aimed to isolate and identify lactic acid bacteria (LAB) from several tropical medicinal plants including Strobilanthes crispus, Gynura procumbens, Justicia gendarussa, Zingiber officinale, Plectranthus aromaticus and Plectranthus amboinicus that grown in the housing area, located at Taman Mahkota Aman, Kuantan, Pahang, Malaysia (GPS coordinate: 3°45'21.4"N 103°11'10.3"E). Leaves of the plants were cut into small pieces, transferred into 20ml of MRS medium and incubated at 30 oC for 48 hours. Eighteen bacterial colonies were isolated from the plant samples. Genomic DNA was extracted from the bacterial cultures and followed by PCR amplification of 16S rRNA genes using 27F and 1492R primers. The PCR products were purified, sequenced using the same primers and entrusted in NCBI GenBank database. BLASTn analysis showed that all the 18 sequences of 16S rRNA genes shared 100% identity with 16S rRNA genes of Lactobacillus plantarum strains. Phylogenetic analysis shown that all the 16 rRNA genes of the bacterial isolates were grouped together with the 16S rRNA genes of other L. plantarum (MG674668.1 and GU138600.1) and L. pentosus (MH844897.1 and GU253891.1) strains that are available in the NCBI GenBank database
Page 110-117
Huai Shin, Khoo, Nashwaan, Ismail, Logis, Joko, Wei Keat, Tana, Kwan Kit, Woo Crystale Siew Ying, Lim and Yong Hui, Tan
CHARATERISATION AND DECOLORISATION CAPACITY OF LACCASE FROM Termitomyces heimii
Abstract
Laccases are enzymes that belong to multi-copper blue oxidases. Laccases show promising application in various industries especially in bioremediation. In this study, laccase from wild edible mushroom, Termitomyces heimii was studied. The fruiting bodies of these mushrooms were homogenised with sodium phosphate buffer, pH 6.0 and centrifuged to obtain the supernatant. The crude extract obtained then underwent ammonium sulphate precipitation at 80% saturation and dialysed against the same phosphate buffer. The dialysed fractions were tested for laccase enzyme activity using ABTS as substrate. Sample was further purified with anion-exchange DEAE-cellulose and gel filtration G-100 Sephadex. T. heimii formed two peaks after DEAE-cellulose. Only D1 fraction of T. heimii was further purified. The specific activity of T. heimii Laccase D1 fraction was 0.13 U/mg. D1 fraction had a final yield of 0.071% and purification fold of 0.72. The optimum pH for maximum activity of laccase from T. heimii was pH 2.5 when ABTS substrate was used, and the optimum temperature was 60oC, with stability up to 55°C. The crude enzyme, as well as the partially purified laccase of T. heimii was able to decolourise dyes: Congo Red at 60% efficiency, Trypan Blue 17.5% and 39% and Coomassie Brilliant blue G-250 at 40% efficiency. Addition of ABTS mediators resulted in increased decolourisation efficiency of dyes. The SDS-PAGE showed that D1 fraction of T. heimii had molecular mass of approximately 85kDa. Native-PAGE of T. heimii was carried out and was stained with ABTS substrates. The band obtained was excised and sent for MALDI-TOF analysis. The proteins were matched to Ludwig database, where it showed 58% identity to glucanase from Volvariella volvacea, however do not show any identity level to any registered laccases or oxidases from basidiomycetes. Further studies need to be carried out to characterise laccase from Termitomyces.
