Amphids are sensory neurons that nematodes use to sense their environment. The IVR-10 strain is an ivermectin (IVM) resistant strain of Caenorhabditis elegans generated in the laboratory by repeated exposure to IVM. We found that the IVR-10 strain is dye filling defective which may be due to shortened amphids. The amphidial pore of the N2 Bristol strain lit up with an IVM antibody, providing direct immunolocalization of IVM and confirming early hypothesis based on functional studies. This suggests that IVM may enter the worms via the amphidial pore. The findings reiterate the importance of amphidial pore as a structure that is exposed to the chemical environment and may be a portal for drug entry.
{"title":"Ivermectin localization at the amphidial pore and dye-filling defects in the IVR-10 strain of Caenorhabditis elegans","authors":"Umer Chaudhry , Mohid Ashraf , Sidra Ashraf , Moneeb Ashraf , Sohaib Ashraf , Ali Raza , Shoaib Ashraf","doi":"10.1016/j.molbiopara.2026.111723","DOIUrl":"10.1016/j.molbiopara.2026.111723","url":null,"abstract":"<div><div>Amphids are sensory neurons that nematodes use to sense their environment. The IVR-10 strain is an ivermectin (IVM) resistant strain of <em>Caenorhabditis elegans</em> generated in the laboratory by repeated exposure to IVM. We found that the IVR-10 strain is dye filling defective which may be due to shortened amphids. The amphidial pore of the N2 Bristol strain lit up with an IVM antibody, providing direct immunolocalization of IVM and confirming early hypothesis based on functional studies. This suggests that IVM may enter the worms via the amphidial pore. The findings reiterate the importance of amphidial pore as a structure that is exposed to the chemical environment and may be a portal for drug entry.</div></div>","PeriodicalId":18721,"journal":{"name":"Molecular and biochemical parasitology","volume":"265 ","pages":"Article 111723"},"PeriodicalIF":1.5,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145977529","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-02-01Epub Date: 2025-12-08DOI: 10.1016/j.molbiopara.2025.111712
Ellen Gomes , Camila Rolemberg Santana Travaglini Berti de Correia , Caroline Torres , Mariele Cristina de Carvalho , Taissa de Oliveira de Castro , Wesley Klaysson Pereira Regatieri , Nayore Tamie Takamiya , Luana Aparecida Rogerio , Adriano Cappellazzo Coelho , Juliana Ide Aoki , Sandra Regina Maruyama , Felipe Roberti Teixeira
LinfCul1 is a key component of the E3 ubiquitin ligase complex (LinfCRL1) in Leishmania infantum, which interacts with LinfSkp1 and LinfRbx1 at the N- and C-termini, respectively. To investigate the role of LinfCul1 in parasite proliferation, rosette formation, and macrophage infection, we generated a mutant LinfCul1 (mLinfCUL1) lacking the LinfSkp1 interaction region. Co-immunoprecipitation assays confirmed that mLinfCul1 exhibited reduced interaction with LinfSkp1, thereby disrupting LinfCRL1 function. Functional assays demonstrated that LinfCUL1 knockout (∆cul1) led to impaired proliferation and enhanced rosette formation, both of which were rescued by LinfCUL1 WT but not by mLinfCUL1 expression, confirming the requirement of the LinfCul1-LinfSkp1 interaction for these processes. Additionally, macrophage infection assays revealed that ∆cul1 parasites exhibited reduced infectivity and amastigote proliferation, which was restored upon LinfCUL1 WT expression in the parasites. Interestingly, mLinfCUL1 exhibited a lower infectivity index than ∆cul1, suggesting that LinfCul1 functions as a LinfCRL1 component that contributes to this process. These findings highlight the essential role of LinfCul1 in parasite proliferation and infectivity and reinforce its canonical function in ubiquitination-dependent parasite biology. Moreover, this study provides valuable insights into the molecular mechanisms governing parasite development and host interactions, thereby contributing to a better understanding of Leishmania infantum biology.
