CURRENT ISSUE

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.

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Syazani Darham, Claudio Gomez-Fuentes, Azham Zulkharnain, Suriana Sabri, Nancy Calisto-Ulloa, Nicolás Ramírez-Moreno and Siti Aqlima Ahmad

 

ISOLATION AND IDENTIFICATION OF MOLYBDENUM-REDUCING COLD-ADAPTED MARINE BACTERIA ISOLATED FROM BERNARDO O’HIGGINS RIQUELME BASE STATION, ANTARCTICA

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.

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M. Ali Sarong , S. Supriyatno, Asiah M.D., Mimie Saputri, S. Safika, Asri Mursawal, Wenni Novita Sari & Agung S. Batubara 

 

DNA HOMOLOGICAL ANALYSIS OF THE GELOINA SHELL IN REULEUNG LEUPUNG MANGROVE ECOSYSTEM, ACEH BESAR DISTRICT, ACEH PROVINCE, INDONESIA

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.

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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.

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Nor Nadia Fadiha Aizuddin, Izyanti Ibrahim, Rusliza Basir, Noor Embi & Hasidah Mohd Sidek 

ANTI-MALARIAL AND CYTOKINE-MODULATING EFFECTS OF SODIUM TUNGSTATE IN MURINE MODELS OF CEREBRAL AND SEVERE MALARIA INVOLVE INHIBITION OF GSK3Β MEDIATED VIA ACTIVATION OF BOTH ERK AND Akt SIGNALING 

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.

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Anton S. Tkachenko, Galina I. Gubina-Vakulyck, Oksana I. Kauk, Anatolii I. Onishchenko, Esmira V. Shekhovtsova & Oksana A. Nakonechna

CHANGES IN S100 PROTEIN EXPRESSION IN THE CEREBELLUM OF RATS ORALLY EXPOSED TO ENERGY DRINKS

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.

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Page 128 - 132

Wira Eka Putra, Wa Ode Salma and Diana Widiastuti

THE POTENCY OF BOESENBERGIN A AND BOESENBERGIN B COMPOUNDS FROM Kaempferia pandurata AS ANTI-METASTATIS AGENT: IN SILICO STUDY

Abstract 

Metastasis is hallmark of the cancer. Recently, targeting molecules that are responsible in promoting metastasis might be one of favorable strategies. In this study, we aimed to evaluate bioactive compounds of Kaempferia pandurata rhizome as anti-metastasis against NF-κB. Molecular docking and molecular interaction among ligands and target protein were applied in this study. Importantly, we found the greatest substances which might possible to become inhibitor to NF-κB. Boesenbergin A and B were predicted to have the favorable interaction scores, -8.2 kcal/mol and -7.1 kcal/mol, respectively. More detail, boesenbergin A has certain interaction to residual amino acids such as GLU160, ASP121, THR153, SER113, LYS149, VAL145, HIS144, THR146, TYR60, ALA62, VAL61, HIS112, ALA111, LEU143, ALA156, ARG157. While the boesenbergin B has interaction to residual amino acids such as SER249, PHE310, LEU272, VAL254, HIS307, ARG308, ALA311, ASP274, CYS273, LYS275, LYS244. In the future, further research is necessary to confirm and validate the biological activities of boesenbergin A and B, especially in metastasis incidence.

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Wira Eka Putra, Wa Ode Salma and Diana Widiastuti

VIRTUAL SCREENING OF POTENTIAL HEPATOPROTECTIVE AGENTS FROM Andrographis panuculata AGAINST MACROPHAGE INFLAMMATORY PROTEIN-1β

