Single-step production of functional OEP24 proteoliposomes

The pea chloroplastic outer envelope protein OEP24 is a voltage-dependent channel that can function as a general solute channel in plants. OEP24 is a close functional homologue of VDAC which, in mammalian cells, modulates the permeability of the outer mitochondrial membrane. Here, we describe the production in a one-step reaction of active OEP24 in proteoliposomes or in soluble form using a cell-free expression system. We combine evidence from electrophysiological experiments, biophysical characterization, and biochemical analysis demonstrating that OEP24 is present as a functional channel in liposomes. Thus, production of OEP-containing proteoliposomes may provide a helpful tool for deciphering the role of the OEP family members.     

Patterns of MADS-box gene expression mark flower-type development in Gerbera hybrida (Asteraceae)

Background  

The inflorescence of the cut-flower crop Gerbera hybrida (Asteraceae) consists of two principal flower types, ray and disc, which form a tightly packed head, or capitulum. Despite great interest in plant morphological evolution and the tractability of the gerbera system, very little is known regarding genetic mechanisms involved in flower type specification. Here, we provide comparative staging of ray and disc flower development and microarray screening for differentially expressed genes, accomplished via microdissection of hundreds of coordinately developing flower primordia.     

Multiplexed CRISPR/CAS9‐mediated engineering of pre‐clinical mouse models bearing native human B cell receptors

B‐cell receptor (BCR) knock‐in (KI) mouse models play an important role in vaccine development and fundamental immunological studies. However, the time required to generate them poses a bottleneck. Here we report a one‐step CRISPR/Cas9 KI methodology to combine the insertion of human germline immunoglobulin heavy and light chains at their endogenous loci in mice. We validate this technology with the rapid generation of three BCR KI lines expressing native human precursors, instead of computationally inferred germline sequences, to HIV broadly neutralizing antibodies. We demonstrate that B cells from these mice are fully functional: upon transfer to congenic, wild type mice at controlled frequencies, such B cells can be primed by eOD‐GT8 60mer, a germline‐targeting immunogen currently in clinical trials, recruited to germinal centers, secrete class‐switched antibodies, undergo somatic hypermutation, and differentiate into memory B cells. KI mice expressing functional human BCRs promise to accelerate the development of vaccines for HIV and other infectious diseases.     

Host‐parasite relationship: occurrence and effect of Ceratothoa parallela (Otto, 1828) on Boops boops (L., 1758) in the Southern Tyrrhenian Sea

The host‐parasite relationship between bogue (Boops boops) and Cerathotoa parallela (Isopoda: Cymothoidae) was studied in the Southern Tyrrhenian Sea (overall prevalence of isopods in the area, age‐dependent and sex‐dependent prevalence). Yearly prevalence ranged from 1.0% in March to 10.7% in October, with a positive correlation between prevalence and mean sea surface temperatures (SST), highest values being related to the period of increasing SST. The first four age‐classes were parasitized, with prevalence decreasing according to the age of the host. The relationship between total length (TL) and total weight (TW) was examined in infested and non‐infested specimens of the same age whereby slight differences were detected, i.e. at a given length non‐infested specimens weighed more than infested fishes. The parasite occurred throughout the study area with wide fluctuations in prevalence, from 0 to 36.4%, the values being higher in the eastern part of the study area. C. parallela parasitize the bogue Boops boops in a manner that does not constitute a serious threat to the life of the host.     

Microbiological quality of drinking water from dispensers in Italy

Background  

Water coolers are popular in office buildings and commercial stores and the quality of this source of drinking water has the potential to cause waterborne outbreaks, especially in sensitive and immunocompromised subjects. The aim of this study was to determine the quality of water plumbed in coolers from commercial stores in comparison with tap water in Italy.