Hepatitis C virus NS5A is a direct substrate of casein kinase I-α, a cellular kinase identified by inhibitor affinity chromatography using specific NS5A hyperphosphorylation inhibitors

The hepatitis C virus encodes a single polyprotein that is processed by host and viral proteases to yield at least 10 mature viral proteins. The nonstructural (NS) protein 5A is a phosphoprotein, and experimental data indicate that the phosphorylation state of NS5A is important for the outcome of viral RNA replication. We were able to identify kinase inhibitors that specifically inhibit the formation of the hyperphosphorylated form of NS5A (p58) in cells. These kinase inhibitors were used for inhibitor affinity chromatography in order to identify the cellular targets of these compounds. The kinases casein kinase I (CKI), p38 MAPK, CIT (Citron Rho-interacting kinase), GAK, JNK2, PKA, RSK1/2, and RIPK2 were identified in the high affinity binding fractions of two NS5A hyperphosphorylation inhibitors (NS5A-p58-i). Even though these kinases are targets of the NS5A-p58-i, the only kinase showing an effect on NS5A hyperphosphorylation was confirmed to be CKI-\u3b1. Although this finding does not exclude the possibility that other kinase(s) might be involved in basal or regulatory phosphorylation of NS5A, we show here that NS5A is a direct substrate of CKI-\u3b1. Moreover, in vitro phosphorylation of NS5A by CKI-\u3b1 resulted for the first time in the production of basal and hyperphosphorylated forms resembling those produced in cells. In vitro kinase reactions performed with NS5A peptides show that Ser-2204 is a preferred substrate residue for CKI-\u3b1 after pre-phosphorylation of Ser-2201

A Bayesian predictive two stage design for phase II clinical trials: the case of continuous endpoints

Single-arm two-stage designs for phase II of clinical trials typically focus on a binary endpoint obtained by dichotomising an underlying continuous measure of treatment efficacy. To avoid the resulting loss of information, we propose a two-stage design based on a Bayesian predictive approach that directly uses the original continuous endpoint. Numerical results are provided with reference to phase II cancer trials aimed at assessing tumour shrinking effect of an experimental treatment

Sharing Italian Botanic Gardens\u2019 living collections: The role of the National Biodiversity Network

This paper presents the role of the Italian National Biodiversity Network in making available biodiversity data from Italian Botanic Gardens at a national and international level. The case study of the Botanic Garden of Rome is presented explaining procedures and methods for collecting georeferenced data on living plant species and making them available through web-based applications

Convergence for varying measures

Some limit theorems of the type ∫Ωfndmn→∫Ωfdm are presented for scalar, (vector), (multi)-valued sequences of mn-integrable functions fn. The convergences obtained, in the vector and multivalued settings, are in the weak or in the strong sense