11 results on '"Brindley, Paul J."'
Search Results
2. Vaccination of hamsters with Opisthorchis viverrini extracellular vesicles and vesicle-derived recombinant tetraspanins induces antibodies that block vesicle uptake by cholangiocytes and reduce parasite burden after challenge infection.
- Author
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Chaiyadet, Sujittra, Sotillo, Javier, Krueajampa, Watchara, Thongsen, Sophita, Brindley, Paul J., Sripa, Banchob, Loukas, Alex, and Laha, Thewarach
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OPISTHORCHIS viverrini ,TREMATODA ,CLONORCHIS sinensis ,HAMSTERS ,RECOMBINANT proteins ,LIVER flukes ,VACCINATION - Abstract
Background: The liver fluke Opisthorchis viverrini infects several million people in Southeast Asia. Adult flukes live in the bile ducts of humans, where they cause hepatobiliary pathology, including cholangiocarcinoma. Here, we investigated the potential of extracellular vesicles (EVs) secreted by the fluke and defined recombinant proteins derived from EVs to generate protective immunity in a hamster vaccination-challenge model. Methodology/Principal findings: EVs isolated from the excretory-secretory products of O. viverrini and two recombinant EV surface proteins encoding the large extracellular loops (LEL) of Ov-TSP-2 (rOv-TSP-2) and Ov-TSP-3 (rOv-TSP-3) were adjuvanted and used to vaccinate hamsters intraperitoneally followed by challenge infection with O. viverrini metacercariae. The number of adult flukes recovered from hamsters immunized with EVs, rOv-TSP-2, rOv-TSP-3 and rOv-TSP-2+rOv-TSP-3 were significantly reduced compared to control animals vaccinated with adjuvant alone. The number of eggs per gram feces was also significantly reduced in hamsters vaccinated with rOv-TSP-2 compared to controls, but no significant differences were found in the other groups. The average length of worms recovered from hamsters vaccinated with EVs, rOv-TSP-2 and rOv-TSP-3 was significantly shorter than that of worms recovered from the control group. Anti-EV IgG levels in serum and bile were significantly higher in hamsters vaccinated with EVs compared to control hamsters both pre- and post-challenge. In addition, levels of anti-rOv-TSP antibodies in the serum and bile were significantly higher than control hamsters both pre- and post-challenge. Finally, antibodies against rOv-TSP-2 and rOv-TSP-3 blocked uptake of EVs by human primary cholangiocyte in vitro, providing a plausible mechanism by which these vaccines exert partial efficacy and reduce the intensity of O. viverrini infection. Conclusion/Significance: Liver fluke EVs and recombinant tetraspanins derived from the EV surface when administered to hamsters induce antibody responses that block EV uptake by target bile duct cells and exert partial efficacy and against O. viverrini challenge. [ABSTRACT FROM AUTHOR]
- Published
- 2019
- Full Text
- View/download PDF
3. The snail Biomphalaria glabrata as a model to interrogate the molecular basis of complex human diseases.
- Author
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Bridger, Joanna M., Brindley, Paul J., and Knight, Matty
- Subjects
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SCHISTOSOMA , *SNAILS , *EUKARYOTIC cells , *PATHOGENIC microorganisms , *GENOMES - Abstract
The article focuses on aspects of the snail or schistosome relationship elucidating common pathways to enable both snails and humans to accommodate parasitism by schistosomes in view of physiological and immunological environments. It notes that the excretory or secretory products (ESPs) of the miracidium include the stress-inducing factor. Gene repositioning in B. glabrata by schistosomes exemplifies how a eukaryotic pathogen influences hijack of the genome behavior of its host.
- Published
- 2018
- Full Text
- View/download PDF
4. The small RNA complement of adult Schistosoma haematobium.
- Author
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Stroehlein, Andreas J., Young, Neil D., Korhonen, Pasi K., Hall, Ross S., Jex, Aaron R., Webster, Bonnie L., Rollinson, David, Brindley, Paul J., and Gasser, Robin B.