{"title":"LinfCul1 interaction with LinfSkp1 affects different cellular processes in Leishmania infantum","authors":"Ellen Gomes , Camila Rolemberg Santana Travaglini Berti de Correia , Caroline Torres , Mariele Cristina de Carvalho , Taissa de Oliveira de Castro , Wesley Klaysson Pereira Regatieri , Nayore Tamie Takamiya , Luana Aparecida Rogerio , Adriano Cappellazzo Coelho , Juliana Ide Aoki , Sandra Regina Maruyama , Felipe Roberti Teixeira","doi":"10.1016/j.molbiopara.2025.111712","DOIUrl":"10.1016/j.molbiopara.2025.111712","url":null,"abstract":"<div><div>LinfCul1 is a key component of the E3 ubiquitin ligase complex (LinfCRL1) in <em>Leishmania infantum</em>, which interacts with LinfSkp1 and LinfRbx1 at the N- and C-termini, respectively. To investigate the role of LinfCul1 in parasite proliferation, rosette formation, and macrophage infection, we generated a mutant LinfCul1 (<em>mLinfCUL1</em>) lacking the LinfSkp1 interaction region. Co-immunoprecipitation assays confirmed that mLinfCul1 exhibited reduced interaction with LinfSkp1, thereby disrupting LinfCRL1 function. Functional assays demonstrated that <em>LinfCUL1</em> knockout (∆<em>cul1</em>) led to impaired proliferation and enhanced rosette formation, both of which were rescued by <em>LinfCUL1</em> WT but not by <em>mLinfCUL1</em> expression, confirming the requirement of the LinfCul1-LinfSkp1 interaction for these processes. Additionally, macrophage infection assays revealed that ∆cul1 parasites exhibited reduced infectivity and amastigote proliferation, which was restored upon <em>LinfCUL1</em> WT expression in the parasites. Interestingly, <em>mLinfCUL1</em> exhibited a lower infectivity index than ∆<em>cul1</em>, suggesting that LinfCul1 functions as a LinfCRL1 component that contributes to this process. These findings highlight the essential role of LinfCul1 in parasite proliferation and infectivity and reinforce its canonical function in ubiquitination-dependent parasite biology. Moreover, this study provides valuable insights into the molecular mechanisms governing parasite development and host interactions, thereby contributing to a better understanding of <em>Leishmania infantum</em> biology.</div></div>","PeriodicalId":18721,"journal":{"name":"Molecular and biochemical parasitology","volume":"265 ","pages":"Article 111712"},"PeriodicalIF":1.5,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145724213","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-02-01Epub Date: 2025-12-20DOI: 10.1016/j.molbiopara.2025.111720
Iman F. Abou-El-Naga
Toxocara canis is unable to synthesize sufficient lipids de novo to meet its biological requirements and therefore depends on host-derived lipids for survival. The parasite expresses a diverse set of lipid transport proteins spanning all major classes, such as pseudocoelomic fluid lipoproteins (vitellogenins), nematode polyprotein antigens/allergens, intracellular carriers (fatty-acid binding proteins, phosphatidylinositol-transfer proteins), secreted lipid-binding proteins (fatty acid-and retinol-binding proteins, venom allergen-like proteins), membrane-associated transporters (Niemann-Pick C, ABC transporters, microsomal triglyceride transfer protein, bridge-like lipid-transfer proteins) and lipid-anchored carriers (phosphatidylethanolamine-binding proteins). These proteins mediate uptake and distribution of dietary and host lipids to drive parasite growth and reproduction, while simultaneously modulating host immune responses. Many of these transporters are released in the parasite’s excretory/secretory products and are found in extracellular vesicles, where they mediate host-parasite interactions and immunomodulation. These specialized lipid-acquisition strategies support parasite survival, drive immune evasion and pathogenesis, and highlight these proteins as candidates for novel diagnostics or therapeutic targets.
