Recombinant Pyruvate Phosphate Dikinase (rPPDK): Development Of Antibody And Antigen Rapid Dipstick Tests For Amoebiasis And Identification Of Potential Anti-Amoebic Compounds

Amebiasis, sejenis penyakit protozoa enterik yang disebabkan oleh Entamoeba histolytica, merupakan satu masalah kesihatan awam di kebanyakan negara membangun Amoebiasis, an enteric protozoan disease caused by Entamoeba histolytica, is a public health problem in many developing countrie

Cloning and Expression of the Nucleocapsid Protein of Newcastle Disease Virus in Pichia Pastoris (Guillierm.) Phaff

Newcastle disease virus (NDV) is the only member of the genus Avulavirus of the family Paramyxoviridae. NDV causes a respiratory disease in birds known as Newcastle disease (ND) which may result in high mortality in susceptible hosts such as chickens leading to substantial loss in the poultry industry. Recent outbreak has been reported in many countries including Malaysia. The continuing treat of ND to the poultry industry requires routine testing through development of better diagnostic tools. Therefore, the objective of the current study was to express the immunogenic nucleocapsid (NP) gene in a Pichia pastoris expression system with a view to develop a potential and cost effective antigen for development of a diagnostic test. In the present study, the gene encoding NP protein of Newcastle disease virus strain AF2240 was cloned into expression vector, pPICZA and placed under the control of methanol inducible alcohol oxidase (AOX) promoter. Then recombinant multi-copy number Pichia cells with Mut+ phenotype were selected for NP protein expression. The optimization of the NP protein production in 50 ml culture was carried out for methanol concentration and different loaded volume in identical shake flask. A time course study for NP production in 250-ml flask with the optimized conditions was performed as well. The result showed that NP protein could be detected after 12 h of methanol induction and the level of protein expression decreased over time. The recombinant NP was purified from the yeast culture using sucrose gradient ultracentrifugation. The high level and intact recombinant nucleocapsid protein expression (570 mg/l) was obtained after 24 h of induction with 1% methanol when 10% of the shake flask was loaded with MMH (minimal methanol with histidine) medium. Western blot analysis using polyclonal NP antibody confirmed the expression of NP with the molecular weight of 53 kDa indicating that NP protein retained its antigenicity. The recombinant NP protein was highly stable in P. pastoris system because there was no degraded product after purification. This result proved that the yeast expression system produces a high yield of recombinant NP protein. The production of recombinant NP protein in bulk as the antigen for diagnostic tools would facilitate the monitoring of NDV infection as well as allowing a more effective control of the disease

Entamoeba histolytica: Membrane and Non-Membrane Protein Structure, Function, Immune Response Interaction, and Vaccine Development

Entamoeba histolytica is a protozoan parasite that is the causative agent of amoebiasis. This parasite has caused widespread infection in India, Africa, Mexico, and Central and South America, and results in 100,000 deaths yearly. An immune response is a body\u27s mechanism for eradicating and fighting against substances it sees as harmful or foreign. E. histolytica biological membranes are considered foreign and immunogenic to the human body, thereby initiating the body\u27s immune responses. Understanding immune response and antigen interaction are essential for vaccine development. Thus, this review aims to identify and understand the protein structure, function, and interaction of the biological membrane with the immune response, which could contribute to vaccine development. Furthermore, the current trend of vaccine development studies to combat amoebiasis is also reviewed

A Review: Natural and Synthetic Compounds Targeting Entamoeba histolytica and Its Biological Membrane

Amoebiasis is the third most common parasitic cause of morbidity and mortality, particularly in countries with poor hygienic settings in central and south America, Africa, and India. This disease is caused by a protozoan parasite, namely Entamoeba histolytica, which infects approximately 50 million people worldwide, resulting in 70,000 deaths every year. Since the 1960s, E. histolytica infection has been successfully treated with metronidazole. However, there are drawbacks to metronidazole therapy: the side effects, duration of treatment, and need for additional drugs to prevent transmission. Previous interdisciplinary studies, including biophysics, bioinformatics, chemistry, and, more recently, lipidomics studies, have increased biomembranes’ publicity. The biological membranes are comprised of a mixture of membrane and cytosolic proteins. They work hand in hand mainly at the membrane part. They act as dedicated platforms for a whole range of cellular processes, such as cell proliferation, adhesion, migration, and intracellular trafficking, thus are appealing targets for drug treatment. Therefore, this review aims to observe the updated trend of the research regarding the biological membranes of E. histolytica from 2015 to 2021, which may help further research regarding the drug targeting the biological membrane