Abstract 

This study aimed to evaluate the bioactive compounds of A. paniculata as an inhibitor against the macrophage inflammatory protein-1β (CCL-4) to avoid liver diseases. Multiple bioactive compounds from A. paniculata were screen through in silico approach. The score of binding affinity and the interaction trends among the ligands and protein were considered to determine potential hepatoprotective agents from those compounds. According to our prediction, there are four compounds of A. paniculata which may have potential as an inhibitor to the protein target, CCL-4 such as andrographidine A, bisandrographidine C, anrographidine C, and neoandrographolide. Furthermore, the ligands – residual amino acid interaction are andrographidine A – CCL4 (Van der Waals, Carbon Hydrogen Bond, Conventional Hydrogen Bond, Pi-Pi Stacked, and Pi-Alkyl); bisandrographidine C – CCL4 (Van der Waals, Conventional Hydrogen Bond, Alkyl/ Pi-Alkyl, and Unfavorable Acceptor-Acceptor); anrographidine C – CCL4 (Van der Waals, Conventional Hydrogen Bond, Carbon Hydrogen Bond, Pi-Pi Stacked, and Unfavorable Acceptor-Acceptor); neoandrographolide – CCL4 (Van der Waals, Conventional Hydrogen Bond, and Alkyl/ Pi-Alkyl). In the future, more experiments based on laboratory works are needed mainly to evaluate the specific mechanism of action of these compounds against the CCL4.

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Nur Fathiah Tahir, Aiman Wadhihah Ruslan and Nadiya Akmal Baharum

DEFINING THE ANTI-FUNGAL POTENTIAL OF PLANT THAUMATIN-LIKE PROTEIN (TLP): A MINI REVIEW

Abstract 

Unrelenting threats of fungal diseases have caused huge losses to farmers around the world. Despite efforts and progress made in strengthening the conventional breeding strategies, a sustainable solution is yet to be discovered. Thus, the development of transgenic crops using anti-fungal genes such as thaumatin-like protein (TLP), a member of class 5 pathogenesis-related protein (PR-5), is deemed as a viable solution to achieve better resistance trait. However, the plant genome usually has multiple TLP copies with varying signature domains and motifs. It is possible that not all of the copies exert anti-fungal activity. In addition, the biological roles of TLPs underlying their ability to improve the plants’ tolerance are still inconclusive. This mini-review will discuss the unique characteristics of TLPs, which might contribute to their anti-fungal property as well as the various biological roles the proteins may play in the plants’ defense mechanism against fungal infection. Several TLPs that have been proven to possess anti-fungal activity will also be discussed. Moreover, the impact of transgenic plants overexpressing TLPs in combating fungal diseases will also be elaborated via several successful transgenic researches. The information presented in this mini-review will greatly highlight the potential of TLPs as an anti-fungal agent, especially in the generation of transgenic plants with improved tolerance against fungal diseases.

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CURRENT ISSUE

December 2020

Malay. J. Biochem. Mol. Biol. (2020) 23 (3)

TABLE OF CONTENTS

Page 1-7

Angelo Mark P. Walag and Romeo M. Del Rosario

INITIAL EVALUATION OF METAL CONTENT OF Acanthaster planci AND Linckia laevigata COLLECTED FROM CARMEN, AGUSAN DEL NORTE, PHILIPPINES

Abstract 

Harmful substances like metals ultimately end in the seafloor and contaminate marine ecosystems. These substances are of particular concern due to their bioavailability and toxicity to marine organisms. In this study, metals (Pb, Cd, Cr, Zn, and Cu) were determined in the body wall of common sea star L. laevigata, and A. planci collected from Carmen, Agusan del Norte, Philippines. The metal content in terms of concentration of L. laevigata follows the order Pb > Cu > Cd > Cr > Zn while for A. planci is Pb > Cu > Cr > Zn > Cd. Both species were found to consistently contain the most lead and copper, which could suggest bioaccumulation of these elements from the environment. It is believed that these metals were sourced from the sediments or their food and are accumulated in their body wall. The differences in metal content for Pb and Cd were believed to be due to the difference in the diet for the two species. Comparable metal content was found in Cd, Cr, and Zn. It is recommended that further studies are conducted, especially in monitoring the metal content in the sediment and bioaccumulation of these metals in different trophic levels.

