Coherent Neutron Scattering and Collective Dynamics in the Protein,GFP

Collective dynamics are considered to be one of the major properties of soft materials, including biological macromolecules. We present coherent neutron scattering studies of the low-frequency vibrations, the so-called boson peak, in fully deuterated green fluorescent protein (GFP). Our analysis revealed unexpectedly low coherence of the atomic motions in GFP. This result implies a low amount of in-phase collective motion of the secondary structural units contributing to the boson peak vibrations and fast conformational fluctuations on the picosecond timescale. These observations are in contrast to earlier studies of polymers and glass-forming systems, and suggest that random or out-of-phase motions of the β-strands contribute greater than two-thirds of the intensity to the low-frequency vibrational spectra of GFP.     

Wiskott-Aldrich Syndrome Protein (WASp) Controls the Delivery of Platelet Transforming Growth Factor-β1

Platelets are immunologically competent cells containing cytokines such as TGF-β1 that regulate cell-mediated immunity. However, the mechanisms underlying cytokine secretion from platelets are undefined. The Wiskott-Aldrich syndrome protein (WASp) regulates actin polymerization in nucleated hematopoietic cells but has other role(s) in platelets. WASp-null (WASp^−/−) platelets stimulated with a PAR-4 receptor agonist had increased TGF-β1 release compared with WT platelets; inhibiting WASp function with wiskostatin augmented TRAP-induced TGF-β1 release in human platelets. TGF-β1 release is dissociated from α-granule secretion (P-selectin up-regulation) and occurs more gradually, with ∼10–15% released after 30–60 min. Blockade of Src family kinase-mediated WASp Tyr-291/Tyr-293 phosphorylation increased TGF-β1 release, with no additive effect in WASp^−/− platelets, signifying that phosphorylation is critical for WASp-limited TGF-β1 secretion. Inhibiting F-actin assembly with cytochalasin D enhanced secretion in WT platelets and further increased TGF-β1 release in WASp^−/− platelets, indicating that WASp and actin assembly independently regulate TGF-β1 release. A permeabilized platelet model was used to test the role of upstream small GTPases in TGF-β1 release. N17Cdc42, but not Rac1 mutants, increased TGF-β1 secretion and abrogated WASp phosphorylation. We conclude that WASp function restricts TGF-β1 secretion in a Cdc42- and Src family kinase-dependent manner and independently of actin assembly.     

Weather impacts on leisure activities in Halifax,Nova Scotia

The aim of this study was to investigate the impact of daily atmospheric weather conditions on daily leisure activity engagement, with a focus on physically active leisure. The methods capitalize on time diary data that were collected in Halifax, Nova Scotia to calculate objective measures of leisure activity engagement. Daily meteorological data from Environment Canada and daily sunrise and sunset times from the National Research Council of Canada are used to develop objective measures of the natural atmospheric environment. The time diary data were merged with the meteorological data in order to quantify the statistical association between daily weather conditions and the type, participation rate, frequency, and duration of leisure activity engagement. The results indicate that inclement and uncomfortable weather conditions, especially relating to thermal comfort and mechanical comfort, pose barriers to physically active leisure engagement, while promoting sedentary and home-based leisure activities. Overall, daily weather conditions exhibit modest, but significant, effects on leisure activity engagement; the strongest associations being for outdoor active sports and outdoor active leisure time budgets. In conclusion, weather conditions influence the type, participation rate, frequency, and duration of leisure activity engagement, which is an important consideration for health-promotion programming.     

FLOW-CYTOMETRIC ANALYSES OF NUCLEAR DNA CONTENT IN FOUR FAMILIES OF NEOTROPICAL BATS

Flow-cytometric analyses of 29 species of microchiropteran bats representing four families and 20 genera revealed that bats possess only 79% (5.43 pg) of the DNA content of a “typical” mammal (e.g., Mus musculus strain C57BL; 7 pg). Chiroptera, the second largest order of mammals, is thus an exception to the prevailing view that mammals possess a minimum nuclear DNA content of 7 pg. Limitations on cell size resulting from a high metabolic rate may have constrained evolution of DNA content and could explain why the extensive heterochromatic additions that are common in some groups of mammals are absent in bats. Chromosomes of bats have been well studied; detailed chromosomal banding data are available for nearly all the species used in this investigation. However, no significant correlations were found between DNA content and karyotypic characteristics such as 2n, fundamental number, and rate or pattern of chromosomal evolution.     

