Three-dimensional electron diffraction of plant light-harvesting complex

Electron diffraction patterns of two-dimensional crystals of light-harvesting chlorophyll a/b-protein complex (LHC-II) from photosynthetic membranes of pea chloroplasts, tilted at different angles up to 60°, were collected to 3.2 Å resolution at -125°C. The reflection intensities were merged into a three-dimensional data set. The Friedel R-factor and the merging R-factor were 21.8 and 27.6%, respectively. Specimen flatness and crystal size were critical for recording electron diffraction patterns from crystals at high tilts. The principal sources of experimental error were attributed to limitations of the number of unit cells contributing to an electron diffraction pattern, and to the critical electron dose. The distribution of strong diffraction spots indicated that the three-dimensional structure of LHC-II is less regular than that of other known membrane proteins and is not dominated by a particular feature of secondary structure.     

Small Angle X-Ray Scattering of the Thylakoid Membranes of Rhodopseudomonas spheroides in Aqueous Suspensions

The diffraction patterns of particles which have the shape of hollow spheres, i.e. vesicles, can be satisfactorily analyzed by means of a new formula of Weick (1974). This formula is used for the small angle X-ray scattering analysis of aqueous suspensions of thylakoids of Rhodopseudomonas spheroides. Some essential results are: (a) The membrane has a rather asymmetric structure with one layer of low electron density at its inner side and two layers of high electron density near the outer surface of the thylakoids. (b) The distance of the electron density maxima of the latter two layers is 45 ± 5 Å. (c) Between the two maxima is a region of an electron density nearly equal to that of water. (d) The sequence of the peaks is - + 0 + with increasing radius. The peaks extend over an interval of 120 ± 10 Å. (e) The thylakoids are strikingly of the same size. Their diameters, if defined by the outmost layer, vary statistically by about 4% and have an average value of approximately 640 Å.     

Influence of Temperature on Multiplication and Egg Hatching of Longidorus africanus

Longidorus africanus multiplication on tomato was highest at 29 °C. Few nematodes were recovered after 6 weeks at soil temperatures of 35 °C or below 23 °C. The time to egg hatching was shortest and the percentage of eggs hatching was highest at 29 °C. The minimum temperature and the heat sum above this temperature required for egg development were calculated to be 14.3 °C and 94.08 degree-days, respectively. The thermal times required for egg development by L. africanus and L. elongatus were nearly identical. For both species the product of the base temperature and the heat sum was near constant, and at a temperature of 22.3 °C the rates of egg development were equal.     

Stable Isotopes in Fish Eye Lenses as Potential Recorders of Trophic and Geographic History

We evaluated eye lenses as potential recorders of stable isotope histories in fish because they consist of metabolically inert optical proteins that are deposited in successive, concentric circles (laminae) much like otolith circuli and tree rings. We conducted four different tests on lenses from red snapper, red grouper, gag, and white grunt. The first test was a low-resolution screening of multiple individuals (4–5 radial groups of laminae per lens, all species except white grunt). Along the radial axis, all individuals exhibited substantial isotopic variability. Red snapper individuals separated into two groups based on δ ¹⁵N and gag separated into two groups based on δ ¹³C. Two gag with the greatest variation were chosen for high-resolution temporal analysis using individual laminae from their second eye lenses. The first-order patterns from the high-resolution analysis generally mimicked patterns from the low-resolution screening of grouped laminae, yet the high-resolution plots revealed early-life details that were not apparent in the low-resolution screenings. For the third test, left- versus right-eye variation was compared using high-resolution methods. White grunt left- and right-eye radial isotopic patterns were almost identical for both δ ¹³C and δ ¹⁵N, suggesting the variations observed among individual fish were not artifacts. The final test evaluated intra-laminar variation; multiple samples were analyzed from different parts of the same lamina. Seven laminae from three individuals of two species were analyzed in this manner; variations among laminae were found to be much higher than variations within laminae. However, nominal intra-laminar variations were comparable to nominal differences between left and right lenses, suggesting intra-laminar variation established measurement precision. Eye lens isotopes appear to be useful for reconstructing the isotopic histories of individual fish; these histories can be compared with spatially-derived isoscapes to reconstruct individual histories for site fidelity, movement and trophic position.     

