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Infection with Toxoplasma gondii remains widespread among humans and animals as water, soil and food continue to serve as the major carriers of the sporulated oocyst. The infection is poorly controlled due to lack of a potent vaccine against the parasite, and the current medication presents severe side effects on the host, less efficacy on the parasite and is accompanied by the potential development of resistance by the parasite. The aim of this study was to evaluate the in vitro activities of ethanolic extract of Tinospora crispa (EETC) on protein kinases involved in the lytic cycle of T. gondii infection in Vero cell line. The EETC was obtained through the maceration of dried stem powder. Vero cells infected with the RH strain of T. gondii were used to evaluate the inhibitory effect of EETC against T. gondii calcium dependent protein kinase (CDPK) genes and microneme proteins (MIC) that are essential for host cell invasion and intracellular replication of the tachyzoite. Gene expression profiling of CDPK genes was determined through quantitative real-time PCR (qPCR) after 24 h of treatment. The expression of microneme protein was determined through western blot technique. The RT-qPCR revealed downregulation of most protein kinase (PK) genes after treatment with EETC. The expression of CDPK1, PKG, CDPK3, CDPK6, CDPK7 genes that participate in the lytic cycle of T. gondii infection was downregulated. The expression of the TgMIC1 and TgMIC2 proteins were observed to have decreased in both 4 and 24 h post-infection treatment models. This study showed that EETC contains promising drug candidates effective against T. gondii that can target the protein kinase genes involved in the lytic cycle of the parasite to prevent disease progression. HIGHLIGHTS This study explored the possible mechanism of action of phytochemicals on Toxoplasma gondii parasite-host cell invasion and intracellular replication The study evaluated the effects of ethanolic extract of Tinospora crispa Miers on protein kinase genes that are involved in host cell invasion The target protein kinase genes were downregulated which prevents the secretion of microneme proteins Inhibition of microneme secretion prevents host cell invasion and intracellular replication by Toxoplasma gondi GRAPHICAL ABSTRACT

Rodent species, such as Rattus rattus diardii and Rattus norvegicus are invasive species of wild rats that serve as potential reservoirs of important human's pathogens. Parasitic zoonosis accounts for over 60% of all human infectious diseases worldwide. This situation arises from the recent changes in the global climate and ecosystem composition, which led to the spread of rodents and rodent-borne pathogens globally. The aim of this study was to determine the occurrence of rodent's parasites and their zoonotic potentials in some selected areas in UPM. Rodents were captured using live-traps and euthanised for helminths and protozoan recovery. Intestinal parasites were detected and identified from stool samples using formalin ethyl-acetate concentration technique (FECT), while tissue parasites were identified by histopathological examination of selected tissue sections of the liver, brain, lungs, and muscle. In this study, a total of 89 wild rats were captured. Twelve species of intestinal and tissue parasites were recorded, of which, Taenia taeniaeformis accounts for the highest infection recorded (28%) followed by Hymenolepis nana (19.5%) and Capillaria hepatica (19.1%), while Toxoplasma gondii was the least parasite (6.7%) identified. Furthermore, other parasites species observed include, Cryptosporidium spp. (21.3%), Entamoeba histolytica/Entamoeba dispar and Moniliformis moniliformis (17.9%), Angiostrongylus cantonensis (16.8%), Hymenolepis diminuta (16.1%), Giardia spp. (14.6%), Trichuris spp. (12.3%), and Sarcocystis spp. (6.74). Based on the results obtained in the present study, 17.1% and 15.4% of the rodents captured were confirmed positive for at least one species of intestinal or tissue parasites, respectively. The presence of these zoonotic parasites in the wild rats suggests the potential risk of rodent-borne zoonotic disease transmission to humans. Hence, the need to improved rats control intervention and public health awareness among the populace., Graphical abstract Image 1, Highlights • This is the first report of endoparasitic infection of wild rats in Serdang, Selangor, Malaysia. • 17.1% of the rodents are positive for intestinal parasites, and 15.4% positive for tissue parasites. • Zoonotic parasites from intestine and tissue parasites were identified. • The likelihood of transmission of zoonotic parasites between wild rats and humans was suggested.

