Intersegmental coordinating system of the lamprey central pattern generator for locomotion

Summary In the lamprey,Ichthyomyzon unicuspis, the wave of activity required for normal swimming movements can be generated by a central pattern generator (CPG) residing in the spinal cord. A constant phase coupling between spinal segments can be organized by intersegmental coordinating neurons intrinsic to the cord. The rostral and caudal segmental oscillators of the CPG have different preferred frequencies when separated from each other. Therefore the system must maintain the segmental oscillators of the locomotor CPG at a single common frequency and with the proper relative timing. Using selective lesions and a split-bath, it is demonstrated that the coordinating system is comprised of at least 3 subsystems, short-axon systems in the lateral and medial tracts and a long axon system in the lateral tracts. Each alone can sustain relatively stable coordinated activity.Abbreviations CPG central pattern generator - NMDA N-methyl-D-aspartate - VR ventral root     

Positive work as a function of eccentric load in maximal leg extension movements

Negative and positive work performed during leg extension movements of 53 well trained subjects was measured with the help of a special dynamometer. The subjects performed four maximal push off trials against five different loads (25-105 kg): two two-legged extensions from a squatting position (SM) with a knee angle of 70 degrees and two trials with a preliminary counter movement (CM) but with the same extension range as in the SM. Positive work differed only by about 4% between CM and SM in spite of large differences in initial forces at the onset of concentric contraction. Based on simulations, it is suggested that in CM the advantage of stored elastic energy can almost completely be nullified by the disadvantage of a limited shortening distance of the contractile elements. It is hypothesised that elastic energy in CM can only cause considerable extra work during concentric contraction compared to the maximal positive work done in SM if the total range of lengthening and shortening of the muscle(s) involved is larger in CM than in SM.     

Domain redistribution within ergosterol-containing model membranes in the presence of the antimicrobial compound fengycin

The cyclic lipopeptide fengycin, produced by Bacillus subtilis, exhibits its antimicrobial capabilities by altering the integrity of the cell membrane of plant pathogens. Previous work has correlated fengycin activity with membrane characteristics, such as sterol content. This work focused on the influence of fengycin on supported lipid bilayers containing varying levels of ergosterol. Total internal reflection fluorescence (TIRF) microscopy was used to visualize and distinguish ordered (L_β/L_o) and disordered (L_α/L_d) domains in the model membranes following exposure to low (50 μg) and high (500 μg) fengycin doses. Application of an initial low dose of fengycin to 0% and 3% ergosterol-containing bilayers resulted in redistribution of L_α/L_β and L_o/L_d domains, respectively, which the bilayers compensated and corrected for over time. These membranes were unable to tolerate a second 50 μg dose or a single high fengycin dose. The 6% ergosterol bilayers were able to tolerate sequential low doses of fengycin. Exposure of these bilayers to the high fengycin dose caused a decrease in the number of L_o domains, albeit less than that seen in the 0% and 3% ergosterol bilayers. Bilayers containing 12% ergosterol, exhibited the least amount of change after fengycin exposure. These were the only bilayer to exhibit an increase in area taken up by ordered domains. These results suggest fengycin may preferentially act on the L_β or L_o phase, the area in which ergosterol resides. Bilayers containing low levels of ergosterol appear to be more sensitive to the lipopeptide, suggesting ergosterol plays a role in buffering perturbations caused by fengycin.     

Specificity and mode of action of the antifungal fatty acid cis-9-heptadecenoic acid produced by Pseudozyma flocculosa

cis-9-Heptadecenoic acid (CHDA), an antifungal fatty acid produced by the biocontrol agent Pseudozyma flocculosa, was studied for its effects on growth and/or spore germination in fungi. Inhibition of growth and/or germination varied considerably and revealed CHDA sensitivity groups within tested fungi. Analysis of lipid composition in these fungi demonstrated that sensitivity was related primarily to a low intrinsic sterol content and that a high level of unsaturation of phospholipid fatty acids was not as involved as hypothesized previously. Our data indicate that CHDA does not act directly with membrane sterols, nor is it utilized or otherwise modified in fungi. A structural mechanism of CHDA, consistent with the other related antifungal fatty acids produced by P. flocculosa, is proposed in light of its activity and specificity. The probable molecular events implicated in the sensitivity of fungi to CHDA are (i) partitioning of CHDA into fungal membranes; (ii) a variable elevation in fluidity dependent on the buffering capability (sterol content) in fungi; and (iii) higher membrane disorder causing conformational changes in membrane proteins, increased membrane permeability and, eventually, cytoplasmic disintegration.     