Page 118-124
Khadijah Nabilah Mohd Zahri, Siti Aqlima Ahmad, Nur Adeela Yasid and Mohd Yunus Shukor
RESPONSE SURFACE METHOD FOR OPTIMISING THE BIODEGRADATION OF ACRYLAMIDE BY Burkholderia sp. PSC19
Abstract
In Malaysia, acrylamide and polyacrylamide are widely used in industries including water treatment. For Sarawak state alone, it has been found out that a large amount of polyacrylamide was used in drinking water treatment for one tonne, which can cause acrylamide pollution. In this study, acrylamide degradation was determined through the optimisation of bacterial growth by Burkholderia sp. PSC19 that can utilise acrylamide as both carbon and nitrogen source. The optimisation was done through one-factor-at-a-time (OFAT) and response surface method (RSM). The growth and degradation study was carried in acrylamide media and supplemented with 0.5 g/L of acrylamide. The OFAT optimisation yields an optimum pH, temperature and acrylamide concentration were at 7.5, 30°C and 0.5 g/L, respectively. Meanwhile, the optimum conditions for bacterial growth through RSM, which is central composite design (CCD) for pH, temperature and acrylamide concentration were at 7, 30°C, and 0.3 g/L, respectively. The bacterial growth rate was seen improved from 47.2% using OFAT to 81.2% after using RSM. The model was shown to be significant as the Prob>F in ANOVA analysis was less than 0.0001. The optimum bacterial growth was obtained through optical density at 600 nm and colony forming unit (CFU/μL) where the acrylamide can be assumed degraded with the increased numbers of biomass. All objectives have been successfully achieved where this bacterium has been able to degrade acrylamide and able to bio-remediate acrylamide pollution from industry sewage.
Page 125-129
Muninathan. N and Selvakumar.C
BIOCHEMICAL STUDIES ON FREE RADICAL SCAVENGING ACTIVITIES OF PACLITAXEL AND DI ALLYL SUFIDE ON 7, 12 DIMETHYL BENZ (A) ANTHRACENE INDUCED SKIN CANCER IN WISTAR RATS
Abstract
Our recent studies have shown that naturally occurring dietary organo sulfure compounds such as di allyl sulfide and Paclitaxel are capable of inhibiting polycyclic aromatic hydrocarbon (PAH) metabolism and subsequent PAH- DNA adduct formation in Wistar rats. In this study these plant phenols were tested for their effects against PAHs and 7,12 Di Methyl Benz (A) Anthracene -induced reactive active species of skin tumorigenesis in rats. Each compound was evaluated as a possible anticarcinogen in an initiation and promotion and a complete skin tumorigenesis protocol. In the two-stage tumor protocol in Wistar rats using 7,12- dimethylbenz(a)anthracene as the initiating agent followed by twice weekly applications of acetone as tumor promoter each plant compounds afforded significant protection against skin tumorigenicity. The protective effects were verified both by prolongation of latency period and by subsequent tumor development. Our results suggest that these plants compounds have substantial though variable potential for modifying the risk of skin tumorigenicity induced by a wide variety of chemicals and of these combinations of Paclitaxel and Di allyl sulfide was shown to have free radical scavenging activities.
Page 130-133
Sahlil Miraz Mohamed Rafie, Darman Nordin and Nurina Anuar
INTERACTION OF ANTIMICROBIAL PEPTIDES AND BACTERIAL CELL MEMBRANE TO COMBAT MDR PROBLEM: AN OVERVIEW
Abstract
The discovery of multi-drug resistant (MDR) microorganisms has created urgency in the demand for research towards the discovery of new antibiotics. There are many ways in which microorganism such as bacteria can become antibiotics resistant, for example: the overuse of antibiotics, evolution in bacterial DNA and the transference of antibiotic-resistant gene from one microorganism to microorganism. In order to combat the problem of MDR, it is important to study the interactions of antibiotics with the bacterial cell membrane. Antimicrobial peptides (AMPs) have been considered as potential novel antibiotics. There are few mechanisms action of AMPs in bacterial membrane have been discussed by previous study such as carpet like, membrane thinning, aggregate, toroidal pore and Barrel-starve.
