In silico characterisation of a mitotic kinesin-related protein from Leishmania donovani KE16 Suad Gazi AL Kufi, Amjed Qays Ibrahim Alqaisi, Mohammad Mahmoud Farhan Al- Halbosiy, Ikhlass Ali Hussain AlHilaly Asia Pacific Journal of Molecular Biology and Biotechnology, 2025 Mitotic kinesin proteins have a significant role in eukaryotic cell mitosis. The kinesin motor domain of Leishmania donovani has been explored as a promising vaccine candidate due to its ability to generate strong immune responses. The current study aimed to fill the knowledge gap and to systematically identify the main characteristic of kinesin related protein in L. donavani strain KE16 (LdonKE16) (AAT40474.1) using TriTrypDB (kinetoplast genomics), NCBI and EMBL database tools. The study was carried out using in silico PCR analysis of LdonKE16 as a potential of mitotic kinesin protein. Amino acid sequence of the kinesin motor domain regions was detected by using BLASTp for AAT40474.1 and showed various identities with other Leishmania spp. The highest sequence identity with L. donovani K39 (98.10%), then with L. infantum (97.09%), and (97%) with L. chagasi, and L. braziliensis respectively. While L. mexicana showed low sequence identity (61%). The conserved domains KISc of this protein contains three typical domains, the N-terminal motor domain (11 to 399 aa), the coiled segment neck domain (429 to 622aa) and the tail region (681 to 858 aa). The current study revealed that the predicted functional structure of LdonKE16 has active state and inactive state by using bioinformatics tool I-TASSER. The PCR product confirmed the identification of the kinesin protein in LdonKE16. Sequence analysis demonstrated strong homology between LdonKE16 and kinesin K39 of visceral Leishmania species, aligning with the kinesin superfamily KIF13 found in parasites and humans. These findings suggest that LdonKE16 is a promising candidate for vaccine development and therapeutic interventions against leishmaniasis.
Assessing the Potentiality of Using CXCL9 as A Predictive Biomarker for Acute and Chronic Toxoplasmosis, and Study the Correlation Between CXCL9, Toxoplasmosis and Thyroid Disorder in These Cases Mariam Ali Najeeb, Amjed Qays Ibrahim Al-Qaisi Iraqi Journal of Science, 2023 Background: Chemokine (C-X-C motif) ligand (CXCL9) has an important role recruiting the T-lymphocytes and immune response after infection by inducing T-cells accumulation around the areas associated with infections. However, this role is poorly known in relation with Toxoplasma gondii infection and also in association with thyroid hormones, which the present study is focused on. Methods: Eighty-seven women were included in this study for the period between September 2021 and February 2022. Blood samples of uninfected healthy pregnant, in addition to aborted and pregnant women infected with toxoplasmosis, were collected. Sera were then obtained and stored at -10°C. Toxo-latex agglutination test was done, followed by detection of IgM and IgG antibodies, which were identified in sera of cases and controls using a commercially available enzyme-linked immunoassay. Finally, an ELISA test for CXCL9 and thyroid hormones tests were performed as well. Results: CXCL9 levels showed a non-significant increase in comparison with the control group. This increase was observed during the first 5 months of abortion and pregnancy. Thyroid hormones T3 and TSH were significantly higher in the aborted and pregnant women than the control group. Whereas T4 results revealed lower levels in the same group in comparison to the control group, specifically during the first 5 months of abortion and pregnancy for most of the groups. Conclusion: Determining CXCL9 chemokine levels in female patients infected with Toxoplasma gondii might play an important role in early diagnosis of Toxoplasmosis, especially during the first 5 months of abortion or pregnancy as well as thyroid hormones levels can be influenced or affected by the parasite infection, which can provide an indicator of thyroid dysfunction in general.