Background The immune modulating potential of IL-35 in multiple human disorders has been reported. Consequent upon the recognition of inflammatory cytokine activation and its preponderance for mediating pathology during malaria infection, the study aimed to characterize the expression and functional contribution(s) of IL-35 in Plasmodium berghei (strain ANKA) infected mice. Methods Plasmodium berghei infection in male ICR mice was used as the rodent model of choice. The time course of IL-35 expression in the systemic circulation and tissues of P. berghei infected mice as well as their healthy control counterparts was assessed by enzyme linked immunosorbent assay and immunohistochemistry respectively. The effect of modulating IL-35 by recombinant IL-35 protein or neutralizing anti-Epstein-Barr virus-induced gene 3 antibody on the cytokine environment during P. berghei infection was assessed by flow cytometry. Furthermore, the influence of modulating IL-35 on histopathological hallmarks of malaria and disease progression was evaluated. Results Interleukin-35 was significantly up regulated in serum and tissues of P. berghei infected mice and correlated with parasitaemia. Neutralization of IL-35 significantly enhanced the release of IFN-γ, decreased the expression of IL-6 and decreased parasitaemia patency. Neutralization of IL-35 was also associated with a tendency towards increased survival as well as the absence of pathological features associated with malaria infection unlike recombinant IL-35 protein administration which sustained a normal course of infection and unfavourable malaria associated histological outcomes in P. berghei infected mice. Conclusion These results indicate the involvement of IL-35 in P. berghei induced malaria infection. IL-35 neutralization strategies may represent viable therapeutic modalities beneficial for the resolution of malaria infection.

Andrographolide (AG) has been shown to have several medicinal and pharmaceutical effects, such as antimicrobial, anti-inflammatory, antioxidant, anti-diabetic, and anti-malarial activities. Moreover, studies to assess the pharmacological effect of AG on the metabolic changes of uninfected red blood cells (uRBCs) have not yet been investigated. This study aims to evaluate the pharmacological effects of AG compared to chloroquine (CQ) on the metabolic variations of uRBCs in vitro using a proton nuclear magnetic resonance (1H-NMR)-based metabolomics approach coupled with multivariate data analysis (MVDA). Forty-one metabolites were successfully identified by 1H-NMR. The results of the unsupervised data analysis principal component analysis (PCA) showed ideal differentiation between AG and CQ. PC1 and PC2 accounted for 71.4% and 17.7% of the explained variation, respectively, with a total variance of 89.10%. Based on S-plot and VIP values, a total of 28 and 32 metabolites were identified as biomarkers in uRBCs-AG and uRBCs-CQ, respectively. In uRBCs treated with AG, ten metabolic pathways were determined to be disturbed, including riboflavin metabolism, d-glutamate and d-glutamine metabolism, phenylalanine metabolism, glutathione metabolism, proline and arginine metabolism, arginine biosynthesis, citrate cycle, glycolysis/gluconeogenesis, and pyruvate metabolism as well as alanine, aspartate, and glutamate metabolism. In contrast, in CQ-treated uRBCs, nine affected metabolic pathways were determined, which involved the same metabolic pathways for uRBCs-AG, except for glutathione metabolism. These findings suggest an evident relationship between AG and CQ associated with metabolic changes in intact RBCs after being exposed to the treatment. The metabolomics results could allow useful comprehensive insights into the underlying mechanism of the action of AG and CQ on red blood cells. Consequently, the 1H-NMR-based metabolomics approach was successfully utilized to identify the pharmacological effects of AG and CQ on the metabolic variations of uRBCs.

