EHD2 mediates trafficking from the plasma membrane by modulating Rac1 activity

EHDs [EH (Eps15 homology)-domain-containing proteins] participate in different stages of endocytosis. EHD2 is a plasma-membrane-associated EHD which regulates trafficking from the plasma membrane and recycling. EHD2 has a role in nucleotide-dependent membrane remodelling and its ATP-binding domain is involved in dimerization, which creates a membrane-binding region. Nucleotide binding is important for association of EHD2 with the plasma membrane, since a nucleotide-free mutant (EHD2 T72A) failed to associate. To elucidate the possible function of EHD2 during endocytic trafficking, we attempted to unravel proteins that interact with EHD2, using the yeast two-hybrid system. A novel interaction was found between EHD2 and Nek3 [NIMA (never in mitosis in Aspergillus nidulans)-related kinase 3], a serine/threonine kinase. EHD2 was also found in association with Vav1, a Nek3-regulated GEF (guanine-nucleotide-exchange factor) for Rho GTPases. Since Vav1 regulates Rac1 activity and promotes actin polymerization, the impact of overexpression of EHD2 on Rac1 activity was tested. The results indicated that wt (wild-type) EHD2, but not its P-loop mutants, reduced Rac1 activity. The inhibitory effect of EHD2 overexpression was partially rescued by co-expression of Rac1 as measured using a cholera toxin trafficking assay. The results of the present study strongly indicate that EHD2 regulates trafficking from the plasma membrane by controlling Rac1 activity.     

Global patterns of guild composition and functional diversity of spiders

The objectives of this work are: (1) to define spider guilds for all extant families worldwide; (2) test if guilds defined at family level are good surrogates of species guilds; (3) compare the taxonomic and guild composition of spider assemblages from different parts of the world; (4) compare the taxonomic and functional diversity of spider assemblages and; (5) relate functional diversity with habitat structure. Data on foraging strategy, prey range, vertical stratification and circadian activity was collected for 108 families. Spider guilds were defined by hierarchical clustering. We searched for inconsistencies between family guild placement and the known guild of each species. Richness and abundance per guild before and after correcting guild placement were compared, as were the proportions of each guild and family between all possible pairs of sites. Functional diversity per site was calculated based on hierarchical clustering. Eight guilds were discriminated: (1) sensing, (2) sheet, (3) space, and (4) orb web weavers; (5) specialists; (6) ambush, (7) ground, and (8) other hunters. Sixteen percent of the species richness corresponding to 11% of all captured individuals was incorrectly attributed to a guild by family surrogacy; however, the correlation of uncorrected vs. corrected guilds was invariably high. The correlation of guild richness or abundances was generally higher than the correlation of family richness or abundances. Functional diversity was not always higher in the tropics than in temperate regions. Families may potentially serve as ecological surrogates for species. Different families may present similar roles in the ecosystems, with replacement of some taxa by other within the same guild. Spiders in tropical regions seem to have higher redundancy of functional roles and/or finer resource partitioning than in temperate regions. Although species and family diversity were higher in the tropics, functional diversity seems to be also influenced by altitude and habitat structure.     

The effect of low- and high-fiber diets on the population of entodiniomorphid ciliates Troglodytella abrassarti in captive chimpanzees (Pan troglodytes)

Troglodytella abrassarti is an intestinal entodiniomorphid ciliate commonly diagnosed in the feces of wild and captive chimpanzees (Pan troglodytes). Entodiniomorphids could be considered to have a mutualistic relationship with the great apes, in that the ciliates benefit from the intestinal ecosystem of the host, while also contributing to the fiber fermentation process. We examined the effect of diet on the infection intensities of T. abrassarti in two captive chimpanzees in the Liberec Zoo, Czech Republic. The chimpanzees were fed a low-fiber diet (LFD) with 14% neutral detergent fiber (NDF) and a high-fiber diet (HFD; 26% NDF) for 10 days with one transition, and two 10-day adaptation periods. Fecal samples were examined coproscopically with the merthiolate-iodine-formaldehyde concentration (MIFC) technique, in order to quantify the number of ciliates per gram of feces. A significant trend of increasing T. abrassarti numbers was observed when the animals were fed the LFD, compared to when they were fed the HFD. Our results suggest, however, that infection intensities of T. abrassarti in captive chimpanzees are not influenced primarily by the amount of fiber in the diet, but rather by the dietary starch concentration (HFD: 1%; LFD: 8%).     

Demonstration of Aminopeptidase Activities Secreted by Amsonia tabernaemontana Walt. Cells

