2H NMR Study of Polymer Segmental Dynamics at Varying Cross-Linking in Poly(N-isopropylacrylamide) Microgels.

A microscopic understanding of the internal structure and dynamics of poly(N-isopropylacrylamide) (PNIPAM) chains, in microgel colloids, is developed using deuterium NMR (2H NMR) to study deuterated PNIPAM suspensions as functions of temperature and pressure for four cross-linker molar fractions (f). The PNIPAM polymers were labeled with deuterons at the backbone (d3-PNIPAM) or on side chains (d7-PNIPAM). 2H NMR spectra of the d3-PNIPAM suspensions for all cross-linker molar fractions indicated freely moving chains at low temperature and a nearly immobilized fraction above ∼35 °C. Polymer segments in the collapsed phase of the d3-PNIPAM suspension were more mobile than those in the dry powder. This is direct microscopic evidence that the polymer remains significantly hydrated in the collapsed phase, consistent with strong, indirect evidence from recent light scattering and rheology measurements from our laboratory. However, the observation of a small fraction of immobilized segments in the swollen phase for higher cross-linker molar fraction suggests that, particularly for high levels of cross-linking, some polymer is nonhydrated even in the swollen phase. Finally, variable-pressure NMR (up to 90 MPa) showed a slight increase in transition temperature with pressure for lower cross-linker molar fractions and a larger increase in transition temperature with pressure for higher cross-linker molar fractions. This is consistent with a previously reported dependence of collapse transition enthalpy on cross-linker molar fraction.

Role of Charge in Lipid Vesicle Binding and Vesicle Surface Saturation by Gaduscidin-1 and Gaduscidin-2.

The histidine-rich antimicrobial peptides Gad-1 and Gad-2, from paralogous genes in cod, provide an opportunity to examine the effect of charge and nonelectrostatic factors on peptide-vesicle interaction and on peptide antimicrobial activity. In this study, the dependence of vesicle ζ-potential on peptide concentration has been used to examine the binding of these peptides to model vesicle surfaces at pH = 5.0, for which the charges of Gad-1 and Gad-2 are +8 and +5, respectively, and at pH = 7.0, where their charges are +3 and +1, respectively. Interpreting the observed ζ-potential behaviors as examples of Langmuir adsorption isotherms, it is possible to infer the equilibrium constant for peptide-vesicle binding, the fraction of the peptide bound at low peptide concentration, and the maximum peptide-to-lipid ratio when the vesicle surface is saturated at high peptide concentration. For both peptides, higher peptide charge is found to be correlated with a lower fraction of the peptide being bound to vesicle surfaces at low peptide concentration and with a smaller maximum bound peptide-to-lipid ratio at high peptide concentration. The equilibrium binding constant, on the other hand, is more strongly correlated with the peptide sequence than with the charge. Gad-1, which has been shown to be more biologically active than Gad-2, displayed a significantly higher equilibrium binding constant. These observations suggest that while the maximum peptide density on the vesicle surface is limited by electrostatic interactions, the free energy of peptide binding, like the observed antimicrobial activities of the Gad peptides, is also sensitive to other peptide factors which might, for example, influence hydrophobic interactions.

Preadolescents' Daily Peer Victimization and Perceived Social Competence: Moderating Effects of Classroom Aggression.

Few studies have assessed children's daily peer experiences, and even fewer have considered their daily self-perceptions. This daily diary study examined relations between preadolescents' daily reports of peer victimization and perceived social competence, along with moderating effects of classroom aggression. A racially diverse sample of 182 children in 5th grade (105 boys; M age = 10.64 years; 35% White, 31% Black, 17% Hispanic, 17% other or not reported) completed daily measures of peer victimization and perceived social competence, with most children completing measures on 8 school days. Teachers completed measures of aggression for each participating pupil. Four types of peer victimization (verbal victimization, social manipulation, social rebuff, and property attacks) predicted decreased daily perceived social competence. Daily social rebuff predicted decreased daily perceived social competence beyond the effects of the other types of victimization. Classroom aggression moderated the relation of verbal victimization with perceived social competence, such that this relation was significant in classrooms with lower aggression and nonsignificant in classrooms with higher aggression. Results indicate that preadolescents' daily self-perceptions fluctuate with daily victimization by peers, particularly with social rebuff. Findings also suggest that the impact of verbal victimization on children's self-views could be exacerbated in classrooms that better manage peer-to-peer aggression. Accordingly, targeted interventions appear critical for children who continue to experience peer victimization in schools with highly effective aggression prevention programs.