Background and Aim: Highly sensitive and specific diagnostic assay for the detection of Strongyloides is needed due to the intermittent and low concentration of eggs, larvae and adult worms that can be found in a faecal specimen. In some cases, repeated sampling of the faecal specimen is required to obtain satisfactory and reliable results. The aim of the study is to develop and evaluates monoclonal antibody-based Sandwich ELISA for the detection of coproantigen associated with Strongyloides infection using S. ratti as a model. Place and Duration of Study: Department of Medical Microbiology and Parasitology, Faculty of Medicine and Health Sciences, University Malaysia, Between September 2018 and March 2019. Methodology: The monoclonal antibody was raised against a soluble antigen of the infective filariform larvae (iL3) of S. ratti. The monoclonal antibody produced (IgG2bMAb) was evaluated for cross-reactivity against homologous and heterologous helminth antigens such as excretory-secretory (ES), infective larvae (iL3) and coproantigen of S. ratti, adult worms of A. caninum, A. suum, T. canis and T. cati. Results: An IgG2bMAb was observed to react with 30 kDa proteins associated with all homologous antigen from iL3, ES and coproantigen of S. ratti and cross-reacted with one heterologous antigen from adult worm of A. caninum at the same molecular weight. There was no cross-reaction observed with other heterologous antigens from adult worms of T. canis, T. cati and A. suum. The sensitivity of IgG2bMAb for the detection of S. ratti was 85% in Sandwich ELISA. Cross- reaction was observed with hookworm antigen that caused by A. caninum in Western immunoblotting. Conclusion: The results indicated that IgG2b have an immunodiagnostic property as IgG2bMAb and was able to detect antigens from coproantigen related to S. ratti with 85% sensitivity based on Sandwich ELISA) even though cross-reaction was observed with A. caninum. These findings will be very useful to tackle many cases of multiple worms’ infections such as both strongyloidiasis and hookworm. Therefore, we recommend that further evaluation and study in the human area where multiple infections can be common should be carried out.

The status of toxoplasmosis management is challenged by drug intolerance, compromised efficacy and potential development of drug resistance. However, currently, there are very limited targets on T. gondii that serve as the site of action of current medications. This review aimed to explore more potential targets that are essential to parasite survival and are absent in humans. Many unexplored targets on T. gondii exist and their specificities to the parasite make them ideal drug targets. The present review had searched relevant English databases such as PubMed, Scopus, Google scholar and Science Direct, for relevant literature on the therapeutic target of T. gondii. Many of the enzyme systems in several pathways are essentially palatable as drug targets. Establishing compounds that can target these enzymes on T. gondii will greatly be beneficial in treating acute and chronic toxoplasmosis in humans.

Background and Aims: Rodents constitute more than 42% of the known mammalian species, with 1700 species which belongs to three different families, include Muridae, Microtidae, and Sigmodontidae. Rodents species such as R. r. diardii and R. norvegicus play an important role as hosts for ectoparasites and reservoirs for various types of viruses, bacteria, rickettsia, protozoa, and helminths which are responsible for causing zoonotic diseases to humans and other vertebrate animals. The aim of this work is to identify the species of mites, ticks, and fleas causing diseases to humans and determined the prevalence of infestation in relation to gender, age, and habitat of the rodents. Place and Duration of the Study: Department of Medical Microbiology and Parasitology, Faculty of Medicine and Health Sciences, University Malaysia, Between September 2018 and March 2019. Methodology: Wild rats were captured using live traps from garbage areas, and places near the cafeteria in the student’s residential colleges at University Putra Malaysia. The rats were humanely euthanised and identified. They were classified as adult or juveniles. Their sex was also determined. Ectoparasites were collected by combing the fur the rodents on to a white plan sheet paper. The ectoparasites collected were washed and mounted with Hoyer’s media on a glass slide. Parasites were identified using a key morphological feature. Results: A total of 89 wild rats were trapped and examined for ectoparasites. Eight different species of ectoparasites that comprised of L. echidnanus, L. nuttalli, O. bacoti, I. granulatus, Heamaphysalis sp., P. spinoluso, H. pacifica, X. cheopis) were identified from the rodents examined. About 55% of the rodents trapped were positive for at least one species of ectoparasite parasites, and about 45.8% of the male rats and 30.8% of female were positive for ectoparasites. Meanwhile, in the adult, 42.9% are positive for at least one species of ectoparasites, whereas 32.2% of the juvenile rodent was also found positive for at least one species of ectoparasites. Conclusion: The results of this study indicated that rodents trapped from the student’s colleges in University Putra Malaysia are infected with various ectoparasites species that might play an important role in the transmission of certain zoonotic diseases to humans. Therefore, we conclude that there is potential risk of rodent-borne zoonotic diseases transmission to humans in the study area. Awareness of prevention and control of rodent-borne diseases should be introduced to educate the students on the importance of zoonotic diseases associated with rodents.