- Subjects
SCHISTOSOMA haematobium ,RNA ,SCHISTOSOMIASIS diagnosis ,TREMATODA ,GENOMES ,HOST-parasite relationships ,SCHISTOSOMA japonicum ,SCHISTOSOMA mansoni ,THERAPEUTICS - Abstract
Background: Blood flukes of the genus Schistosoma cause schistosomiasis—a neglected tropical disease (NTD) that affects more than 200 million people worldwide. Studies of schistosome genomes have improved our understanding of the molecular biology of flatworms, but most of them have focused largely on protein-coding genes. Small non-coding RNAs (sncRNAs) have been explored in selected schistosome species and are suggested to play essential roles in the post-transcriptional regulation of genes, and in modulating flatworm-host interactions. However, genome-wide small RNA data are currently lacking for key schistosomes including Schistosoma haematobium—the causative agent of urogenital schistosomiasis of humans. Methodology: MicroRNAs (miRNAs) and other sncRNAs of male and female adults of S. haematobium and small RNA transcription levels were explored by deep sequencing, genome mapping and detailed bioinformatic analyses. Principal findings: In total, 89 transcribed miRNAs were identified in S. haematobium—a similar complement to those reported for the congeners S. mansoni and S. japonicum. Of these miRNAs, 34 were novel, with no homologs in other schistosomes. Most miRNAs (n = 64) exhibited sex-biased transcription, suggestive of roles in sexual differentiation, pairing of adult worms and reproductive processes. Of the sncRNAs that were not miRNAs, some related to the spliceosome (n = 21), biogenesis of other RNAs (n = 3) or ribozyme functions (n = 16), whereas most others (n = 3798) were novel (‘orphans’) with unknown functions. Conclusions: This study provides the first genome-wide sncRNA resource for S. haematobium, extending earlier studies of schistosomes. The present work should facilitate the future curation and experimental validation of sncRNA functions in schistosomes to enhance our understanding of post-transcriptional gene regulation and of the roles that sncRNAs play in schistosome reproduction, development and parasite-host cross-talk. [ABSTRACT FROM AUTHOR]
- Published
- 2018
- Full Text
- View/download PDF
5. Genomes of Fasciola hepatica from the Americas Reveal Colonization with Neorickettsia Endobacteria Related to the Agents of Potomac Horse and Human Sennetsu Fevers.
- Author
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McNulty, Samantha N., Rosa, Bruce A., Choi, Young-Jun, Tyagi, Rahul, Hallsworth-Pepin, Kymberlie, Mitreva, Makedonka, Tort, Jose F., Smircich, Pablo, Fontenla, Santiago, Dell'Oca, Nicolas, Dominguez, Fernanda, Rinaldi, Gabriel, Mann, Victoria H., Brindley, Paul J., Fischer, Kerstin, Fischer, Peter U., Kammili, Lakshmi, Latham, Patricia S., and Carmona, Carlos
- Subjects
FASCIOLA hepatica ,NEORICKETTSIA ,PLATYHELMINTHES ,PATHOGENIC microorganisms ,FOODBORNE diseases - Abstract
Food borne trematodes (FBTs) are an assemblage of platyhelminth parasites transmitted through the food chain, four of which are recognized as neglected tropical diseases (NTDs). Fascioliasis stands out among the other NTDs due to its broad and significant impact on both human and animal health, as Fasciola sp., are also considered major pathogens of domestic ruminants. Here we present a reference genome sequence of the common liver fluke, Fasciola hepatica isolated from sheep, complementing previously reported isolate from cattle. A total of 14,642 genes were predicted from the 1.14 GB genome of the liver fluke. Comparative genomics indicated that F. hepatica Oregon and related food-borne trematodes are metabolically less constrained than schistosomes and cestodes, taking advantage of the richer milieus offered by the hepatobiliary organs. Protease families differentially expanded between diverse trematodes may facilitate migration and survival within the heterogeneous environments and niches within the mammalian host. Surprisingly, the sequencing of Oregon and Uruguay F. hepatica isolates led to the first discovery of an endobacteria in this species. Two contigs from the F. hepatica Oregon assembly were joined to complete the 859,205 bp genome of a novel Neorickettsia endobacterium (nFh) closely related to the etiological agents of human Sennetsu and Potomac horse fevers. Immunohistochemical studies targeting a Neorickettsia surface protein found nFh in specific organs and tissues of the adult trematode including the female reproductive tract, eggs, the Mehlis’ gland, seminal vesicle, and oral suckers, suggesting putative routes for fluke-to-fluke and fluke-to-host transmission. The genomes of F. hepatica and nFh will serve as a resource for further exploration of the biology of F. hepatica, and specifically its newly discovered trans-kingdom interaction with nFh and the impact of both species on disease in cattle and humans. [ABSTRACT FROM AUTHOR]
- Published
- 2017
- Full Text
- View/download PDF
6. P53 and Cancer-Associated Sialylated Glycans Are Surrogate Markers of Cancerization of the Bladder Associated with Schistosoma haematobium Infection.
