Lipid Binding Defects and Perturbed Synaptogenic Activity of a Collybistin R290H Mutant That Causes Epilepsy and Intellectual Disability

Signaling at nerve cell synapses is a key determinant of proper brain function, and synaptic defects—or synaptopathies—are at the basis of many neurological and psychiatric disorders. In key areas of the mammalian brain, such as the hippocampus or the basolateral amygdala, the clustering of the scaffolding protein Gephyrin and of γ-aminobutyric acid type A receptors at inhibitory neuronal synapses is critically dependent upon the brain-specific guanine nucleotide exchange factor Collybistin (Cb). Accordingly, it was discovered recently that an R290H missense mutation in the diffuse B-cell lymphoma homology domain of Cb, which carries the guanine nucleotide exchange factor activity, leads to epilepsy and intellectual disability in human patients. In the present study, we determined the mechanism by which the Cb^R290H mutation perturbs inhibitory synapse formation and causes brain dysfunction. Based on a combination of biochemical, cell biological, and molecular dynamics simulation approaches, we demonstrate that the R290H mutation alters the strength of intramolecular interactions between the diffuse B-cell lymphoma homology domain and the pleckstrin homology domain of Cb. This defect reduces the phosphatidylinositol 3-phosphate binding affinity of Cb, which limits its normal synaptogenic activity. Our data indicate that impairment of the membrane lipid binding activity of Cb and a consequent defect in inhibitory synapse maturation represent a likely molecular pathomechanism of epilepsy and mental retardation in humans.     

Ecological patterns strongly impact the biogeography of western Palaearctic longhorn beetles (Coleoptera: Cerambycoidea)

We aim to unravel the biogeographic structuring of western Palaearctic longhorn beetles with focus on the location of different refugia, barriers to dispersal and postglacial range expansions with their particular filters. The interaction of different ecological features with these structures is analysed. The western Palaearctic was divided into 95 geographic entities. We produced presence-only matrices for all 955 Cerambycoidea species autochthonous to this area and derived species richness distributions and extracted faunal regions and faunal elements by cluster analyses and principal component analyses. Similar analyses were performed for sub-families and ecological groups. Longhorn beetles show a strong biogeographic structuring in the western Palaearctic. Species numbers strongly decrease to the north and west. Less mobile species and root feeders mostly contribute to the fauna of the Mediterranean region, whilst mobile species are more widespread. Feeders on broad-leaved trees dominate in western Europe, whilst feeders on coniferous trees are most important in northern Europe. Our results support multiple refugia in the Mediterranean region and underline the importance of Provence, Crimea and Crete as such refugia. Crete even might be an area of old endemism. The Atlanto- and the Ponto-Mediterranean regions are more strongly structured than assumed in classical biogeography. Mediterranean assemblages are mostly composed of non-flying species, root feeders and species with small distributions not found outside their glacial refugia. Tree feeders left their glacial retreats with their host plants. These range dynamics result in biogeographic structures with several dispersal barriers and filters composed of mountains, sea straits and climatic conditions.     

Comparison of different culture conditions for smooth muscle cell differentiation of human umbilical cord vein CD146+ perivascular cells

Pericytes are CD146+ perivascular cells (PCs) that have multipotential differentiation capacity as mesenchymal stem cells. Beside their crucial roles in vascular development and blood flow regulation, they have ability to differentiate into vascular cell types in vivo. These properties make pericytes preferred cells in the field of vascular tissue engineering. Culture medium for in vitro differentiation of pericytes to vascular smooth muscle cells (SMCs) has not been defined yet. The aim of this study is to try different culture media for SMC differentiation of CD146+ PCs. For this purpose, CD146+ PCs were isolated from human umbilical cord vein. Then they were characterized by immunofluorescence staining and flow cytometric analysis. Three different culture media including; (1) Transforming growth factor beta 1 (TGF-β1)+ bone morphogenic protein 4, (2) TGF-β1+ l-ascorbic acid (l-AA) and (3) Horse serum, were compared for differentiation of CD146+ PCs to SMCs by IFS and real time polymerase chain reaction. As a result, in the case of SMC differentiation of CD146+ PCs, second culture medium including TGF-β1 and l-AA was found to be more effective than other two media. These results are important for establishing proper culture conditions for in vitro SMC differentiation of CD146+ PCs.     