Effects of Amphipathic Polypeptides on Membrane Organization Inferred from Studies Using Bicellar Lipid Mixtures.

SP-B63-78, a lung surfactant protein fragment, and magainin 2, an antimicrobial peptide, are amphipathic peptides with the same overall charge but different biological functions. Deuterium nuclear magnetic resonance has been used to compare the interactions of these peptides with dispersions of 1,2-dimyristoyl- sn-glycero-3-phophocholine (DMPC)/1,2-dihexanoyl- sn-glycero-3-phophocholine (DHPC) (4:1) and DMPC/1,2-dimyristoyl- sn-glycero-3-phopho-(1'-rac-glycerol) (DMPG)/DHPC (3:1:1), two mixtures of long-chain and short-chain lipids that display bicellar behavior. This study exploited the sensitivity of a bicellar system structural organization to factors that modify partitioning of their lipid components between different environments. In small bicelle particles formed at low temperatures, short-chain components preferentially occupy curved rim environments around bilayer disks of the long-chain components. Changes in chain order and lipid mixing, on heating, can drive transitions to more extended assemblies including a magnetically orientable phase at intermediate temperature. In this work, neither peptide had a substantial effect on the behavior of the zwitterionic DMPC/DHPC mixture. For bicellar mixtures containing the anionic lipid DMPG, the peptide SP-B63-78 lowered the temperature at which magnetically orientable particles coalesced into more extended lamellar structures. SP-B63-78 did not promote partitioning of the zwitterionic and anionic long-chain lipid components into different environments. Magainin 2, on the other hand, was found to promote separation of the anionic lipid, DMPG, and the zwitterionic lipid, DMPC, into different environments for temperatures above 34 °C. The contrast between the effects of these two peptides on the lipid mixtures studied appears to be consistent with their functional roles in biological systems.

The simultaneous assessment of and relations between children's sympathetic and parasympathetic psychophysiology and their reactive and proactive aggression.

The goal of the current study was to examine the link between children's psychophysiology and aggression when both constructs were assessed simultaneously in scenarios designed to provide the opportunity to aggress for either a reactive reason or a proactive reason. Both sympathetic nervous system (SNS) activity (skin conductance) and parasympathetic nervous system (PNS) activity (respiratory sinus arrhythmia or RSA), as well as their interaction, were included as physiological measures. Participants were 35 5th-grade children who were placed in two virtual-peer scenarios; one scenario provided the opportunity to aggress in response to peer provocation (i.e., reactive aggression) and the other scenario provided the opportunity to aggress for instrumental gain (i.e., proactive aggression). Both skin conductance and RSA were assessed at the time that children were given the opportunity to aggress; this simultaneous assessment of psychophysiology and aggression allowed for an examination of in-the-moment relations between the two constructs. For the reactive scenario, RSA moderated the in-the-moment relation between skin conductance and aggression such that the association was positive at low RSA but negative at high RSA. For the proactive scenario, skin conductance negatively predicted aggression in-the-moment, and RSA positively predicted aggression in-the-moment, but their interaction was not a significant predictor of aggression. Theoretical implications for reactive and proactive aggression and underlying physiological processes are discussed.

Nonconscious priming enhances the therapy relationship: An experimental analog study.