- Author
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Santos, Júlio, Fernandes, Elisabete, Ferreira, José Alexandre, Lima, Luís, Tavares, Ana, Peixoto, Andreia, Parreira, Beatriz, Correia da Costa, José Manuel, Brindley, Paul J., Lopes, Carlos, and Santos, Lúcio L.
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P53 protein ,GLYCANS ,BLADDER cancer diagnosis ,BLADDER cancer patients ,BLADDER cancer treatment - Abstract
Background: Bladder cancer is a significant health problem in rural areas of Africa and the Middle East where Schistosoma haematobium is prevalent, supporting an association between malignant transformation and infection by this blood fluke. Nevertheless, the molecular mechanisms linking these events are poorly understood. Bladder cancers in infected populations are generally diagnosed at a late stage since there is a lack of non-invasive diagnostic tools, hence enforcing the need for early carcinogenesis markers. Methodology/Principal Findings: Forty-three formalin-fixed paraffin-embedded bladder biopsies of S. haematobium-infected patients, consisting of bladder tumours, tumour adjacent mucosa and pre-malignant/malignant urothelial lesions, were screened for bladder cancer biomarkers. These included the oncoprotein p53, the tumour proliferation rate (Ki-67>17%), cell-surface cancer-associated glycan sialyl-Tn (sTn) and sialyl-Lewis
a/x (sLea /sLex ), involved in immune escape and metastasis. Bladder tumours of non-S. haematobium etiology and normal urothelium were used as controls. S. haematobium-associated benign/pre-malignant lesions present alterations in p53 and sLex that were also found in bladder tumors. Similar results were observed in non-S. haematobium associated tumours, irrespectively of their histological nature, denoting some common molecular pathways. In addition, most benign/pre-malignant lesions also expressed sLea . However, proliferative phenotypes were more prevalent in lesions adjacent to bladder tumors while sLea was characteristic of sole benign/pre-malignant lesions, suggesting it may be a biomarker of early carcionogenesis associated with the parasite. A correlation was observed between the frequency of the biomarkers in the tumor and adjacent mucosa, with the exception of Ki-67. Most S. haematobium eggs embedded in the urothelium were also positive for sLea and sLex . Reinforcing the pathologic nature of the studied biomarkers, none was observed in the healthy urothelium. Conclusion/Significance: This preliminary study suggests that p53 and sialylated glycans are surrogate biomarkers of bladder cancerization associated with S. haematobium, highlighting a missing link between infection and cancer development. Eggs of S. haematobium express sLea and sLex antigens in mimicry of human leukocytes glycosylation, which may play a role in the colonization and disease dissemination. These observations may help the early identification of infected patients at a higher risk of developing bladder cancer and guide the future development of non-invasive diagnostic tests. [ABSTRACT FROM AUTHOR]- Published
- 2014
- Full Text
- View/download PDF
7. Urinary Estrogen Metabolites and Self-Reported Infertility in Women Infected with Schistosoma haematobium.
- Author
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Santos, Júlio, Gouveia, Maria João, Vale, Nuno, Delgado, Maria de Lurdes, Gonçalves, Ana, da Silva, José M. Teixeira., Oliveira, Cristiano, Xavier, Pedro, Gomes, Paula, Santos, Lúcio L., Lopes, Carlos, Barros, Alberto, Rinaldi, Gabriel, Brindley, Paul J., da Costa, José M. Correia, Sousa, Mário, and Botelho, Mónica C.