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Galina Gubina-Vakulyck, Anatolii Onishchenko, Oksana Nakonechna, Andrii Lupyr, Tatyana Gorbach,
Alla Bondareva, Svetlana Stetsenko, Viktoriya Tkachenko, Hanna Polikarpova and Anton Tkachenko

 

LONG-TERM EXPOSURE TO CAFFEINATED ENERGY DRINKS PROMOTES APOPTOSIS OF CARDIOMYOCYTES AND RESULTS IN ENDOTHELIAL DYSFUNCTION IN MYOCARDIAL BLOOD VESSELS

Abstract 

This study aimed to assess the apoptosis rate and endothelial function of the heart of rats that were orally exposed to a caffeinated energy drink (CED). The rats were orally exposed to CED for two weeks. The content of caspase-3, endothelial NO-synthase (eNOS), endothelin-1 (ET-1), vascular endothelial growth factor A (VEGFA), the activity of poly-(ADP-ribose) polymerase (PARP) was determined in the heart homogenates. Expression of heat shock protein 90α (HSP90α) and proliferation-associated Ki-67 protein in the heart were evaluated. Biochemical analysis demonstrated the activation of apoptosis in the heart of rats exposed to CED with the evidence of the elevated concentration of caspase-3 and the reduced activity of PARP. Consumption of CED was accompanied by the development of endothelial dysfunction in the cardiac vasculature, which was confirmed by a higher content of ET-1 and VEGFA in heart homogenates and HSP90α overexpression in endothelial and endocardial cells. Oral exposure to CED for two weeks is associated with enhanced apoptosis of cardiomyocytes and the development of endothelial dysfunction in the cardiac blood vessels.

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Ghaidaa Raheem Lateef Al-Awsi, Gulboy Nasir, Majida M. Meteab Alshammari and Abdolmajid Ghasemian

 

ANTIBACTERIAL AND ANTIBIOFILM EFFECTS OF BISMUTH NANOPARTICLES PRODUCED BY BACILLUS SUBTILIS AGAINST MULTIDRUG-RESISTANT Pseudomonas aeruginosa

Abstract 

This study aimed to determine the effect of green bismuth oxide (BiO) NPs against multidrug-resistant (MDR) Pseudomonas aeruginosa (P. aeruginosa) from wound infections. Among 450 wound samples collected from patients admitted to the hospital, 200 P. aeruginosa isolates were identified. MDR strains of P. aeruginosa were detected by disc diffusion method. BiO NPs were synthesized using wild Bacillus subtilis (B. subtilis) strain and infrared spectroscopy, X-ray diffraction and scanning electron microscopy techniques. The antibacterial effect of the NPs compared to antibiotics against MDR strains was evaluated using a standard disk diffusion method. BiO NPs were synthesized at 0.005 M concentration of solution. According to the SEM images, the BiO NPs were irregular in size and ranged from 23-41 nm. According to the disk diffusion method, the highest inhibitory effect of BiO NPs was observed against MDR P. aeruginosa at 2000 ppm. The MIC and MBC of Bi-O NPs at 2000 ppm and >2000 ppm was observed against 97/200 (47.5%) of MDR P. aeruginosa. Eighty percent and 20% of P. aeruginosa produced strong and moderate-level biofilms, respectively. In the presence of sub-inhibitory concentration (1200 ppm) of Bi-ONPs, 52 (26%) isolates produced strong-level biofilms (p=0.001) and 22 (11%) produced moderate-level biofilm (p=0.029), while 53% of remaining isolates produced weak biofilms. The results exhibited a significant decrease in biofilm formation in the presence of Bi-O NPs. BiO NPs exerted an antibacterial effect that 2000 ppm and had a significant inhibitory effect against P. aeruginosa biofilms.