Page 134-141
Sharifah Fathiyah Sy Mohamad, Farhan Mohd Said, Mimi Sakinah Abdul Munaim, Shahril Mohamad and Wan Mohd. Azizi Wan Sulaiman
BACKWARD EXTRACTION OF JACALIN-LOADED REVERSE MICELLES INTO NEW STRIPPING PHASE AND TOXICITY EVALUATION ON EMBRYONIC DEVELOPMENT OF ZEBRAFISH
Abstract
This study demonstrates the effects of aqueous phase pH, KCl concentration and IPA amount on backward extraction efficiency of jacalin from AOT reverse micellar phase into a fresh stripping phase solution. The best condition for backward extraction of jacalin was achieved at the stripping phase pH of 9, 0.75 M KCl and with the addition of 30% v/v IPA. This work also investigates the possible toxicity caused by the backward-extracted jacalin in vivo using zebrafish embryo as a model organism. Our results showed that within the tested concentrations, the purified jacalin had a potentially harmful effect on the embryonic development of zebrafish embryos, as demonstrated by the surviving rates and morphological examination. The survival rate of embryos was found to decrease in concentration and time-dependent manner. The computed LC50 were 20.35, 18.86 and 14.81 μg/ml after exposures of 48, 72 and 96 hours, respectively. Major morphological malformations observed in jacalin-treated embryos were pericardial edema and body curvature for the concentration of jacalin above 12.5 μg/ml. Hence, it can be concluded that the reverse micellar extraction is a suitable method for purification of jacalin from jackfruit seeds, the use of jacalin for therapeutic applications, however, should be done with caution.
Page 142-147
Siti Yatimah Mohamad, Navindra Kumari Palanisamy, Jamal Housaini, Musalmah Mazlan, Norazah Ahmad and Zaini Mohd-Zain
CONJUGAL TRANSFER OF TRIMETHOPRIM-SULPHONAMIDE RESISTANCE GENES FROM NON-TYPEABLE Haemophilus influenzae TO Haemophilus influenzae TYPE B
Abstract
Non-typeable Haemophilus influenzae (NTHi) is an opportunistic pathogen causing a variety of respiratory tract infections in human. Co-trimoxazole (SXT), being one of the drugs for the treatment, loses its importance due to emergence of SXT-resistant strains. This study aims to determine whether conjugal transfer of SXT-resistant genes from an NTHi strain to a typeable strain could occur when both strains co-exist in human upper respiratory tract. Conjugal transfer of the resistance genes from an NTHi (strain H152, donor) to a Haemophilus influenzae type b (Hib strain H582, recipient) were performed by using three different methods (filter, solid and broth mating) in ratios of 1:1 and 1:10. Repeated experiments showed that all attempts failed to produce any transconjugants. Whole genome sequencing of both strains revealed 11 amino acid changes in dihydrofolate reductase (DHFR) and a 15-bp insertion sequence in dihydropteroate synthetase (DHPS) in the donor strain, which attributed to the SXT resistance in strain H152. The tRNALeu sequences for DNA site-specific recombination were detected only in the recipient strain H582 but absent in donor strain H152. Furthermore, it was shown that type IV secretory pathway, the VirB4/VirD4, which is vital for conjugation was only present in H582 strain. One of the possibilities of the inability for SXT-resistance genes in this strain of NTHi to transfer to Hib by conjugation was due to the inadequate presence of the necessary genes components for transfer.
Page 148-151
Varashree BS, Vijetha Shenoy Belle, Krishnananda Prabhu RV and Pragna Rao
INFLUENCE OF OBESITY ON FIRST TRIMESTER SCREENING MARKERS IN SINGLETON PREGNANT WOMEN ENROLLED AT TERTIARY CARE HOSPITAL
Abstract
Pregnancy is diagnosed by the detection of beta human Chorionic Gonadotropin (beta hCG) in maternal serum or urine after implantation of the embryo. Primary function of hCG is to maintain the corpus luteum in early weeks of gestation. Pregnancy associated plasma protein A is a glycoprotein exhibits insulin like growth factor binding protein 4 protease activity, which regulates bioavailability of Insulin like Growth Factor-II. Maternal obesity increases the likelihood of serious pregnancy complications both for mother and infant. The altered maternal environment in women with a raised BMI could have adverse consequences for placental development and function thus we hypothesized if maternal body mass index is associated with alterations in serum levels of free β- HCG and PAPP-A After obtaining approval from institutional ethics committee, Kasturba Hospital Manipal, 676 singleton pregnant women of 19-45 years, in 10-14 weeks of gestations were enrolled for the study. Serum free beta hCG and PAPP-A by ECLIA method. BMI was calculated using the formula weight (in kgs)/height in m2. Data were compiled, statistical analysis was done. Non parametric tests were used for analysis. Median and MoM levels of both free beta hCG and PAPP-A showed a significant negative correlation with BMI (p<0.001). To conclude obese women have a larger extracellular volume, larger distribution volume for hCG and PAPPA. Low hCG in women with high BMI may be sequestration of hCG by adipose tissue–resident macrophages. Inverse association of HCG concentration with BMI may be a result of impaired synthesis of HCG in obese women.