- Subjects
ESTROGEN receptors ,SCHISTOSOMIASIS ,SCHISTOSOMA haematobium ,TROPICAL medicine ,CATECHOL ,LIQUID chromatography - Abstract
Background: Schistosomiasis is a neglected tropical disease, endemic in 76 countries, that afflicts more than 240 million people. The impact of schistosomiasis on infertility may be underestimated according to recent literature. Extracts of Schistosoma haematobium include estrogen-like metabolites termed catechol-estrogens that down regulate estrogen receptors alpha and beta in estrogen responsive cells. In addition, schistosome derived catechol-estrogens induce genotoxicity that result in estrogen-DNA adducts. These catechol estrogens and the catechol-estrogen-DNA adducts can be isolated from sera of people infected with S. haematobium. The aim of this study was to study infertility in females infected with S. haematobium and its association with the presence of schistosome-derived catechol-estrogens. Methodology/Principal Findings: A cross-sectional study was undertaken of female residents of a region in Bengo province, Angola, endemic for schistosomiasis haematobia. Ninety-three women and girls, aged from two (parents interviewed) to 94 years were interviewed on present and previous urinary, urogenital and gynecological symptoms and complaints. Urine was collected from the participants for egg-based parasitological assessment of schistosome infection, and for liquid chromatography diode array detection electron spray ionization mass spectrometry (LC/UV-DAD/ESI-MSn) to investigate estrogen metabolites in the urine. Novel estrogen-like metabolites, potentially of schistosome origin, were detected in the urine of participants who were positive for eggs of S. haematobium, but not detected in urines negative for S. haematobium eggs. The catechol-estrogens/ DNA adducts were significantly associated with schistosomiasis (OR 3.35; 95% CI 2.32–4.84; P≤0.001). In addition, presence of these metabolites was positively associated with infertility (OR 4.33; 95% CI 1.13–16.70; P≤0.05). Conclusions/Significance: Estrogen metabolites occur widely in diverse metabolic pathways. In view of the statistically significant association between catechol-estrogens/ DNA adducts and self-reported infertility, we propose that an estrogen-DNA adduct mediated pathway in S. haematobium-induced ovarian hormonal deregulation could be involved. In addition, the catechol-estrogens/ DNA adducts described here represent potential biomarkers for schistosomiasis haematobia. [ABSTRACT FROM AUTHOR]
- Published
- 2014
- Full Text
- View/download PDF
8. Elevated Plasma IL-6 Associates with Increased Risk of Advanced Fibrosis and Cholangiocarcinoma in Individuals Infected by Opisthorchis viverrini.
- Author
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Sripa, Banchob, Thinkhamrop, Bandit, Mairiang, Eimorn, Laha, Thewarach, Kaewkes, Sasithorn, Sithithaworn, Paiboon, Periago, Maria Victoria, Bhudhisawasdi, Vajarabhongsa, Yonglitthipagon, Ponlapat, Mulvenna, Jason, Brindley, Paul J., Loukas, Alex, and Bethony, Jeffrey M.
- Subjects
FIBROSIS ,CHOLANGIOCARCINOMA ,INTERLEUKIN-6 ,BILE ducts ,PREVENTIVE medicine ,BIOCHEMISTRY ,CANCER ,DISEASE risk factors - Abstract
Opisthorchis viverrini is considered among the most important of the food-borne trematodes due to its strong association with advanced periductal fibrosis and bile duct cancer (cholangiocarcinoma). We investigated the relationship between plasma levels of Interleukin (IL)-6 and the risk of developing advanced fibrosis and bile duct cancer from chronic Opisthorchis infection. We show that IL-6 circulates in plasma at concentrations 58 times higher in individuals with advanced fibrosis than age, sex, and nearest-neighbor matched controls and 221 times higher in individuals with bile duct cancer than controls. We also observed a dose-response relationship between increasing levels of plasma IL-6 and increasing risk of advanced fibrosis and bile duct cancer; for example, in age and sex adjusted analyses, individuals with the highest quartiles of plasma IL-6 had a 19 times greater risk of developing advanced periductal fibrosis and a 150 times greater risk of developing of bile duct cancer than individuals with no detectable level of plasma IL-6. Finally, we show that a single plasma IL-6 measurement has excellent positive