Structural and dynamic characterization of a vinculin binding site in the talin rod

Talin is a key protein involved in linking integrins to the actin cytoskeleton. The long flexible talin rod domain contains a number of binding sites for vinculin, a cytoskeletal protein important in stabilizing integrin-mediated cell-matrix junctions. Here we report the solution structure of a talin rod polypeptide (residues 1843-1973) which contains a single vinculin binding site (VBS; residues 1944-1969). Like other talin rod polypeptides, it consists of a helical bundle, in this case a four-helix bundle with a right-handed topology. The residues in the VBS important for vinculin binding were identified by studying the binding of a series of VBS-related peptides to the vinculin Vd1 domain. The key binding determinants are buried in the interior of the helical bundle, suggesting that a substantial structural change in the talin polypeptide is required for vinculin binding. Direct evidence for this was obtained by NMR and EPR spectroscopy. [1H,15N]-HSQC spectra of the talin fragment indicate that vinculin binding caused approximately two-thirds of the protein to adopt a flexible random coil. For EPR spectroscopy, nitroxide spin labels were attached to the talin polypeptide via appropriately located cysteine residues. Measurements of inter-nitroxide distances in doubly spin-labeled protein showed clearly that the helical bundle is disrupted and the mobility of the helices, except for the VBS helix, is markedly increased. Binding of vinculin to talin is thus a clear example of the unusual phenomenon of protein unfolding being required for protein/protein interaction.     

Detection of QTL for flowering time in multiple families of elite maize

Flowering time is a fundamental quantitative trait in maize that has played a key role in the postdomestication process and the adaptation to a wide range of climatic conditions. Flowering time has been intensively studied and recent QTL mapping results based on diverse founders suggest that the genetic architecture underlying this trait is mainly based on numerous small-effect QTL. Here, we used a population of 684 progenies from five connected families to investigate the genetic architecture of flowering time in elite maize. We used a joint analysis and identified nine main effect QTL explaining approximately 50?% of the genotypic variation of the trait. The QTL effects were small compared with the observed phenotypic variation and showed strong differences between families. We detected no epistasis with the genetic background but four digenic epistatic interactions in a full 2-dimensional genome scan. Our results suggest that flowering time in elite maize is mainly controlled by main effect QTL with rather small effects but that epistasis may also contribute to the genetic architecture of the trait.     

The dynamics of food chains under climate change and nutrient enrichment

Warming has profound effects on biological rates such as metabolism, growth, feeding and death of organisms, eventually affecting their ability to survive. Using a nonlinear bioenergetic population-dynamic model that accounts for temperature and body-mass dependencies of biological rates, we analysed the individual and interactive effects of increasing temperature and nutrient enrichment on the dynamics of a three-species food chain. At low temperatures, warming counteracts the destabilizing effects of enrichment by both bottom-up (via the carrying capacity) and top-down (via biological rates) mechanisms. Together with increasing consumer body masses, warming increases the system tolerance to fertilization. Simultaneously, warming increases the risk of starvation for large species in low-fertility systems. This effect can be counteracted by increased fertilization. In combination, therefore, two main drivers of global change and biodiversity loss can have positive and negative effects on food chain stability. Our model incorporates the most recent empirical data and may thus be used as the basis for more complex forecasting models incorporating food-web structure.     

Erythropoietin attenuates the sequels of ischaemic spinal cord injury with enhanced recruitment of CD34+ cells in mice

Erythropoietin has been shown to promote tissue regeneration after ischaemic injury in various organs. Here, we investigated whether Erythropoietin could ameliorate ischaemic spinal cord injury in the mouse and sought an underlying mechanism. Spinal cord ischaemia was developed by cross-clamping the descending thoracic aorta for 7 or 9 min. in mice. Erythropoietin (5000 IU/kg) or saline was administrated 30 min. before aortic cross-clamping. Neurological function was assessed using the paralysis score for 7 days after the operation. Spinal cords were histologically evaluated 2 and 7 days after the operation. Immunohistochemistry was used to detect CD34(+) cells and the expression of brain-derived neurotrophic factor and vascular endothelial growth factor. Each mouse exhibited either mildly impaired function or complete paralysis at day 2. Erythropoietin-treated mice with complete paralysis demonstrated significant improvement of neurological function between day 2 and 7, compared to saline-treated mice with complete paralysis. Motor neurons in erythropoietin-treated mice were more preserved at day 7 than those in saline-treated mice with complete paralysis. CD34(+) cells in the lumbar spinal cord of erythropoietin-treated mice were more abundant at day 2 than those of saline-treated mice. Brain-derived neurotrophic factor and vascular endothelial growth factor were markedly expressed in lumbar spinal cords in erythropoietin-treated mice at day 7. Erythropoietin demonstrated neuroprotective effects in the ischaemic spinal cord, improving neurological function and attenuating motor neuron loss. These effects may have been mediated by recruited CD34(+) cells, and enhanced expression of brain-derived neurotrophic factor and vascular endothelial growth factor.     