Background: The increasing incidence of drug resistance among various strains of Plasmodium falciparum has compelled researchers to search for new improved therapeutic alternatives to current antimalarials. Consequently, the study aimed to investigate the effect of varying the duration of andrographolide exposure on its anti-plasmodial effect against intra erythrocytic stages of the P. falciparum 3D7 parasite. Although andrographolide has demonstrated prior anti-plasmodial effect against P. falciparum 3D7, its time-dependent effect subsequent to different durations of drug exposure in addition to the impact of relevant pharmacologically active concentrations on the cellular morphology of various intraerythrocytic stages of the P. falciparum 3D7 parasite cycle are limited.Methods: P. falciparum 3D7 parasites cultivated in vitro in blood cultures were individually incubated with different concentrations of andrographolide, chloroquine and drug-free parasite culture which served as the representative control. Suppression of parasite growth was determined by parasite lactate dehydrogenase (pLDH) based drug sensitivity assay. The inhibition of parasite growth and changes in morphology of intraerythrocytic parasites subsequent to treatment initiation with andrographolide or chloroquine were assessed upon commencement of a synchronized cycle at 12, 24 and 48 h respectively. Results: Andrographolide showed satisfactory growth inhibitory effect however its inhibitory activity was substantially lower when compared to that of chloroquine. Unlike chloroquine which showed maximal inhibitory activity within the first 12 h of the cycle, suppression of parasite growth by andrographolide was most prominent during the development of early trophozoites (viz the second 12 hours). Andrographolide failed to produce any effect on the morphology of ring stage parasites, it however produced a noticeable change in the morphological appearance and sizes of mature trophozoites. Whereas, with chloroquine notable changes to ring and trophozoite stages of the parasites were evident. Conclusion: The data obtained indicates the potential role of andrographolide as an adjunctive treatment in malaria subject to further clinical evaluations.

Toxoplasmosis is a disease caused by the protozoan parasite Toxoplasma gondii (T. gondii). Human toxoplasmosis seroprevalence in Malaysia has increased since it was first reported in 1973 as shown in previous reviews of 1991 and 2007. However, over a decade since the last review, comprehensive data on toxoplasmosis in Malaysia is lacking. This work aimed at reviewing articles on toxoplasmosis research in Malaysia in order to identify the research gaps, create public awareness, and efforts made so far and proffer management options on the disease. The present review examines the available published research articles from 2008 to 2018 related to toxoplasmosis research conducted in Malaysia. The articles reviewed were retrieved from nine credible databases such as Web of Science, Google Scholar, ScienceDirect, PubMed, Scopus, Springer, Wiley online library, Ovid, and Cochrane using the keywords; Malaysia, toxoplasmosis, Toxoplasma gondii, toxoplasma encephalitis, seroprevalence, human immunodeficiency virus (HIV) patients, pregnant women, genotype strain, anti-toxoplasma antibodies, felines, and vaccine. The data highlighted seropositive cases from healthy community members in Pangkor Island (59.7%) and among migrant workers (57.4%) at alarming rates, as well as 42.5% in pregnant women. Data on animal seroprevalence were limited and there was no information on cats as the definitive host. Genetic characterization of Toxoplasma gondii from HIV patients; pregnant women, and domestic cats is lacking. This present review on toxoplasmosis is beneficial to researchers, health workers, animal health professionals, and policymakers. Therefore, attention is required to educate and enlighten health workers and the general public about the risk factors associated with T. gondii infection in Malaysia.

The major route for Toxoplasma gondii (T. gondii) infection is through the ingestion of foods contaminated with oocyst from cat faeces. The microscopic detection of T. gondii oocysts in cat faeces is challenging, which contributes to the failure of detecting or differentiating it from other related coccidian parasites. This study aims to detect T. gondii oocysts in cat faeces using two multicopy-target PCR assays and to evaluate their genetic diversity. Cat faecal (200) samples were collected from pet cats (PCs, 100) and free-roaming cats (FRCs, 100) within Klang Valley, Malaysia, and screened for coccidian oocysts by microscopy using Sheather’s sucrose floatation. PCR assays were performed on each faecal sample, targeting a B1 gene and a repetitive element (REP) gene to confirm T. gondii oocysts. Additionally, the PCR amplicons from the REP gene were sequenced to further confirm T. gondii-positive samples for phylogenetic analysis. Microscopy detected 7/200 (3.5%) T. gondii-like oocysts, while both the B1 gene and the REP gene detected 17/200 (8.5%) samples positive for T. gondii. All samples that were microscopically positive for T. gondii-like oocysts were also shown to be positive by both B1 and REP genes. The BLAST results sequenced for 16/200 (8.0%) PCR-positive T. gondii samples revealed homology and genetic heterogeneity with T. gondii strains in the GenBank, except for only one positive sample that did not show a result. There was almost perfect agreement (k = 0.145) between the two PCR assays targeting the B1 gene and the REP gene. This is the first report on microscopic, molecular detection and genetic diversity of T. gondii from cat faecal samples in Malaysia. In addition, the sensitivities of either the B1 gene or REP gene multicopy-target PCR assays are suitable for the accurate detection of T. gondii from cat faeces.