Using synthetic substrates, an uncomplicated and sensitive procedure for the determination of extracellular aminopeptidase was developed. The studied enzyme produced by the tested plant material (calli, cell suspension culture and roots of Amsonia tabernaemontana Walt. seedlings) hydrolyzed the substrates β‐naphthylamides (βNA) and 4‐(phenylazo) phenylamides (PAP‐amide) of the amino acids to β‐naphthylamine and 4‐(phenylazo) aniline, respectively, and amino acid. The β‐naphthylamides of the amino acids were applied for the identification of extracellular aminopeptidase, whereas the 4‐(phenylazo) phenylamides of the amino acids were used for the determination of intra‐ and extracellular aminopeptidase activity. By simultaneous azocoupling of β‐naphthol with Fast Garnet GBC salt on agar plates a corresponding brown‐red hardly water‐soluble azo‐dye was produced. The evaluation of dyed zones allowed the extracellular aminopeptidase activity to be assessed. No coloration of the agar medium was observed without inoculum, with heat‐inactivated cells (10 min at 100 °C) or in medium inoculated without substrate. On the agar plates with substrate and sterile Amsonia seedlings, changes in coloration were observed indicating a release of aminopeptidase from the roots during germination. The results show a 91.0 % intracellular and 9.0 % extracellular distribution of aminopeptidase activity, when a cell suspension culture of A. tabernaemontana Walt. as the plant material was used. The agar plate method described permits the rapid, uncomplicated and specific detection of plant producers of extracellular aminopeptidase, which could be particularly useful in future inhibitory and/or biotechnological studies.     

Coreus marginatus (Heteroptera: Coreidae) as a natural enemy of Rumex obtusifolius (Polygonaceae)

Rumex obtusifolius is the preferred host plant of Coreus marginatus. The survival of nymphal C. marginatus, its development on a diet of R. obtusifolius and the effects of herbivory on Rumex seed were investigated. Although the diet consisting solely of seed did not allow complete development, growth rates and survival of nymphs were proportionate to the quantity of provided seed. Nymphal feeding did not noticeably decrease seed mass but significantly decreased the proportion of germinating seed. In this way C. marginatus affects Rumex reproduction. Germination ability decreased with decreasing seed availability and increasing duration of seed predation. Nymphs confined to Rumex inflorescences in the field-completed development and significantly reduced both seed mass and germination ability. The mortality of seed overwintering on standing dry shoots of dock plants was moderately affected by feeding of C. marginatus.     

Capture efficiency and trophic adaptations of a specialist and generalist predator: A comparison

Specialist true predators are expected to exhibit higher capture efficiencies for the capture of larger and dangerous prey than generalist predators due to their possession of specialized morphological and behavioral adaptations. We used an araneophagous spider (Lampona murina) and a generalist spider (Drassodes lapidosus) as phylogenetically related model species and investigated their realized and fundamental trophic niches and their efficacy with respect to prey capture and prey handling. The trophic niche of both species confirmed that Lampona had a narrow trophic niche with a predominance of spider prey (including conspecifics), while the niche of Drassodes was wide, without any preference. DNA analysis of the gut contents of Lampona spiders collected in the field revealed that spiders form a significant part of its natural diet. Lampona captured significantly larger prey than itself and the prey captured by Drassodes. As concerns hunting strategy, Lampona grasped the prey with two pairs of legs possessing scopulae, whereas Drassodes immobilized prey with silk. Lampona possess forelegs equipped with scopulae and a thicker cuticle similar to other nonrelated araneophagous spiders. Lampona fed for a longer time and extracted more nutrients than Drassodes. We show that specialized behavioral and morphological adaptations altogether increase the hunting efficiency of specialists when compared to generalists.     

The correct approach to community-acquired pneumonia in immunocompetent adults: review of current guidelines

Community-acquired pneumonia (CAP) often represents a clinical emergency requiring prompt and adequate antimicrobial treatment. The choice of antimicrobials, however, is difficult due to the variety of potential pathogens and to the spread of drug-resistance. Hence, a correct therapeutic approach should be based on the knowledge of the most frequently reported etiologies for the different clinical conditions, specific patient risk factors and the treatment setting (home, hospital, intensive or non intensive care unit) chosen accordingly. The awareness of the local drug-resistance epidemiology and individual patient characteristics, such as age, history of antibiotic treatments and related adverse events, underlying diseases, concurrent therapies and expected adherence to treatment should also be considered. Lastly, an adequate CAP management should address other issues, including therapy duration, monitoring of its efficacy and adverse effects, and supportive measures. The guidelines for CAP management aim to provide the physician with the necessary knowledge and criteria to assist him in these crucial decisions, and their adoption result in a significant reduction of mortality, frequency and length of hospitalization, and costs. Herein, the authors review and discuss some of the main current guidelines for CAP management, highlighting their differences and similarities.     

Hybrid insulin cocrystals for controlled release delivery

The ability to tailor the release profile of a drug by manipulating its formulation matrix offers important therapeutic advantages. We show here that human insulin can be cocrystallized at preselected ratios with the fully active lipophilically modified insulin derivative octanoyl-N(epsilon)-LysB29-human insulin (C8-HI). The cocrystal is analogous to the NPH (neutral protamine Hagedorn) crystalline complex formed with human insulin, which is commonly used as the long-acting insulin component of diabetes therapy. The in vitro and in vivo release rates of the cocrystal can be controlled by adjusting the relative proportions of the two insulin components. We identified a cocrystal composition comprising 75% C8-HI and 25% human insulin that exhibits near-ideal basal pharmacodynamics in somatostatin-treated beagle dogs. The dependence of release rate on cocrystal ratio provides a robust mechanism for modulating insulin pharmacodynamics. These findings show that a crystalline protein matrix may accommodate a chemical modification that alters the dissolution rate of the crystal in a therapeutically useful way, yet that is structurally innocuous enough to preserve the pharmaceutical integrity of the original microcrystalline entity and the pharmacological activity of the parent molecule.