Metastable ripple phase of fully hydrated dipalmitoylphosphatidylcholine as studied by small angle x-ray scattering

Fully hydrated dipalmitoylphosphatidylcholine (DPPC) undergoes liquid crystalline to metastable P_β, phase transition in cooling. A small angle x-ray scattering study has been performed for obtaining further evidence about the structure of this phase. From a high-resolution observation of x-ray diffraction profiles, a distinct multipeak pattern has become obvious. Among them the (01) reflection in the secondary ripple structure is identified clearly. There are peaks assigned straightforwardly to (10) and (20) reflections in the primary ripple structure and peaks assigned to (10) and (20) reflections in the secondary ripple structure. Therefore the multipeak pattern is due to superposition of the reflections cause by the primary and secondary ripple structures. The lattice parameters are estimated as follows: for the primary ripple structure a = 7.09 nm, b = 13.64 nm, and γ = 95°, and for the secondary ripple structure a = 8.2 nm, b = 26.6 nm, and γ = 90°. The lattice parameters thus obtained for the secondary ripple structure are not conclusive, however. The hydrocarbon chains in the primary ripple structure have been reported as being tilted against the bilayer plane and, on the other hand, the hydrocarbon chains in the secondary ripple structure are likely to be perpendicular to the bilayer plane. This fact seems to be related to a sequential mechanism of phase transitions. On heating from the L_β, phase where the hydrocarbon chains are tilted the primary ripple structure having tilted hydrocarbon chains takes place and on cooling from the L_α phase where the hydrocarbon chains are not tilted the secondary ripple structure with untilted chains tends to be stabilized. It appears that the truly metastable ripple phase is expressed by the second ripple structure although in the course of the actual cooling transition both the secondary and primary ripple structures form and coexist.     

ELECTRON MICROSCOPE STUDIES ON THE HAEMOGLOBIN MOLECULES

Haemoglobin molecules isolated from normal human subjects have been directly micrographed under the electron microscope following in general Hall's technique. The average height (h) and the widths along (w₁₁) and perpendicular (w) to the shadow direction of the molecules have been measured as 56.5 ± 6.6 A, 122.7 ± 15 A, and 120.9 ± 20 A, respectively. The exaggeration in the molecular widths due to the deposition of metal cap ranges between 60 to 70 A. The probable resolution of the substructure of the molecule, e.g., presence of "holes" and dimples, in the present electron microscopic evidence has been discussed. The electron microscopic results on the size of the individual haemoglobin molecules are in satisfactory agreement with the recent x-ray diffraction model of Perutz and his associates for horse haemoglobin.     

Analysis of emergent patterns in crossing flows of pedestrians reveals an invariant of ‘stripe’ formation in human data

When two streams of pedestrians cross at an angle, striped patterns spontaneously emerge as a result of local pedestrian interactions. This clear case of self-organized pattern formation remains to be elucidated. In counterflows, with a crossing angle of 180°, alternating lanes of traffic are commonly observed moving in opposite directions, whereas in crossing flows at an angle of 90°, diagonal stripes have been reported. Naka (1977) hypothesized that stripe orientation is perpendicular to the bisector of the crossing angle. However, studies of crossing flows at acute and obtuse angles remain underdeveloped. We tested the bisector hypothesis in experiments on small groups (18-19 participants each) crossing at seven angles (30° intervals), and analyzed the geometric properties of stripes. We present two novel computational methods for analyzing striped patterns in pedestrian data: (i) an edge-cutting algorithm, which detects the dynamic formation of stripes and allows us to measure local properties of individual stripes; and (ii) a pattern-matching technique, based on the Gabor function, which allows us to estimate global properties (orientation and wavelength) of the striped pattern at a time T. We find an invariant property: stripes in the two groups are parallel and perpendicular to the bisector at all crossing angles. In contrast, other properties depend on the crossing angle: stripe spacing (wavelength), stripe size (number of pedestrians per stripe), and crossing time all decrease as the crossing angle increases from 30° to 180°, whereas the number of stripes increases with crossing angle. We also observe that the width of individual stripes is dynamically squeezed as the two groups cross each other. The findings thus support the bisector hypothesis at a wide range of crossing angles, although the theoretical reasons for this invariant remain unclear. The present results provide empirical constraints on theoretical studies and computational models of crossing flows.     