{"title":"Lipid transport proteins in Toxocara canis: Host lipid acquisition and immune modulation","authors":"Iman F. Abou-El-Naga","doi":"10.1016/j.molbiopara.2025.111720","DOIUrl":"10.1016/j.molbiopara.2025.111720","url":null,"abstract":"<div><div><em>Toxocara canis</em> is unable to synthesize sufficient lipids de novo to meet its biological requirements and therefore depends on host-derived lipids for survival. The parasite expresses a diverse set of lipid transport proteins spanning all major classes, such as pseudocoelomic fluid lipoproteins (vitellogenins), nematode polyprotein antigens/allergens, intracellular carriers (fatty-acid binding proteins, phosphatidylinositol-transfer proteins), secreted lipid-binding proteins (fatty acid-and retinol-binding proteins, venom allergen-like proteins), membrane-associated transporters (Niemann-Pick C, ABC transporters, microsomal triglyceride transfer protein, bridge-like lipid-transfer proteins) and lipid-anchored carriers (phosphatidylethanolamine-binding proteins). These proteins mediate uptake and distribution of dietary and host lipids to drive parasite growth and reproduction, while simultaneously modulating host immune responses. Many of these transporters are released in the parasite’s excretory/secretory products and are found in extracellular vesicles, where they mediate host-parasite interactions and immunomodulation. These specialized lipid-acquisition strategies support parasite survival, drive immune evasion and pathogenesis, and highlight these proteins as candidates for novel diagnostics or therapeutic targets.</div></div>","PeriodicalId":18721,"journal":{"name":"Molecular and biochemical parasitology","volume":"265 ","pages":"Article 111720"},"PeriodicalIF":1.5,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145810646","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Candida albicans poses a serious health threat, contributing to approximately 1.5 million deaths each year. Although azole drugs have been used to manage this pathogen, their effectiveness has been compromised by the emergence of drug-resistant strains. Therefore, silver nanoparticles (nano-Ag) and vitamin D₃ are being explored as complementary rather than direct antifungal agents. The present study aims to investigate the effects of microbially synthesized nano-Ag alone and with vitamin D3 against fluconazole-susceptible and fluconazole-resistant C. albicans. The broth microdilution method, checkerboard microdilution assay, hyphal formation inhibition, and gene expression analysis of key virulence genes were performed on C. albicans treated with microbially synthesized nano-Ag, either alone or combined with vitamin D3. Furthermore, the survival rate, haemocyte density, and microbial load in haemolymph were assessed in C. albicans-infected Galleria mellonella larvae after treatment with microbially synthesized nano-Ag alone and with vitamin D3. The results demonstrated that microbially synthesized nano-Ag exhibited synergistic and additive interactions with vitamin D3 against C. albicans. This study also revealed that the combination of microbially synthesized nano-Ag and vitamin D3 effectively inhibited hyphal formation and significantly downregulated the expression of SAP and HWP1 genes in C. albicans. In vivo experiments further demonstrated that this combined treatment enhanced larval survival, increased haemocyte density, and reduced microbial load in the haemolymph. Taken together, these findings underscore the potential of microbially synthesized nano-Ag with vitamin D3 as a promising synergistic treatment for C. albicans infections, particularly those resistant to fluconazole.
{"title":"Potential synergistic antifungal activity of microbially synthesized silver nanoparticles and vitamin D3 against Candida albicans: vitro and Galleria mellonella model studies","authors":"Zainab Saberi Moqaddam , Pouria Khodavandi , Alireza Khodavandi , Fahimeh Alizadeh","doi":"10.1016/j.molbiopara.2025.111710","DOIUrl":"10.1016/j.molbiopara.2025.111710","url":null,"abstract":"<div><div><em>Candida albicans</em> poses a serious health threat, contributing to approximately 1.5 million deaths each year. Although azole drugs have been used to manage this pathogen, their effectiveness has been compromised by the emergence of drug-resistant strains. Therefore, silver nanoparticles (nano-Ag) and vitamin D<sub>₃</sub> are being explored as complementary rather than direct antifungal agents. The present study aims to investigate the effects of microbially synthesized nano-Ag alone and with vitamin D<sub>3</sub> against fluconazole-susceptible and fluconazole-resistant <em>C. albicans</em>. The broth microdilution method, checkerboard microdilution assay, hyphal formation inhibition, and gene expression analysis of key virulence genes were performed on <em>C. albicans</em> treated with microbially synthesized nano-Ag, either alone or combined with vitamin D<sub>3</sub>. Furthermore, the survival rate, haemocyte density, and microbial load in haemolymph were assessed in <em>C. albicans</em>-infected <em>Galleria mellonella</em> larvae after treatment with microbially synthesized nano-Ag alone and with vitamin D<sub>3</sub>. The results demonstrated that microbially synthesized nano-Ag exhibited synergistic and additive interactions with vitamin D<sub>3</sub> against <em>C. albicans</em>. This study also revealed that the combination of microbially synthesized nano-Ag and vitamin D<sub>3</sub> effectively inhibited hyphal formation and significantly downregulated the expression of <em>SAP</em> and <em>HWP1</em> genes in <em>C. albicans</em>. <em>In vivo</em> experiments further demonstrated that this combined treatment enhanced larval survival, increased haemocyte density, and reduced microbial load in the haemolymph. Taken together, these findings underscore the potential of microbially synthesized nano-Ag with vitamin D<sub>3</sub> as a promising synergistic treatment for <em>C. albicans</em> infections, particularly those resistant to fluconazole.</div></div>","PeriodicalId":18721,"journal":{"name":"Molecular and biochemical parasitology","volume":"264 ","pages":"Article 111710"},"PeriodicalIF":1.5,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145416513","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-01Epub Date: 2025-11-26DOI: 10.1016/j.molbiopara.2025.111711
Raghda R. Qadir , Hamin J. Mohammed , Samir M. Hamad , Yousef Mirzaei , Mukhtar H. Ahmed
Background
Coccidiosis is a significant parasitic disease affecting poultry, resulting in substantial economic losses due to its impact on growth, increased mortality, and compromised bird health.