OBJECTIVE: We examined whether nonconscious priming could enhance client perceptions of the therapist in an experimental analog study. METHOD: Sixty undergraduate participants each played the part of client in a brief scripted role-play of a therapy intake session. Sessions lasted about 3 min. The first author played the role of therapist. Participants were randomly assigned to view a line drawing depicting two individuals pointing either in the same direction (empathy prime) or opposite directions (non-empathy prime). The drawing appeared as a watermark on initial screening forms. The experimenter was blind to group assignment and participants received a cover story designed to mask the purpose of the primes. RESULTS: Participants in the empathy priming condition gave the therapist higher ratings of empathy and congruence, spoke to him longer, and rated their likelihood of future progress higher compared to participants in the non-empathy group. None of the participants expressed awareness of the priming manipulation during a funneled debriefing. CONCLUSIONS: These results demonstrate that client ratings of the therapy relationship can be experimentally manipulated without awareness and open the door to experimental studies of the association between relationship factors and treatment outcome.

REVIEW OF MORTALITY AND EFFECTIVENESS OF NEONATAL TREATMENT IN CAPTIVE ATTWATER'S PRAIRIE CHICKENS ( TYMPANUCHUS CUPIDO ATTWATERI).

A retrospective study of mortality and neonatal treatment for 975 Attwater's prairie chickens ( Tympanuchus cupido attwateri) was conducted from 2009-2015. Gross necropsy, histopathology, and medical records for chicks less than 8 wk of age at the Houston Zoo were reviewed and summarized. The total mortality rate was 36% ( n = 352). Yolk sac infection was the most common cause of death (21%; n = 73) followed by maladaptation (19%; n = 68), musculoskeletal abnormalities (19%; n = 66), necrotic enteritis (13%; n = 44), and mucoid enteritis (11%; n = 39). Gavage feeding of a highly digestible formula was associated with a higher proportion of necrotic enteritis and a reduced occurrence of both mucoid enteritis and maladaptation. Meloxicam administration had a significantly reduced incidence of mucoid enteritis. Survival past 10 days of age, prevention of neonatal bacterial infection, and supportive treatment with meloxicam are important factors to increase chick survival and improve captive breeding success for Attwater's prairie chickens. Gavage feeding of a diet low in digestible carbohydrates has the potential for reducing disease due to necrotic enteritis in this species.

Divide & Conquer: Surfactant Protein SP-C and Cholesterol Modulate Phase Segregation in Lung Surfactant.

Lung surfactant (LS) is an essential system supporting the respiratory function. Cholesterol can be deleterious for LS function, a condition that is reversed by the presence of the lipopeptide SP-C. In this work, the structure of LS-mimicking membranes has been analyzed under the combined effect of SP-C and cholesterol by deuterium NMR and phosphorus NMR and by electron spin resonance. Our results show that SP-C induces phase segregation at 37°C, resulting in an ordered phase with spectral features resembling an interdigitated state enriched in dipalmitoylphosphatidylcholine, a liquid-crystalline bilayer phase, and an extremely mobile phase consistent with small vesicles or micelles. In the presence of cholesterol, POPC and POPG motion seem to be more hindered by SP-C than dipalmitoylphosphatidylcholine. The use of deuterated cholesterol did not show signs of specific interactions that could be attributed to SP-C or to the other hydrophobic surfactant protein SP-B. Palmitoylation of SP-C had an indirect effect on the extent of protein-lipid perturbations by stabilizing SP-C structure, and seemed to be important to maximize differences among the lipids participating in each phase. These results shed some light on how SP-C-induced lipid perturbations can alter membrane structure to sustain LS functionality at the air-liquid interface.

Relations Between Actual Group Norms, Perceived Peer Behavior, and Bystander Children's Intervention to Bullying.

The goals of the study were (a) to predict children's intervention in bullying situations from class-level norms for intervention, as well as child-level perceptions of the number of peers who would intervene, and (b) to determine whether these predictions held when accounting for children's levels of empathy, prosocial behavior, and callous-unemotional traits. Participants were 751 racially and ethnically diverse fourth- and fifth-grade students (53.8% female) in 43 classes. Participants completed peer nominations about which classmates they perceived would intervene during bullying situations. Empathy and callous-unemotional traits were assessed via self report, whereas prosocial behavior was measured through peer report. Using multilevel modeling, each child's intervention in bullying was positively predicted from class-level norms for intervention (class means for the percentage of children who nominated each child as intervening) but negatively predicted from child-level perceptions of the number of peers who would intervene, after accounting for the 3 child traits. Class-level findings support past research on group norms which suggest that children are more likely to display a behavior if their peers display the same behavior. Child-level findings support the presence of the "bystander effect" in children's bullying episodes, in which children are less likely to intervene if they believe that more peers will do so. Thus, although children were more likely to intervene in classrooms with cultures that made intervention more normative, within the context of each class's culture, children were more likely to intervene if they perceived that fewer peers would do so.

