Retromer Regulates Postendocytic Sorting of β-Secretase in Polarized Madin-Darby Canine Kidney Cells

β-Amyloid (Aβ) peptides are generated from the successive proteolytic processing of the amyloid precursor protein (APP) by the β-APP cleaving enzyme (BACE or β-secretase) and the γ-secretase complex. Initial cleavage of APP by BACE leads into the amyloidogenic pathway, causing or exacerbating Alzheimer's disease. Therefore, their intracellular traffic can determine how easily and frequently BACE has access to and cleaves APP. Here, we have used polarized Madin-Darby canine kidney (MDCK) cells stably expressing APP and BACE to examine the regulation of their polarized trafficking by retromer, a protein complex previously implicated in their endosome-to-Golgi transport. Our data show that retromer interacts with BACE and regulates its postendocytic sorting in polarized MDCK cells. Depleting retromer, inhibiting retromer function, or preventing BACE interaction with retromer, alters trafficking of BACE, which thereby increases its localization in the early endocytic compartment. As a result, this slows endocytosis of apically localized BACE, promoting its recycling and apical-to-basolateral transcytosis, which increases APP/BACE interaction and subsequent cleavage of APP toward generation and secretion of Aβ peptides.     

Effect of VIP on the balance between cytokines and master regulators of activated helper T cells

CD4T helper cells are decisive in the struggle against pathogens and in maintaining immune homeostasis. Nevertheless, they also drive immune-mediated disease. Recently, emerging evidence suggests that seemingly committed Th cells possess plasticity and may convert into other types of effector cells. Vasoactive Intestinal Peptide (VIP) is an immunomodulator neuropeptide, which is able to promote or inhibit individually the differentiation or function of some T-helper subsets. We conducted ex vivo study with erythrocyte-depleted spleen cells from healthy mice to check the balance between cytokines and master regulators of different T-helper subsets. This neuropeptide adversely affected the differentiation and functionality phases of Th17 cells and had a negative influence on cytokines related to Th1 function, increasing Th17 cells over those of the Th1 cell subset. With respect to Th2 subsets, VIP augmented the interleukin (IL)-4/IL-9 mRNA ratio, and a negative correlation between IL-4 and IL-9 was observed in culture supernatants. VIP augmented Th2 relative to Th1 in cell subsets. VIP decreased the iTreg/Th17 balance. Regarding the induced T-regulatory (iTreg)/Th1 balance, VIP increased the presence of immunoregulatory cytokines in relation to IFNγ. Although additional studies are needed to clarify the role of VIP on the balance between cytokines and master regulators during T-helper differentiation, our data show that VIP reduces Th17/Th1 and Th1/Th2 ratios. However, the iTreg/Th17 ratio was differently counterbalanced, probably because of culture conditions. Finally, this is the first study showing that VIP also modulates Th2/Th9 and iTreg/Th1 ratios.     

Structure and Sites of Phosphorylation of 14-3-3 Protein: Role in Coordinating Signal Transduction Pathways

The 14-3-3 family are homo- and heterodimeric proteins whose biological role has been unclear for some time, although they are now gaining acceptance as a novel type of adaptor protein that modulates interactions between components of signal transduction pathways, rather than by direct activation or inhibition. It is becoming apparent that phosphorylation of the binding partner and possibly also the 14-3-3 proteins may regulate these interactions. 14-3-3 isoforms interact with a novel phosphoserine (Sp) motif on many proteins, RSX_1,2SpXP. The two isoforms that interact with Raf-1 are phosphorylated in vivo on Ser185 in a consensus sequence motif for proline-directed kinases. The crystal structure of 14-3-3 indicates that this phosphorylation could regulate interaction of 14-3-3 with its target proteins. We have now identified a number of additional phosphorylation sites on distinct mammalian and yeast isoforms.     

Parasitism by isopods in sandy beach fish assemblages: role of urbanization and environmental characteristics

Parasitism by isopods is a common phenomenon in marine fishes and may induce negative effects, especially in juvenile stages. The surf-zones of sandy beaches are an important nursery habitat for many fish species, including commercially important ones, and are structured by beach characteristics and human activities, which are suggested to influence the fishes’ susceptibility to parasitism. Here, we analyze the early infestation and prevalence of parasitism in surf zones fishes of 27 beaches in southeastern Brazil with different characteristics. Parasites were recorded in only seven of the beaches. A high infestation was found in a single site, the highly urbanized Itaguá beach, where the prevalence of parasitism on juveniles and early adults of the Atlantic anchoveta Cetengraulis edentulus was high (68.11%). Four species of parasitic isopods were found, all belonging to the Cymothoidae family, but Livoneca desterroensis was the most abundant (78.7%). No effects of parasitism were detected on the size, weight and physiological condition of fishes, but presence of parasites and prevalence of infestation were positively affected by urbanization. Overall, our results indicate that the surf-zones have low parasite infestation, yet, they suggest that anthropic effects such as urbanization may enhance the prevalence of isopod parasitism.