Reciprocal Recurrent Selection Compared to within-Strain Selection for Increasing Rate of Egg Lay of Tribolium under Optimal and Stress Conditions

A replicated comparison of reciprocal recurrent selection (rrs) based on crossbred performance and within strain-selection (wss) based on purebred performance was made in three diverse environments over ten generations for the improvement of a heterotic trait, 4-day virgin egg lay of Tribolium castaneum. A selection intensity of 20% based on performance in either an optimum (33°), a mild stress (38°), or a severe stress (28°) environment was applied uniformly. Periodically, the performance of each population was measured in all three environments to provide both direct and correlated responses.—Heritability of egg lay in the base population ranged from 0.36 ± 0.03 in optimum to 0.26 ± 0.03 in severe stress. Estimates of dominance effects assumed significant proportions in severe stress only. Genetic correlations for egg lay in diverse environments were large and positive (.6 to.8).—Only in severe stress did the rrs response significantly exceed that for wss. Quadratic adjustments fitted to response curves revealed that small initial genetic gains under rrs were followed by significantly increasing rates of gain in late generations of selection. The reverse was true for wss. This and evidence from realized heritabilities and genetic correlations suggested that rrs had utilized both additive and dominance effects, but wss response was limited to additive effects.—These results agree with selection theory in demonstrating that purebred selection is more efficient than crossbred selection in utilizing additive gene effects. The latter method has merit when non-additive effects assume significant proportions, and this is the more probable case for severe stress conditions.     

Copper-Containing Anti-Biofilm Nanofiber Scaffolds as a Wound Dressing Material

Copper particles were incorporated into nanofibers during the electrospinning of poly-D,L-lactide (PDLLA) and poly(ethylene oxide) (PEO). The ability of the nanofibers to prevent Pseudomonas aeruginosa PA01 and Staphylococcus aureus (strain Xen 30) to form biofilms was tested. Nanofibers containing copper particles (Cu-F) were thinner (326 ± 149 nm in diameter), compared to nanofibers without copper (CF; 445 ± 93 nm in diameter). The crystalline structure of the copper particles in Cu-F was confirmed by X-ray diffraction (XRD). Copper crystals were encapsulated, but also attached to the surface of Cu-F, as shown scanning transmission electron microscopy (STEM) and transmission electron microscopy (TEM), respectively. The copper particles had no effect on the thermal degradation and thermal behaviour of Cu-F, as shown by thermogravimetric analysis (TGA) and differential scanning calorimeter (DSC). After 48 h in the presence of Cu-F, biofilm formation by P. aeruginosa PA01 and S. aureus Xen 30 was reduced by 41% and 50%, respectively. Reduction in biofilm formation was ascribed to copper released from the nanofibers. Copper-containing nanofibers may be incorporated into wound dressings.     

Linear Dichroism and Orientation of the Phycomyces Photopigment

The greater sensitivity of a cylindrical Phycomyces sporangiophore to blue light polarized transversely rather than longitudinally is a consequence of the dichroism and orientation of the receptor pigment. The abilities of wild type and several carotene mutants to distinguish between the two directions of polarization are the same. The E-vector angle for maximum response relative to the transverse direction is 42 ± 4° at 280 nm, 7° ± 3° at 456 nm, and 7° ± 8° at 486 nm. The in vivo attenuation of polarized light at these wavelengths is very small. The polarized light effect in Phycomyces cannot arise from reflections at the cell surface or from differential attenuations due to internal screening or scattering.     

The accelerating action of illumination in recovery of Arbacia eggs from exposure to ultraviolet radiation

Light of wave lengths 0.30µ to 0.50µ, accelerates return of the cleavage rate of Arbacia eggs to normal, after delay by exposure to ultraviolet radiation (wave lengths 0.238µ to 0.31µ). Recovery is apparently complete. Wave lengths 0.30µ to 0.50µ have no effect on the cleavage rate of normal eggs, nor does such illumination previous to dosage with ultraviolet radiation influence subsequent recovery. Acceleration of recovery of the egg occurs before fertilization as well as after. The effects of ultraviolet radiation and recovery therefrom are essentially the same in nucleated "white halves" as in the intact eggs. This phenomenon in the Arbacia egg seems basically comparable to photoreactivation of bacteria and fungi.     

