Biocompatible wound dressings based on chemically degradable triblock copolymer hydrogels

The synthesis of a series of thermo-responsive ABA triblock copolymers in which the outer A blocks comprise poly(2-hydroxypropyl methacrylate) and the central B block is poly(2-(methacryloyloxy)ethyl phosphorylcholine) is achieved using atom transfer radical polymerization. These novel triblock copolymers form thermo-reversible physical gels with critical gelation temperatures and mechanical properties that are highly dependent on the copolymer composition and concentration. TEM studies on dried dilute copolymer solutions indicate the presence of colloidal aggregates, which is consistent with micellar gel structures. This hypothesis is consistent with the observation that incorporating a central disulfide bond within the B block leads to thermo-responsive gels that can be efficiently degraded using mild reductants such as dithiothreitol (DTT) over time scales of minutes at 37 degrees C. Moreover, the rate of gel dissolution increases at higher DTT/disulfide molar ratios. Finally, these copolymer gels are shown to be highly biocompatible. Only a modest reduction in proliferation was observed for monolayers of primary human dermal fibroblasts, with no evidence for cytotoxicity. Moreover, when placed directly on 3D tissue-engineered skin, these gels had no significant effect on cell viability. Thus, we suggest that these thermo-responsive biodegradable copolymer gels may have potential applications as wound dressings.     

Enhancement of tumor-TRAIL susceptibility by modulation of autophagy

Autophagy has recently been recognized as an important cellular response to stress. However, the prospect of manipulating the autophagic process for the enhancement of cancer therapy remains unresolved. This lack of resolution stems from the current controversy regarding the fundamental function of autophagy in tumor stress response: Does it have a positive or negative impact on tumor survival capability? Our studies were designed to investigate the role of autophagy in the response to TRAIL of tumor cells with various apoptotic defects. Based on our findings, we propose that divergent mechanisms of resistance to TRAIL can be reversed by a common approach of targeting specific components of the autophagic process for inhibition. This concept may have significant implications for the development of new strategies to circumvent TRAIL resistance in tumors.     

Clathrin, AP-2, and the NPXY-binding subset of alternate endocytic adaptors facilitate FimH-mediated bacterial invasion of host cells

The FimH adhesin, localized at the distal tips of type 1 pili, binds mannose-containing glycoprotein receptors like alpha3beta1 integrins and stimulates bacterial entry into target host cells. Strains of uropathogenic Escherichia coli (UPEC), the major cause of urinary tract infections, utilize FimH to invade bladder epithelial cells. Here we set out to define the mechanism by which UPEC enters host cells by investigating four of the major entry routes known to be exploited by invasive pathogens: caveolae, clathrin, macropinocytosis and secretory lysosomes. Using pharmacological inhibitors in combination with RNA interference against specific endocytic pathway components, mutant host cell lines and a mouse infection model system, we found that type 1 pili-dependent bacterial invasion of host cells occurs via a cholesterol- and dynamin-dependent phagocytosis-like mechanism. This process did not require caveolae or secretory lysosomes, but was modulated by calcium levels, clathrin, and cooperative input from the primary clathrin adaptor AP-2 and a subset of alternate adaptors comprised of Numb, ARH and Dab2. These alternate clathrin adaptors recognize NPXY motifs, as found within the cytosolic tail of beta1 integrin, suggesting a functional link between the engagement of integrin receptors by FimH and the clathrin-dependent uptake of type 1-piliated bacteria.     

Floral diversity, parasitoids and hyperparasitoids – A laboratory approach

Adding floral resources to agro-ecosystems to improve biological control can enhance the survival, egg load, and parasitism rate of insect parasitoids. However, this may not always be the case because the herbivore may benefit from the added resource as much as, or more than the third-trophic level. In addition, the natural enemies of those in the third-trophic level may also derive improved fitness from the added resources. Both these processes will dampen trophic cascades, leading to less-effective biological control. In this study, the effect of adding different flowering plants on the longevity, egg load, aphid parasitism rates and hyperparasitism of Aphidius ervi Haliday (Hymenoptera: Braconidae) by its hyperparasitoid Dendrocerus aphidum Rondani (Hymenoptera: Megaspilidae) were investigated, using the pea aphid Acyrthosiphon pisum Harris (Homoptera: Aphididae) as the herbivore. Parasitoids exposed to buckwheat survived, on average, between four to five times as long as those in the control (water) and those in phacelia, alyssum and coriander treatments survived three to four times as long. Hyperparasitoids exposed to buckwheat survived five to six times as long as those in the control and three to five times longer with the other plants compared with the control. Almost all flower species significantly increased parasitoid and hyperparasitoid egg loads and the number of parasitised aphids and parasitised mummies compared with control. Understanding the factors influencing the dynamics of multitrophic interactions involving flowering plants, herbivores, parasitoids and hyperparasitoids is a fertile area for future research. One of the most challenging areas in contemporary ecology concerns the relative importance of different types of biodiversity mediating trophic interactions and thereby influencing the structure of communities and food webs. This paper begins to explore this using an experimental, laboratory-based approach.     