Toxoplasmosis is a disease caused by the protozoan parasite Toxoplasma gondii (T. gondii). Human toxoplasmosis seroprevalence in Malaysia has increased since it was first reported in 1973 as shown in previous reviews of 1991 and 2007. However, over a decade since the last review, comprehensive data on toxoplasmosis in Malaysia is lacking. This work aimed at reviewing articles on toxoplasmosis research in Malaysia in order to identify the research gaps, create public awareness, and efforts made so far and proffer management options on the disease. The present review examines the available published research articles from 2008 to 2018 related to toxoplasmosis research conducted in Malaysia. The articles reviewed were retrieved from nine credible databases such as Web of Science, Google Scholar, ScienceDirect, PubMed, Scopus, Springer, Wiley online library, Ovid, and Cochrane using the keywords; Malaysia, toxoplasmosis, Toxoplasma gondii, toxoplasma encephalitis, seroprevalence, human immunodeficiency virus (HIV) patients, pregnant women, genotype strain, anti-toxoplasma antibodies, felines, and vaccine. The data highlighted seropositive cases from healthy community members in Pangkor Island (59.7%) and among migrant workers (57.4%) at alarming rates, as well as 42.5% in pregnant women. Data on animal seroprevalence were limited and there was no information on cats as the definitive host. Genetic characterization of Toxoplasma gondii from HIV patients; pregnant women, and domestic cats is lacking. This present review on toxoplasmosis is beneficial to researchers, health workers, animal health professionals, and policymakers. Therefore, attention is required to educate and enlighten health workers and the general public about the risk factors associated with T. gondii infection in Malaysia.

Identification of drug target in protozoan T. gondii is an important step in the development of chemotherapeutic agents. Likewise, exploring phytochemical compounds effective against the parasite can lead to the development of new drug agent that can be useful for prophylaxis and treatment of toxoplasmosis. In this review, we searched for the relevant literature on the herbs that were tested against T. gondii either in vitro or in vivo, as well as different phytochemicals and their potential activities on T. gondii. Potential activities of major phytochemicals, such as alkaloid, flavonoid, terpenoids and tannins on various target sites on T. gondii as well as other related parasites was discussed. It is believed that the phytochemicals from natural sources are potential drug candidates for the treatment of toxoplasmosis with little or no toxicity to humans.

Background. Toxoplasmosis remains widely distributed globally and is one of the major neglected parasitic zoonotic infections. The infection is still endemic in most parts of the world due to poor control as well as challenges of the currently used medications which can be overcome by using natural products. This study evaluated the effect of ethanolic extract from the stem of Tinospora crispa (EETC) on host cell invasion and intracellular replication of Toxoplasma gondii. Method. The stem powder of T. crispa was soaked in absolute ethanol for 72 hours. The resulting ethanolic extract was screened for the presence of phytochemicals. Vero cells monolayer in 96-well plate was infected with RH strain of T. gondii and treated with concentrations of the EETC, Veratrine alkaloid, and clindamycin ranging from 1.56 to 200 μg/mL. MTT assay was conducted after 24 hours to evaluate the cytotoxicity and antiparasitic activities of the EETC. Four and 24 hours treatment models were adapted to assess the infection index and intracellular proliferation of T. gondii in vitro. Microscopic analysis was conducted after 24 and 48 hours exposure to EETC in both treatment models. Results. The study revealed that the EETC had no cytotoxic effects on Vero cells with IC50 = 179 μg/mL, as compared to clindamycin (IC50 = 116.5 μg/mL) and Veratrine alkaloid (IC50 = 60.4 μg/mL). The EETC had good anti-toxoplasma activities with IC50 = 6.31 μg/mL in comparison with clindamycin (IC50 = 8.33 μg/mL) and Veratrine alkaloid (IC50 = 14.25 μg/mL). The EETC caused more than 70% and 80% reduction in infection index and intracellular proliferation in both treatment models, respectively. Conclusion. This in vitro study showed that the EETC contains promising phytochemicals effective against T. gondii and safe to the host cells.