Specific alleles at immune genes, rather than genome-wide heterozygosity, are related to immunity and survival in the critically endangered Attwater's prairie-chicken.

The negative effects of inbreeding on fitness are serious concerns for populations of endangered species. Reduced fitness has been associated with lower genome-wide heterozygosity and immune gene diversity in the wild; however, it is rare that both types of genetic measures are included in the same study. Thus, it is often unclear whether the variation in fitness is due to the general effects of inbreeding, immunity-related genes or both. Here, we tested whether genome-wide heterozygosity (20 990 SNPs) and diversity at nine immune genes were better predictors of two measures of fitness (immune response and survival) in the endangered Attwater's prairie-chicken (Tympanuchus cupido attwateri). We found that postrelease survival of captive-bred birds was related to alleles of the innate (Toll-like receptors, TLRs) and adaptive (major histocompatibility complex, MHC) immune systems, but not to genome-wide heterozygosity. Likewise, we found that the immune response at the time of release was related to TLR and MHC alleles, and not to genome-wide heterozygosity. Overall, this study demonstrates that immune genes may serve as important genetic markers when monitoring fitness in inbred populations and that in some populations specific functional genes may be better predictors of fitness than genome-wide heterozygosity.

Effect of an Anionic Lipid on the Barotropic Behavior of a Ternary Bicellar Mixture.

Dispersions of lipid mixtures comprising long- and short-chain phospholipids (bicellar mixtures) can form small isotropically reorienting particles (bilayered micelles), magnetically orientable stuctures, or unorientable lamellar structures. Application of hydrostatic pressure can also induce interdigitation of the long-chain lipid components. In this work, variable-pressure 2H NMR was used to study the effect of head group charge on the barotropic behavior of bicellar mixtures. Observations at pressures up to 152 MPa and temperatures up to 64 °C were combined with earlier observations at lower pressure and lower temperature to obtain a pressure-temperature phase diagram for DMPC-d54/DMPG/DHPC (3:1:1). In this phase diagram, a region corresponding to small, isotropically reorienting particles at lower temperature and higher pressure is separated from a region corresponding to unorientable lamellar organization, at higher temperature and lower pressure, by a band in which the magnetically orientable phase is stable below ∼100 MPa and in which an interdigitated gel phase is stable above ∼120 MPa. From ∼46 to ∼52 °C, the dispersion transforms directly from the unorientable lamellar to isotropically reorienting particle phases upon isothermal pressurization. The extent to which this behavior reflects the presence of anionic lipid in the long-chain fraction of this mixture is illustrated by comparison with spectral series obtained during isothermal pressurization of DMPC-d54/DHPC (4:1) and DMPC-d54/DMPG/DHPC (2.7:1.3:1) at selected temperatures. These observations show how electrostatic interactions at a bilayer surface can affect the balance between hydrophobic and hydrophilic interactions that is reflected by a dispersion's barotropic phase behavior.

Decreased small mammal and on-host tick abundance in association with invasive red imported fire ants (Solenopsis invicta).

Invasive species may impact pathogen transmission by altering the distributions and interactions among native vertebrate reservoir hosts and arthropod vectors. Here, we examined the direct and indirect effects of the red imported fire ant (Solenopsis invicta) on the native tick, small mammal and pathogen community in southeast Texas. Using a replicated large-scale field manipulation study, we show that small mammals were more abundant on treatment plots where S. invicta populations were experimentally reduced. Our analysis of ticks on small mammal hosts demonstrated a threefold increase in the ticks caught per unit effort on treatment relative to control plots, and elevated tick loads (a 27-fold increase) on one common rodent species. We detected only one known human pathogen (Rickettsia parkeri), present in 1.4% of larvae and 6.7% of nymph on-host Amblyomma maculatum samples but with no significant difference between treatment and control plots. Given that host and vector population dynamics are key drivers of pathogen transmission, the reduced small mammal and tick abundance associated with S. invicta may alter pathogen transmission dynamics over broader spatial scales.