Factors influencing the short-term variation in phytoplankton composition and biomass in coral reef waters

The short-term temporal dynamics of phytoplankton composition was compared among coral reef waters, the adjacent ocean and polluted harbour water from July until October along the south-western coast of Curaçao, southern Caribbean. Temporal variations in phytoplankton pigment 'fingerprints' (zeaxanthin, chlorophyll b, 19'-hexanoyloxyfucoxanthin, fucoxanthin, 19'-butanoyloxyfucoxanthin, chlorophyll c₂ and c₃ relative to chlorophyll a) in the ocean were also observed in waters overlying the reef. However, with respect to specific pigments and algal-size distribution, the algal composition in reef waters was usually slightly different from that in the oceanic water. Phytoplankton biomass (chlorophyll a) was either higher or lower than in the oceanic water. The relative amount of fucoxanthin and peridinin was usually higher, and the relative and absolute amount of zeaxanthin was significantly lower than in oceanic water. Zeaxanthin-containing Synechococci were significantly reduced in reef water. Average algal cell size increased from the open water to the reef and the harbour entrance. Large centric diatoms (>20 m Ø) were better represented in reef than in oceanic water. In reef-overlying waters, the nitrate and nitrite concentrations were higher than in oceanic water. In front of the town, anthropogenic eutrophication (sewage discharge and ground water seepage) resulted in higher NH₄, NO₃ and PO₄ concentrations than at other reef stations. This concurred with significantly enhanced phytoplankton biomass (chlorophyll a), chlorophyll c₂ and peridinin amounts at Town Reef compared with the other reef stations. Polluted harbour water usually showed the highest phytoplankton biomass of all stations, dominated by diatoms and dinoflagellates. Conditions in reef waters and harbour water promoted the occurrence and the relative abundance of diatoms and dinoflagellates. Harbour water did not influence the phytoplankton composition and biomass at reef stations situated >5 km away from the harbour entrance. We conclude that phytoplankton undergoes a shift in algal composition during transit over the reef. The dominant processes appear to be selective removal of zeaxanthin-containing Synechococcus (by the reef benthos) and (relative) increase in diatoms and dinoflagellates. The difference in the phytoplankton composition between reef and oceanic waters tends to increase with decreasing dilution of reef water with ocean water.     

Studies of the deformability of preserved red blood cells using a simple filtration method]

The applied method to measure the filtrability of red blood cells is accurate enough for the study of the effects of the conditions of storage on their deformability. In this work the standard deviation of the method, the mean biological deviation of the deformability of human red blood cells and their deformability in the course of storage in the ACD-AG-medium have been investigated. Considerable loss of deformability occurs towards the end of storage time of the cells, which is not reversible be restitution of ATP alone. The applicability of this method as a simple test of the vitatity of preserved erythrocytes if to be tested.     

Spectral shifting of UV-A wavelengths to blue light for enhancing growth rate of cyanobacteria

This study investigates the modification of UV-A radiation for enhancing the growth rate of Synechococcus species in a simulated photobioreactor made of UV-stabilized polycarbonate (PC) plates. Uvitex OB as was dissolved in a poly(methyl methacrylate) (PMMA)-based resin and the mixture was applied as a coating to shift the UV-A radiation to photosynthetically active radiation (PAR). The feasibility of applying the Uvitex OB in a PMMA-based resin, which was used as fluorescent layer to shift UV-A to PAR range, was explored. For this purpose, a variety of concentrations of Uvitex OB in resin were prepared and coated on PC sheets which were placed between the radiation source and the culture flask. Compared to the uncoated PC sheet, the PC panels coated with a few number of the wavelength shifter layer caused an approximately 4-fold increase in cell number during the same culture period.     

Untersuchungen über die Haftfähigkeit von Duftstoffen am Bienenkörper

Ohne ZusammenfassungHerrn Prof. v. Frisch zum 60. Geburtstag gewidmet.     

Membrane Region M2C2 in Subunit KtrB of the K+ Uptake System KtrAB from Vibrio alginolyticus Forms a Flexible Gate Controlling K+ Flux: AN ELECTRON PARAMAGNETIC RESONANCE STUDY*

Transmembrane stretch M_2C from the bacterial K⁺-translocating protein KtrB is unusually long. In its middle part, termed M_2C2, it contains several small and polar amino acids. This region is flanked by the two α-helices M_2C1 and M_2C3 and may form a flexible gate at the cytoplasmic side of the membrane controlling K⁺ translocation. In this study, we provide experimental evidence for this notion by using continuous wave and pulse EPR measurements of single and double spin-labeled cysteine variants of KtrB. Most of the spin-labeled residues in M_2C2 were shown to be immobile, pointing to a compact structure. However, the high polarity revealed for the microenvironment of residue positions 317, 318, and 327 indicated the existence of a water-accessible cavity. Upon the addition of K⁺ ions, M_2C2 residue Thr-318R1 (R1 indicates the bound spin label) moved with respect to M_2B residue Asp-222R1 and M_2C3 residue Val-331R1 but not with respect to M_2C1 residue Met-311R1. Based on distances determined between spin-labeled residues of double-labeled variants of KtrB in the presence and absence of K⁺ ions, structural models of the open and closed conformations were developed.     

Influence of fluorescent coating at rear and front side of a flat panel photobioreactor on algal growth

The focus of this study is the enhancement of microalgae growth rate using spectral conversion of green light. For this purpose, three reactors were considered and fluorescent pigment Rhodamine 6G was dissolved in a thermoplastic acrylic resin, the mixture was then applied on the front side of the first reactor, and on a mirror located at the rear side of the second one. Comparing their maximum specific growth rate (μ _max) of Chlorella sp. to that in the third (uncoated) reactor, the former resulted in an increase up to 15% while the latter in decrease to at least 30%. Also, the rear side coated reactor showed up to 50% increase in biomass productivity rate (P) in early 4 days of experiment. However, this value decreased over time and the uncoated reactor in 12 days exhibited higher biomass productivity rate.