Validation of a CE-IVD, urine exosomal RNA expression assay for risk assessment of prostate cancer prior to biopsy.

Improved risk stratification of patients suspected of prostate cancer prior to biopsy continues to be an unmet clinical need. ExoDx Prostate (IntelliScore) "EPI" is a non-invasive urine test utilizing RNA from exosomes to provide a risk score that correlates with the likelihood of finding high grade prostate cancer at biopsy. Here, we present the results from a prospective clinical validation study of EPI-CE, a CE-marked in-vitro diagnostic (IVD) assay, specifically developed for use in European clinical laboratories. The study (NCT04720599) enrolled patients with ≥ 50 years, PSA 2-10 ng/mL, prior to MRI, who were scheduled for initial biopsy. First catch urine samples were collected from participants without prior digital rectal examination or prostate massage. Exosomal RNA was isolated and expression levels of three biomarkers ERG, PCA3 and SPDEF were analyzed according to the EPI-CE Instructions For Use. In the study cohort of N = 109 patients, EPI-CE was validated to have a Negative Predictive Value of 89%, a Sensitivity of 92% and a superior performance to two commonly used multiparametric risk calculators (PCPT and ERSPC) in both Receiver Operating Characteristics with a higher Area Under the Curve for EPI-CE 0.67 (95% CI 0.56-0.77) versus PCPT 0.59 (95% CI 0.47-0.71) and ERSPC 0.60 (95% CI 0.49-0.72) and higher Net Benefits analysis across a wide range of risk acceptance levels. This is the first clinical study reporting on the performance of EPI-CE. We demonstrate that EPI-CE provides information beyond standard clinical parameters and provides a better risk assessment prior to MRI, of patients suspected of prostate cancer, than the commonly used multiparametric risk calculators.

MitoSegNet: Easy-to-use Deep Learning Segmentation for Analyzing Mitochondrial Morphology.

While the analysis of mitochondrial morphology has emerged as a key tool in the study of mitochondrial function, efficient quantification of mitochondrial microscopy images presents a challenging task and bottleneck for statistically robust conclusions. Here, we present Mitochondrial Segmentation Network (MitoSegNet), a pretrained deep learning segmentation model that enables researchers to easily exploit the power of deep learning for the quantification of mitochondrial morphology. We tested the performance of MitoSegNet against three feature-based segmentation algorithms and the machine-learning segmentation tool Ilastik. MitoSegNet outperformed all other methods in both pixelwise and morphological segmentation accuracy. We successfully applied MitoSegNet to unseen fluorescence microscopy images of mitoGFP expressing mitochondria in wild-type and catp-6 ATP13A2 mutant C. elegans adults. Additionally, MitoSegNet was capable of accurately segmenting mitochondria in HeLa cells treated with fragmentation inducing reagents. We provide MitoSegNet in a toolbox for Windows and Linux operating systems that combines segmentation with morphological analysis.

Lewis acidic alkaline earth metal complexes with a perfluorinated diphenylamide ligand.

Alkaline earth metal (Ae) chemistry with the anion [N(C6F5)2]- has been explored. Deprotonation of the amine (C6F5)2NH, abbreviated in here as NFH, with 0.5 equivalent of AeN''2 (N'' = N(SiMe3)2) is fast and gave, dependent on the solvent, the complexes AeNF2, AeNF2·(THF)2 and AeNF2·(Et2O)2 (Ae = Mg, Ca, Sr). Using a 1/1 ratio, mixed amide complexes were obtained: NFAeN'' (Ae = Mg, Ca, Sr). Crystal structures of the monomers AeNF2·(THF)2 (Ae = Mg, Ca, Sr) and AeNF2·(Et2O)2 (Ae = Mg, Ca) are presented and compared with those of AeN''2·(THF)2. In addition, crystal structures of the homoleptic dimer (MgNF2)2 and the heteroleptic dimers (NFAeN'')2 (Ae = Mg, Ca, Sr) are discussed. All structures are strongly influenced by very short AeF contacts down to circa 2.11 Å (Mg), 2.50 Å (Ca) and 2.73 Å (Sr). AIM analysis illustrates that, although AeF contacts are short, there is no bond-critical-point along this axis, indicating an essentially electrostatic interaction. The monomeric complexes feature strong C6F5C6F5π-stacking, resulting in unusually acute NF-Ae-NF angles as small as 95°. Heteroleptic (NFAeN'')2 complexes retain their dimeric structure in C6D6 solution and there is no indication of ligand scrambling by the Schlenk equilibrium, suggesting that an electron withdrawing ligand may stabilize heteroleptic complexes. According to DFT calculations, the heteroleptic arrangement is 70 kJ mol-1 more stable than the homoleptic dimers. The Lewis acidity of MgNF2 has been quantified with the Gutmann-Beckett method and by calculation of the Fluoride-Ion-Affinity. The latter calculations show that the Lewis acidity of MgNF2 and CaNF2 is comparable to that of B(C6F5)3. Dimeric (MgNF2)2 fully abstracts Et3PO from Et3PO·B(C6F5)3 and may have potential in Lewis acid catalysis.

Nucleophilic Aromatic Substitution at Benzene with Powerful Strontium Hydride and Alkyl Complexes.

Key to the isolation of the first alkyl strontium complex was the synthesis of a strontium hydride complex that is stable towards ligand exchange reactions. This goal was achieved by using the super bulky ß-diketiminate ligand DIPeP BDI (CH[C(Me)N-DIPeP]2 , DIPeP=2,6-diisopentylphenyl). Reaction of DIPeP BDI-H with Sr[N(SiMe3 )2 ]2 gave (DIPeP BDI)SrN(SiMe3 )2 , which was converted with PhSiH3 into [(DIPeP BDI)SrH]2 . Dissolved in C6 D6 , the strontium hydride complex is stable up to 70 °C. At 60 °C, H-D isotope exchange gave full conversion into [(DIPeP BDI)SrD]2 and C6 D5 H. Since H-D exchange with D2 is facile, the strontium hydride complex served as a catalyst for the deuteration of C6 H6 by D2 . Reaction of [(DIPeP BDI)SrH]2 with ethylene gave [(DIPeP BDI)SrEt]2 . The high reactivity of this alkyl strontium complex is demonstrated by facile ethylene polymerization and nucleophilic aromatic substitution with C6 D6 , giving alkylated aromatic products and [(DIPeP BDI)SrD]2 .

Bora-amidinate as a cooperative ligand in group 2 metal catalysis.

Syntheses and crystal structures of the monomeric bora-amidinate (bam) complexes DIPPNBN-Mg·(THF)3 and DIPPNBN-Ca·(THF)4 are presented; DIPPNBN = HB[N(2,6-iPr2-C6H3)]2. The simplicity of their 1H NMR spectra in THF-d8 suggest that their monomeric solid state structures are retained in solution. DIPPNBN-Mg·(THF)3 in C6D6, however, is in equilibrium with a dimeric species. Calculations (B3PW91/6-311++G**) reveal a very high localized negative charge (NPA: -1.103) on the N atoms in DIPPNBN-Mg. The strongly basic properties of the bam ligand are in agreement with catalytic activity of these complexes in the intramolecular alkene hydroamination. A mechanism is proposed in which the bam ligand is non-innocent and cooperative, playing an active role in substrate deprotonation and product protonation.