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Manaf A. Guma, Hekmat Ahmed Owaid, Kiffah Shaddhan Hamad, Hazim Abdul Rahman Alhiti, Mustafa Abdulrahman Jumaah Alhiti, Tahir Rissan Dikheel and Bilal Aldahham

ASSOCIATION OF Helicobacter pylori INFECTION WITH THE MOST COMMON AFFECTED AGE: A STATISTICAL STUDY

Abstract 

Helicobacter pylori (H. pylori) is a bacterium that causes infections, and the most prevalent symptom is upper abdominal pain. However, the age group of people who are most susceptible to H.pylori infection is still a matter of debate. In this study, we aim to analyse and report the results of H. pylori tests for patients amongst the Iraqi population. The descriptive and retrospective studies involve 224 Iraqi patients of both genders, aged between 18-67 years who gave oral consent and displayed upper abdominal pain. All patients were interviewed and clinically examined before being tested with a qualitative serological test and stool examination. The population of the study consisted of 114 males and 125 females with mean age 25 and 26 years, respectively. The results showed that there were no negative serum tests for both genders: the peak age of infection was between 30-39 years and 50 patients showed positive for both tests. Results showed that the middle-aged population is significantly the more affected group according to serological tests which were positive in all tested patients. Moreover, the study also showed that the antigen detection of H. pylori by IgG, IgM and IgA antibodies using the Rapid Test strips is the most trustable biomarker and is recommended as the initial test for any work involving H. pylori.

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Muna H. Jankeer

THE PREVENTIVE ROLE OF Thymus vulgaris ALCOHOLIC LEAF EXTRACT AGAINST CARBON TETRACHLORIDE INDUCED HEPATOTOXICITY IN MALE ALBINO RABBITS

Abstract 

This study aimed to evaluate the preventive role of Thymus vulgaris alcoholic leaves extract against carbon tetrachloride (CCl4) hepatotoxicity in male albino rabbits. Thirty rabbits were randomly divided into 5 groups (6 rabbits/group). They were dosed daily for 21 days as follows: The 1st group was given 1.5 ml olive oil/kg of body weight (BW) orally days/week as a negative control. The 2nd group was treated with 3 ml orally (1 CCl4:1 olive oil) as a singular dose every 3 days/week as a control. The 3rd, 4th and 5th groups were treated with 3 ml CCl4/kg BW, and after 30 min of the ingestion, these groups were treated once with Thymus vulgaris alcoholic (200, 400, 600 mg/kg) daily, respectively. The results showed that treatment with CCl4 caused a significant decrease in the level of protein, albumin, globulin and high-density lipoprotein-cholesterol, but a significant increase in the level of bilirubin, cholesterol, triglycerides, very low-density lipoprotein-cholesterol, low-density lipoprotein-cholesterol, alkaline phosphatase, alanine aminotransferase and aspartate aminotransferase in serum of positive group as compared with the negative group. A significant decrease in the level of the glutathione, an increase in the level of malondialdehyde and protein were observed in liver tissue, which indicates the ability of CCl4 to induce hepatotoxicity. The results showed that the rabbits that dosed with Thymus vulgaris extract caused an increase in some of the liver function tests and lipid profile. It is concluded that the Thymus vulgaris possesses liver protective activity against CCl4 induced hepatotoxicity in rabbits.

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N.A. Hasan, M.Y. Rafii, A.R. Harun, F. Ahmad, S. Hussein, Kogeethavani Ramachandran, A.I. Akmal Shukri and Ab. M. Haslinda

RADIOSENSITIVITY OF RICE GENOTYPES TO ION BEAM IRRADIATION BASED ON SEEDLING TRAITS AND PHYSIOLOGICAL INDICES

Abstract 

Three Malaysian modern rice genotypes viz., MR84, MR219 and MR284 were examined for varietal differences in radiosensitivity to ion beam irradiations. Dry healthy seeds were exposed to variable doses of ion beam radiations at 10-100 Gy with 10 Gy intervals. The result in this study demonstrated that LD50 for MR84, MR219 and MR284 were 70.9331 Gy, 69.3927 Gy and 52.78Gy, respectively. Shoulder dose for MR84, MR219, MR284 were ranged between 40Gy- 60Gy. This study found highly significant differences among the genotypes (p<0.05) for all traits studied allowing the distinction of three genotype categories. The differences among radiation treatments were highly significant (p<0.05) for germination percentage, shoot and root length, chlorophyll contents (a, b), plant height and panicle fertility. Furthermore, Duncan Multiple Range Test was applied to compare the mean values of all genotypes and doses. MR84 and MR219 showed no significant differences with respect to root length. Increasing in ion beam irradiation doses caused a significant reduction in shoot length and chlorophyll A content. The genotype × dose interaction for all tested traits exhibited significant differences indicating the effect of different radiation levels in performance for characters. In general, genotypes displayed variable response towards ion beam irradiations.

