Iron and Ferritin Modulate MHC Class I Expression and NK Cell Recognition

The ability of pathogens to sequester iron from their host cells and proteins affects their virulence. Moreover, iron is required for various innate host defense mechanisms as well as for acquired immune responses. Therefore, intracellular iron concentration may influence the interplay between pathogens and immune system. Here, we investigated whether changes in iron concentrations and intracellular ferritin heavy chain (FTH) abundance may modulate the expression of Major Histocompatibility Complex molecules (MHC), and susceptibility to Natural Killer (NK) cell cytotoxicity. FTH downregulation, either by shRNA transfection or iron chelation, led to MHC surface reduction in primary cancer cells and macrophages. On the contrary, mouse embryonic fibroblasts (MEFs) from NCOA4 null mice accumulated FTH for ferritinophagy impairment and displayed MHC class I cell surface overexpression. Low iron concentration, but not FTH, interfered with IFN-γ receptor signaling, preventing the increase of MHC-class I molecules on the membrane by obstructing STAT1 phosphorylation and nuclear translocation. Finally, iron depletion and FTH downregulation increased the target susceptibility of both primary cancer cells and macrophages to NK cell recognition. In conclusion, the reduction of iron and FTH may influence the expression of MHC class I molecules leading to NK cells activation

Waveguiding and SERS Simplified Raman Spectroscopy on Biological Samples

Biomarkers detection at an ultra-low concentration in biofluids (blood, serum, saliva, etc.) is a key point for the early diagnosis success and the development of personalized therapies. However, it remains a challenge due to limiting factors like (i) the complexity of analyzed media, and (ii) the aspecificity detection and the poor sensitivity of the conventional methods. In addition, several applications require the integration of the primary sensors with other devices (microfluidic devices, capillaries, flasks, vials, etc.) where transducing the signal might be difficult, reducing performances and applicability. In the present work, we demonstrate a new class of optical biosensor we have developed integrating an optical waveguide (OWG) with specific plasmonic surfaces. Exploiting the plasmonic resonance, the devices give consistent results in surface enhanced Raman spectroscopy (SERS) for continuous and label-free detection of biological compounds. The OWG allows driving optical signals in the proximity of SERS surfaces (detection area) overcoming spatial constraints, in order to reach places previously optically inaccessible. A rutile prism couples the remote laser source to the OWG, while a Raman spectrometer collects the SERS far field scattering. The present biosensors were implemented by a simple fabrication process, which includes photolithography and nanofabrication. By using such devices, it was possible to detect cell metabolites like Phenylalanine (Phe), Adenosine 5-triphosphate sodium hydrate (ATP), Sodium Lactate, Human Interleukin 6 (IL6), and relate them to possible metabolic pathway variation

Sublime communion: The theology of the natural world in Laudato Si\u27

Laudato Si’ offers a profound and in some respects new theology of the natural world. In the analysis offered here, it is proposed that three threads can be discerned in the theology of nature contained in Laudato Si’: first, other creatures have intrinsic value; second, they express and reveal God; third, they form with human beings a sublime communion of creation in God. The article concludes with a brief theological reflection on a theological development of the concept of sublime communion

Proteins identified by nanoLC-MS/MS analysis of IP bands.

<p>(*) The Mascot score for each individual peptide is reported in parenthesis. Significance threshold (p<0.05) corresponded to a Mascot score of 15. Protein identifications based on a single peptide above the significance threshold were validated by careful visual inspection of MS/MS data and western blot analysis.</p

Expression of activated Capn3 in urothelial tumors.

<p>Detection of IS1 and IS2 inserts in Capn3 transcript. PCR analysis was performed on the cDNAs synthesized from bladders isolated from pathological animals or human PBMC as reported in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0010299#s2" target="_blank">Methods</a>. The primer pair utilized to detect Capn3 IS1 insert was Sn831/Asn1845 (A) and that utilized to detect IS2 insert was Sn1884/Asn2669 (B). PCR conditions are reported in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0010299#s2" target="_blank">Methods</a>. PCR products were separated by electrophoresis on 1.2% agarose gel. Marker sizes: GeneRuler 100 bp DNA ladder (Fermentas). cDNA amounts are normalized with respect to equal GAPDH levels. The expected sizes for the PCR fragments in (A) are 1018 bp with IS1, and 829 bp without IS1. The expected sizes for the PCR fragments in (B) are 805 bp with IS2, and 577 bp without IS2.</p

Expression of activated Capn3 in urothelial tumors.

<p>Western blot analysis to characterize the expression of E2F3. Lane 1, healthy animal as control; lanes 2–7, E2F3 protein appears to be overexpressed in all examined tumor samples.</p

Expression of activated Capn3 in urothelial tumors.

<p>Capn3 expression is also detected in some nuclei. Streptavidin-biotin-peroxidase, Mayer's hematoxylin counterstain.</p

Expression of activated Capn3 in urothelial tumors.

<p>A weak immunoreactivity can be seen in urothelial cells of the grossly unaffected mucosa. Streptavidin-biotin-peroxidase, Mayer's hematoxylin counterstain.</p