Indirect effects of red imported fire ants on Attwater's prairie-chicken brood survival.

The invasive red imported fire ant (Solenopsis invicta) has negatively affected a host of taxonomic groups throughout its acquired North American range. Many studies have hypothesized indirect trophic impacts, but few documented those impacts. We evaluated invertebrate abundance as a factor limiting juvenile survival of the endangered Attwater's prairie-chicken (Tympanuchus cupido attwateri), and whether fire ants reduce invertebrate numbers and biomass. From 2009-2013, we monitored survival of Attwater's prairie-chicken broods (n = 63) with radio telemetry during the first 2 weeks post-hatch and collected daily invertebrate samples at brood sites. Broods located in areas with the highest median invertebrate count (338 invertebrates/25 sweeps) had a survival probability of 0.83 at 2 weeks post-hatch compared to 0.07 for broods located in areas with the lowest median invertebrate count (18 invertebrates/25 sweeps). During 2011-2012, we evaluated the reduction of fire ants on invertebrate numbers and biomass by aerially treating areas with Extinguish Plus™ in an impact-reference study design. Treated fields had 27% more individual invertebrates and 26% higher invertebrate biomass than reference fields. Our results clearly document that invertebrate abundance affects Attwater's prairie-chicken brood survival and that fire ants may indirectly contribute to low brood survival by suppressing invertebrate abundance. We posit that within the fire ant's acquired North American range, fire ants are likely contributing to declines of other insectivorous species. Published 2015. This article is a U.S. Government work and is in the public domain in the USA.

Evaluation of wireless sensor networks (WSNs) for remote wetland monitoring: design and initial results.

Here, we describe and evaluate two low-power wireless sensor networks (WSNs) designed to remotely monitor wetland hydrochemical dynamics over time scales ranging from minutes to decades. Each WSN (one student-built and one commercial) has multiple nodes to monitor water level, precipitation, evapotranspiration, temperature, and major solutes at user-defined time intervals. Both WSNs can be configured to report data in near real time via the internet. Based on deployments in two isolated wetlands, we report highly resolved water budgets, transient reversals of flow path, rates of transpiration from peatlands and the dynamics of chromophoric-dissolved organic matter and bulk ionic solutes (specific conductivity)-all on daily or subdaily time scales. Initial results indicate that direct precipitation and evapotranspiration dominate the hydrologic budget of both study wetlands, despite their relatively flat geomorphology and proximity to elevated uplands. Rates of transpiration from peatland sites were typically greater than evaporation from open waters but were more challenging to integrate spatially. Due to the high specific yield of peat, the hydrologic gradient between peatland and open water varied with precipitation events and intervening periods of dry out. The resultant flow path reversals implied that the flux of solutes across the riparian boundary varied over daily time scales. We conclude that WSNs can be deployed in remote wetland-dominated ecosystems at relatively low cost to assess the hydrochemical impacts of weather, climate, and other perturbations.

A comparison of pedigree- and DNA-based measures for identifying inbreeding depression in the critically endangered Attwater's Prairie-chicken.

The primary goal of captive breeding programmes for endangered species is to prevent extinction, a component of which includes the preservation of genetic diversity and avoidance of inbreeding. This is typically accomplished by minimizing mean kinship in the population, thereby maintaining equal representation of the genetic founders used to initiate the captive population. If errors in the pedigree do exist, such an approach becomes less effective for minimizing inbreeding depression. In this study, both pedigree- and DNA-based methods were used to assess whether inbreeding depression existed in the captive population of the critically endangered Attwater's Prairie-chicken (Tympanuchus cupido attwateri), a subspecies of prairie grouse that has experienced a significant decline in abundance and concurrent reduction in neutral genetic diversity. When examining the captive population for signs of inbreeding, variation in pedigree-based inbreeding coefficients (f(pedigree)) was less than that obtained from DNA-based methods (f(DNA)). Mortality of chicks and adults in captivity were also positively correlated with parental relatedness (r(DNA)) and f(DNA), respectively, while no correlation was observed with pedigree-based measures when controlling for additional variables such as age, breeding facility, gender and captive/release status. Further, individual homozygosity by loci (HL) and parental rDNA values were positively correlated with adult mortality in captivity and the occurrence of a lethal congenital defect in chicks, respectively, suggesting that inbreeding may be a contributing factor increasing the frequency of this condition among Attwater's Prairie-chickens. This study highlights the importance of using DNA-based methods to better inform management decisions when pedigrees are incomplete or errors may exist due to uncertainty in pairings.