Enterobius vermicularis infections in Iraq H. S. Al-Warid, A. Q. I. Alqaisi, I. M. Al Saqur, H. S. Al-Bahadely Helminthologia Poland, 2022 Summary Enterobiasis continues to be among the highest parasitic infections affecting the human population worldwide. A study was conducted between 2011 – 2015 in Iraq to evaluate the enterobiasis reported by the Communicable Diseases Control Center (n=220,607 cases) in relation to demographic (age, sex, rural population and family size) and spatial variables (local and regional sources). Females were more parasitized than males, as well as children and youth ages 4 to 15. Approximately 40 % of cases are from the South region provinces (Thiqar, Miasan, Basrah and Wassit). However, most cases occurred in regions with high rural populations and a high family size average. The results may provide insights for researchers assessing management approaches to control enterobiasis in Iraq.
Laminin Is a Promising Predictive Biomarker for Acute and Chronic Toxoplasmosis Mariam Ali Najeeb, Amjed Qays Ibrahim Alqaisi Egyptian Journal of Hospital Medicine, 2022 Background: Laminin (LN) is an important extracellular matrix glycoprotein plays an important role in early embryonic development by promoting the cell adhesion and angiogenesis. It regulates many functions in the cell including proliferation, invasion, and signaling. However; the role of laminin is poorly known in relation with Toxoplasma gondii infection. Objective: The current study aimed to examine the possibility of using the laminin as an indicator of an early infection of toxoplasmosis. Methods: Eighty-seven women aged from 15-45 years. They were included in this study from September 2021 to February 2022. Blood samples were collected from healthy pregnant women and aborted and pregnant women infected with toxoplasmosis. Toxo-latex agglutination test was done followed by the detection of IgM and IgG antibodies, which were determined in sera from cases and controls using a commercially available enzyme-linked immunoassay. Finally, an ELISA test for laminin was performed as well. Results: The seroprevalence of anti- T. gondii antibodies IgM or IgG were 14.94% (13/87) and 62.07% (54/87) respectively. Levels of laminin showed a significant decrease in the serum of 1-5 months aborted women with acute and chronic Toxoplasma Gondii infection in comparison with the control. Conclusion: The levels of laminin in female patients infected with Toxoplasma G ondii might play an important role in early diagnosis of toxoplasmosis especially during the first 5 months of abortion or pregnancy because laminin is an important glycoprotein in the extracellular matrix component, which involved in embryogenesis, implantation, and placentation.
Evaluating the in vitro anti-Leishmanial activity of essential oil extracted from Cymbopogon Citratus against Leishmania Donovani Mahmmoud Shakir Abdullah, Amjed Qays Ibrahim Alqaisi Iraqi Journal of Science, 2022 Leishmania is one of the protozoan parasites that are transferred to human by infected sand flies and gives rise to a range of diseases entitled as Leishmaniasis. More than 20 known species of Leishmania can infect humans and cause various clinical symptoms. Three most known clinical manifestations are Cutaneous Leishmaniasis (CL), Mucocutaneous Leishmaniasis (MCL) and Visceral Leishmaniasis (VL) (kala-azar or black fever). The difference in the clinical form dependent on several factors: species of Leishmania, type of vector that transmits the Leishmania, and the immune status of an infected individual. The current drugs which are used as anti-leishaminial treatment are characterized by enormous side effects, including their toxicity to human, long term treatment, liver problems and huge cost. Therefore, there is a necessity to find an alternative treatment marked as low cost, more specific against parasite’s stages, and metabolic pathways, and non-toxicity to human. Plants are considered one of the important sources for the remedy of the tropical diseases caused by bacteria, viruses, and parasites. Cymbopogon Citratus (lemon grass) is a herbal medicine used as anti-inflammatory, antibacterial, antifungal, anti-malarial, anti-protozoal and for gastrointestinal problems remedy. In order to detect the effects of C. Citratus against Leishmaniasis, in this study, serial dilution for the essential oil of C.citratus (1000, 500, 250, 125, 62.5 and 31.25) µg/ml were used against L. donovani Promastigotes. Viability was also evaluated at 24, 48 and 72 hours post-treatment. The results revealed that high concentrations (1000, 500 and 250) µg/ml were more effective than other concentrations during all time intervals, and IC50 values were 640, 492 and 442 µg/ml at 24, 48 and 72 hours respectively. In conclusion, this current study is one of a persistent search to find new treatments characterized by its high activity and low adverse effects to treat protozoa parasites for instance, Leishmania, and displays the effectiveness of the essential oils as a promising alternative