predictive value for the detection of both advanced bile duct fibrosis and bile duct cancer in regions with high O. viverrini transmission. These data support our hypothesis that common mechanisms drive bile duct fibrosis and bile duct tumorogenesis from chronic O. viverrini infection. Our study also adds a unique aspect to the literature on circulating levels of IL-6 as an immune marker of hepatobiliary pathology by showing that high levels of circulating IL-6 in plasma are not related to infection with O. viverrini, but to the development of the advanced and often lethal pathologies resulting from chronic O. viverrini infection. [ABSTRACT FROM AUTHOR]
- Published
- 2012
- Full Text
- View/download PDF
9. Tumor Necrosis Factor and Schistosoma mansoni egg antigen omega-1 shape distinct aspects of the early egg-induced granulomatous response
- Author
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Antonio J. Pagán, Paul J. Brindley, Kevin K. Takaki, Gabriele Schramm, Ruud H. P. Wilbers, Francisco J. Roca, Matthew Berriman, Wannaporn Ittiprasert, Lalita Ramakrishnan, Gabriel Rinaldi, Takaki, Kevin K [0000-0002-4790-4598], Roca, Francisco J [0000-0002-6744-148X], Wilbers, Ruud HP [0000-0003-1149-9268], Ittiprasert, Wannaporn [0000-0001-9411-8883], Brindley, Paul J [0000-0003-1765-0002], Rinaldi, Gabriel [0000-0002-7767-4922], Berriman, Matthew [0000-0002-9581-0377], Ramakrishnan, Lalita [0000-0003-0692-5533], Pagán, Antonio J [0000-0002-0280-1800], Apollo - University of Cambridge Repository, Takaki, Kevin K. [0000-0002-4790-4598], Roca, Francisco J. [0000-0002-6744-148X], Wilbers, Ruud H. P. [0000-0003-1149-9268], Brindley, Paul J. [0000-0003-1765-0002], and Pagán, Antonio J. [0000-0002-0280-1800]
- Subjects
0301 basic medicine ,Physiology ,Hydrolases ,Eggs ,RC955-962 ,Biochemistry ,White Blood Cells ,0302 clinical medicine ,Animal Cells ,Reproductive Physiology ,Arctic medicine. Tropical medicine ,Immune Physiology ,Medicine and Health Sciences ,Parasite hosting ,Zebrafish ,Innate Immune System ,Granuloma ,biology ,Eukaryota ,Brain ,Animal Models ,Helminth Proteins ,Schistosoma mansoni ,Enzymes ,Infectious Diseases ,Experimental Organism Systems ,Osteichthyes ,Receptors, Tumor Necrosis Factor, Type I ,Larva ,Vertebrates ,Granulomas ,Schistosoma ,Cytokines ,Tumor necrosis factor alpha ,Public aspects of medicine ,RA1-1270 ,Cellular Types ,Anatomy ,Research Article ,Nucleases ,Immune Cells ,Immunology ,Hindbrain ,Research and Analysis Methods ,03 medical and health sciences ,Model Organisms ,Ribonucleases ,Antigen ,Helminths ,DNA-binding proteins ,medicine ,Life Science ,Animals ,Laboratorium voor Nematologie ,Glycoproteins ,Ovum ,Blood Cells ,Tumor Necrosis Factor-alpha ,Macrophages ,Public Health, Environmental and Occupational Health ,Organisms ,Biology and Life Sciences ,Proteins ,Chemotaxis ,Cell Biology ,Molecular Development ,biology.organism_classification ,medicine.disease ,Invertebrates ,Schistosomiasis mansoni ,030104 developmental biology ,Fish ,Immune System ,Antigens, Helminth ,Mutation ,Animal Studies ,Enzymology ,EPS ,Laboratory of Nematology ,Zoology ,030217 neurology & neurosurgery ,Developmental Biology - Abstract
Infections by schistosomes result in granulomatous lesions around parasite eggs entrapped within the host tissues. The host and parasite determinants of the Schistosoma mansoni egg-induced granulomatous response are areas of active investigation. Some studies in mice implicate Tumor Necrosis Factor (TNF) produced in response to the infection whereas others fail to find a role for it. In addition, in the mouse model, the S. mansoni secreted egg antigen omega-1 is found to induce granulomas but the underlying mechanism remains unknown. We have recently developed the zebrafish larva as a model to study macrophage recruitment and granuloma formation in response to Schistosoma mansoni eggs. Here we use this model to investigate the mechanisms by which TNF and omega-1 shape the early granulomatous response. We find that TNF, specifically signaling through TNF receptor 1, is not required for macrophage recruitment to the egg and granuloma initiation but does mediate granuloma enlargement. In contrast, omega-1 mediates initial macrophage recruitment, with this chemotactic activity being dependent on its RNase activity. Our findings further the understanding of