STICCS Reveals Matrix-Dependent Adhesion Slipping and Gripping in?Migrating Cells

Two-color spatio-temporal image cross-correlation spectroscopy (STICCS) is a new, to our knowledge, image analysis method that calculates space-time autocorrelation and cross-correlation functions from fluorescence intensity fluctuations. STICCS generates cellular flow and diffusion maps that reveal interactions and cotransport of two distinct molecular species labeled with different fluorophores. Here we use computer simulations to map the capabilities and limitations of STICCS for measurements in complex heterogeneous environments containing micro- and macrostructures. We then use STICCS to analyze the co-flux of adhesion components in migrating cells imaged using total internal reflection fluorescence microscopy. The data reveal a robust, time-dependent co-fluxing of certain integrins and paxillin in adhesions in protrusions when they pause, and in adhesions that are sliding and disassembling, demonstrating that the molecules in these adhesions move as a complex. In these regions, both α6β1- or αLβ2-integrins, expressed in CHO.B2 cells, co-flux with paxillin; an analogous cotransport was seen for α6β1-integrin and α-actinin in U2OS. This contrasts with the behavior of the α5β1-integrin and paxillin, which do not co-flux. Our results clearly show that integrins can move in complexes with adhesion proteins in protrusions that are retracting.     

Visualizing biomolecular electrostatics in virtual reality with UnityMol‐APBS

Virtual reality is a powerful tool with the ability to immerse a user within a completely external environment. This immersion is particularly useful when visualizing and analyzing interactions between small organic molecules, molecular inorganic complexes, and biomolecular systems such as redox proteins and enzymes. A common tool used in the biomedical community to analyze such interactions is the Adaptive Poisson‐Boltzmann Solver (APBS) software, which was developed to solve the equations of continuum electrostatics for large biomolecular assemblages. Numerous applications exist for using APBS in the biomedical community including analysis of protein ligand interactions and APBS has enjoyed widespread adoption throughout the biomedical community. Currently, typical use of the full APBS toolset is completed via the command line followed by visualization using a variety of two‐dimensional external molecular visualization software. This process has inherent limitations: visualization of three‐dimensional objects using a two‐dimensional interface masks important information within the depth component. Herein, we have developed a single application, UnityMol‐APBS, that provides a dual experience where users can utilize the full range of the APBS toolset, without the use of a command line interface, by use of a simple graphical user interface (GUI) for either a standard desktop or immersive virtual reality experience.     

Development of High‐Level Omega‐3 Eicosapentaenoic Acid (EPA) Production from Phaeodactylum tricornutum

Phaeodactylum tricornutum is a lipid‐rich marine diatom that contains a high level of omega‐3 polyunsaturated fatty acids, especially eicosapentaenoic acid (EPA). In an effort to reduce costs for large‐scale cultivation of this microalga, this study first established a New BBM medium (0.3 x strength BBM with only 3% of the initial phosphate level) to replace the traditional F/2 medium. Phaeodactylum tricornutum could grow in extremely low phosphate concentrations (25 µM), without compromising the EPA content. In the presence of sea salts, silicate addition was not necessary for high rate growth, high EPA content, or lipid accumulation in this species. Using urea as the sole nitrogen source tended to increase EPA contents per dry biomass (by 24.7%) while not affecting growth performance. The use of sea salts, rather than just sodium chloride, led to significantly improved biomass yields (20% increase) and EPA contents of total fatty acid (46–52% increase), most likely because it supplied sufficient essential elements such as magnesium. A salinity level of 35 led to significantly higher biomass yields compared with 20, but salinity had no significant influence on EPA content. EPA became the dominant fatty acid with average levels of 51.8% of total fatty acids during the exponential growth phase at 20 ppt in New BBM medium with sea salts.     