Endemic diversification in the mountains: genetic,morphological, and geographical differentiation of the <Emphasis Type="Italic">Hemidactylus</Emphasis> geckos in southwestern Arabia

In this study, we provide genetic, morphological, and geographical comparisons for 11 species of the southwestern Arabian radiation of Hemidactylus geckos, nine of which are endemic to the region. By using a coalescence-based species-tree reconstruction in combination with divergence time estimations and speciation probability testing, we show that most of the speciation events occurred in the Pliocene, which is more recent than previously thought based on calibrations of concatenated data sets. The current dating indicates that the changing climate at the beginning of the Pliocene, from hot and dry to cold and wet, is likely responsible for increased speciation in Hemidactylus. Analyses of geographic and altitudinal overlap of the species and their morphological differentiation show that most species do not occur in sympatry. Those that overlap geographically are usually differentiated by their altitudinal preference, head shape, body size, or their combination. Our results indicate that the topographically complex mountains of southwestern Arabia support a significant radiation of Hemidactylus geckos by allowing multiple allopatric speciation events to occur in a relatively small area. Consequently, we describe two new species endemic to the Asir Mountains of Saudi Arabia, H. alfarraji sp. n. and H. asirensis sp. n., and elevate two former subspecies of H. yerburii to a species level, H. montanus and H. pauciporosus.     

Multistep Photoluminescence Decay Reveals Dissociation of Geminate Charge Pairs in Organolead Trihalide Perovskites

Charge carrier dynamics in organolead iodide perovskites is analyzed by employing time‐resolved photoluminescence spectroscopy with several ps time resolution. The measurements performed by varying photoexcitation intensity over five orders of magnitude enable separation of photoluminescence components related to geminate and nongeminate charge carrier recombination and to address the dynamics of an isolated geminate electron–hole pair. Geminate recombination dominates at low excitation fluence and determines the initial photoluminescence decay. This decay component is remarkably independent of the material structure and experimental conditions. It is demonstrated that dependences of the geminate and nongeminate radiative recombination components on excitation intensity, repetition rate, and temperature, are hardly compatible with carrier trapping and exciton dissociation models. On the basis of semiclassical and quantum mechanical numerical calculation results, it is argued that the fast photoluminescence decay originates from gradual spatial separation of photogenerated weakly bound geminate charge pairs.     

New Type of Papillomavirus and Novel Circular Single Stranded DNA Virus Discovered in Urban Rattus norvegicus Using Circular DNA Enrichment and Metagenomics

Rattus norvegicus (R. norvegicus) are ubiquitous and their presence has several effects on the human populations in our urban areas on a global scale. Both historically and presently, this close interaction has facilitated the dissemination of many pathogens to humans, making screening for potentially zoonotic and emerging viruses in rats highly relevant. We have investigated faecal samples from R. norvegicus collected from urban areas using a protocol based on metagenomic enrichment of circular DNA genomes and subsequent sequencing. We found a new type of papillomavirus, with a L1 region 82% identical to that of the known R. norvegicus Papillomavirus 2. Additionally, we found 20 different circular replication associated protein (Rep)-encoding single stranded DNA (CRESS-DNA) virus-like genomes, one of which has homology to the replication-associated gene of Beak and feather disease virus. Papillomaviruses are a group of viruses known for their carcinogenic potential, and although they are known to infect several different vertebrates, they are mainly studied and characterised in humans. CRESS-DNA viruses are found in many different environments and tissue types. Both papillomaviruses and CRESS-DNA viruses are known to have pathogenic potential and screening for novel and known viruses in R. norvegicus could help identify viruses with pathogenic potential.     

Purification of the Chlorella HUP1 hexose—proton symporter to homogeneity and its reconstitution in vitro

A prokaryotic biotin acceptor domain was fused to the carboxy terminal end of the Chlorella hexose—proton sym- porter. The plant symporter is biotinylated in vivo when expressed in Schizosaccharomyces pombe. The extended biotinylated transport protein is fully active, catalyzes accumulation of d -glucose analogs and restores growth of a glucose-uptake-deficient yeast strain. Crude membranes were solubilized with octyl-β-d -glucoside in the presence of Escherichia colil -α-phosphatidylethanolamine. Biotinylated symporter was purified to homogeneity by biotinavidin affinity chromatography. The symporter protein was reconstituted together with cytochrome-c oxidase prepared from beef heart mitochondria into proteo-liposomes. Cytochrome-c oxidase is a redox-driven H⁺-pump generating a proton motive force (inside negative and alkaline) while transferring electrons from cytochrome-c to oxygen; this energy is used by the symporter to accumulate d -glucose at least 30-fold. In the absence of the driving force the transport protein facilitates diffusion of d -glucose until the concentration equilibrium is reached. It was shown that maximal transport activity depends highly on the amount of co-reconstituted cytochrome-c oxidase and that the symporter possesses 10% of its in vivo turnover number under optimized in vitro transport conditions.