the role of these host- and parasite-derived factors and show that they impact distinct facets of the granulomatous response to the schistosome egg., Author summary Schistosomiasis is a disease caused by parasitic flatworms which lay eggs within the veins of their human host. Upon sensing the parasite egg, macrophages, the first line defense cells, aggregate tightly around the egg to encapsulate it within an immune structure known as a granuloma. These granulomas are the key pathological structures which determine both host disease outcome and parasite transmission. Studies in mice have implicated omega-1, a secreted parasite protein. Omega-1 is an RNase, an enzyme that degrades host RNA. Mouse studies have also suggested that a host defense protein, Tumor Necrosis Factor (TNF), is required to form granulomas around the egg. We used the small and transparent zebrafish larva to examine the requirement of omega-1 and TNF for granuloma formation. We find that omega-1 induces rapid macrophage migration and that its RNase activity is required for this. In contrast, TNF is not involved in the initial recruitment of macrophages. Rather, it enlarges granulomas after they are initiated. These findings improve our understanding of the role of omega-1 and TNF, and show that they impact distinct facets of granuloma formation around Schistosoma eggs.
- Published
- 2021
10. Urinary Estrogen Metabolites and Self-Reported Infertility in Women Infected with Schistosoma haematobium.
- Author
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Santos, Júlio, Gouveia, Maria João, Vale, Nuno, Delgado, Maria de Lurdes, Gonçalves, Ana, da Silva, José M. Teixeira., Oliveira, Cristiano, Xavier, Pedro, Gomes, Paula, Santos, Lúcio L., Lopes, Carlos, Barros, Alberto, Rinaldi, Gabriel, Brindley, Paul J., da Costa, José M. Correia, Sousa, Mário, and Botelho, Mónica C.
- Subjects
- *
ESTROGEN receptors , *SCHISTOSOMIASIS , *SCHISTOSOMA haematobium , *TROPICAL medicine , *CATECHOL , *LIQUID chromatography - Abstract
Background: Schistosomiasis is a neglected tropical disease, endemic in 76 countries, that afflicts more than 240 million people. The impact of schistosomiasis on infertility may be underestimated according to recent literature. Extracts of Schistosoma haematobium include estrogen-like metabolites termed catechol-estrogens that down regulate estrogen receptors alpha and beta in estrogen responsive cells. In addition, schistosome derived catechol-estrogens induce genotoxicity that result in estrogen-DNA adducts. These catechol estrogens and the catechol-estrogen-DNA adducts can be isolated from sera of people infected with S. haematobium. The aim of this study was to study infertility in females infected with S. haematobium and its association with the presence of schistosome-derived catechol-estrogens. Methodology/Principal Findings: A cross-sectional study was undertaken of female residents of a region in Bengo province, Angola, endemic for schistosomiasis haematobia. Ninety-three women and girls, aged from two (parents interviewed) to 94 years were interviewed on present and previous urinary, urogenital and gynecological symptoms and complaints. Urine was collected from the participants for egg-based parasitological assessment of schistosome infection, and for liquid chromatography diode array detection electron spray ionization mass spectrometry (LC/UV-DAD/ESI-MSn) to investigate estrogen metabolites in the urine. Novel estrogen-like metabolites, potentially of schistosome origin, were detected in the urine of participants who were positive for eggs of S. haematobium, but not detected in urines negative for S. haematobium eggs. The catechol-estrogens/ DNA adducts were significantly associated with schistosomiasis (OR 3.35; 95% CI 2.32–4.84; P≤0.001). In addition, presence of these metabolites was positively associated with infertility (OR 4.33; 95% CI 1.13–16.70; P≤0.05). Conclusions/Significance: Estrogen metabolites occur widely in diverse metabolic pathways. In view of the statistically significant association between catechol-estrogens/ DNA adducts and self-reported infertility, we propose that an estrogen-DNA adduct mediated pathway in S. haematobium-induced ovarian hormonal deregulation could be involved. In addition, the catechol-estrogens/ DNA adducts described here represent potential biomarkers for schistosomiasis haematobia. [ABSTRACT FROM AUTHOR]