TMEM41B and VMP1 are scramblases and regulate the distribution of cholesterol and phosphatidylserine

TMEM41B and VMP1 are integral membrane proteins of the endoplasmic reticulum (ER) and regulate the formation of autophagosomes, lipid droplets (LDs), and lipoproteins. Recently, TMEM41B was identified as a crucial host factor for infection by all coronaviruses and flaviviruses. The molecular function of TMEM41B and VMP1, which belong to a large evolutionarily conserved family, remains elusive. Here, we show that TMEM41B and VMP1 are phospholipid scramblases whose deficiency impairs the normal cellular distribution of cholesterol and phosphatidylserine. Their mechanism of action on LD formation is likely to be different from that of seipin. Their role in maintaining cellular phosphatidylserine and cholesterol homeostasis may partially explain their requirement for viral infection. Our results suggest that the proper sorting and distribution of cellular lipids are essential for organelle biogenesis and viral infection.     

The incidence of turnip yellows virus in oilseed rape crops (Brassica napus L.) in three different regions of England over three consecutive growing seasons and the relationship with the abundance of flying Myzus persicae

Turnip yellows virus (TuYV) is the most important virus infecting oilseed rape in the United Kingdom. The incidence and spatial distribution of TuYV in winter oilseed rape (WOSR) crops in three regions of England were determined over three growing seasons. Leaf samples were collected from three fields in each region, in autumn (November–December) and spring (April) of the three crop seasons and tested for virus presence by enzyme-linked immunosorbent assay. Infection was detected in all fields except one. Higher TuYV incidences were recorded in 2007–2008 (≤89%) and 2009–2010 (≤100%) crop seasons than in 2008–2009 (≤24%). Highest incidences were recorded in Lincolnshire (≤100%), followed by Warwickshire (≤88%), with lowest incidences in Yorkshire (1–74%). There was a significant increase in incidence detected between autumn and spring sampling in eight fields, a significant decrease in one field and no significant change in 18 fields. Rothamsted Insect Survey suction trap data for the aphid Myzus persicae in Lincolnshire, Warwickshire and Yorkshire revealed two peaks of flight activity in most years (2007–2009). The second peak (September–November) coincided with emergence of WOSR. The highest cumulative (August–November) trap catches in the three regions during the three crop seasons occurred in Lincolnshire and the lowest in Yorkshire; catches in autumn 2009 were highest and lowest in autumn 2008. Regression analysis revealed a highly significant association between the cumulative numbers of M. persicae caught in the suction traps closest to the crops between August and November each year and the incidence of TuYV detected in the WOSR crops in the autumn of each year. Results are discussed in the light of factors affecting the spread of TuYV and future possibilities for control.     

Toxic effects of mercury(II), cadmium(II) and lead(II) porphyrins on Trypanosoma brucei brucei growth

The effects of free mercury(II), cadmium(II) and lead(II) ions and their metalloporphyrin-derivatives on Trypanosoma brucei brucei growth in culture were studied. All experiments were conducted in the dark. IC(50) values on growth obtained in 24-h time-course experiments were 1.5 x 10(-7), 2.4 x 10(-6), 4.4 x 10(-6) and 2.6 x 10(-5) M for mercury(II) porphyrin, cadmium(II) porphyrin, lead(II) porphyrin and free base porphyrin, respectively. While the IC50 values for Hg2+, Cd2+ and Pb2+ were 3.6 x 10(-6), 1.5 x 10(-5) and 1.6 x 10(-5) M, respectively. These results clearly indicate that the toxicity of the metalloporphyrin complexes of mercury(II), cadmium(II) and lead(II) to T. b. brucei parasites was much higher compared to their free metal ions and free base porphyrin at low concentrations. It was also observed after 8 h incubation that the metalloporphyrins were effective in inhibiting the division of the parasites at concentrations >1.25 x 10(-7) M for mercury(II) porphyrin, concentrations >1.2 x 10(-6) M for cadmium(II) and lead(II) porphyrins and at concentrations >3.6 x 10(-6) M for Hg2+ ion. These observations were not detected in samples treated with the free metal ions and the free base porphyrin at the same concentrations. Interestingly, trypanosomes treated with metalloporphyrin complexes displayed different morphological features from those cells treated with free base porphyrin or metal ions. The chemotherapeutic potential of the metalloporphyrins of H2TMPyP for treatment of African trypanosomiasis is discussed.     