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Noor Dina Muhd Noor, Nor Hafizah Ahmad Kamarudin, Mohd Shukuri Mohamad Ali and Raja Noor Zaliha Raja Abd. Rahman

CHAPERONE CO-EXPRESSION OF INDUSTRIALLY IMPORTANT ENZYMES

Abstract 

Overproduction of recombinant proteins particularly the industrial important enzymes continues to be an inevitable step in their biochemical and structural characterization. Along with other methods, chaperones highly contribute to the improvements in protein overexpression. Due to its versatility, molecular chaperones play a great role to inhibit protein denaturation, suppress the accumulation of disabled protein in the cell and assist in refolding and degradation of denatured proteins. This review focuses on recent advances utilizing chaperones in promoting and regulating protein folding among industrial enzymes; amylases, lipases, proteases and catalases. This review emphasizes the function and role of molecular chaperones which facilitate the production of recombinant enzymes. We presented the effect of chaperone and co-chaperone expression for the soluble expression of recombinant enzymes which would be advantageous for the developments and industrial applications of these enzymes in the future.

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Noor Masyitah Jumahat, Navindra Kumari Palanisamy, Annamalai Chandra Mouli and Jamal Hussaini

IN VITRO STUDY AND MOLECULAR ANALYSIS OF babA AND sabA ADHESIN GENES OF CLINICAL AND ENVIRONMENTAL H. pylori ISOLATES AND THEIR ROLE IN H. pylori COLONIZATION AND PATHOGENESIS

Abstract 

Helicobacter pylori (H. pylori) colonization in the gastric mucosa is the initial stage of infection to occur and ultimately causing gastric diseases such as chronic gastritis, peptic ulcer, and gastric cancer. H. pylori produce several virulence factors, including babA and sabA, that are essential for colonization of the stomach. This study aims to determine the expression of babA and sabA of different H. pylori isolates in vitro and its role in H. pylori colonization and pathogenesis. H. pylori S3 (cagA-), S5 (cagA+), C7 (cagA-) and C8 (cagA+) strains from humans and cockroaches were co-cultured into AGS cells for 6h. Bacterial adherence was determined using Giemsa staining and dilution plating. The expression of babA and sabA was conducted using quantitative real-time PCR (qRT-PCR). Fold differences in the gene expression were determined using the 2-ΔΔCt method. Bacteria adherence rate was higher in infected AGS cells with H. pylori S5 (cagA+) and C8 (cagA+). The expression of the babA gene was significantly upregulated in H. pylori cagA+. However, babA gene expression was downregulated in H. pylori cagA- strains after interaction with the AGS cells. In contrast, the expression of the sabA gene was upregulated in all H. pylori strains. Higher expression of the babA and sabA gene in H. pylori cagA+ increased the bacteria adherence rate in which permit the H. pylori to invade the epithelial cells. Hence, this study postulates that babA and sabA gene plays a vital role in H. pylori colonization. The adhesion of H. pylori to the gastric epithelium is mediated by the adhesin and allows the release of cagA into the host cells that facilitates colonization, induces mucosal inflammation and enable persistent infection that can lead to severe clinical outcomes.