Response to hydrostatic pressure of bicellar dispersions containing an anionic lipid: pressure-induced interdigitation.

Bicellar dispersions of chain perdeuterated 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC-d54), 1,2-dimyristoyl-sn-glycero-3-phospho-(1'-rac-glycerol) (DMPG), and 1,2-dihexanoyl-sn-glycero-3-phosphocholine (DHPC), with molar ratios of 3:1:1, were studied using variable-pressure (2)H NMR spectroscopy at hydrostatic pressures up to 125 MPa. Upon warming of the dispersions, spectra at ambient pressure indicated a progressive coalescence from small bilayered disks undergoing isotropic reorientation to more extended micellar structures in which spectra indicated anisotropic reorientation and, under some conditions, magnetic orientation and finally to randomly oriented lamellae or multilamellar vesicles. Temperatures for the onsets of anisotropic reorientation and random lamellar orientation increased with pressure at rates of 0.22 and 0.15 °C/MPa, respectively. In the 3.5-T magnetic field used for this work, magnetic orientation within the intermediate phase was not observed at 83 MPa or higher pressures. Comparison of spectra obtained at fixed pressure showed significant asymmetry between behaviors upon warming and cooling. For samples of DMPC-d54/DMPG/DHPC (3:1:1), but not DMPC-d54/DHPC (4:1), a persistent interdigitated phase was formed after repeated cooling from high temperature at 83 MPa. This is likely a metastable phase and might reflect kinetic trapping of the short-chain lipid component, DHPC, in a nonequilibrium spatial distribution as temperature is lowered at high pressure. Bicellar dispersions typically behave differently upon warming and cooling, and these observations could provide some insight into the observed behaviors in such systems. This work also suggests the possibility of trapping bicellar dispersions in persistent nonequilibrium morphologies.

Dependence of bicellar system phase behavior and dynamics on anionic lipid concentration.

Bicellar dispersions of chain perdeuterated 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC-d54) and 1,2-dihexanoyl-sn-glycero-3-phosphocholine (DHPC) were prepared with the molar fraction of DHPC held fixed at 20% and varying amounts of DMPC replaced by the anionic lipid 1,2-dimyristoyl-sn-glycero-3-phospho-(1'-rac-glycerol) (DMPG). (2)H NMR spectra were examined to assess the effect of added DMPG on mixture phase behavior and morphology. Quadrupole echo decay and quadrupole-Carr-Purcell-Mieboom-Gill echo train measurements provided information about slow motions contributing to echo decay in the high temperature phases. The spectra and quadrupole echo decay properties of DMPC-d54/DHPC (4:1) and DMPC-d54/DMPG/DHPC (3:1:1) were qualitatively similar. With increasing DMPG concentration, the transition between the magnetically orientable phase and the higher temperature phase became increasingly distinct, and the spectral shape and echo decay characteristics of the high temperature bicellar phase became increasingly similar to those of DMPC-d54 in the liquid crystalline phase. The observation that DMPG changes spectra in the orientable phase incrementally while increasing the distinction between the orientable and high temperature bicellar phases provides new insights into how DMPG influences bicellar mixture morphology.

²H solid-state nuclear magnetic resonance investigation of whole Escherichia coli interacting with antimicrobial peptide MSI-78.