Characterization of a novel organelle in Toxoplasma gondii with similar composition and function to the plant vacuole

Toxoplasma gondii belongs to the phylum Apicomplexa and is an important cause of congenital disease and infection in immunocompromised patients. Like most apicomplexans, T. gondii possesses several plant‐like features, such as the chloroplast‐like organelle, the apicoplast. We describe and characterize a novel organelle in T. gondii tachyzoites, which is visible by light microscopy and possesses a broad similarity to the plant vacuole. Electron tomography shows the interaction of this vacuole with other organelles. The presence of a plant‐like vacuolar proton pyrophosphatase (TgVP1), a vacuolar proton ATPase, a cathepsin L‐like protease (TgCPL), an aquaporin (TgAQP1), as well as Ca²⁺/H⁺ and Na⁺/H⁺ exchange activities, supports similarity to the plant vacuole. Biochemical characterization of TgVP1 in enriched fractions shows a functional similarity to the respective plant enzyme. The organelle is a Ca²⁺ store and appears to have protective effects against salt stress potentially linked to its sodium transport activity. In intracellular parasites, the organelle fragments, with some markers colocalizing with the late endosomal marker, Rab7, suggesting its involvement with the endocytic pathway. Studies on the characterization of this novel organelle will be relevant to the identification of novel targets for chemotherapy against T. gondii and other apicomplexan parasites as well.     

Adding value to cocoa (Theobroma cacao L.) germplasm information with domestication history and admixture mapping

A sound understanding of crop history can provide the basis for deriving novel genetic information through admixture mapping. We confirmed this, by using characterization data from an international collection of cocoa, collected 25 years ago, and from a contemporary plantation. We focus on the trees derived from three centuries of admixture between Meso-American Criollo and South American Forastero genomes. In both cacao sets of individuals, linkage disequilibrium extended over long genetic distances along chromosome regions, as expected in populations derived from recent admixture. Based on loose genome scans, genomic regions involved in useful traits were identified. Fifteen genomic regions involved in seed and fruit weight variation were highlighted. They correspond to ten previously identified QTLs and five novel ones. Admixture mapping can help to add value to genetic resources and thus, help to encourage investment in their conservation. Maria Marcano and Tatiana Pugh contributed equally to this work.     

Skeletal mass in adolescent male athletes and nonathletes: relationships with high-impact sports

Dias Quiterio, AL, Canero, EA, Baptista, FM, and Sardinha, LB. Skeletal mass in adolescent male athletes and nonathletes: relationships with high-impact sports. J Strength Cond Res 25(12): 3439-3447, 2011-This study examined the relationships between the practice of different categories of sports (high-impact vs. nonimpact) and bone status in adolescent male athletes and investigated differences from an age-matched control group. A total of 54 adolescent male athletes and 26 adolescent nonathletes were evaluated. Bone mineral density, bone mineral content (BMC), and bone area at the whole-body, limbs, and lumbar spine were determined by dual-energy x-ray absorptiometry, along with total and regional fat-free mass and body fat. The high-impact group included 34 athletes: 9 gymnasts, 18 basketball players, and 7 handball players (age: 15.7 ± 1.6 years; weight: 72.0 ± 15.0 kg; height: 178.5 ± 12.5 cm). The nonimpact group consisted of 20 swimmers (age: 16.4 ± 2.5 years; weight: 66.9 ± 10.4 kg; height: 173.7 ± 10.9 cm). The nonathletic control group included 26 male adolescents (age: 15.9 ± 2.8 years; weight: 64.7 ± 16.3 kg; height: 168.6 ± 15.1 cm). No differences were observed between the nonimpact and the control group in all bone variables, before and after adjustments for maturation level, body weight, and height (p > 0.05). After adjustments for these variables, the high-impact group displayed greater bone mass in most of the measured sites when compared to the other 2 groups (p < 0.001). Subjects in the nonimpact group showed lower values of BMC, particularly in the lower limbs, than both the high-impact and the nonathletic control groups (p < 0.05) after adjustments for maturation, high, and fat-free mass. This study reinforces the positive associations between high-impact physical activities and skeletal health in adolescent boys.