Aim
This study aimed to evaluate the in vitro anticoccidial effects of a novel green-synthesised ZnO-Ag-CuO nanocomposite, using Thymus vulgaris extract.
Methods
The nanocomposite was synthesised through an eco-friendly method employing T. vulgaris as a stabilising and reducing agent. Characterisation was performed using UV-Vis spectroscopy, FTIR, XRD, SEM, and EDX, confirming its high crystallinity, nanoscale size, and the successful integration of ZnO, Ag, and CuO phases. Anticoccidial activity was assessed via a sporulation inhibition assay against Eimeria spp. Oocysts isolated from broiler chickens.
Results
The nanocomposite significantly reduced oocyst sporulation and increased the proportion of damaged and unpopulated oocysts in a dose-dependent manner (0.1–1 mg/mL) (p < 0.0001). ZnO–Ag–CuO NCs showed a dose-dependent anticoccidial effect, reducing sporulated oocysts to 56.41 %, 33.63 % and 22.9 % at 0.1, 0.5 and 1.0 mg/mL (control 88.62 %; p < 0.0001). Unsporulated oocysts increased to 15.9–62.22 % (control 13.33 %), while damaged oocysts reached up to 14.82 % (control 0 %).
Conclusion
The green-synthesised ZnO-Ag-CuO nanocomposite demonstrated strong in vitro anticoccidial activity; however, further studies are needed to evaluate the nanocomposite’s potential toxicity, formulation, stability under biological conditions, safety before practical applications and potential environmental impact within a One Health framework.
Future plans
In vivo studies are recommended to validate the efficacy and safety of these approaches for large-scale applications.
{"title":"Green-synthesised ZnO-Ag-CuO nanocomposites from Thymus vulgaris and their in vitro anticoccidial activity","authors":"Raghda R. Qadir , Hamin J. Mohammed , Samir M. Hamad , Yousef Mirzaei , Mukhtar H. Ahmed","doi":"10.1016/j.molbiopara.2025.111711","DOIUrl":"10.1016/j.molbiopara.2025.111711","url":null,"abstract":"<div><h3>Background</h3><div>Coccidiosis is a significant parasitic disease affecting poultry, resulting in substantial economic losses due to its impact on growth, increased mortality, and compromised bird health.</div></div><div><h3>Aim</h3><div>This study aimed to evaluate the <em>in vitro</em> anticoccidial effects of a novel green-synthesised ZnO-Ag-CuO nanocomposite, using <em>Thymus vulgaris</em> extract.</div></div><div><h3>Methods</h3><div>The nanocomposite was synthesised through an eco-friendly method employing <em>T. vulgaris</em> as a stabilising and reducing agent. Characterisation was performed using UV-Vis spectroscopy, FTIR, XRD, SEM, and EDX, confirming its high crystallinity, nanoscale size, and the successful integration of ZnO, Ag, and CuO phases. Anticoccidial activity was assessed via a sporulation inhibition assay against <em>Eimeria</em> spp. Oocysts isolated from broiler chickens.</div></div><div><h3>Results</h3><div>The nanocomposite significantly reduced oocyst sporulation and increased the proportion of damaged and unpopulated oocysts in a dose-dependent manner (0.1–1 mg/mL) (p < 0.0001). ZnO–Ag–CuO NCs showed a dose-dependent anticoccidial effect, reducing sporulated oocysts to 56.41 %, 33.63 % and 22.9 % at 0.1, 0.5 and 1.0 mg/mL (control 88.62 %; p < 0.0001). Unsporulated oocysts increased to 15.9–62.22 % (control 13.33 %), while damaged oocysts reached up to 14.82 % (control 0 %).