Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.1.25.1 (deoxyribonuclease)
 1,471 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

We studied the fundamental instrumental issues relevant to a capillary-based integrated system to measure expression of a specific gene directly from cells. Samples were introduced into a capillary by use of a syringe pump. All reactions were carried out in a microthermocycler, where a part of the capillary having 1 microL inner volume was used as a reaction vessel. First, cells were lysed by heating to release RNA, followed by deoxyribonuclease (DNase) treatment. Then, reverse transcription-polymerase chain reaction (RT-PCR) was performed to obtain amplified products from the targeted mRNA. Finally, the product was verified by capillary electrophoresis (CE) with laser-induced fluorescence detection. The whole protocol was completed in the system in 3 h. PCR product from beta-actin mRNA in 16 human lymphoblast cells was obtained with a signal-to-noise ratio (S/N) of 3400 +/- 730 (n = 3). Therefore, the system is reproducible and sensitive enough to measure gene expression from a single cell. We show that the amplified fragment from breast cancer-specific mRNA was obtained from cells of breast cancer cell line, but was not obtained from cells of hepatoma cell line. These results therefore lay the foundations for future CE or microchip instrumentation for high-throughput automated gene-expression analysis.
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PMID:Integrated on-capillary instrumentation for gene expression measurement directly from cells. 1256 37

Periodontitis is an inflammatory disease initiated by host-parasite interactions which contributes to connective tissue destruction and alveolar bone resorption. Porphyromonas gingivalis (P.g.), a black-pigmented Gram-negative anaerobic bacterium, is a major pathogen in the development and progression of periodontitis. To characterize the role that P. gingivalis and its cell surface components play in disease processes, we investigated the differential expression of proteins induced by live P.g., P.g. LPS, and P.g. FimA, using two-dimensional gel electrophoresis in combination with mass spectrometry. We have tested whether, at the level of protein expression, unique signaling pathways are differentially induced by the bacterial components P.g. LPS and P.g. FimA, as compared to live P.g. We found that P.g. LPS stimulation of THP-1 up-regulated the expression of a set of proteins compared to control: deoxyribonuclease, actin, carbonic anhydrase 2, alpha enolase, adenylyl cyclase-associated protein (CAP1), protein disulfide isomerase (PDI), glucose regulated protein (grp78), and 70-kDa heat shock protein (HSP70), whereas FimA treatment did not result in statistically significant changes to protein levels versus the control. Live P.g. stimulation resulted in 12 differentially expressed proteins: CAP1, tubulin beta-2 chain, ATP synthase beta chain, tubulin alpha-6 chain, PDI, vimentin, 60-kDa heat shock protein, and nucleolin were found to be up-regulated, while carbonic anhydrase II, beta-actin, and HSP70 were down-regulated relative to control. These differential changes by the bacteria and its components are interpreted as preferential signal pathway activation in host immune/inflammatory responses to P.g. infection.
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PMID:Proteomic mapping of stimulus-specific signaling pathways involved in THP-1 cells exposed to Porphyromonas gingivalis or its purified components. 1747 57