A Novel Labeling Approach Identifies Three Stability Levels of Acetylcholine Receptors in the Mouse Neuromuscular Junction In Vivo

The turnover of acetylcholine receptors at the neuromuscular junction is regulated in an activity-dependent manner. Upon denervation and under various other pathological conditions, receptor half-life is decreased., in our setup the same animals are used throughout the whole measurement period, thereby permitting a dramatic reduction of animal numbers at increased data quality. We identified three stability levels of acetylcholine receptors depending on the presence or absence of innervation: one pool of receptors with a long half-life of ∼13 days, a second with an intermediate half-life of ∼8 days, and a third with a short half-life of ∼1 day. Data were highly reproducible from animal to animal and followed simple exponential terms. The principal outcomes of these measurements were reproduced by an optical pulse-labeling assay introduced recently.A novel assay to determine kinetics of acetylcholine receptor turnover with small animal numbers is presented. Our data show that nerve activity acts on muscle acetylcholine receptor stability by at least two different means, one shifting receptor lifetime from short to intermediate and another, which further increases receptor stability to a long lifetime. We hypothesize on possible molecular mechanisms

On the Hyperbolicity of Lorenz Renormalization

We consider infinitely renormalizable Lorenz maps with real critical exponent $\alpha>1$ and combinatorial type which is monotone and satisfies a long return condition. For these combinatorial types we prove the existence of periodic points of the renormalization operator, and that each map in the limit set of renormalization has an associated unstable manifold. An unstable manifold defines a family of Lorenz maps and we prove that each infinitely renormalizable combinatorial type (satisfying the above conditions) has a unique representative within such a family. We also prove that each infinitely renormalizable map has no wandering intervals and that the closure of the forward orbits of its critical values is a Cantor attractor of measure zero.Comment: 63 pages; 10 figure

Dynamics of the Universal Area-Preserving Map Associated with Period Doubling: Hyperbolic Sets

It is known that the famous Feigenbaum-Coullet-Tresser period doubling universality has a counterpart for area-preserving maps of {\fR}^2. A renormalization approach has been used in \cite{EKW1} and \cite{EKW2} in a computer-assisted proof of existence of a "universal" area-preserving map $F_*$ -- a map with orbits of all binary periods 2^k, k \in \fN. In this paper, we consider maps in some neighbourhood of $F_*$ and study their dynamics. We first demonstrate that the map $F_*$ admits a "bi-infinite heteroclinic tangle": a sequence of periodic points $\{z_k\}$, k \in \fZ, |z_k| \converge{{k \to \infty}} 0, \quad |z_k| \converge{{k \to -\infty}} \infty, whose stable and unstable manifolds intersect transversally; and, for any N \in \fN, a compact invariant set on which $F_*$ is homeomorphic to a topological Markov chain on the space of all two-sided sequences composed of $N$ symbols. A corollary of these results is the existence of {\it unbounded} and {\it oscillating} orbits. We also show that the third iterate for all maps close to $F_*$ admits a horseshoe. We use distortion tools to provide rigorous bounds on the Hausdorff dimension of the associated locally maximal invariant hyperbolic set: $$ 0.7673 \ge {\rm dim}_H(\cC_F) \ge \varepsilon \approx 0.00044 e^{-1797}.$

Проведение липидной «он лайн» гемодиафильтрации (экспериментальное исследование)

Objective: to develop technology for the transmembrane mass transfer of fat-soluble substances with emulsified solution of the essential lipids exposed to dialysis. Materialaadmethods. The pharmacopoeia lipofundin MCT/LCT (20% intravenous fat emulsion) (B. Braun, Germany) was used at all study stages. Coaclusioa. Bicarbonate hemodialysis-modified lipofundin solution contains no triglycerides; its osmolarity is normalized and the solution is ready to use for lipid on-line hemodiafiltration, which allows fat-soluble substances that accumulate in acute liver failure to be removed. Key words: lipid hemodi-afiltration, lipofundin, acute liver failure.Цель работы — разработка технологии трансмембранного массопереноса жирорастворимых веществ эмульгированным раствором эссенциальных липидов, подвергшихся диализу. Материал и методы. Для всех этапов исследования использовали фармакопейный липофундин МСТ/ЛСТ (B.Braun, Германия) — 20% жировую эмульсию для внутривенного введения. Выводы. Модифицированный бикарбонатным гемодиализом раствор липофундина не содержит триглицеридов, нормализуется его осмолярность и он готов к применению для проведение липидной on line гемодиа-фильтрации, что позволяет удалять жирорастворимые вещества, накапливающиеся при острой печеночной недостаточности. Ключевые слова: липидная гемодиафильтрация, липофундин, острая печеночная недостаточность