Biogenic proficient synthesis of (Au-NPs) via aqueous extract of Red Dragon Pulp and seed oil: Characterization,antioxidant, cytotoxic properties,anti-diabetic anti-inflammatory,anti-Alzheimer and their anti-proliferative potential against cancer cell lines

Gold nanoparticles with tiny sizes and biostability are particularly essential and are employed in a variety of biomedical applications. Using a reducing agent and a stabilising agent to make gold nanoparticles has been reported in a number of studies. Gold nanoparticles with a particle size of 25.31 nm were synthesized in this study utilising Hylocereus polyrhizus (Red Pitaya) extract, which functions as a reducing and stabilising agent. The extract of Red Pitaya is said to be a powerful antioxidant and anti-cancer agent. Because of its substantial blood biocompatibility and physiological stability, green production of gold nanoparticles with H. polyrhizus fruit extract is an alternative to chemical synthesis and useful for biological and medical applications. The formation and size distribution of gold nanoparticles were confirmed by HPLC, UV-Vis spectrophotometer, X-ray diffraction (XRD), Dynamic light scattering (DLS), Zeta potential, Transmission electron microscopy (TEM), Fourier transformed infrared spectroscopy (FTIR), Energy dispersive X-ray (EDX) and X-ray photoelectron spectroscopy (XPS). The well-analysed NPs were used in various biological assays, including anti-diabetic, anti-inflammatory, anti-Alzheimer, and antioxidant (DPPH), and cytotoxic investigations. The NPs also showed a dose-dependent cytotoxic activity against HCT-116, HepG2 and MCF-7 cell lines, with IC₅₀ of 100 µg/mL for HCT-116 cells, 155 µg/mL for HepG2, and for MCF-7 cells the value was 165 µg/mL respectively. Finally, the outstanding biocompatibility of Au-NPs has led to the conclusion that they are a promising choice for various biological applications.     

Electric Field-Induced Changes in Lipids Investigated by Modulated Excitation FTIR Spectroscopy

The effect of electric fields on dry oriented multibilayers of dimyristoylphosphatidylcholine (DMPC) was investigated by transmission Fourier transform infrared electric field modulated excitation (E-ME) spectroscopy. A periodic rectangular electric potential (0–150 V, 1.25 Hz, 28.4°C ± 0.2°C) was applied across the sample. To discriminate electric field-induced effects from possible temperature-induced effects resulting from a current flow (<1 pA) across the sample, corresponding temperature-modulated excitation (T-ME) measurements within the temperature uncertainty limits of ±0.2°C at 28.4°C were performed. T-ME induced reversible gauche defects in the hydrocarbon chains, whereas E-ME resulted in reversible compression of dry DMPC bilayers. Periodic variation of the tilt angle of the hydrocarbon chains is suggested. The degree of absorbance modulation in the CH-stretching region was found to be in the order of 1:700, corresponding to a variation of the bilayer thickness of Δz = 0.0054 nm. Using a series connection of capacitors as equivalent circuit of the cell resulted in E = (1.2 ± 0.7) × 10⁷ V/m for the electric field in DMPC. Young's elasticity modulus of DMPC could be calculated to be E = 2.2 × 10⁶ Pa ± 1.8 × 10⁶ Pa, which is in good agreement with published data obtained by electric field-dependent capacitance measurements.     

Effect of Initial Nematode Density on Managing Globodera rostochiensis with Resistant Cultivars and Nonhosts

Cropping systems in which resistant potato cultivars were grown at different frequencies in rotation with susceptible cultivars and a nonhost (oats) were evaluated at four initial nematode population densities (Pi) for their ability to maintain Globodera rostochiensis at a target level of <0.2 egg/cm³ of soil. At a Pi of 0.1 to 1 egg/cm³ of soil, cropping systems with 2 successive years of a resistant cultivar every 3 years of potato production reduced and maintained G. rostochiensis at <0.2 egg/cm³ of soil. At a Pi of 1 to 4 eggs/cm³ of soil, 2 successive years of a resistant cultivar followed by 1 year of oats for every 4 years of production were necessary to reduce and maintain G. rostochiensis populations at <0.2 egg/cm³ of soil. At a Pi greater than 4 eggs/cm³ of soil, 2 successive years of a resistant cultivar plus 1 year of oats reduced G. rostochiensis densities to <0.2 egg/cm³ of soil, but the population increased above that density after cropping 1 year to a susceptible cultivar. The numbers of cysts and eggs per cyst in the final population (Pf) of G. rostochiensis were influenced by initial density and the frequency of growing a susceptible cultivar in a cropping system. The lowest number of cysts and eggs per cyst in the final G. rostochiensis population occurred with a cropping system consisting of 2 successive years of a resistant cultivar followed by oats with a susceptible cultivar grown the fourth year of production.     