</div></div><div><h3>Conclusion</h3><div>The green-synthesised ZnO-Ag-CuO nanocomposite demonstrated strong <em>in vitro</em> anticoccidial activity; however, further studies are needed to evaluate the nanocomposite’s potential toxicity, formulation, stability under biological conditions, safety before practical applications and potential environmental impact within a One Health framework.</div></div><div><h3>Future plans</h3><div>In vivo studies are recommended to validate the efficacy and safety of these approaches for large-scale applications.</div></div>","PeriodicalId":18721,"journal":{"name":"Molecular and biochemical parasitology","volume":"264 ","pages":"Article 111711"},"PeriodicalIF":1.5,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145620183","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-01Epub Date: 2025-09-03DOI: 10.1016/j.molbiopara.2025.111699
Muhammad Adnan Sabir Mughal , Muhammad Kasib Khan , He Lan , Rao Zahid Abbas , Muhammad Imran , Zaheer Abbas , Muhammad Shahid Mehmood , Sultan Ali
Parasitic diseases caused by Leishmania spp. create considerable health concerns in animals, resulting in a considerable financial impact. They causes a complex infection in equines, affecting weight gain, skin, liver, and spleen. To date, there is a lack of reports on the occurrence of Leishmania in equines in Southern Punjab, Pakistan, highlighting the need for molecular epidemiological surveillance. The current study focused on determining the prevalence of Leishmania in the equine population from District Rahim Yar Khan, Southern Punjab, Pakistan, through amplification of mitochondrial (Cytochrome b) and nuclear (18S rRNA) genes of the parasite. For this purpose, a total of 384 equine - i.e. horses, mules, and donkeys - blood specimens, determined by calculation of the sample size formula, were obtained from District Rahim Yar Khan. The parasite was examined through the Microhematocrit method under the microscope. Leishmania was detected from the buffy coat layer after centrifugation of blood-filled microhematocrit tubes. To detect and characterize Leishmania spp.at the molecular level, DNA extraction from blood samples was carried out using standardized commercial kits, followed by PCR amplification. Information on potential risk factors was gathered through a structured questionnaire. The overall prevalence of Leishmania infection was observed to be 2.1 % via microscopy and 7.3 % and 8.8 % by amplification of the 18S rRNA and Cytochrome b genes using molecular methods. A significantly higher infection percentage was observed in female animals compared to males, and in older and underweight animals compared to younger and healthier ones. Additionally, the infection was non-significantly (P ≥ 0.05) more prevalent in gestating, non-dewormed, symptomatic, and poor body condition animals. Phylogenetic and sequence analyses confirmed that the identified gene sequences clustered within the Leishmania (Leishmania) infantum clade, consistent with strains reported in different animal hosts from various regions. In conclusion, the nuclear gene, i.e., 18S rRNA proved to be a more sensitive molecular marker for detecting Leishmania infection in equines compared to the mitochondrial gene, i.e., Cytochrome b.