Novel Mouse Model Reveals Distinct Activity-Dependent and –Independent Contributions to Synapse Development

The balanced action of both pre- and postsynaptic organizers regulates the formation of neuromuscular junctions (NMJ). The precise mechanisms that control the regional specialization of acetylcholine receptor (AChR) aggregation, guide ingrowing axons and contribute to correct synaptic patterning are unknown. Synaptic activity is of central importance and to understand synaptogenesis, it is necessary to distinguish between activity-dependent and activity-independent processes. By engineering a mutated fetal AChR subunit, we used homologous recombination to develop a mouse line that expresses AChR with massively reduced open probability during embryonic development. Through histological and immunochemical methods as well as electrophysiological techniques, we observed that endplate anatomy and distribution are severely aberrant and innervation patterns are completely disrupted. Nonetheless, in the absence of activity AChRs form postsynaptic specializations attracting motor axons and permitting generation of multiple nerve/muscle contacts on individual fibers. This process is not restricted to a specialized central zone of the diaphragm and proceeds throughout embryonic development. Phenotypes can be attributed to separate activity-dependent and -independent pathways. The correct patterning of synaptic connections, prevention of multiple contacts and control of nerve growth require AChR-mediated activity. In contrast, myotube survival and acetylcholine-mediated dispersal of AChRs are maintained even in the absence of AChR-mediated activity. Because mouse models in which acetylcholine is entirely absent do not display similar effects, we conclude that acetylcholine binding to the AChR initiates activity-dependent and activity-independent pathways whereby the AChR modulates formation of the NMJ

Successive Increases in the Resistance of Drosophila to Viral Infection through a Transposon Insertion Followed by a Duplication

To understand the molecular basis of how hosts evolve resistance to their parasites, we have investigated the genes that cause variation in the susceptibility of Drosophila melanogaster to viral infection. Using a host-specific pathogen of D. melanogaster called the sigma virus (Rhabdoviridae), we mapped a major-effect polymorphism to a region containing two paralogous genes called CHKov1 and CHKov2. In a panel of inbred fly lines, we found that a transposable element insertion in the protein coding sequence of CHKov1 is associated with increased resistance to infection. Previous research has shown that this insertion results in a truncated messenger RNA that encodes a far shorter protein than the susceptible allele. This resistant allele has rapidly increased in frequency under directional selection and is now the commonest form of the gene in natural populations. Using genetic mapping and site-specific recombination, we identified a third genotype with considerably greater resistance that is currently rare in the wild. In these flies there have been two duplications, resulting in three copies of both the truncated allele of CHKov1 and CHKov2 (one of which is also truncated). Remarkably, the truncated allele of CHKov1 has previously been found to confer resistance to organophosphate insecticides. As estimates of the age of this allele predate the use of insecticides, it is likely that this allele initially functioned as a defence against viruses and fortuitously “pre-adapted” flies to insecticides. These results demonstrate that strong selection by parasites for increased host resistance can result in major genetic changes and rapid shifts in allele frequencies; and, contrary to the prevailing view that resistance to pathogens can be a costly trait to evolve, the pleiotropic effects of these changes can have unexpected benefits

Maternal Perception of Reduced Fetal Movements Is Associated with Altered Placental Structure and Function

Maternal perception of reduced fetal movement (RFM) is associated with increased risk of stillbirth and fetal growth restriction (FGR). DFM is thought to represent fetal compensation to conserve energy due to insufficient oxygen and nutrient transfer resulting from placental insufficiency. To date there have been no studies of placental structure in cases of DFM.To determine whether maternal perception of reduced fetal movements (RFM) is associated with abnormalities in placental structure and function.Placentas were collected from women with RFM after 28 weeks gestation if delivery occurred within 1 week. Women with normal movements served as a control group. Placentas were weighed and photographs taken. Microscopic structure was evaluated by immunohistochemical staining and image analysis. System A amino acid transporter activity was measured as a marker of placental function. Placentas from all pregnancies with RFM (irrespective of outcome) had greater area with signs of infarction (3.5% vs. 0.6%; p<0.01), a higher density of syncytial knots (p<0.001) and greater proliferation index (p<0.01). Villous vascularity (p<0.001), trophoblast area (p<0.01) and system A activity (p<0.01) were decreased in placentas from RFM compared to controls irrespective of outcome of pregnancy.This study provides evidence of abnormal placental morphology and function in women with RFM and supports the proposition of a causal association between placental insufficiency and RFM. This suggests that women presenting with RFM require further investigation to identify those with placental insufficiency