Effect of magnesium ions on the tertiary structure of the hepatitis C virus IRES and its affinity for the cyclic peptide antibiotic viomycin

A key ion-dependent folding unit within the hepatitis C IRES comprises the IIIef junction and pseudoknot. This region is also important in recruitment of the 40S ribosomal subunit. Here, circular dichroism is used to study the influence of metal ions on the structure and stability of this region. Comparison of the thermal stability of an IRES fragment encompassing subdomains IIIe/f and IV (named 3EF4) with that of a larger fragment also possessing subdomain IIId (3DEF4) indicates an additional stabilizing effect of Mg(2+) ions on the latter fragment. Magnesium and potassium ions stabilize both fragments through nonspecific counterion effects. The additional effect of magnesium on 3DEF4, observed in the absence or presence of 100 mM KCl, is attributed to a nonspecific but high-affinity site for metal ions created by a region of unusual high charge density. Subdomain IIId presumably participates in tertiary packing interactions that provide such a site. Viomycin binds to the full-length IRES and RNA fragments with K(d) values of 25-55 microM. Interestingly, viomycin binding to the two fragments is affected differently by Mg(2+); noncompetitive inhibition of binding to 3DEF4 is observed, whereas binding to 3EF4 is not impaired. Formation of a Mg(2+)-stabilized tertiary fold, involving subdomain IIId, may thereby hinder viomycin binding to 3DEF4 indirectly. Mutational and deletion studies locate viomycin binding within subdomains IIIe/f rather than within the pseudoknot. In pseudoknot mutants, Mg(2+) ions have different effects on viomycin binding and thermal stability, suggesting altered tertiary interactions involving subdomain IIId.     

Unusual presentation of choriocarcinoma

Background  

Choriocarcinoma is an aggressive neoplasm arising in the body of the uterus. The disease normally spreads to lung and brain.     

Estimating the Strength and Direction of Functional Coupling in the Lamprey Spinal Cord

A method of estimating coupling strength between two neural oscillators based on their spikes trains (Kiemel and Cohen, J. Comput. Neurosci. 5: 267–284, 1998) is tested using simulated data and then applied to experimental data from the central pattern generator (CPG) for swimming in the lamprey. The method is tested using a model of two connectionist oscillators and a model of two endogenously bursting cells. For both models, the method provides useful estimates of the relative strength of coupling in each direction, as well as estimates of total strength. The method is applied to pairs of motor-nerve recordings from isolated 50-segment pieces of spinal cords from adult silver lampreys (Ichthyomyzon unicuspus). The strength and direction of coupling is estimated under control conditions and conditions in which intersegmental coupling between the two recording locations is weakened by hemisections of the spinal cords and/or chambers containing an inhibitory solution that blocks firing in postsynaptic cells. The relevance of these measures in constraining models of the CPG is discussed.     

Inhibition of the Inositol Kinase Itpkb Augments Calcium Signaling in Lymphocytes and Reveals a Novel Strategy to Treat Autoimmune Disease

Emerging approaches to treat immune disorders target positive regulatory kinases downstream of antigen receptors with small molecule inhibitors. Here we provide evidence for an alternative approach in which inhibition of the negative regulatory inositol kinase Itpkb in mature T lymphocytes results in enhanced intracellular calcium levels following antigen receptor activation leading to T cell death. Using Itpkb conditional knockout mice and LMW Itpkb inhibitors these studies reveal that Itpkb through its product IP4 inhibits the Orai1/Stim1 calcium channel on lymphocytes. Pharmacological inhibition or genetic deletion of Itpkb results in elevated intracellular Ca²⁺ and induction of FasL and Bim resulting in T cell apoptosis. Deletion of Itpkb or treatment with Itpkb inhibitors blocks T-cell dependent antibody responses in vivo and prevents T cell driven arthritis in rats. These data identify Itpkb as an essential mediator of T cell activation and suggest Itpkb inhibition as a novel approach to treat autoimmune disease.