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1.
PLoS Negl Trop Dis ; 11(6): e0005691, 2017 Jun.
Article in English | MEDLINE | ID: mdl-28650976

ABSTRACT

Schistosomiasis is a chronic parasitic disease caused by sexually dimorphic blood flukes of the genus Schistosoma. Praziquantel (PZQ) is the only drug widely available to treat the disease but does not kill juvenile parasites. Here we report the use of next generation sequencing to study the transcriptional effect of PZQ on murine hepatic inflammatory, immune and fibrotic responses to Schistosoma mansoni worms and eggs. An initial T helper cell 1 (Th1) response is induced against schistosomes in mice treated with drug vehicle (Vh) around the time egg laying begins, followed by a T helper cell 2 (Th2) response and the induction of genes whose action leads to granuloma formation and fibrosis. When PZQ is administered at this time, there is a significant reduction in egg burden yet the hepatic Th1, Th2 and fibrotic responses are still observed in the absence of granuloma formation suggesting some degree of gene regulation may be induced by antigens released from the dying adult worms. Quantitative real-time PCR was used to examine the relative expression of 16 juvenile and adult S. mansoni genes during infection and their response to Vh and PZQ treatment in vivo. While the response of stress genes in adult parasites suggests the worms were alive immediately following exposure to PZQ, they were unable to induce transcription of any of the 9 genes encoding ATP-binding cassette (ABC) transporters tested. In contrast, juvenile schistosomes were able to significantly induce the activities of ABCB, C and G family members, underscoring the possibility that these efflux systems play a major role in drug resistance.


Subject(s)
ATP-Binding Cassette Transporters/analysis , Anthelmintics/administration & dosage , Gene Expression Profiling , Liver/pathology , Praziquantel/administration & dosage , Schistosomiasis mansoni/drug therapy , Schistosomiasis mansoni/pathology , Animals , Disease Models, Animal , Drug Resistance , High-Throughput Nucleotide Sequencing , Liver Cirrhosis/pathology , Mice , Real-Time Polymerase Chain Reaction , Th1 Cells/immunology , Th2 Cells/immunology
2.
Sci Rep ; 5: 11136, 2015 Jun 15.
Article in English | MEDLINE | ID: mdl-26073445

ABSTRACT

Honey bee colonies exhibit an age-related division of labor, with worker bees performing discrete sets of behaviors throughout their lifespan. These behavioral states are associated with distinct brain transcriptomic states, yet little is known about the regulatory mechanisms governing them. We used CAGEscan (a variant of the Cap Analysis of Gene Expression technique) for the first time to characterize the promoter regions of differentially expressed brain genes during two behavioral states (brood care (aka "nursing") and foraging) and identified transcription factors (TFs) that may govern their expression. More than half of the differentially expressed TFs were associated with motifs enriched in the promoter regions of differentially expressed genes (DEGs), suggesting they are regulators of behavioral state. Strikingly, five TFs (nf-kb, egr, pax6, hairy, and clockwork orange) were predicted to co-regulate nearly half of the genes that were upregulated in foragers. Finally, differences in alternative TSS usage between nurses and foragers were detected upstream of 646 genes, whose functional analysis revealed enrichment for Gene Ontology terms associated with neural function and plasticity. This demonstrates for the first time that alternative TSSs are associated with stable differences in behavior, suggesting they may play a role in organizing behavioral state.


Subject(s)
Bees/genetics , Brain/metabolism , Early Growth Response Transcription Factors/genetics , Gene Expression Regulation, Developmental , Insect Proteins/genetics , Transcription, Genetic , Animals , Basic Helix-Loop-Helix Transcription Factors/genetics , Basic Helix-Loop-Helix Transcription Factors/metabolism , Bees/growth & development , Bees/metabolism , Behavior, Animal , Brain/growth & development , Early Growth Response Transcription Factors/metabolism , Eye Proteins/genetics , Eye Proteins/metabolism , Gene Expression Profiling , Homeodomain Proteins/genetics , Homeodomain Proteins/metabolism , Insect Proteins/metabolism , Multigene Family , NF-kappa B/genetics , NF-kappa B/metabolism , Neuronal Plasticity/genetics , PAX6 Transcription Factor , Paired Box Transcription Factors/genetics , Paired Box Transcription Factors/metabolism , Promoter Regions, Genetic , Repressor Proteins/genetics , Repressor Proteins/metabolism , Signal Transduction
3.
Arch Androl ; 53(4): 207-18, 2007.
Article in English | MEDLINE | ID: mdl-17852045

ABSTRACT

Alus and B1s are short interspersed repeat elements (SINEs) derived from the 7SL RNA gene. Alus and B1s exist in the cytoplasm as non-coding RNA indicating that they are actively transcribed, but their function, if any, is unknown. Transcription of individual SINEs is a prerequisite for retroposition, but it is also possible that individual Alu and B1 elements have some cellular functions. Previous studies suggest that transcription of Alu elements depends on the presence of an RNA polymerase-III bipartite promoter and the poly-A tail. Sequencing of small RNAs has demonstrated that the members of the Y and S subfamily are expressed. We analyzed almost one million Alu sequences longer than 200 nucleotides for the presence of RNA polymerase-III bipartite promoter sequences. More than half contained a promoter indicating some potential for expression. We searched 7.7 million human EST sequences in dbEST for the presence of Alu non-coding RNAs and found evidence for the expression of 452. Analysis of mouse spermatogenic dbEST libraries revealed an apparent relationship between the level of differentiation and the level of B1-related sequences in the EST library.


Subject(s)
Alu Elements/physiology , Databases, Genetic , Expressed Sequence Tags/chemistry , Animals , Humans , Male , Mice , Poly A/analysis , Promoter Regions, Genetic
4.
J Cell Biochem ; 102(1): 110-21, 2007 Sep 01.
Article in English | MEDLINE | ID: mdl-17407136

ABSTRACT

Alus and B1s are short interspersed repeat elements (SINEs) indirectly derived from the 7SL RNA gene. While most researchers recognize that there exists extensive variability between individual elements, the extent of this variability has never been systematically tested. We examined all Alu elements over 200 nucleotides and all B1 elements over 100 nucleotides in the human and mouse genomes, and analyzed the number of copies of each element at various stringencies from 22 nucleotides to full length. Over 98% of 923,277 Alus and 365,377 B1s examined were unique when queried at full length. When the criterion was reduced to half the length of the repeat, 97% of the Alus and 73% of the B1s were still found to be a single copy. All single and multi-copy sequences have been mapped and documented. Access to the data is possible using the AluPlus website http://www.ibr.hawaii.edu.


Subject(s)
Alu Elements , Short Interspersed Nucleotide Elements , Animals , Chromosomes, Mammalian/chemistry , Databases, Nucleic Acid , GC Rich Sequence , Genetic Variation , Genomics , Humans , Internet , Mice , Sequence Deletion
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