Specific features of neuronal size and shape are regulated by tropomyosin isoforms

Spatially distinct populations of microfilaments, characterized by different tropomyosin (Tm) isoforms, are present within a neuron. To investigate the impact of altered tropomyosin isoform expression on neuronal morphogenesis, embryonic cortical neurons from transgenic mice expressing the isoforms Tm3 and Tm5NM1, under the control of the beta-actin promoter, were cultured in vitro. Exogenously expressed Tm isoforms sorted to different subcellular compartments with Tm5NM1 enriched in filopodia and growth cones, whereas the Tm3 was more broadly localized. The Tm5NM1 neurons displayed significantly enlarged growth cones accompanied by an increase in the number of dendrites and axonal branching. In contrast, Tm3 neurons displayed inhibition of neurite outgrowth. Recruitment of Tm5a and myosin IIB was observed in the peripheral region of a significant number of Tm5NM1 growth cones. We propose that enrichment of myosin IIB increases filament stability, leading to the enlarged growth cones. Our observations support a role for different tropomyosin isoforms in regulating interactions with myosin and thereby regulating morphology in specific intracellular compartments.     

A Rab requirement is not bypassed in SLY1-20 suppression

The Rab GTPase Ypt1p and the large homodimer Uso1p are both required for tethering endoplasmic reticulum-derived vesicles to early Golgi compartments in yeast. Loss-of-function ypt1 and uso1 mutations are suppressed by SLY1-20, a dominant allele that encodes the Sed5p-associated protein, Sly1p. Here, we investigate the mechanism of SLY1-20 suppression. In wild-type strains, Ypt1p can be coimmunoprecipitated with Uso1p; however, in a ypt1Delta/SLY1-20 strain, which lacks this complex, membrane binding of Uso1p was reduced. In spite of Ypt1p depletion, Uso1p-dependent vesicle tethering was not bypassed under the ypt1Delta/SLY1-20 condition. Moreover, tethering and fusion assays with ypt1Delta/SLY1-20 membranes remained sensitive to Rab GDP dissociation inhibitor. These results indicate that an alternative Rab protein satisfies the Ypt1p requirement in Uso1p-dependent tethering when SLY1-20 is expressed. Further genetic and biochemical tests revealed that a related Rab protein, Ypt6, might substitute for Ypt1p in ypt1Delta/SLY1-20 cells. Additional experimentation to address the mechanism of SLY1-20 suppression in a cog2Delta [sec35Delta] strain indicated that the Cog2p subunit of the conserved oligomeric Golgi complex is either functionally redundant or is not directly required for anterograde transport to the Golgi complex.     

Oral albuterol dosing during the latter stages of a resistance exercise program

Subjects performed isoload variable resistance exercise (REX) 3 days per week. After 10 weeks, they received a double-blind albuterol (n = 11) or placebo (n = 11) capsule assignment with no crossover and continued training. During the first week of capsule administration, dosages were increased from 4 mg to 16 mg daily and then maintained for 14 days. At weeks 0, 10, and 13, we measured upper arm and thigh cross-sectional area, knee and elbow extensor and flexor (KE, KF, EE, EF) strength at 3 angular velocities, and lean body mass. Data after 10 weeks showed insignificant between-group differences. From weeks 10-13, as subjects continued REX training, albuterol evoked higher (p < 0.05) KE-KF strength gains at multiple velocities versus placebo dosing. A higher lean body mass trend also occurred with albuterol from weeks 10-13. Results suggest that albuterol augments REX to provide greater strength gains from hypertrophic factors than an REX-placebo assignment.     

Reversing systemic inflammatory response syndrome with chemokine receptor pepducins

We describe a new therapeutic approach for the treatment of lethal sepsis using cell-penetrating lipopeptides-termed pepducins-that target either individual or multiple chemokine receptors. Interleukin-8 (IL-8), a ligand for the CXCR1 and CXCR2 receptors, is the most potent endogenous proinflammatory chemokine in sepsis. IL-8 levels rise in blood and lung fluids to activate neutrophils and other cells, and correlate with shock, lung injury and high mortality. We show that pepducins derived from either the i1 or i3 intracellular loops of CXCR1 and CXCR2 prevent the IL-8 response of both receptors and reverse the lethal sequelae of sepsis, including disseminated intravascular coagulation and multi-organ failure in mice. Conversely, pepducins selective for CXCR4 cause a massive leukocytosis that does not affect survival. CXCR1 and CXCR2 pepducins conferred nearly 100% survival even when treatment was postponed, suggesting that our approach might be beneficial in the setting of advanced disease.     