Micronization of a Soft Material: Air-Jet and Micro-Ball Milling

The air-jet and ball-mill are frequently used in fine micronization of active pharmaceutical ingredients to the order of 1–5 μm, which is important for increasing dissolution rates, and also for pulmonary delivery. In this study, we investigated the ability of air-jet and ball-mill to achieve adequate micronization on the lab scale using a model soft material, Pluronic® F-68. Material mechanical properties were characterized using the nanometer 600. Pluronic® F-68 was ball-milled in a micro-mill at different material weights and durations in liquid nitrogen vapor. In comparison, a lab scale air-jet mill was used at various milling parameters according to a full factorial design, where the response factors were particle yield and particle size distribution, which was analyzed using laser diffraction and scanning electron microscopy. The yield achieved with the micro-ball mill was 100% but was ~80% for the air-jet mill, which reduced the size of Pluronic® F-68 from 70 μm to sizes ranging between 23–39 μm median diameters. Ball milling produced particles less than 10 μm after 15 min. Although air-jet milling proved capable of particle size reduction of the relatively soft material Pluronic® F-68, limitations to the lower size range achievable were observed. The feed rate of the material into the air jet mill was a significant factor and slower feed rates lead to smaller sizes by allowing more time for particle collisions and subsequent particle breakage to occur. Micro-ball milling under cold condition was more successful at achieving a lower range particle size reduction of soft materials.     

Balance between maternal and paternal alleles sets the timing of resource accumulation in the maize endosperm

Key aspects of seed development in flowering plants are held to be under epigenetic control and to have evolved as a result of conflict between the interests of the male and female gametes (kinship theory). Attempts to identify the genes involved have focused on imprinted sequences, although imprinting is only one mechanism by which male or female parental alleles may be exclusively expressed immediately post-fertilization. We have studied the expression of a subset of endosperm gene classes immediately following interploidy crosses in maize and show that departure from the normal 2 : 1 ratio between female and male genomes exerts a dramatic effect on the timing of expression of some, but not all, genes investigated. Paternal genomic excess prolongs the expression of early genes and delays accumulation of reserves, while maternal genomic excess foreshortens the expression period of early genes and dramatically brings forward endosperm maturation. Our data point to a striking interdependence between the phases of endosperm development, and are consonant with previous work from maize showing progression from cell proliferation to endoreduplication is regulated by the balance between maternal and paternal genomes, and from Arabidopsis suggesting that this ‘phasing’ is regulated by maternally expressed imprinted genes. Our findings are discussed in context of the kinship theory.     

Stabilized bubbles in the body: pressure-radius relationships and the limits to stabilization

Van Liew, Hugh D., and Soumya Raychaudhuri. Stabilizedbubbles in the body: pressure-radius relationships and the limits tostabilization. J. Appl. Physiol.82(6): 2045-2053, 1997.We previously outlined the fundamentalprinciples that govern behavior of stabilized bubbles, such as themicrobubbles being put forward as ultrasound contrast agents. Ourpresent goals are to develop the idea that there are limits to thestabilization and to provide a conceptual framework for comparison ofbubbles stabilized by different mechanisms. Gases diffuse in or out ofstabilized bubbles in a limited and reversible manner in response tochanges in the environment, but strong growth influences will cause thebubbles to cross a threshold into uncontrolled growth. Also, bubblesstabilized by mechanical structures will be destroyed if outsideinfluences bring them below a critical small size. The in vivo behaviorof different kinds of stabilized bubbles can be compared by using plotsof bubble radius as a function of forces that affect diffusion of gasesin or out of the bubble. The two ends of the plot are the limits forunstabilized growth and destruction; these and the curve's slopepredict the bubble's practical usefulness for ultrasonic imaging orO₂ carriage to tissues.

     

Comparison of ageing methods and growth rates for largemouth bass,Micropterus salmoides Lacépède,from northern latitudes

Synopsis Ages determined by counts of apparent annuli on scales, sagittae, vertebrae, pectoral fin ray and dorsal fin spine cross sections of largemouth bass from northern populations, which are older and slower growing fish than in the southern parts of its native range, were compared to establish the accuracy of each method. Linear regression techniques indicated strong agreement (r> 0.9) among ages assigned from the examination of scales, sagittae, and vertebrae. The pattern of growth zones on pectoral fin ray and dorsal fm spine cross sections proved too variable for accurate age determination. Limited data suggest that ages greater than 7+ assigned from scales were more likely to underestimate true ages than the other body parts used, although none of these methods gave satisfactory results. Examination of scales from recovered tagged fish, and the similarity between back-calculated lengths of fish through age 7+ to annulus I and observed lengths of juvenile largemouth bass near the end of their first growing season, support the validity of ages determined from scales. Despite a very limited amount of habitat suitable for largemouth bass and severe climatic conditions, growth of this species in Tadenac Lake was similar to growth in other waters north of the Great Lakes. Differences in physical characteristics among these waters does not appear to influence growth rates of largemouth bass, but probably affects production and biomass.