     

Ingression Progression Complexes Control Extracellular Matrix Remodelling during Cytokinesis in Budding Yeast

Eukaryotic cells must coordinate contraction of the actomyosin ring at the division site together with ingression of the plasma membrane and remodelling of the extracellular matrix (ECM) to support cytokinesis, but the underlying mechanisms are still poorly understood. In eukaryotes, glycosyltransferases that synthesise ECM polysaccharides are emerging as key factors during cytokinesis. The budding yeast chitin synthase Chs2 makes the primary septum, a special layer of the ECM, which is an essential process during cell division. Here we isolated a group of actomyosin ring components that form complexes together with Chs2 at the cleavage site at the end of the cell cycle, which we named ‘ingression progression complexes’ (IPCs). In addition to type II myosin, the IQGAP protein Iqg1 and Chs2, IPCs contain the F-BAR protein Hof1, and the cytokinesis regulators Inn1 and Cyk3. We describe the molecular mechanism by which chitin synthase is activated by direct association of the C2 domain of Inn1, and the transglutaminase-like domain of Cyk3, with the catalytic domain of Chs2. We used an experimental system to find a previously unanticipated role for the C-terminus of Inn1 in preventing the untimely activation of Chs2 at the cleavage site until Cyk3 releases the block on Chs2 activity during late mitosis. These findings support a model for the co-ordinated regulation of cell division in budding yeast, in which IPCs play a central role.     

Selective Serotonin Reuptake Inhibitors and Violent Crime: A Cohort Study

BackgroundAlthough selective serotonin reuptake inhibitors (SSRIs) are widely prescribed, associations with violence are uncertain.ConclusionsThe association between SSRIs and violent crime convictions and violent crime arrests varied by age group. The increased risk we found in young people needs validation in other studies.     

Isolation and characterization of a new suite of microsatellite markers in the European White Stork, Ciconia ciconia

The White Stork (Ciconia ciconia) has an extensive European breeding distribution. There have been significant demographic changes since the 1930s, with country-specific extinctions throughout the western distribution since the mid-1940s. Following various reintroduction programs significant concerns have been raised about the geographic origins of introduced storks, and the effect this may have on individual biology, particularly breeding success. Here we report on seven new microsatellite markers developed for C. ciconia that have been successfully combined in a series of multiplex PCR’s with six markers previously described for the Wood Stork.     

Evaluation of restricted access media for the purification of cell culture-derived Orf viruses

Recently, multimodal chromatography using restricted access media (RAM) for the purification of nanoparticles, such as viruses has regained increasing attention. These chromatography resins combine size exclusion on the particle shell and adsorptive interaction within the core. Accordingly, smaller process-related impurities, for example, DNA and proteins, can be retained, while larger product viruses can pass unhindered. We evaluated a range of currently available RAM, differing in the shells’ pore cut-off and the core chemistry, for the purification of a cell culture-derived clarified model virus, namely the Orf virus (ORFV). We examined impurity depletion and product recovery as relevant criteria for the evaluation of column performance, as well as scale-up robustness and regeneration potential for evaluating a multiple use application. The results indicate that some columns, for example, the Capto Core, enable both a high DNA and protein removal, while others, for example, the Monomix Core 60 (MC60), are more suitable for DNA depletion. Furthermore, column regeneration is facilitated by using columns with larger shell pores (5000 vs. 700 kDa) and weaker binding interactions (anion exchange vs. multimodal). According to these findings, the choice of RAM resins should be selected according to the respective feed sample composition and the planned number of application cycles.     

Sexual behavior,insemination and development of the freshwater leech Helobdella stagnalis (Annelida,Hirudinea, Glossiphoniidae)

The reproductive biology of the Glossiphoniidae leech Helobdella stagnalis, under laboratory conditions, the structure of its eggs and its developmental stages were studied. Sperm transfer and insemination are made by hypodermic injection: one or numerous spermatophores were attached to the skin of the partner during copulation and sperm is injected through it. The leeches can copulate repeatedly with several partners. Each leech produces 4–8 cocoons containing 20–60 eggs whose are attached to the ventral side of the parent and carried around. At 22 °C, the developmental duration is 24 days from the oviposition until the juveniles leave the parent leech. Three (3) major stages have been distinguished: Eggs cleavage, germinal band generation, and juveniles hatching. The sexual behavior of Hellobdella stagnalis was described with a special attention given to parental care.     

Metallothionein and brain inflammation

Since the seminal discoveries of Bert Vallee regarding zinc and metallothioneins (MTs) more than 50 years ago, thousands of studies have been published concerning this fascinating story. One of the most active areas of research is the involvement of these proteins in the inflammatory response in general, and in neuroinflammation in particular. We describe the general aspects of the inflammatory response, highlighting the essential role of the major cytokine interleukin-6, and review briefly the expression and function of MTs in the central nervous system in the context of neuroinflammation. Particular attention is paid to the Tg2576 Alzheimer disease mouse model and the preliminary results obtained in mice into which human Zn₇MT-2A was injected, which suggest a reversal of the behavioral deficits while enhancing amyloid plaque load and gliosis.