Impact of Collection Volume and DNA Extraction Method on the Detection of Biomarkers and HPV DNA in First-Void Urine

The potential of first-void (FV) urine as a non-invasive liquid biopsy for detection of human papillomavirus (HPV) DNA and other biomarkers has been increasingly recognized over the past decade. In this study, we investigated whether the volume of this initial urine stream has an impact on the analytical performance of biomarkers. In parallel, we evaluated different DNA extraction protocols and introduced an internal control in the urine preservative. Twenty-five women, diagnosed with high-risk HPV, provided three home-collected FV urine samples using three FV urine collection devices (Colli-Pee) with collector tubes that differ in volume (4, 10, 20 mL). Each collector tube was prefilled with Urine Conservation Medium spiked with phocine herpesvirus 1 (PhHV-1) DNA as internal control. Five different DNA extraction protocols were compared, followed by PCR for GAPDH and PhHV-1 (qPCR), HPV DNA, and HBB (HPV-Risk Assay), and ACTB (methylation-specific qPCR). Results showed limited effects of collection volume on human and HPV DNA endpoints. In contrast, significant variations in yield for human endpoints were observed for different DNA extraction methods (p < 0.05). Additionally, the potential of PhHV-1 as internal control to monitor FV urine collection, storage, and processing was demonstrated.     

On the Poles of Maximal Order of the Topological Zeta Function

The global and local topological zeta functions are singularityinvariants associated to a polynomial f and its germ at 0, respectively.By definition, these zeta functions are rational functions inone variable, and their poles are negative rational numbers.In this paper we study their poles of maximal possible order.When f is non-degenerate with respect to its Newton polyhedron,we prove that its local topological zeta function has at mostone such pole, in which case it is also the largest pole; wegive a similar result concerning the global zeta function. Moreover,for any f we show that poles of maximal possible order are alwaysof the form –1/N with N a positive integer. 1991 MathematicsSubject Classification 14B05, 14E15, 32S50.     

Arrangements with one bounded component andp-adic integrals

In this article we study arrangementsA, such that ℝ ⁿ \A has exactly one bounded component. We obtain a result about their structure which gives us a method to construct all combinatorially different such arrangements in a given dimension. (A complete list for dimensions 1,2,3 and 4 is included). Furthermore we associate ap-adic integral to each such arrangement and proof that this integral can be written as a product ofp-adic beta functions. This is analogous to results of Varchenko and Loeser for integrals over ℝ and character sums over finite fields respectively.     

Nanobody‐Enabled Reverse Pharmacology on G‐Protein‐Coupled Receptors

The conformational complexity of transmembrane signaling of G‐protein‐coupled receptors (GPCRs) is a central hurdle for the design of screens for receptor agonists. In their basal states, GPCRs have lower affinities for agonists compared to their G‐protein‐bound active state conformations. Moreover, different agonists can stabilize distinct active receptor conformations and do not uniformly activate all cellular signaling pathways linked to a given receptor (agonist bias). Comparative fragment screens were performed on a β₂‐adrenoreceptor–nanobody fusion locked in its active‐state conformation by a G‐protein‐mimicking nanobody, and the same receptor in its basal‐state conformation. This simple biophysical assay allowed the identification and ranking of multiple novel agonists and permitted classification of the efficacy of each hit in agonist, antagonist, or inverse agonist categories, thereby opening doors to nanobody‐enabled reverse pharmacology.