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Prajna P Shetty, Thriveni. G.K and Nalini K

 

INHIBITION ASSAY OF ALA DEHYDRATASE BY SUCCINYL ACETONE IN TYROSINEMIA TYPE 1: A RAPID HT1 SCREENING TECHNIQUE FOR RESOURCE CONSTRAINED DIAGNOSTIC SETUPS

Abstract 

Succinyl acetone is a specific marker of tyrosinemia type I. The present study was undertaken to develop a simple and cost-effective quantitation method for succinyl acetone in dried blood spot (DBS) implementable in resource-constrained diagnostic settings. Spectrophotometric microassay based on the inhibitory effect of succinyl acetone on δ- aminolevulinate dehydratase (ALA-D) in erythrocytes, which catalyzes the formation of porphobilinogen from δ-aminolevulinic acid and is measured at 550nm. The reliability and suitability of the method for routine laboratory practice was assessed before the method was used for screening. Greater inhibition of ALA-D activity was observed when succinyl acetone was spiked to blood before spotting. The stability of succinyl acetone in DBS progressively deteriorated when stored at -800C. Among the 71 samples processed; an inhibition ranging from 2.75% to 55.47% was observed. The maximum inhibition of 63 - 73.47% was observed for a couple of the tyrosinemia true positive cases. The proposed method is useful for the mass screening of newborn babies for Tyrosinemia type I. This could ensure early detection and intervention while addressing the challenge of expenses involved in high-end technology, till the newborn becomes a universal reality.

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Ragaa F. Fathy, Shallan M.A, Wagdy K.B. Khalil, Samah M. Bassem, Fagr Kh. Abdel-Gawad and Abdel- Rahim E.A

 

GENOTOXICOLOGICAL AND BIOCHEMICAL STUDIES ON Holothuria atra FOR ALLEVIATING REACTIVE TEXTILE DYES CONTAINING WASTEWATER IMPACTS IN NILE TILAPIA, Oreochromis niloticus (L.)

Abstract 

Anthropogenic organic dyestuffs and their derivatives have been enormously used in various manufactures such as textiles, pharmaceuticals, food, cosmetics and leather. Such environmental chemicals can readily be discharged into the aquatic ecosystem through industrial effluent drainage. Several research studies have indicated that synthetic organic dyes and their derivatives could cause reproductive impairment in aquatic fauna. However, the endocrine-disrupting impacts of such dyestuffs and colorants containing wastewaters have been scarcely explored. Herein, we examined the potential of three different sorts of reactive textile dyes (RTDs), i.e., reactive red 195 (RR195), reactive yellow 18 (RY18), and reactive blue 4 (RB4) to interrupt sex steroids synthesis. Furthermore, our study investigated the biological role of Holothuria atra (H. atra) in the attenuation of the deleterious effects induced by such organic dyes. For this purpose, tilapia fish juveniles of Oreochromis niloticus (O. niloticus) were used. Regulatory fluctuations have been observed in three substantial hypothalamic-pituitary-gonadal axis (HPG axis) genes by all tested reactive dyes, i.e., downregulation of FSHβ, LHβ (sex steroids) and CYP19A1 (steroidogenic gene) and up-regulation of them in the presence of H. atra. Our outputs suggest that some RTDs could act as endocrine disruptors and may consequently severely impact the aquatic environment.

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Redzuan Nul Hakim Abdul Razak, Azantee Yazmie Abdul Wahab, Roszaman Ramli, Nik Mohammad Faez Nik Osman, Afzan Mat Yusof, Nik Fakhuruddin Nik Hassan, Hussin Muhammad, Muhammad Alif Mazlan and Muhammad Lokman Md Isa

IDENTIFICATION OF BRCA1 TUMOR SUPPRESSOR GENE EXPRESSION AS A DNA REPAIR GENE IN FROZEN CLEAVAGE EMBRYO CELL AT IIUM FERTILITY CENTRE

Abstract 

BRCA1 is known as the essential tumor suppressor gene in human that expresses a protein that called breast cancer type 1 susceptibility protein. It has two main functions which are repairing the damaged DNA and also inducing the apoptotic mechanism to eliminate the cell when mutated DNA could not be repaired. In this study, all the embryo samples that have been used were considered as grade 1 according to Embryo Grading. This study aimed to investigate BRCA1 gene expression in relation to the human embryo quality. The RNA from the cleavage embryo was extracted and was tested by Reverse-Transcriptase Polymerase Chain Reaction (rt-PCR) and gel electrophoresis. The results showed that BRCA1 gene expression was expressed in frozen cleavage embryos. Furthermore, the statistical analysis revealed that there are no significant differences among the fragmentation rates and also in Pre-embryo Genome Activation (EGA) and Embryo Genome Activation (EGA) phases. This study would be beneficial in terms of controlling the DNA repair gene activity during the pre-implantation phase. The expression of BRCA1 could be a useful health assessment indicator for embryos.