- Published
- 2014
- Full Text
- View/download PDF
11. Targeted mutagenesis in a human-parasitic nematode
- Author
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Matteo Pellegrini, Michelle L. Castelletto, Nicholas Mancuso, Astra S. Bryant, Elissa A. Hallem, Jacqueline B. Lopez, Emily Yang, Spencer S. Gang, and Brindley, Paul J
- Subjects
0301 basic medicine ,Nematoda ,Mutant ,Muscle Proteins ,Artificial Gene Amplification and Extension ,Synthetic Genome Editing ,Biochemistry ,Polymerase Chain Reaction ,Genome Engineering ,Animals, Genetically Modified ,0302 clinical medicine ,Strongyloides ,Medicine and Health Sciences ,2.1 Biological and endogenous factors ,Clustered Regularly Interspaced Short Palindromic Repeats ,Biology (General) ,Aetiology ,Nematode Infections ,Gene Editing ,Genetics ,Plasmid Vectors ,biology ,Crispr ,Eukaryota ,Strongyloides ratti ,Animal Models ,3. Good health ,Infectious Diseases ,Experimental Organism Systems ,Ribonucleoproteins ,Caenorhabditis Elegans ,Medical Microbiology ,Engineering and Technology ,Synthetic Biology ,Genetic Engineering ,Research Article ,Biotechnology ,QH301-705.5 ,DNA repair ,030231 tropical medicine ,Immunology ,Bioengineering ,Genetically Modified ,Locus (genetics) ,Research and Analysis Methods ,Microbiology ,Article ,Strongyloides stercoralis ,03 medical and health sciences ,Model Organisms ,Virology ,Parasitic Diseases ,Animals ,Humans ,Molecular Biology Techniques ,Molecular Biology ,Gene ,Organisms ,Biology and Life Sciences ,Proteins ,Synthetic Genomics ,RC581-607 ,biology.organism_classification ,Invertebrates ,Rats ,030104 developmental biology ,Nematode ,Synthetic Bioengineering ,Mutagenesis ,Caenorhabditis ,Calmodulin-Binding Proteins ,Parasitology ,Immunologic diseases. Allergy ,Digestive Diseases - Abstract
Parasitic nematodes infect over 1 billion people worldwide and cause some of the most common neglected tropical diseases. Despite their prevalence, our understanding of the biology of parasitic nematodes has been limited by the lack of tools for genetic intervention. In particular, it has not yet been possible to generate targeted gene disruptions and mutant phenotypes in any parasitic nematode. Here, we report the development of a method for introducing CRISPR-Cas9-mediated gene disruptions in the human-parasitic threadworm Strongyloides stercoralis. We disrupted the S. stercoralis twitchin gene unc-22, resulting in nematodes with severe motility defects. Ss-unc-22 mutations were resolved by homology-directed repair when a repair template was provided. Omission of a repair template resulted in deletions at the target locus. Ss-unc-22 mutations were heritable; we passed Ss-unc-22 mutants through a host and successfully recovered mutant progeny. Using a similar approach, we also disrupted the unc-22 gene of the rat-parasitic nematode Strongyloides ratti. Our results demonstrate the applicability of CRISPR-Cas9 to parasitic nematodes, and thereby enable future studies of gene function in these medically relevant but previously genetically intractable parasites., Author summary Parasitic worms are a widespread public health burden, yet very little is known about the cellular and molecular mechanisms that contribute to their parasitic lifestyle. One of the major barriers to better understanding these mechanisms is that there are currently no available methods for making targeted gene knockouts in any parasitic worm species. Here, we describe the first mutant phenotype in a parasitic worm resulting from a targeted gene disruption. We applied CRISPR-Cas9-mediated mutagenesis to parasitic worms in the genus Strongyloides and developed a method that overcomes many of the challenges that have previously inhibited generating mutant parasitic worms. We characterize heritable mutant phenotypes and outline a toolkit that will be applicable to many other genes with potential roles in parasitism. Importantly, we developed our method for gene knockouts in a human-parasitic worm. By directly investigating the genes and molecular pathways that enable worms to parasitize humans, we may be able to develop novel anthelmintic therapies or other measures for preventing nematode infections.
- Published
- 2017
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