{"title":"Decoding Leishmania in equines: A comparative analysis of molecular targets","authors":"Muhammad Adnan Sabir Mughal , Muhammad Kasib Khan , He Lan , Rao Zahid Abbas , Muhammad Imran , Zaheer Abbas , Muhammad Shahid Mehmood , Sultan Ali","doi":"10.1016/j.molbiopara.2025.111699","DOIUrl":"10.1016/j.molbiopara.2025.111699","url":null,"abstract":"<div><div>Parasitic diseases caused by <em>Leishmania</em> spp. create considerable health concerns in animals, resulting in a considerable financial impact. They causes a complex infection in equines, affecting weight gain, skin, liver, and spleen. To date, there is a lack of reports on the occurrence of <em>Leishmania</em> in equines in Southern Punjab, Pakistan, highlighting the need for molecular epidemiological surveillance. The current study focused on determining the prevalence of <em>Leishmania</em> in the equine population from District Rahim Yar Khan, Southern Punjab, Pakistan, through amplification of mitochondrial (<em>Cytochrome b</em>) and nuclear (<em>18S rRNA</em>) genes of the parasite. For this purpose, a total of 384 equine - i.e. horses, mules, and donkeys - blood specimens, determined by calculation of the sample size formula, were obtained from District Rahim Yar Khan. The parasite was examined through the Microhematocrit method under the microscope. <em>Leishmania</em> was detected from the buffy coat layer after centrifugation of blood-filled microhematocrit tubes. To detect and characterize <em>Leishmania</em> spp.at the molecular level, DNA extraction from blood samples was carried out using standardized commercial kits, followed by PCR amplification. Information on potential risk factors was gathered through a structured questionnaire. The overall prevalence of <em>Leishmania</em> infection was observed to be 2.1 % via microscopy and 7.3 % and 8.8 % by amplification of the <em>18S rRNA</em> and <em>Cytochrome b</em> genes using molecular methods. A significantly higher infection percentage was observed in female animals compared to males, and in older and underweight animals compared to younger and healthier ones. Additionally, the infection was non-significantly (P ≥ 0.05) more prevalent in gestating, non-dewormed, symptomatic, and poor body condition animals. Phylogenetic and sequence analyses confirmed that the identified gene sequences clustered within the <em>Leishmania</em> (<em>Leishmania</em>) <em>infantum</em> clade, consistent with strains reported in different animal hosts from various regions. In conclusion, the nuclear gene, i.e., <em>18S rRNA</em> proved to be a more sensitive molecular marker for detecting <em>Leishmania</em> infection in equines compared to the mitochondrial gene, i.e., <em>Cytochrome b</em>.</div></div>","PeriodicalId":18721,"journal":{"name":"Molecular and biochemical parasitology","volume":"264 ","pages":"Article 111699"},"PeriodicalIF":1.5,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145006396","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The emergence of multidrug resistance in Plasmodium falciparum poses a serious threat to antimalarial treatment, particularly with growing resistance to artemisinin-based combination therapies (ACTs) and partner drugs like piperaquine. Mutations in key proteins, such as PfCRT (P. falciparum chloroquine resistance transporter) and PfDHFR (P. falciparum dihydrofolate reductase), play a critical role in this resistance. Understanding these molecular mechanisms is essential for the development of effective antimalarial therapies. This study aimed to investigate the structural and functional impact of polymorphisms on drug-target interactions and resistance mechanisms in P. falciparum. Molecular docking and molecular dynamics (MD) simulations were performed to analyze interactions of the mutated PfCRT and PfDHFR proteins with nine antimalarial drugs, including piperaquine. The PfCRT-K76A piperaquine complex strong binding affinity (-9.5 kcal/mol) with moderate structural deviation (0.970 ± 0.202 nm) and greater solvent accessibility (246.01 ± 6.135 nm²), suggesting favourable binding conditions. The PfDHFR-N51I–piperaquine complex showed even stronger binding (-10.8 kcal/mol) but higher structural fluctuation (RMSD: 4.491 ± 1.462 nm) and increased compactness (1.861 ± 0.029 nm), which may reflect restricted ligand accommodation and possible resistance. Overall, the findings provide valuable insights into how PfCRT and PfDHFR mutations contribute to drug resistance and establish a foundation for designing more effective antimalarial strategies. Future research should integrate experimental validation and explore additional resistance-associated mutations to develop targeted therapies for combating multidrug-resistant P. falciparum.
