Measurement of serum DNA binding in chronic active hepatitis and systemic lupus erythematosus using the Farr assay

Pollard, K. Michael; Steele, Richard; Hogg, Sister S; Webb, John

doi:10.1007/bf00270351

Cited by 4 publications

(1 citation statement)

References 36 publications

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“…In addition, Akt inhibits the transcriptional activity of FOXO3a by phosphorylation [26] followed by inhibition of MAFbx expression. AMPK can also phosphorylate FOXO3a, but unlike Akt, AMPKmediated phosphorylation increases the activity of this target molecule [37,38]. Here, we observed that LA overload for 24 h significantly increased the activity of AMPK but suppressed Akt activity, indicating that LA accumulation for longer times can initiate the canonical anti-myogenic signaling pathways.…”

Section: Discussionmentioning

confidence: 61%

Lactate overload inhibits myogenic activity in C2C12 myotubes

Oh¹,

Kim

Moon

et al. 2019

Open Life Sciences

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Lactate (LA), an endogenous metabolite produced from pyruvate, can accumulate in skeletal muscle in certain conditions including major diseases, as well as during intensive exercise. Using differentiated C2C12 myotubes, we evaluated the early (1-h) and delayed (24-h) effects of LA (8 mM) on mechanisms involved in myogenesis or muscle atrophy, including 5'-adenosine monophosphate-activated protein kinase (AMPK)-mediated inhibition of protein synthesis through the mTOR/P70-S6K pathway, Akt-mediated inhibition of expression of the MAFbx atrophic factor by FOXO3a and expression of the myogenic transcription factors, MyoD, myogenin and myosin heavy chain. Although the early effects of LA overload were not significant on myogenic or atrophic mechanisms, LA treatment for 24 h significantly activated atrophic mechanisms but suppressed myogenesis in myotubes. In addition, LA overload for 24 h significantly suppressed the expression of Sirtuin 1 and peroxisome proliferator-activated receptor gamma coactivator-1 alpha. Consistent with LA-induced activation of atrophic mechanisms, the diameter of C2C12 myotubes treated with LA for 24 h, but not for 1 h, was significantly lower than in control myotubes. Thus, a sustained, but not a transient, LA overload could induce muscle atrophy through the regulation of AMPK- and Akt-mediated pathways, although further in vivo studies are needed to confirm this.

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Section: Discussionmentioning

confidence: 61%

Lactate overload inhibits myogenic activity in C2C12 myotubes

Oh¹,

Kim

Moon

et al. 2019

Open Life Sciences

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Immunohistologically definable light chain restriction in autoimmune disease

Pollard¹,

Toumasatos²,

Furphy³

et al. 1988

The Journal of Pathology

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Structural requirements of DNA used in the Farr assay to detect antibodies directed against double-stranded DNA

Pollard

Webb

1987

Rheumatol Int

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The measurement of antibodies to DNA in SLE requires the use of double-stranded DNA (dsDNA), demonstrably free of single-stranded regions. Such dsDNA preparations can, however, contain other structural components. In this study DNA preparations with defined structure, both secondary (single- and double-stranded and random base-paired) and tertiary (superhelical and open circular), were used in the Farr assay to measure the DNA binding of sera from patients with SLE and related connective-tissue diseases. The presence of true single-stranded DNA regions in denatured DNA, native DNA, and dsDNA containing single-stranded regions increased the DNA binding measured in all sera. DsDNA, whether intact or containing small regions of random base-pairing, was bound by sera from the majority of patients with SLE but not by non-SLE sera. Superhelical dsDNA from bacteriophage PM2 was bound by SLE sera to a greater extent than linear dsDNA was. Inhibition experiments suggested that this difference in binding to DNA according to tertiary, as opposed to secondary, structure is because there are fewer available binding sites on superhelical dsDNA. DNA binding, as measured by the Farr assay, can thus be influenced by both secondary and tertiary DNA structure. Using superhelical DNA, advantage can be taken of the dsDNA form plus tertiary structure to enhance DNA binding of SLE sera beyond the levels achieved using linear dsDNA.

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Measurement of serum DNA binding in chronic active hepatitis and systemic lupus erythematosus using the Farr assay

Cited by 4 publications

References 36 publications

Lactate overload inhibits myogenic activity in C2C12 myotubes

Lactate overload inhibits myogenic activity in C2C12 myotubes

Immunohistologically definable light chain restriction in autoimmune disease

Structural requirements of DNA used in the Farr assay to detect antibodies directed against double-stranded DNA

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