Malay. J. Biochem. Mol. Biol. (2019) 22 (1)
TABLE OF CONTENTS
Tengku Athirrah Tengku Mazuki, Mohd Yunus Shukor and Siti Aqlima Ahmad
BIOREMEDIATION OF PHENOL IN ANTARCTIC: A MINI REVIEW
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.
AbdulMushin M. Shami
EFFECT OF PEPTIDES EXTRACTED FROM Lactobacillus casei AGAINST PATHOGENIC
BACTERIA WITH ANTIOXIDANTS
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.
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
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.
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
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.
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
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.
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
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.
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
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.
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
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.
Diajeng Sekar Adisuri and Adeline Chia Yoke Yin
HUMAN PAPILLOMAVIRUS: RECENT ADVANCEMENTS IN STUDY MODELS AND PIPELINE PROPHYLACTICS AND THERAPEUTICS FOR ONCOGENIC HPV
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.
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
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.
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
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.
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
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.
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
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.
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
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.
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
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.
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
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.
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
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.
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
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
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
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.
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
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.
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
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.
Sahlil Miraz Mohamed Rafie, Darman Nordin and Nurina Anuar
INTERACTION OF ANTIMICROBIAL PEPTIDES AND BACTERIAL CELL MEMBRANE TO COMBAT MDR PROBLEM: AN OVERVIEW
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.
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
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.
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
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.
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
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.
Wira Eka Putra, Firda Agustin, Lailiyavina Rochmatika and Wa Ode Salma
POTENTIAL OF INDONESIAN MEDICINAL PLANTS AS ANTI-CANCER: IN SILICO STUDY
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
Mahmmoud Ismail Mohammed, Muwafaq Ayesh Rabeea and Yazen S. Alnajjar
INFLUENCE FASTING ON PHYSIOLOGICAL AND BIOCHEMICAL VARIABLES IN THE HUMAN BODY
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%.