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Background: There are different types of computed tomography (CT) contrast enhancement patterns of the uterus. It is not known whether these are hormonally dependent. Purpose: To assess the relationship between these patterns and the menstrual cycle in non-users of hormonal contraception, and the possible impact of hormonal contraception. Material and Methods: Prospective observational study of abdominal CT scans of 53 premenopausal women of whom 28 were non-users and 25 users of hormonal contraception. The non-users were divided according to menstrual cycle phase: follicular (n = 12); ovulatory (n = 1); and luteal (n = 12). The pattern and intensity of contrast enhancement of the uterine myometrium were assessed. Results: The dominant pattern of contrast enhancement of the myometrium was the diffuse homogeneous type in both non-users and users. The intensity of the enhancement measured in Hounsfield units (HU) was higher in the follicular phase (median 102, range 73–130) compared to the luteal phase in non-users (median 92, range 57–130); however this was not statistically significant (P = 0.2). The HU values observed in users (median 95, range 45–160) were at the same levels compared to those of the luteal phase in non-users. Conclusion: The dominant pattern of contrast enhancement in the portal venous phase of the myometrium in fertile ages is the diffuse homogeneous type and is independent of menstrual cycle phase or the use of hormonal contraception. However, these factors seem to play a role in the intensity of contrast enhancement, with a tendency of higher HU values in the follicular phase of non-users. © The Foundation Acta Radiologica 2020.

Conventional dendritic cell (DC) vaccine strategies, in which DCs are loaded with antigens ex vivo, suffer biological issues such as impaired DC migration capacity and laborious GMP production procedures. In a promising alternative, antigens are targeted to DC-associated endocytic receptors in vivo with antibody-antigen conjugates co-administered with toll-like receptor (TLR) agonists as adjuvants. To combine the potential advantages of in vivo targeting of DCs with those of conjugated TLR agonists, we generated a multifunctional antibody construct integrating the DC-specific delivery of viral- or tumor-associated antigens and DC activation by TLR ligation in one molecule. We validated its functionality in vitro and determined if TLR ligation might improve the efficacy of such a molecule. In proof-of-principle studies, an alpha CD40 antibody containing a CMV pp65-derived peptide as an antigen domain (alpha CD40(CMV)) was genetically fused to the TLR5-binding D0/D1 domain of bacterial flagellin (alpha CD40.Flg(CMV)). The analysis of surface maturation markers on immature DCs revealed that fusion of flagellin to alpha CD40(CMV) highly increased DC maturation (3.4-fold elevation of CD80 expression compared to alpha CD40(CMV) alone) by specifically interacting with TLR5. Immature DCs loaded with alpha CD40.Flg(CMV) induced significantly higher CMVNLV-specific T cell activation and proliferation compared to alpha CD40(CMV) in co-culture experiments with allogeneic and autologous T cells (1.8-fold increase in % IFN-gamma/TNF-alpha(+) CD8(+) T cells and 3.9-fold increase in % CMVNLV-specific dextramer(+) CD8(+) T cells). More importantly, we confirmed the beneficial effects of flagellin-dependent DC stimulation using a tumor-specific neoantigen as the antigen domain. Specifically, the acute myeloid leukemia (AML)-specific mutated NPM1 (mNPM1)-derived neoantigen CLAVEEVSL was delivered to DCs in the form of alpha CD40(mNPM1) and alpha CD40.Flg(mNPM1) antibody constructs, making this study the first to investigate mNPM1 in a DC vaccination context. Again, alpha CD40.Flg(mNPM1)-loaded DCs more potently activated allogeneic mNPM1(CLA)-specific T cells compared to alpha CD40(mNPM1). These in vitro results confirmed the functionality of our multifunctional antibody construct and demonstrated that TLR5 ligation improved the efficacy of the molecule. Future mouse studies are required to examine the T cell-activating potential of alpha CD40.Flg(mNPM1) after targeting of dendritic cells in vivo using AML xenograft models.

To improve light propagation through the retina, the rod nuclei of nocturnal mammals are uniquely changed compared to the nuclei of other cells. In particular, the main classes of chromatin are segregated in them and form regular concentric shells in order; inverted in comparison to conventional nuclei. A broad study of the epigenetic landscape of the inverted and conventional mouse retinal nuclei indicated several differences between them and several features of general interest for the organization of the mammalian nuclei. In difference to nuclei with conventional architecture, the packing density of pericentromeric satellites and LINE-rich chromatin is similar in inverted rod nuclei; euchromatin has a lower packing density in both cases. A high global chromatin condensation in rod nuclei minimizes the structural difference between active and inactive X chromosome homologues. DNA methylation is observed primarily in the chromocenter, Dnmt1 is primarily associated with the euchromatic shell. Heterochromatin proteins HP1-alpha and HP1-beta localize in heterochromatic shells, whereas HP1-gamma is associated with euchromatin. For most of the 25 studied histone modifications, we observed predominant colocalization with a certain main chromatin class. Both inversions in rod nuclei and maintenance of peripheral heterochromatin in conventional nuclei are not affected by a loss or depletion of the major silencing core histone modifications in respective knock-out mice, but for different reasons. Maintenance of peripheral heterochromatin appears to be ensured by redundancy both at the level of enzymes setting the epigenetic code (writers) and the code itself, whereas inversion in rods rely on the absence of the peripheral heterochromatin tethers (absence of code readers).

We show systematic electrical impedance measurements of single motile cells on microelectrodes. Wild-type cells and mutant strains were studied that differ in their cell-substrate adhesion strength. We recorded the projected cell area by time-lapse microscopy and observed irregular oscillations of the cell shape. These oscillations were correlated with long-term variations in the impedance signal. Superposed to these long-term trends, we observed fluctuations in the impedance signal. Their magnitude clearly correlated with the adhesion strength, suggesting that strongly adherent cells display more dynamic cell-substrate interactions.