The emergence of drug-resistant strains of Plasmodium falciparum is the worst catastrophe that has ever confronted the dedicated efforts to eradicate malaria. This urged for searching other alternatives or sensitizers that reverse chloroquine resistance. In this experiment, the potential of andrographolide to inhibit plasmodial growth and reverse CQ resistance was tested in vitro using the SYBRE green-1-based drug sensitivity assay and isobologram technique, respectively. Its safety level toward mammalian cells was screened as well against Vero cells and RBCs using MTT-based drug sensitivity and RBC hemolysis assays, respectively. Its effect against hemozoin formation was screened using β-hematin formation and heme fractionation assays. Its molecular characters were determined using the conventional tests for the antioxidant effect measurement and the in silico molecular characterization using the online free chemi-informatic Molinspiration software. Results showed that andrographolide has a moderate antiplasmodium effect that does not entitle it to be a substituent for chloroquine. Furthermore, andrographolide ameliorated the sensitivity of the parasite to chloroquine. Besides, it showed an indirect inhibitory effect against hemozoin formation within the parasite and augmented the chloroquine-induced inhibition of hemozoin formation. The study suggests that its chloroquine resistance reversal effect may be due to inhibition of chloroquine accumulation or due to its impact on the biological activity of the parasite. Overall, this in vitro study is a clue for the reliability of andrographolide to be added with chloroquine for reversal of chloroquine resistance and tolerance, but further in vivo studies are recommended to confirm this notion. In spite of its prominent and safe in vitro and in vivo growth inhibitory effect and its in vitro chloroquine resistance reversing effect, it is inapplicable to implement it in malaria chemotherapy to substitute chloroquine or to reverse its resistance.

The recently identified cytokines-interleukin (IL)-35 and interleukin (IL)-37-have been described for their anti-inflammatory and immune-modulating actions in numerous inflammatory diseases, auto-immune disorders, malignancies, infectious diseases and sepsis. Either cytokine has been reported to be reduced and in some cases elevated and consequently contributed towards disease pathogenesis. In view of the recent advances in utilizing cytokine profiles for the development of biological macromolecules, beneficial in the management of certain intractable immune-mediated disorders, these recently characterized cytokines (IL-35 and IL-37) offer potential as reasonable targets for the discovery of novel immune-modulating anti-inflammatory therapies. A detailed comprehension of their sophisticated regulatory mechanisms and patterns of expression may provide unique opportunities for clinical application as highly selective and target specific therapeutic agents. This review seeks to summarize the recent advancements in discerning the dynamics, mechanisms, immunoregulatory and anti-inflammatory actions of IL-35 and IL-37 as they relate to disease pathogenesis.

Interleukin-33 (IL-33) is an IL-1 family member, which exhibits both pro- and anti-inflammatory properties solely based on the type of the disease itself. Generally, IL-33 is expressed by both endothelial and epithelial cells and mediates its function based on the interaction with various receptors, mainly with ST2 variants. IL-33 is a potent inducer for the Th2 immune response which includes defence mechanism in brain diseases. Thus, in this paper, we review the biological features of IL-33 and the critical roles of IL-33/ST2 pathway in selected neurological disorders including Alzheimer’s disease, multiple sclerosis, and malaria infection to discuss the involvement of IL-33/ST2 pathway during these brain diseases and its potential as future immunotherapeutic agents or for intervention purposes.

Monoclonal antibodies (mAbs) have proven to be effective biological reagents in the immunodiagnostic assays. This is due to their binding accuracy to many pathogens, thus, making them valuable research tools. Since the discovery of hybridoma technology by Kohler and Milstein, the use of monoclonal antibodies produced by the hybrid cells have been employed in diagnosis of several diseases. Monoclonal antibody production has several procedures with considerable variations, but the principles remain the same. Improvements in the field of cell culture technology have led to the production of improved qualities of monoclonal antibodies. In general, these antibodies are important biomedical reagents used in research, especially in the field of laboratory diagnostics for a number of different types of diseases in humans and animals. Some of the areas where application of monoclonal antibodies triumph are herein discussed. This review is aimed to assess various diagnostic assay techniques where monoclonal antibodies are applied in order to provide a first-hand information, especially for beginners in monoclonal antibody production, characterisation, evaluation and /or its applications in research and diagnosis.Keywords: Monoclonal antibodies, immunodiagnostic, assays, recent applications