A key aspect of the activity of antimicrobial peptides (AMPs) is their interaction with membranes. Efforts to elucidate their detailed mechanisms have focused on applying biophysical methods, including nuclear magnetic resonance (NMR), to AMPs in model lipid systems. However, these highly simplified systems fail to capture many of the features of the much more complex cell envelopes with which AMPs interact in vivo. To address this issue, we have designed a procedure to incorporate high levels of (2)H NMR labels specifically into the cell membrane of Escherichia coli and used this approach to study the interactions between the AMP MSI-78 and the membranes of intact bacteria. The (2)H NMR spectra of these membrane-deuterated bacteria can be reproduced in the absence and presence of MSI-78. Because the (2)H NMR data provide a quantitative measure of lipid disorder, they directly report on the lipid bilayer disruption central to the function of AMPs, in the context of intact bacteria. Addition of MSI-78 to the bacteria leads to decreases in the order of the lipid acyl chains. The molar peptide:lipid ratios required to observe the effects of MSI-78 on acyl chain order are approximately 30 times greater than the ratios needed to observe effects in model lipid systems and approximately 100 times less than the ratios required to observe inhibition of cell growth in biological assays. The observations thus suggest that MSI-78 disrupts the bilayer even at sublethal AMP levels and that a large fraction of the peptide does not actually reach the inner membrane.

Dynamic properties of bicellar lipid mixtures observed by rheometry and quadrupole echo decay.

In bicellar dispersions of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) and 1,2-dihexanoyl-sn-glycero-3-phosphocholine (DHPC), the transition from isotropic reorientation to partial orientational order, on warming, is known to coincide with a sharp increase in viscosity. In this work, cone-and-plate rheometry, (2)H NMR spectroscopy, and quadrupole echo decay observations have been used to obtain new insights into the dynamics of phases observed in bicellar DMPC/DHPC mixtures. Samples with 25% of the DMPC component deuterated were used to correlate rheological measurements with phase behavior observed by (2)H NMR spectroscopy. Mixtures containing only normal DMPC (DMPC/DHPC) or only chain perdeuterated DMPC (DMPC-d(54)/DHPC) were used to refine rheology and quadrupole echo decay measurements respectively. The viscosity peaked at 4-9 Pa·s, just above the isotropic-to-nematic transition, and then dropped as samples were warmed through the nematic-to-lamellar transition. Quadrupole echo decay times above the nematic-to-lamellar transition were significantly longer than typically observed in the liquid crystalline phase of saturated lipid multilamellar vesicles. This may indicate a damping of slow bilayer undulations resulting from the coupling of opposite bilayer surfaces by DHPC-lined pores.

Effects of the lung surfactant protein B construct Mini-B on lipid bilayer order and topography.

The hydrophobic lung surfactant protein, SP-B, is essential for survival. Cycling of lung volume during respiration requires a surface-active lipid-protein layer at the alveolar air-water interface. SP-B may contribute to surfactant layer maintenance and renewal by facilitating contact and transfer between the surface layer and bilayer reservoirs of surfactant material. However, only small effects of SP-B on phospholipid orientational order in model systems have been reported. In this study, N-terminal (SP-B(8-25)) and C-terminal (SP-B(63-78)) helices of SP-B, either linked as Mini-B or unlinked but present in equal amounts, were incorporated into either model phospholipid mixtures or into bovine lipid extract surfactant in the form of vesicle dispersions or mechanically oriented bilayer samples. Deuterium and phosphorus nuclear magnetic resonance (NMR) were used to characterize effects of these peptides on phospholipid chain orientational order, headgroup orientation, and the response of lipid-peptide mixtures to mechanical orientation by mica plates. Only small effects on chain orientational order or headgroup orientation, in either vesicle or mechanically oriented samples, were seen. In mechanically constrained samples, however, Mini-B and its component helices did have specific effects on the propensity of lipid-peptide mixtures to form unoriented bilayer populations which do not exchange with the oriented fraction on the timescale of the NMR experiment. Modification of local bilayer orientation, even in the presence of mechanical constraint, may be relevant to the transfer of material from bilayer reservoirs to a flat surface-active layer, a process that likely requires contact facilitated by the formation of highly curved protrusions.