BAP31 interacts with Sec61 translocons and promotes retrotranslocation of CFTRDeltaF508 via the derlin-1 complex

BAP31 is an endoplasmic reticulum protein-sorting factor that associates with newly synthesized integral membrane proteins and controls their fate (i.e., egress, retention, survival, or degradation). BAP31 is itself an integral membrane protein and a constituent of several large protein complexes. Here, we show that a part of the BAP31 population interacts with two components of the Sec61 preprotein translocon, Sec61beta and TRAM. BAP31 associates with the N terminus of one of its newly synthesized client proteins, the DeltaF508 mutant of CFTR, and promotes its retrotranslocation from the ER and degradation by the cytoplasmic 26S proteasome system. Depletion of BAP31 reduces the proteasomal degradation of DeltaF508 and permits a significant fraction of the surviving protein to reach the cell surface. Of note, BAP31 also associates physically and functionally with the Derlin-1 protein disclocation complex in the DeltaF508 degradation pathway. Thus, BAP31 operates at early steps to deliver newly synthesized CFTRDeltaF508 to its degradation pathway.     

Mechanisms regulating seedling emergence of orchardgrass (Dactylis glomerata L.) and western wheatgrass (Pascopyrum smithii [Rydb.] L.): Dormancy change,seed fate and seeding date

Seed germination and seedling emergence of ‘Arctic’ and ‘Lineta’ orchardgrass (Dactylis glomerata L.) and ‘Walsh’ and ‘LC9078a’ western wheatgrass (Pascopyrum smithii [Rydb.] L.) were studied both in the field and laboratory. Four seeding dates were conducted each year over 2 years and seedling emergence and seed fate in the soil were monitored. The effects of alternating temperature and light on germination were quantified and correlated with seedling emergence from soil and in the field. Orchardgrass seeds were less dormant than western wheatgrass as indicated by the disparity in germination percentage between constant and alternating temperatures. Seed germination percentage was usually higher than seedling emergence in the field for orchardgrass but lower for western wheatgrass, and temperature was not responsible for the difference. Exposing orchardgrass seeds to light during germination check helped break dormancy in orchardgrass when temperature was unfavorable (low and/or constant temperatures), while favorable temperatures (optimal, alternating temperatures) conditions overcame the inhibiting effect of light in western wheatgrass. The final seedling emergence of orchardgrass was either similar among the four seeding dates or decreased slightly from early May to early June. For western wheatgrass, however, final seedling emergence increased with seeding dates from early to late May and decreased in early June. Soil temperatures of the first 2 weeks after seeding increased from the early May to late May and then decreased. These temperatures were below or near the optimal temperatures for western wheatgrass seeds to release dormancy and germinate. Germination of the previously buried seeds indicated that orchardgrass and western wheatgrass had the potential for a high germination percentage under field conditions for all seeding dates. While soil temperatures close to the optimal temperature for dormancy breaking and germination promoted germination of orchardgrass, the same conditions could cause deterioration of seeds if they failed to germinate. For western wheatgrass, deeper dormancy reduced seed mortality.     

Nocturnal activity positively correlated with auditory sensitivity in noctuoid moths

We investigated the relationship between predator detection threshold and antipredator behaviour in noctuoid moths. Moths with ears sensitive to the echolocation calls of insectivorous bats use avoidance manoeuvres in flight to evade these predators. Earless moths generally fly less than eared species as a primary defence against predation by bats. For eared moths, however, there is interspecific variation in auditory sensitivity. At the species level, and when controlling for shared evolutionary history, nocturnal flight time and auditory sensitivity were positively correlated in moths, a relationship that most likely reflects selection pressure from aerial-hawking bats. We suggest that species-specific differences in the detection of predator cues are important but often overlooked factors in the evolution and maintenance of antipredator behaviour.