Many modern-day diagnostic tests for parasitic diseases rely on conventional labour-intensive technologies such as serology and microscopy. Although major advances have been recorded in the diagnosis of infectious diseases in humans, parasitic diseases continue to present challenges, particularly in resource-poor countries, and this is mainly attributable to war and famine. Factors such as poverty, deteriorated health facilities and destruction of infrastructure are the consequence of the lack of suitable sanitary practices and proper hygiene, especially in refugee camps, that adversely promote infectious diseases to migrants, particularly among vulnerable children. Generally, the gastrointestinal tract is the predilection site for most helminths and protozoa. They are therefore regarded as a serious public-health problem, as they cause malabsorption, malnutrition and blood loss, leading to anaemia or even death. In addition to their health effects, parasitic infections cause physical and mental impairment in children, retard their educational achievements and hinder economic development. Keywords: Prevalence, Parasitic diseases, Intestinal parasite, Food-borne, Water-borne, Pathogens, Post-war, Iraq

The increasing demand for monoclonal antibodies (mAbs) used for diagnostic and therapeutic applications has led to the development of large scale manufacturing processes, with improvements in production achieved through continuous optimization of the inherent systems. The number of monoclonal antibodies (mAbs) that have already been approved for therapeutic applications and for use in clinical trials have significantly increased in the past few years. In view of the side effects and limitations of mAbs, several improvements and modifications to monoclonal antibodies have been developed. These modifications have facilitated the use of mAbs in various forms of therapeutic applications such as treatment of infectious diseases caused by bacterial, viral, fungal and parasitic organisms. Monoclonal antibodies have also been applied in the treatment of non-infectious diseases such as cancer, immune diseases, arthritis and other disorders resulting from organ transplantation. This review highlights mAbs applications in biomedicine, and discusses state-of-the-art technologies related to their potential uses. Keywords: Monoclonal antibodies, Therapeutic application, Infectious diseases, Cancer, Auto-immune diseases, Metabolic disorders

Background: Cytochrome b mutations confer atovaquone resistance, resulting in antimalarial drug failures. Results: Mutation Y268S reduces bc1 catalytic turnover and stability. Conclusion: Reduction of catalytic turnover and iron-sulfur protein content in parasite Y268S bc1 confers a fitness cost. These results were not predicted using yeast models. Significance: Data will aid novel bc1 inhibitor design and inform epidemiological studies of atovaquone resistance., Atovaquone is an anti-malarial drug used in combination with proguanil (e.g. MalaroneTM) for the curative and prophylactic treatment of malaria. Atovaquone, a 2-hydroxynaphthoquinone, is a competitive inhibitor of the quinol oxidation (Qo) site of the mitochondrial cytochrome bc1 complex. Inhibition of this enzyme results in the collapse of the mitochondrial membrane potential, disruption of pyrimidine biosynthesis, and subsequent parasite death. Resistance to atovaquone in the field is associated with point mutations in the Qo pocket of cytochrome b, most notably near the conserved Pro260-Glu261-Trp262-Tyr263 (PEWY) region in the ef loop). The effect of this mutation has been extensively studied in model organisms but hitherto not in the parasite itself. Here, we have performed a molecular and biochemical characterization of an atovaquone-resistant field isolate, TM902CB. Molecular analysis of this strain reveals the presence of the Y268S mutation in cytochrome b. The Y268S mutation is shown to confer a 270-fold shift of the inhibitory constant (Ki) for atovaquone with a concomitant reduction in the Vmax of the bc1 complex of ∼40% and a 3-fold increase in the observed Km for decylubiquinol. Western blotting analyses reveal a reduced iron-sulfur protein content in Y268S bc1 suggestive of a weakened interaction between this subunit and cytochrome b. Gene expression analysis of the TM902CB strain reveals higher levels of expression, compared with the 3D7 (atovaquone-sensitive) control strain in bc1 and cytochrome c oxidase genes. It is hypothesized that the observed differential expression of these and other key genes offsets the fitness cost resulting from reduced bc1 activity.