Elastic-Mathematical Theory of Cells and Mitochondria in Swelling Process: II. Effect of Temperature upon Modulus of Elasticity of Membranous Material of Egg Cells of Sea Urchin, Strongylocentrotus purpuratus, and of Oyster, Crassostrea virginica

The elastic behavior of the cell wall as a function of the temperature has been studied with particular attention being given to the swelling of egg cells of Strongylocentrotus purpuratus and Crassostrea virginica in different sea water concentrations at different temperatures. It was found that the modulus of elasticity is a nonlinear function of temperature. At about 12-13°C the modulus of elasticity (E) is constant, independent of the stress (σ) and strain (ε_ν) which exist at the cell wall; the membranous material follows Hooke's law, and E ≈ 3 × 10⁷ dyn/cm² for S. purpuratus and C. virginica. When the temperature is higher or lower than 12-13°C, the modulus of elasticity increases, and the membranous material does not follow Hooke's law, but is almost directly proportional to the stresses existing at the cell wall. On increasing the stress, the function E_σ = E(σ) approaches saturation. The corresponding stress-strain diagrams, σ = σ(ε_ν), and the graphs, E_σ = E(σ) and E_σ = E(t) are given. The cyto-elastic phenomena at the membrane are discussed.     

Preliminary X-ray crystallographic data on phospholipase C from Bacillus cereus.

Low-angle X-ray diffraction shows that, despite the well-defined regular axially projected structure, there is no long-range lateral order in the packing of molecules in native (undried) or dried elastoidin spicules from the fin rays of the spurhound Squalus acanthias. The equatorial intensity distribution of the X-ray diffraction pattern from native elastoidin indicates a molecular diameter of 1.1 nm and a packing fraction for the structure projected on to a plane perpendicular to the spicule (fibril) axis of 0.31 (the value for tendon is much higher at around 0.6). Density measurements support this interpretation. When the spicule dries the packing fraction increases to 0.43 but there is still no long-range order in the structure. The X-ray diffraction patterns provide no convincing evidence for any microfibrils or subfibrils in elastoidin. Gel electrophoresis shows that the three chains in the elastoidin molecule are identical. The low packing fraction for collagen molecules in elastoidin explains the difference in appearance between electron micrographs of negatively stained elastoidin and tendon collagen. In elastoidin, but not in tendon collagen, an appreciable proportion of the stain is able to penetrate between the collagen molecules.     

Elastic-Mathematical Theory of Cells and Mitochondria in Swelling Process: The Membranous Stresses and Modulus of Elasticity of the Egg Cell of Sea Urchin, Strongylocentrotus purpuratus

To the revolution-ellipsoidal and spherical membranous shell (cell mitochondrion) are introduced the equations for the calculation of both the modulus of elasticity (Young's modulus) and the stresses, which exist at the membrane. The existing pressure difference between the inner and outer surface of the membrane is calculated in the dilution of seawater media in the osmotic steady state. The experimental results are obtained by using egg cells of the sea urchin, Strongylocentrotus purpuratus. Up to the specific volume of the egg cell (V_E ≈ 35·10^-8 cm³) Boyle-van't Hoff's law is valid (defined as the subelastic range) beyond that the elastic stresses exist (elastic range). For the maximum value of the stresses existing at the cell wall one obtains σ ≈ 5.5·10⁶ dyne/cm² and for the modulus of elasticity E = 1.0·10⁷ dyne/cm², which is constant when the value of relative strain ε_ν > 15%. The breaking limit by an approximate calculation is σ_U ≈ 11·10⁶ dyne/cm². The membrane is assumed to be convoluted and its hypothetical degree of folding was calculated [unk]_a = 34%. The results are compared with the values existing in the literature and other types of cells are found to have values of elasticity in the same range as values of the membrane of S. purpuratus. Both compression and cell elastometer methods are criticized and in certain cases results of these methods are considered to belong to the subelastic domain.     