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Redzuan Nul Hakim Abdul Razak, Muhammad Alif Mazlan, Azmir Ahmad, Mohd. Azri Abd. Jalil, Abdul Halim Abdul Jalil and Muhammad Lokman Md. Isa

 

VIABILITY OF FRESHLY PREPARED CELLS DERIVED FROM FOETAL BRAIN OF RABBIT AT DIFFERENT TEMPERATURE SETTINGS DURING TRANSPORTATION

Abstract 

The characteristics of rabbit cells, i.e., high proliferative activity, high passage numbers, and morphologically similar to human cells renders it a potential source of a cell line. Nonetheless, they are not widely characterised for research purposes. This study aimed to observe the effects of different temperature settings on the viability of rabbit foetal brain (RFB) cells at different time points. The harvested RFB cells were divided into room temperature (RT), ice packs (ICE), and refrigerator (4 °C) groups. Cell viability was determined at 0, 6, 12, 24, 48, and 72 h. Following cell viability determination, the cells were recultured for 24 h to observe cell viability and morphology post-transportation. The cell viability was analysed statistically for within and more than 24 h duration using the one-way ANOVA test, and the cell morphology was observed under the microscope. For the within 24 h analysis, the ICE group showed the best condition for 24 h transportation due to significantly higher cell viability (p<0.05) compared to the RT group. Meanwhile, for the more than 24 h analysis, the RT group showed the best condition for more than 24 h transportation due to higher cell viability compared to the ICE and 4 °C groups, albeit the non-significance. The reculturing of RFB cells showed no remarkable difference in the cell viability and morphology in all transportation time points at all temperature settings. In conclusion, the study found that the RFB cells are suitable to be transported using ice packs for transportation within 24 h and room temperature for transportation of more than 24 hours.

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Page 98 - 102

Sujatha R. and Kailash K

PENTRAXIN-3 AS AN EMERGING RELIABLE NON-INVASIVE SERUM MARKER IN THE DETECTION OF ALCOHOLIC FIBROSIS

Abstract 

Alcoholic cirrhosis is the end spectrum of alcoholic liver disease, which includes fatty liver, alcoholic hepatitis, fibrosis, cirrhosis and superimposed hepatocellular carcinoma. Fibrosis is central to the pathology of all types of chronic liver disease. Pentraxin-3 is involved in numerous roles like regulation of inflammation and innate resistance to pathogens. It is strongly expressed on collagen bundles and fibrotic areas. After ethical clearance, demographic and laboratory details were collected from medical records. Leftover samples were used for serum pentraxin-3 estimation using an ELISA kit. The study included a total of 40 participants who were males 20 known cases of alcoholic liver disease and the control group included 20 healthy controls. The average value of serum pentraxin-3 in alcoholic liver disease cases was significantly (p=0.006) elevated as compared with healthy volunteers (0.51 ± 0.13ng/ml vs 0.27 ± 0.1ng/ml).This study concludes that serum pentraxin-3 could emerge as one of the novel non-invasive biomarkers in predicting the progression of fibrosis in alcoholic liver disease patients.

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Page 103 - 118

Suzana Adenan, Chee Fah Wong, Saripah Salbiah Syed Abdul Azziz, Som Cit Si Nang, Rosmilah Misnan, Iffah Izzati Zakaria, Mardiana Mohd Ashaari, Dhilia Udie Lamasudin and Raja Noor Zaliha Raja Abd. Rahman