Microbiologic consequences of new approaches to managing hematologic malignancies

Opportunistic infections have always been pitfalls on the road of progress in the treatment of diseases that are accompanied by compromised host defences. Because of the severe morbidity and mortality associated with these infections, they have become substantial challenges for the clinicians who offer such patients care. With medical progress, the number of immunocompromised patients is still steadily climbing and it has become evident that deficiencies in host defences mechanisms are multiple as well as changing in harmony with alterations in treatment modalities for underlying diseases. Under normal circumstances, the intact epithelial surfaces of the gastrointestinal tract will prohibit invasion by micro-organisms and the mucociliary barrier of the respiratory tract prevents aspiration of fungal cells and spores, while, in contrast, dead or damaged tissue creates a nidus for infection. It is, however, questionable whether transmigration of organisms inevitably leads to infection. With the growing use of potent immunosuppressive purine analogues, fludarabine, pentostatin and cladibrine, and anti-T and anti-B cell antibodies, such as rituximab and campath, in the management of lymphoreticular malignancies, in combination with increasing emphasis on dose intensity, the number of patients at risk has almost reached levels encountered in recipients of allogenic stem cell grafts as a consequence of long-lasting deficiencies in the cellular immunity. The spectrum of opportunistic pathogens are shifting as anti-leukemic and anti-lymphoma therapy become more intensive and bone marrow transplant practices evolve. Recent studies demonstrate, that patients treated with nonmyeloablative allogeneic transplantation (or "minitransplants") to reduce transplant-related toxicity, are at high risk of contracting a serious infections. Initially bacterial infections were most problematic. However, as strategies to control bacterial infections improved, viruses demanded more attention from the clinicians but the associated morbidity declined due to advances in rapid diagnostics and the introduction of effective antivirals such as acyclovir and ganciclovir. Next to viruses, resistant bacteria, particularly Gram-positive organisms like enterococci and methicillin-resistant staphylococci urged to vigilance. It was obvious that enhanced use of antibacterials inevitably will be accompanied by selection and induction of resistant organisms. Today, opportunistic fungi have become the most frequent and dangerous pathogens. Since the 1980's the rate of nosocomial invasive fungal diseases has doubled without any sign of slowing at the turn of the millenium. During the past decades we have even observed an increased incidence of invasive fungal infections in patients who are not in an end stage of their underlying disease. Yeasts and moulds rank amongst the most frequently isolated pathogens. The relative incidence of the various fungal infections depends on geography as well as on medical practices and local conditions. Candida Aspergillus species remain the prominent fungal pathogens but more rare species are increasingly cultured.     

Chloroplast genome phylogenetics: why we need independent approaches to plant molecular evolution

The traditional approach to plant molecular phylogenetics involves amplifying, sequencing and analyzing one or a few genes from many species and is conducive to broad taxon sampling. An independent approach involves chloroplast genome sequencing, providing much larger amounts of data per taxon but for a smaller number of species. In principle, the two strategies can inform each other but in practice their results sometimes conflict for reasons that are currently debated. An Opinion article published in the October 2004 issue of Trends in Plant Science cautioned against the pursuit of genome-based phylogenies. Here, we provide a different perspective on issues at the heart of the current debate and defend the use of chloroplast genome phylogenetics for crucial species because it provides an independent test of hypotheses generated by the traditional approach.     

Ca2+-dependent lipid binding and membrane integration of PopA, a harpin-like elicitor of the hypersensitive response in tobacco

PopA is released by type III secretion from the bacterial plant pathogen Ralstonia solanacearum and triggers the hypersensitive response (HR) in tobacco. The function of PopA remains obscure, mainly because mutants lacking this protein are not altered in their ability to interact with plants. In an attempt to identify the site of PopA activity in plant cells, we generated transgenic tobacco plants expressing the popA gene under the control of an inducible promoter. Immunocytologic analysis revealed that the HR phenotype of these plants correlated with the presence of PopA at the plant plasma membrane. Membrane localization was observed irrespective of whether the protein was designed to accumulate in the cytoplasm or to be secreted by the plant cell, suggesting a general lipid-binding ability. We found that the protein had a high affinity for sterols and sphingolipids in vitro and that it required Ca2+ for both lipid binding and oligomerization. In addition, the protein was integrated into liposomes and membranes from Xenopus laevis oocytes where it formed ion-conducting pores. These characteristics suggest that PopA is part of a system that aims to attach the host cell plasma membrane and to allow molecules cross this barrier.     

Structure-activity relationship studies on a series of cyclohexylpiperazines bearing a phanylacetamide as ligands of the human melanocortin-4 receptor

Synthesis and structure-activity relationship studies of a series of cyclohexylpiperazines bearing an amide side chain as ligands of the MC4 receptor are discussed. Compounds such as 11i from this series are potent agonists (EC(50)=33nM, IA=96%).