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.     

<Emphasis Type="Italic">Artemisia lerchiana</Emphasis> as a producer of essential oil

Abstrac  The composition of essential oil of Artemisia lerchiana Web. plants growing in Volgograd oblast was studied. Sampling was performed from plots contrasting in climatic and soil characteristics. Essential oil was obtained by hydrodistillation. The content of essential oil in shoot biomass increased gradually during shoot formation, flower bud formation, and flowering beginning and then decreased. The highest content of essential oil varied from 1.1 to 1.5% of plant dry weight at the stage of flower bud formation. More than thirty compounds were identified by gas chromatography-mass spectrometry. The following major components were found: camphor, borneol, bornylacetate, camphene, and 1,8-cineole. Some of compounds (sesquiterpenes and sesquiterpenoids) were identified for the first time. The time-course of accumulation of essential oil components strongly depended on habitat edaphic factors and climatic conditions during the year of sampling. The results permit a conclusion that A. lerchiana is a valuable producer of essential oils. Original Russian Text ? E.B. Kirichenko, Yu.V. Orlova, D.V. Kurilov, 2008, published in Fiziologiya Rastenii, 2008, Vol. 55, No. 6, pp. 934–941.     

Residue Histidine 50 Plays a Key Role in Protecting α-Synuclein from Aggregation at Physiological pH

α-Synuclein (αSyn) aggregation is involved in the pathogenesis of Parkinson disease (PD). Recently, substitution of histidine 50 in αSyn with a glutamine, H50Q, was identified as a new familial PD mutant. Here, nuclear magnetic resonance (NMR) studies revealed that the H50Q substitution causes an increase of the flexibility of the C-terminal region. This finding provides direct evidence that this PD-causing mutant can mediate long range effects on the sampling of αSyn conformations. In vitro aggregation assays showed that substitution of His-50 with Gln, Asp, or Ala promotes αSyn aggregation, whereas substitution with the positively charged Arg suppresses αSyn aggregation. Histidine carries a partial positive charge at neutral pH, and so our result suggests that positively charged His-50 plays a role in protecting αSyn from aggregation under physiological conditions.     

N-terminal Acetylation Stabilizes N-terminal Helicity in Lipid- and Micelle-bound α-Synuclein and Increases Its Affinity for Physiological Membranes

The Parkinson disease protein α-synuclein is N-terminally acetylated, but most in vitro studies have been performed using unacetylated α-synuclein. Binding to lipid membranes is considered key to the still poorly understood function of α-synuclein. We report the effects of N-terminal acetylation on α-synuclein binding to lipid vesicles of different composition and curvature and to micelles composed of the detergents β-octyl-glucoside (BOG) and SDS. In the presence of SDS, N-terminal acetylation results in a slightly increased helicity for the N-terminal ∼10 residues of the protein, likely due to the stabilization of N-terminal fraying through the formation of a helix cap motif. In the presence of BOG, a detergent used in previous isolations of helical oligomeric forms of α-synuclein, the N-terminally acetylated protein adopts a novel conformation in which the N-terminal ∼30 residues bind the detergent micelle in a partly helical conformation, whereas the remainder of the protein remains unbound and disordered. Binding of α-synuclein to lipid vesicles with high negative charge content is essentially unaffected by N-terminal acetylation irrespective of curvature, but binding to vesicles of lower negative charge content is increased, with stronger binding observed for vesicles with higher curvature. Thus, the naturally occurring N-terminally acetylated form of α-synuclein exhibits stabilized helicity at its N terminus and increased affinity for lipid vesicles similar to synaptic vesicles, a binding target of the protein in vivo. Furthermore, the novel BOG-bound state of N-terminally acetylated α-synuclein may serve as a model of partly helical membrane-bound intermediates with a role in α-synuclein function and dysfunction.     

Polymorphism in sexual versus non-sexual disease transmission

Pathogens causing sexually transmitted diseases (STDs) often consist of related strains that cause non-sexually transmitted, or ''ordinary infectious'', diseases (OIDs). We use differential equation models of single populations to derive conditions under which a genetic variant with one (e.g. sexual) transmission mode can invade and successfully displace a genetic variant with a different (e.g. non-sexual) transmission mode. Invasion by an STD is easier if the equilibrium population size in the presence of an OID is smaller; conversely an OID can invade more easily if the equilibrium size of the population with the STD is larger. Invasion of an STD does not depend on the degree of sterility caused by the infection, but does depend on the added mortality caused by a resident OID. In contrast, the ability of an OID to invade a population at equilibrium with an STD decreases as the degree of sterility caused by the STD increases. When equilibrium population sizes for a population infected with an STD are above the point at which non-sexual contacts exceed sexual contacts (the sexual–social crossover point) and when equilibrium population sizes for an OID are below this point, there can be a stable genetic polymorphism for transmission mode. This is most likely when the STD is mildly sterilizing, and the OID causes low or intermediate levels of added mortality. Because we assume the strains are competitively equivalent and there are no heterogeneities associated with the transmission process, the polymorphism is maintained by density-dependent selection brought about by pathogen effects on population size.     