Page 152-154
Wira Eka Putra, Firda Agustin, Lailiyavina Rochmatika and Wa Ode Salma
POTENTIAL OF INDONESIAN MEDICINAL PLANTS AS ANTI-CANCER: IN SILICO STUDY
Abstract
Cancer is the biggest causes of death in the world and lung cancer take the greatest prevalence compared with all cancer incidences. The most dominant type of lung cancer is non-small cell lung cancer (NSCLC) which is generally caused by changes in the genetic composition of epidermal growth factor receptor (EGFR)-tyrosine kinase. Furthermore, the genetic changes of EGFR-tyrosine kinase contribute in developing resistance to anti-cancer drugs. Indonesian medicinal plants exert massive therapeutic advantages; one of them is anti-cancer potency. This research aimed to evaluate anti-cancer potency of Indonesian medicinal plants as lung-cancer therapeutic strategy through in silico. In this study, the practical methods were divided into three major parts including materials retrieval, molecular docking process by using AutoDock Tools, and visualization and docking analysis by using Discovery Studio 4.0. According to the molecular docking result, there were three bioactive compounds of Indonesian medicinal plants which have lower Gibbs free energy than gefitinib as control. Those three bioactive compounds consist of berberin (∆G= -8.1 kcal/mol), physcionin (∆G= -7.9 kcal/mol), and pinostrobin (∆G=-7.4 kcal/mol) which predicted to have anti-cancer properties greater than gefitinib (∆G= -5.9
kcal/mol).
Page 155-160
Mahmmoud Ismail Mohammed, Muwafaq Ayesh Rabeea and Yazen S. Alnajjar
INFLUENCE FASTING ON PHYSIOLOGICAL AND BIOCHEMICAL VARIABLES IN THE HUMAN BODY
Abstract
The present study aims to understand the relationship between fasting and some of the blood biochemical variables of 40 persons at different ages who fasted the month of Ramadan with different fasting hours. Also, urine samples were collected in this study. Some blood and serum samples were performed to calculate concentration of Hemoglobin (Hb), Pack Cells Volume (PCV), Total Count of Red Blood Cells (RBC*106/mm3), Total Protein concentration (T.P), Blood Glucose concentration (B.S), Albumin concentration (ALb), Lipids and Liver Enzymes. In addition to the measuring of some salts concentration like Sodium, Chloride, Potassium, and Calcium. These results were compared with 20 non-fasted people as a control group. The results showed clear changes in the concentration of the above variables. The concentration of Hb showed a decrease of 15.11 ∓ 0.17 and 15.41 ∓ 0.35 according to the time of fasting 18 and 16 hours respectively compared with the control group (15.65 ∓ 0.04). RBC showed a decrease according to the ages 50 and 30 years with values 4.92±0.33 and 5.02±0.06 compared with control group (5.28±0.11) respectively. Furthermore, T.P showed a decrease of 5.16±0.14 and 5.36±0.27 according to the fasting time 18 and 16 hours compared to control group (6.45±0.13) respectively. Then, the concentration of cholesterol reduced to 4.75±0.15 and 5.31+0.13 according to the ages 50 and 30 years compared with control group (5.16±0.11) respectively. Concentrations of electrolytes were high by fasting hours, hence, Sodium ion showed a clear rise of 146.85±10.27 and 142.33±0.35 according to the fasting time 18 and 16 hours respectively, compared to control group (136.25±0.38). General test of urine showed different types of cells with different rates. Red Blood Cells in urine showed the highest rate of 50% followed by purified cells which reached 25.5% and then Calcium Oxalate reached 24.5%.