Antileishmanial Chemotherapy through Clemastine Fumarate Mediated Inhibition of the Leishmania Inositol Phosphorylceramide Synthase John G. M. Mina, Rebecca L. Charlton, Edubiel Alpizar-Sosa, Douglas O. Escrivani, Christopher Brown, Amjed Alqaisi, Maria Paula G. Borsodi, Claudia P. Figueiredo, Emanuelle V. de Lima, Emily A. Dickie, Wenbin Wei, Robson Coutinho-Silva, Andy Merritt, Terry K. Smith, Michael P. Barrett, Bartira Rossi-Bergmann, Paul W. Denny, Patrick G. Steel ACS Infectious Diseases, 2021 Current chemotherapeutics for leishmaniasis have multiple deficiencies, and there is a need for new safe, efficacious, and affordable medicines. This study describes a successful drug repurposing approach that identifies the over-the-counter antihistamine, clemastine fumarate, as a potential antileishmanial drug candidate. The screening for inhibitors of the sphingolipid synthase (inositol phosphorylceramide synthase, IPCS) afforded, following secondary screening against Leishmania major (Lmj) promastigotes, 16 active compounds. Further refinement through the dose response against LmjIPCS and intramacrophage L. major amastigotes identified clemastine fumarate with good activity and selectivity with respect to the host macrophage. On target engagement was supported by diminished sensitivity in a sphingolipid-deficient L. major mutant (ΔLmjLCB2) and altered phospholipid and sphingolipid profiles upon treatment with clemastine fumarate. The drug also induced an enhanced host cell response to infection indicative of polypharmacology. The activity was sustained across a panel of Old and New World Leishmania species, displaying an in vivo activity equivalent to the currently used drug, glucantime, in a mouse model of L. amazonensis infection. Overall, these data validate IPCS as an antileishmanial drug target and indicate that clemastine fumarate is a candidate for repurposing for the treatment of leishmaniasis.
Molecular characterization of Contracaecum rudolphii Hartwich, 1964 (Nematoda: Anisakidae) from the cormorant phalacrocorax carbo in Iraq Amjed Qays Alqaisi, , Harith Saeed Al-Warid, Azhar A. Al-Moussawi, , and Bulletin of the Iraq Natural History Museum, 2020 Contracaecum rudolphii Hartwich, 1964 is a nematode which causes major concerns to human and wildlife animal’s health. However, the population genetics of C. rudolphii has been poorly studied in Iraq. In order to gain a deeper understanding in the outline of the genetic diversity of the nematode C. rudolphii that were isolated from its host cormorant Phalacrocorax carbo (Linnaeus, 1758), in the middle areas of Iraq, twenty specimens of C. rudolphii adults were isolated from nine individuals of P. carbo. The first (ITS-1) internal transcribed spacers (ITS) of ribosomal DNA (rDNA) of C. rudolphii were amplified using conventional polymerase chain reaction (PCR); then, the amplicons were subjected to sequencing. Concatenation of ITS-1 (rDNA) sequences resulted in four unique genotypes that have not been previously recorded in Iraq. The present study showed that the most common genotype occurred in 85% of C. rudolphii, and in 88.9% of cormorants. Furthermore, the infrapopulation difference in the genotypes was fairly high, with an average of 1.3 ± 0.48 genotypes per host of those with ≥two nematodes. All the sequences of the current study were distributed into two different populations. The sequences of ITS-1 for the first population had the highest similarity to ITS-1 sequence of C. rudolphii B, while the sequences of ITS-1 for the second population had the highest similarity to ITS-1 sequence of C. rudolphii A. This study provides an insight about the genetic divergence of C. rudolphii among P. carbo in Iraq. As well, the results likely support the hypothesis that C. rudolphii represents a complex of at least two sibling species.