POLYCYCLIC AROMATIC HYDROCARBONS: CHARACTERISTICS AND ITS DEGRADATION BY BIOCATALYSIS REMEDIATION

Abstract 

An excessive released of polycyclic aromatic hydrocarbons (PAHs) to surroundings is one of the major factors that cause environmental pollution to increase globally. This issue had gained scientist’s attention to study PAHs biodegradation pathways and their toxicity towards humans and the environment. They found that the major mechanism responsible for the ecological recovery of PAH-contaminated sites happened to be from the microbial degradation process. However, there are a few limitations faced by the PAHs degrading bacteria where the bacteria die due to extremely polluted areas. This leads the researchers to utilize genetic engineering to produce enzymes that can withstand and survive in extreme environments. Recent information and technology such as path sources, properties and biochemical pathways by means to produce the simplest and less harmful components in polluted ecosystems are discussed in this review. In-depth studies in regards to bacteria biocatalysis involving bacterial-produced-enzymes to degrade PAHs help develop new methods to enhance the bioremediation effectiveness in the future.

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Page 119 - 127

Tajudeen A Owolarafe, Chimaobi J. Ononamadu, Godwin O. Ihegboro, Kailani Salawu Muhammad K Atiku, Adamu J. Alhassan and Alhassan M.Wudil

TOXICOLOGICAL EVALUATION OF ACUTE AND SUBCHRONIC ORAL ADMINISTRATION OF ETHYL ACETATE EXTRACT OF Ziziphus mauritiana LEAF

Abstract 

This study aimed at evaluating the effect of ethyl acetate extract of Ziziphus mauritiana leaves on biochemical parameters in Wistar rats. Twenty-five albino rats equally divided into five experimental groups were used. One group served as control and received the carrier solvent treatment. Four test groups were treated with Z. mauritiana extract at 200, 400, 600 and 1000 mg/kg body weight respectively. The experiment lasted for 21 days after which the rats were sacrificed and blood collected for biochemical and haematological evaluation. Liver and Kidney-body weight ratio was computed. histoarchitecture of the Liver and Kidney were also investigated. The results showed no death at 5000mg/kg body weight while some haematological parameters were significantly (P<0.05) affected at 400, 600 and 1000mg/kg body weight for haemoglobin, red blood cell, parked cell volume, white blood cell and platelet concentrations. There were also significant (P<0.05) alterations in activities of gamma-glutamyl transferase, alanine aminotransferase, and aspartate aminotransferase, as well as the levels of total protein, albumin and globulin in the serum. Significant (P<0.05) increase were observed in the computed liver-body weight ratio with marked alterations in histoarchitecture of the liver cells. Significant (P<0.05) alterations were observed at all doses administered for creatinine, urea, sodium, chloride and biocarbonate ions with marked difference in kidney-body weight ratio and kidney cell architecture. These alterations in haematological parameters, liver function enzymes, kidney function indices and histological evaluation suggest toxicity of the extract on the animals at 400, 600 and 1000mg/kg body weight despite its non-toxic classification of acute administration of the extract.

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Page 138 - 149

Yow San Chan and Khim Phin Chong

PHYTOCHEMICAL INVESTIGATION AND ANTIOXIDANT ACTIVITY OF Ganoderma boninense

Abstract 

Ganoderma lucidum is known as a functional mushroom and is traditionally being used as medicine but G. boninense is well known as the causal pathogen of basal stem rot disease of oil palm. Numerous secondary metabolites of mushrooms from this genus have been reported with various biological activities. However, there are not many reports report on the medicinal benefits of G. boninense. Hence, this study was designed to investigate the phytochemical constituents and antioxidant activities of the fruiting body G. boninense extracts. Various groups of phytochemicals were extracted using solvents with different polarities such as chloroform, ethyl acetate, acetone, methanol, ethanol and distilled water. Results confirmed the presence of numerous phytochemicals in G. boninense fruiting body. Ethanolic extract of G. boninense has the highest phenolic (33.05 ± 1.374 mg GAE/g DW) and total flavonoid (8.20 ± 0.059 mg QE/g DW) content. Furthermore, ethanolic extract of G. boninense also exhibited the greatest potency in antiradical activity with IC50 of 113.9 ± 2.78 μg/ml. Correlation coefficient, R2 = 0.97 suggested phenolic acids of ethanol extract was contributed to its radical scavenging activity.

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