{"title":"Computational investigation of mutations in PfCRT and PfDHFR proteins for emerging resistance of Plasmodium falciparum to antimalarial drugs","authors":"Sushruta Ghosh , Deepesh Joshi , Chandra Sekar Ponnusamy , Bhavani Sridharan , Mahesh Velusamy","doi":"10.1016/j.molbiopara.2025.111700","DOIUrl":"10.1016/j.molbiopara.2025.111700","url":null,"abstract":"<div><div>The emergence of multidrug resistance in <em>Plasmodium falciparum</em> poses a serious threat to antimalarial treatment, particularly with growing resistance to artemisinin-based combination therapies (ACTs) and partner drugs like piperaquine. Mutations in key proteins, such as PfCRT (<em>P. falciparum</em> chloroquine resistance transporter) and PfDHFR (<em>P. falciparum</em> dihydrofolate reductase), play a critical role in this resistance. Understanding these molecular mechanisms is essential for the development of effective antimalarial therapies. This study aimed to investigate the structural and functional impact of polymorphisms on drug-target interactions and resistance mechanisms in <em>P. falciparum</em>. Molecular docking and molecular dynamics (MD) simulations were performed to analyze interactions of the mutated PfCRT and PfDHFR proteins with nine antimalarial drugs, including piperaquine. The PfCRT-K76A piperaquine complex strong binding affinity (-9.5 kcal/mol) with moderate structural deviation (0.970 ± 0.202 nm) and greater solvent accessibility (246.01 ± 6.135 nm²), suggesting favourable binding conditions. The PfDHFR-N51I–piperaquine complex showed even stronger binding (-10.8 kcal/mol) but higher structural fluctuation (RMSD: 4.491 ± 1.462 nm) and increased compactness (1.861 ± 0.029 nm), which may reflect restricted ligand accommodation and possible resistance. Overall, the findings provide valuable insights into how PfCRT and PfDHFR mutations contribute to drug resistance and establish a foundation for designing more effective antimalarial strategies. Future research should integrate experimental validation and explore additional resistance-associated mutations to develop targeted therapies for combating multidrug-resistant <em>P. falciparum</em>.</div></div>","PeriodicalId":18721,"journal":{"name":"Molecular and biochemical parasitology","volume":"264 ","pages":"Article 111700"},"PeriodicalIF":1.5,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145011010","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-01Epub Date: 2025-10-24DOI: 10.1016/j.molbiopara.2025.111706
Sundas Afresham , Muhammad Kasib Khan , Muhammad Adnan Sabir Mughal , Muhammad Shahid Mehmood , Sultan Ali , Maryam Bashir , Zaheer Abbas , Abdullah Azeem , Waqar Ahmed , Muhammad Imran , Rao Zahid Abbas , Zia-ud-Din Sindhu , Muhammad Sohail Sajid
Parasitic infections present a significant health risk to the public, affecting millions of people, particularly in underdeveloped and developing countries. In developing countries, these infections are also responsible for causing significant economic challenges due to elevated healthcare expenditure. Accurate diagnosis and effective treatment methods are essentially required to combat this global issue. For decades, traditional diagnostic methods such as microscopy, serological testing, histopathology, and culturing have been used for the diagnosis of these parasitic infections. While these methods can be effective and helpful in many ways, they often consume a lot of time, require an elevated level of expertise, and have limited applications particularly in endemic regions having issues like poor infrastructure and limited access to healthcare facilities. This review aims to highlight the urgent need for a revolution to replace these conventional techniques with more affordable, quick, and field-adjustable tools such as rapid diagnostic tests (RDTs) and molecular methods and provides a comprehensive picture of advanced diagnostic tools used in the identification of parasites. With the advancements in science and technology, molecular methods such as Polymerase chain reaction, Next generation sequencing, and isothermal loop-mediated amplification have remarkably enhanced the sensitivity and accuracy of parasite detection and identification. The range of these diagnostic methods has further extended by advanced serological methods, imaging techniques, and immunological methods. Moreover, the innovations in nanotechnology, CRISPR-Cas methods, and multi-omics techniques for identification of parasite DNA, antigens, metabolites, and host responses are invaluable for diagnostic accuracy, comprehensive understanding of parasite biology, and for the discovery of new therapeutic targets and diagnostic biomarkers. However, further research and developments are required for an effective and long-lasting impact of these advancements.