Prevalence of gastrointestinal parasites in Iraq during 2015 Ihsan Mahdi Al-Saqur, Harith Saeed Al-Warid, Amjed Qays Al-Qaisi, Hussin Salman Al-Bahadely Aip Conference Proceedings, 2020 Infections with gastro-intestinal parasites are associated with poor sanitation, unsafe human waste disposal, inadequate of safe drinking water, low socioeconomic status, demographic factors and spatial. The aims of this survey were to identify possible associations of nine gastro-intestinal parasites with patient demographics (age,gender) and spatial localities (provincial sources). This study is retrospective, including reported cases of infections using an available surveillance database from January 2015 to December 2015 of all provinces of Iraq by the Ministry of Health. Entamoeba histolytica/dispar, Enterobius vermicularis and Giardia lamblia are found to be the predominant parasites. Generally, males showed higher risk for infections than females, and the majority of cases were recorded among 5-14- and 15-45-year age groups. Approximately 79% of cases derived from Baghdad, Basra, Thi-qar, Najaf, Diyala, Babil and Qadisiyah each showed total cases ˂ 10000. These findings may provide simple insights for public health specialist assessing management approaches for the control of gastro-intestinal parasites in Iraq especially for the top three parasites.
Complex interplay between sphingolipid and sterol metabolism revealed by perturbations to the leishmania metabolome caused by miltefosine Emily G. Armitage, Amjed Q. I. Alqaisi, Joanna Godzien, Imanol Peña, Alison J. Mbekeani, Vanesa Alonso-Herranz, Ángeles López-Gonzálvez, Julio Martín, Raquel Gabarro, Paul W. Denny, Michael P. Barrett, Coral Barbas Antimicrobial Agents and Chemotherapy, 2018 With the World Health Organization reporting over 30,000 deaths and 200,000 to 400,000 new cases annually, visceral leishmaniasis is a serious disease affecting some of the world's poorest people. As drug resistance continues to rise, there is a huge unmet need to improve treatment. Miltefosine remains one of the main treatments for leishmaniasis, yet its mode of action (MoA) is still unknown. Understanding the MoA of this drug and parasite response to treatment could help pave the way for new and more successful treatments for leishmaniasis. A novel method has been devised to study the metabolome and lipidome ofLeishmania donovaniaxenic amastigotes treated with miltefosine. Miltefosine caused a dramatic decrease in many membrane phospholipids (PLs), in addition to amino acid pools, while sphingolipids (SLs) and sterols increased.Leishmania majorpromastigotes devoid of SL biosynthesis through loss of the serine palmitoyl transferase gene (ΔLCB2) were 3-fold less sensitive to miltefosine than wild-type (WT) parasites. Changes in the metabolome and lipidome of miltefosine-treatedL. majormirrored those ofL. donovani. A lack of SLs in the ΔLCB2 mutant was matched by substantial alterations in sterol content. Together, these data indicate that SLs and ergosterol are important for miltefosine sensitivity and, perhaps, MoA.
The antifungal Aureobasidin A and an analogue are active against the protozoan parasite Toxoplasma gondii but do not inhibit sphingolipid biosynthesis A. Q. I. ALQAISI, A. J. MBEKEANI, M. BASSAS LLORENS, A. P. ELHAMMER, P. W. DENNY Parasitology, 2018 SUMMARYToxoplasma gondiiis an obligate intracellular protozoan parasite of the phylum Apicomplexa, and toxoplasmosis is an important disease of both humans and economically important animals. With a limited array of drugs available there is a need to identify new therapeutic compounds. Aureobasidin A (AbA) is an antifungal that targets the essential inositol phosphorylceramide (IPC, sphingolipid) synthase in pathogenic fungi. This natural cyclic depsipeptide also inhibitsToxoplasmaproliforation, with the protozoan IPC synthase orthologue proposed as the target. The data presented here show that neither AbA nor an analogue (Compound 20), target the protozoan IPC synthase orthologue or total parasite sphingolipid synthesis. However, further analyses confirm that AbA exhibits significant activity against the proliferative tachyzoite form ofToxoplasma, and Compound 20, whilst effective, has reduced efficacy. This difference was more evident on analyses of the direct effect of these compounds against isolatedToxoplasma, indicating that AbA is rapidly microbicidal. Importantly, the possibility of targeting the encysted, bradyzoite, form of the parasite with AbA and Compound 20 was demonstrated, indicating that this class of compounds may provide the basis for the first effective treatment for chronic toxoplasmosis.