BMP signaling in vascular biology and dysfunction

The vascular system is critical for developmental growth, tissue homeostasis and repair but also for tumor development. Bone morphogenetic protein (BMP) signaling has recently emerged as a fundamental pathway of the endothelium by regulating cardiovascular and lymphatic development and by being causative for several vascular dysfunctions. Two vascular disorders have been directly linked to impaired BMP signaling: pulmonary arterial hypertension and hereditary hemorrhagic telangiectasia. Endothelial BMP signaling critically depends on the cellular context, which includes among others vascular heterogeneity, exposure to flow, and the intertwining with other signaling cascades (Notch, WNT, Hippo and hypoxia). The purpose of this review is to highlight the most recent findings illustrating the clear need for reconsidering the role of BMPs in vascular biology.     

Recruitment patterns of Daphnia: a key for understanding midsummer declines?

The population dynamics of D. galeata was studied during the period May–July of 5 successive years in Bautzen Reservoir (Germany). Sampling intervals of 3–4 days and knowledge of the egg–age distribution made it possible to calculate recruitment of the population for each sampling interval. In every year, high Daphnia densities and high Secchi depth readings (clear-water stage) were recorded in spring. In two of the 5 years, Daphnia abundance remained on a high level throughout summer, whereas in the remaining 3 years a midsummer decline of Daphnia was observed. Birth rates declined while the abundance increased in spring in every year but the timing of this decline in relation to abundance and the onset of the clear-water stage revealed no obvious pattern, distinguishing between years with or without a midsummer decline. However, cumulative recruitment did reveal a clear pattern: in years without a midsummer decline, cumulative recruitment increased slowly, but continually, starting from a low level in early May. By contrast, in years with a midsummer decline, recruitment increased strongly during one week in May, starting from a relatively high density level (>30 ind. l^–1). This quick increase was followed by extended periods with low recruitment. Consequently, in these years the population was dominated by a strong peak-cohort of about the same age during the clear-water stage. If all members of this cohort, while approaching their maximum life-span, die off during a short time interval, this may have a pronounced effect on the initiation of a midsummer decline. From an extensive literature search, evidence is provided that the findings of this study represent a demographic mechanism which is of general importance for the initiation of a midsummer decline in temperate waters.     

Porcine Cloned Embryos Reconstructed with the Cell Nuclei of Tetraploid M-phase Fibroblast Cells Can Restore Normal Diploidy at the Blastocyst Stage

The cell cycle of donor cells as a major factor that affects cloning efficiency remains debatable. G2/M phase cells as a donor can successfully produce cloned animals, but a minimal amount is known regarding nuclear remodeling events. In this study, porcine fetal fibroblasts (PFFs) were carefully synchronized at G1 or M phase as donor cells. Most of the cloned embryos reconstructed from PFFs at G1 (G1-embryos) or M (M-embryos) phase formed a pronucleus-like nucleus (PN) within 6-h post fusion (hpf), but the M-embryos formed PN earlier than the G1-embryos did. Moreover, 77.4% of the M-embryos formed two PNs, whereas the G1-embryos formed a single PN. The rate of extrusion of polar body-like structures by the M-embryos was significantly lower than that extruded by the G1-embryos (26.3% vs. 37.1%, P?相似文献   

Focus: Addiction: Relapse Prevention and the Five Rules of Recovery

There are four main ideas in relapse prevention. First, relapse is a gradual process with distinct stages. The goal of treatment is to help individuals recognize the early stages, in which the chances of success are greatest. Second, recovery is a process of personal growth with developmental milestones. Each stage of recovery has its own risks of relapse. Third, the main tools of relapse prevention are cognitive therapy and mind-body relaxation, which are used to develop healthy coping skills. Fourth, most relapses can be explained in terms of a few basic rules. Educating clients in these rules can help them focus on what is important: 1) change your life (recovery involves creating a new life where it is easier to not use); 2) be completely honest; 3) ask for help; 4) practice self-care; and 5) don’t bend the rules.