{"title":"Recent advancements in the diagnosis of parasitic diseases","authors":"Sundas Afresham , Muhammad Kasib Khan , Muhammad Adnan Sabir Mughal , Muhammad Shahid Mehmood , Sultan Ali , Maryam Bashir , Zaheer Abbas , Abdullah Azeem , Waqar Ahmed , Muhammad Imran , Rao Zahid Abbas , Zia-ud-Din Sindhu , Muhammad Sohail Sajid","doi":"10.1016/j.molbiopara.2025.111706","DOIUrl":"10.1016/j.molbiopara.2025.111706","url":null,"abstract":"<div><div>Parasitic infections present a significant health risk to the public, affecting millions of people, particularly in underdeveloped and developing countries. In developing countries, these infections are also responsible for causing significant economic challenges due to elevated healthcare expenditure. Accurate diagnosis and effective treatment methods are essentially required to combat this global issue. For decades, traditional diagnostic methods such as microscopy, serological testing, histopathology, and culturing have been used for the diagnosis of these parasitic infections. While these methods can be effective and helpful in many ways, they often consume a lot of time, require an elevated level of expertise, and have limited applications particularly in endemic regions having issues like poor infrastructure and limited access to healthcare facilities. This review aims to highlight the urgent need for a revolution to replace these conventional techniques with more affordable, quick, and field-adjustable tools such as rapid diagnostic tests (RDTs) and molecular methods and provides a comprehensive picture of advanced diagnostic tools used in the identification of parasites. With the advancements in science and technology, molecular methods such as Polymerase chain reaction, Next generation sequencing, and isothermal loop-mediated amplification have remarkably enhanced the sensitivity and accuracy of parasite detection and identification. The range of these diagnostic methods has further extended by advanced serological methods, imaging techniques, and immunological methods. Moreover, the innovations in nanotechnology, CRISPR-Cas methods, and multi-omics techniques for identification of parasite DNA, antigens, metabolites, and host responses are invaluable for diagnostic accuracy, comprehensive understanding of parasite biology, and for the discovery of new therapeutic targets and diagnostic biomarkers. However, further research and developments are required for an effective and long-lasting impact of these advancements.</div></div>","PeriodicalId":18721,"journal":{"name":"Molecular and biochemical parasitology","volume":"264 ","pages":"Article 111706"},"PeriodicalIF":1.5,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145362934","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-01Epub Date: 2025-10-10DOI: 10.1016/j.molbiopara.2025.111703
Francois Korbmacher , Manuel Rauch , Sanketha Kenthirapalan , Taco W.A. Kooij , Alexander G. Maier , Kai Matuschewski
Plasmodium parasites encode a chloroquine resistance transporter (CRT), which is an integral membrane protein of the digestive vacuole and transports the antimalarial compound chloroquine out of this organelle. Here, we profiled the spatio-temporal expression of CRT during life cycle progression employing CRT-mCherry Plasmodium berghei parasites. We show that CRT is expressed during asexual blood stage growth and localizes to the hemozoin-containing digestive vacuole. The compartmentalized CRT-mCherry signal is also abundant in gametocytes and ookinetes, indicating that CRT continues to exert important functions in this digestive organelle up until mosquito midgut colonization. Expression is switched off during sporogony and early liver infection but CRT-mCherry is present again in mature liver stages, likely in preparation for blood infection. Together, visualization of the P. berghei digestive vacuole by endogenous tagging of PbCRT revealed expression of this transport protein and the presence of this cellular compartment beyond asexual propagation inside erythrocytes.
{"title":"Stage-dependent expression and vacuolar localization of Plasmodium berghei chloroquine resistance transporter (CRT)","authors":"Francois Korbmacher , Manuel Rauch , Sanketha Kenthirapalan , Taco W.A. Kooij , Alexander G. Maier , Kai Matuschewski","doi":"10.1016/j.molbiopara.2025.111703","DOIUrl":"10.1016/j.molbiopara.2025.111703","url":null,"abstract":"<div><div><em>Plasmodium</em> parasites encode a chloroquine resistance transporter (CRT), which is an integral membrane protein of the digestive vacuole and transports the antimalarial compound chloroquine out of this organelle. Here, we profiled the spatio-temporal expression of CRT during life cycle progression employing CRT-mCherry <em>Plasmodium berghei</em> parasites. We show that CRT is expressed during asexual blood stage growth and localizes to the hemozoin-containing digestive vacuole. The compartmentalized CRT-mCherry signal is also abundant in gametocytes and ookinetes, indicating that CRT continues to exert important functions in this digestive organelle up until mosquito midgut colonization. Expression is switched off during sporogony and early liver infection but CRT-mCherry is present again in mature liver stages, likely in preparation for blood infection. Together, visualization of the <em>P. berghei</em> digestive vacuole by endogenous tagging of <em>Pb</em>CRT revealed expression of this transport protein and the presence of this cellular compartment beyond asexual propagation inside erythrocytes.</div></div>","PeriodicalId":18721,"journal":{"name":"Molecular and biochemical parasitology","volume":"264 ","pages":"Article 111703"},"PeriodicalIF":1.5,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145280680","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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