Proteomics of broiler’s liver

In this study, the differences in prote in expression from chicken liver between commercialbroilers (CB) and indigenous chicken (IC) have been analysed by using two-dimensional gel electrophoresis (2D-PAGE). The study was conducted adjacent to consumer experience in order to find out the differences of the protein profile in liver tissues between the two popular types of chickens that have been offered to the Malaysians consumer for over 50 years. In 2D-PAGE, six spots were successfully identified differentially expressed in term of intensity. Those proteins are phosphoenolpyruvate carboxykinase (PEPCK) (2 spots), delta-1-pyrolline-5-carboxylate (P5CDH), soluble epoxide hydrolase (sEH), heat shock protein 90 (HSP90) and γ-actin. PEPCK and P5CDH enzymes were expressed higher in CB compared to IC, suggesting the utilization of alternative metabolic pathways for broilers’ energy metabolism. The sEH expression was found higher in CB compared to IC probably due to the process of xenobiotic metabolic activity to eliminate harmful epoxide inside the CB’s liver. HSP90 was found less in CB compared to IC, which may indicate the lack of regulation of heat stress response in CB. The higher expression of actin in IC liver suggested a good form of structure integrity in IC liver compared to CB, which may happen due to the age differences. Most of the differentially expressed proteins are involved in energy metabolism, stress regulation, and cellular structure. These differences could arise from many variables involved which may include breeds, age, feed regimen,drug treatment and rearing environment

The near consumer insight on liver proteomics of Malaysian broiler in comparison with indigenous chicken

In this study, the differences in protein expression from chicken liver between commercial broilers (CB) and indigenous chicken (IC) have been analysed by using two-dimensional gel electrophoresis (2D-PAGE). The study was conducted adjacent to consumer experience in order to find out the differences of the protein profile in liver tissues between the two popular types of chickens that have been offered to the Malaysians consumer for over 50 years. In 2D-PAGE, six spots were successfully identified deferentially expressed in term of intensity. Those proteins are phosphoenol pyruvate carboxykinase (PEPCK) (2 spots), delta-1-pyrolline-5-carboxylate (P5CDH), soluble epoxide hydrolase (sEH), heat shock protein 90 (HSP90) and γ-actin. PEPCK and P5CDH enzymes were expressed higher in CB compared to IC, suggesting the utilization of alternative metabolic pathways for broilers’ energy metabolism. The sEH expression was found higher in CB compared to IC probably due to the process of xenobiotic metabolic activity to eliminate harmful epoxide inside the CB’s liver. HSP90 was found less in CB compared to IC, which may indicate the lack of regulation of heat stress response in CB. The higher expression of actin in IC liver suggested a good form of structure integrity in IC liver compared to CB, which may happen due to the age differences.Most of the differentially expressed proteins are involved in energy metabolism, stress regulation, and cellular structure.These differences could arise from many variables involved which may include breeds, age, feed regimen, drug treatment and rearing environment

Production of Toxocara cati TES-120 Recombinant Antigen and Comparison with its T. canis Homolog for Serodiagnosis of Toxocariasis

Photograph of a scene at Ed Galloway's Totem Pole Park

Molecular assessment of native fish diversity in UNESCO heritage site, Tasik Raban, using DNA barcoding

Freshwater species are the most threatened group to be assessed to date by the IUCN. Freshwater fish has enormous importance as animal protein supplies for human, and it is estimated that 6% of the world annual animal protein supplies come from freshwater fishes. Barcoding projects have been initiated all over the world and the field is constantly growing. In Malaysia however, the field has not been deeply investigated and not many barcoding projects have been undertaken especially for freshwater fishes. The aim of our study is to support the progress of DNA barcode project, especially for inland reservoirs like Tasik Raban, Perak. A Standard methodology using Cytochrome c oxidase subunit 1 (COI) marker was developed to ensure native fishes are barcoded taxonomically and  molecularly and ready to be accessed through online databases. Such public references can help increase awareness on local fish diversity management. Information on taxonomy and molecular characterization can be used to plan further conservation programmed especially for depleted, unrecognized, and cryptic native species