Exocytosis Precedes and Predicts the Increase in Growth in Oscillating Pollen Tubes

We examined exocytosis during oscillatory growth in lily (Lilium formosanum and Lilium longiflorum) and tobacco (Nicotiana tabacum) pollen tubes using three markers: (1) changes in cell wall thickness by Nomarski differential interference contrast (DIC), (2) changes in apical cell wall fluorescence in cells stained with propidium iodide (PI), and (3) changes in apical wall fluorescence in cells expressing tobacco pectin methyl esterase fused to green fluorescent protein (PME-GFP). Using PI fluorescence, we quantified oscillatory changes in the amount of wall material from both lily and tobacco pollen tubes. Measurement of wall thickness by DIC was only possible with lily due to limitations of microscope resolution. PME-GFP, a direct marker for exocytosis, only provides information in tobacco because its expression in lily causes growth inhibition and cell death. We show that exocytosis in pollen tubes oscillates and leads the increase in growth rate; the mean phase difference between exocytosis and growth is –98° ± 3° in lily and –124° ± 4° in tobacco. Statistical analyses reveal that the anticipatory increase in wall material predicts, to a high degree, the rate and extent of the subsequent growth surge. Exocytosis emerges as a prime candidate for the initiation and regulation of oscillatory pollen tube growth.     

Photo-mediated optimized synthesis of silver nanoparticles using the extracts of outer shell fibre of Cocos nucifera L. fruit and detection of its antioxidant,cytotoxicity and antibacterial potential

Presently, photo-mediated optimized synthesis of SNPs (CS-AgNPs) was carried out with the help of aqueous extracts of coconut (Cocos nucifera) outer shell fibre. Green synthesis of CS-AgNPs was undertaken under laboratory light conditions and characterized by several standard techniques such as UV–visible spectrophotometer (UV–Vis), X-ray diffraction pattern (XRD), Fourier transform infrared (FT-IR), and scanning electron microscopy (SEM) images and energy dispersive spectroscopy (EDX). UV–Vis spectra displayed a surface plasmon resonance peak at 468 nm equivalent to CS-AgNPs, and the FT-IR spectra confirmed the association of biological molecules from the extract in the synthesis process. The SEM image data confirmed the round and circular nature of CS-AgNPs. The EDX data presented the elemental configuration with a solid peak at 3 KeV that matched with the Ag. The synthesized CS-AgNPs exhibited substantial cytotoxicity potential against the HepG2 cells with (effective concentration (IC₅₀) value of 15.28 µg/ml along with robust antioxidant potential, with respect to its 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging (IC₅₀ of 96.39 µg/ml) and reducing assay (IC_0.5 of 209.96 µg/ml). The CS-AgNPs demonstrated encouraging antimicrobial potential against four different pathogenic bacteria and one Candida sp. with inhibition zone diameter ranged between 8.87 and 13.07 mm. Overall, the existing investigation suggested that CS-AgNPs can be an attractive, cost-effective, and environment-friendly candidate for its possible uses in the food, cosmetics, and therapeutic fields.     

Dynamics of the Serine Chemoreceptor in the Escherichia coli Inner Membrane: A High-Speed Single-Molecule Tracking Study

We investigated the mobility of the polar localized serine chemoreceptor, Tsr, labeled by the fluorescent protein Venus in the inner membrane of live Escherichia coli cells at observation rates up to 1000 Hz. A fraction (7%) of all Tsr molecules shows free diffusion over the entire cell surface with an average diffusion coefficient of 0.40 ± 0.01 μm² s⁻¹. The remaining molecules were found to be ultimately confined in compartments of size 290 ± 15 nm and showed restricted diffusion at an inner barrier found at 170 ± 10 nm. At the shortest length-scales (<170 nm), all Tsr molecules diffuse equally. Disruption of the cytoskeleton and rounding of the cells resulted in an increase in the mobile fraction of Tsr molecules and a fragmenting of the previously polar cluster of Tsr consistent with a curvature-based mechanism of Tsr cluster maintenance.