Molecularly specific detection of bacterial lipoteichoic acid for diagnosis of prosthetic joint infection of the bone

Discriminating sterile inflammation from infection, especially in cases of aseptic loosening versus an actual prosthetic joint infection, is challenging and has significant treatment implications. Our goal was to evaluate a novel human monoclonal antibody（mAb） probe directed against the Gram-positive bacterial surface molecule lipoteichoic acid（LTA）. Specificity and affinity were assessed in vitro. We then radiolabeled the anti-LTA mAb and evaluated its effectiveness as a diagnostic imaging tool for detecting infection via immuno PET imaging in an in vivo mouse model of prosthetic joint infection（PJI）. In vitro and ex vivo binding of the anti-LTA mAb to pathogenic bacteria was measured with Octet, ELISA, and flow cytometry. The in vivo PJI mouse model was assessed using traditional imaging modalities, including positron emission tomography（PET） with [^（18）F]FDG and [^（18）F]Na F as well as X-ray computed tomography（CT）, before being evaluated with the zirconium-89-labeled antibody specific for LTA（[^（89）Zr]SAC55）.The anti-LTA mAb exhibited specific binding in vitro to LTA-expressing bacteria. Results from imaging showed that our model could reliably simulate infection at the surgical site by bioluminescent imaging, conventional PET tracer imaging, and bone morphological changes by CT. One day following injection of both the radiolabeled anti-LTA and isotype control antibodies, the anti-LTA antibody demonstrated significantly greater（P 〈 0.05） uptake at S. aureus-infected prosthesis sites over either the same antibody at sterile prosthesis sites or of control non-specific antibody at infected prosthesis sites. Taken together, the radiolabeled anti-LTA mAb, [^（89）Zr]SAC55, may serve as a valuable diagnostic molecular imaging probe to help distinguish between sterile inflammation and infection in the setting of PJI. Future studies are needed to determine whether these findings will translate to human PJI.

Identification and characterization of long non-coding RNAs in porcine granulosa cells exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin

Background: Long non-coding RNAs(lncRNAs) may regulate gene expression in numerous biological processes including cellular response to xenobiotics.The exposure of living organisms to 2,3,7,8-tetrachlorodibenzo-p-dioxin(TCDD),a persistent environmental contaminant,results in reproductive defects in many species including pigs.The aims of the study were to identify and characterize lncRNAs in porcine granulosa cells as well as to examine the effects of TCDD on the lncRNA expression profile in the cells.Results: One thousand six hundred sixty-six lncRNAs were identified and characterized in porcine granulosa cells.The identified lncRNAs were found to be shorter than mRNAs.In addition,the number of exons was lower in lncRNAs than in m RNAs and their exons were longer.TCDD affected the expression of 22 lncRNAs(differentially expressed lncRNAs [DELs]; log2 fold change ≥ 1,P-adjusted < 0.05) in the examined cells.Potential functions of DELs were indirectly predicted via searching their target cis-and trans-regulated protein-coding genes.The coexpression analysis revealed that DELs may influence the expression of numerous genes,including those involved in cellular response to xenobiotics,dioxin metabolism,endoplasmic reticulum stress and cell proliferation.Aryl hydrocarbon receptor(AhR) and cytochrome P450 1 A1(CYP1 A1) were found among the trans-regulated genes.Conclusions: These findings indicate that the identified lncRNAs may constitute a part of the regulatory mechanism of TCDD action in granulosa cells.To our knowledge,this is the first study describing lncRNAs in porcine granulosa cells as well as TCDD effects on the lncRNA expression profile.These results may trigger new research directions leading to better understanding of molecular processes induced by xenobiotics in the ovary.