Identification of specific proteins and glycoproteins associated with membrane fractions isolated from Zea mays L. coleoptiles

The aim of this work was to identify proteins specific for plant cell membranes which could then be used as unique markers. A crude membrane fraction was isolated from corn coleoptiles and separated on non-linear sucrose density gradients. Separation of endoplasmic reticulum (NADH-cytochrome c reductase), mitochondria (cytochrome c oxidase), golgi (inosine diphosphatase), and plasma membranes (N-1-naphthylphthalamic acid-binding) was achieved. The membrane proteins from the gradient fractions were separated using sodium dodecyl sulphate-poly-acrylamide gel electrophoresis and the gels stained with coomassie blue or with concanavalin A/peroxidase to detect glycoproteins. Proteins specific for the various membranes were identified. Five proteins including two glycoproteins were plasma membrane markers. Protoplasts were isolated and iodinated using lactoperoxidase/glucose oxidase covalently attached to beads. Eleven iodinated proteins were found and three of these corresponded to proteins specifically associated with plasma membranes in the density gradients. Two methods for detecting Ca²⁺-binding proteins following sodium dodecylsulphate polyacrylamide gel electrophoresis were employed. The majority of such proteins were found in the endoplasmatic reticulum and one was specific for plasma membranes. In vitro and in vivo phosphorylation of membrane proteins was examined and the majority of proteins phosphorylated were glycoproteins. Two of the phosphorylated proteins (M_r=110,000 and 20,000) were also iodinated on protoplasts and may be part of the plasma membrane ATPases.Abbreviations ER endoplasmic reticulum - IDP inosine diphosphate - NPA N-1-naphthylphthalamic acid     

Real space refinement of neutron diffraction data from sperm whale carbonmonoxymyoglobin

Neutron diffraction data from crystals of sperm whale carbonmonoxymyoglobin have been refined by the real space refinement technique. Estimates of the neutron occupancies at the end of the refinement show that the mean for each atom type (including hydrogen and deuterium) is close to the expected value and has a standard deviation from the mean of about 5%. Mean neutron occupancies of main-chain atoms involved in deuterium bonds versus those not involved in deuterium bonds demonstrate that the hydrogen/deuterium exchange of the latter group is higher. The oxygen and deuterium co-ordinates for 40 water molecules have been determined: 27 of these water molecules were involved in bridges between protein atoms, and nine were involved in deuterium bonds with main-chain atoms. The deuterium-bond angles in helical regions show significant deviations from linearity. The mean ND … O angle was 154(3) °² and the mean CO … D angle was 145(3) °.     

Bounded Rationality and Voting Decisions over 160 Years: Voter Behavior and Increasing Complexity in Decision-Making

Using a quasi-natural voting experiment encompassing a 160-year period (1848–2009) in Switzerland, we investigate whether a higher level of complexity leads to increased reliance on trusted parliamentary representatives. We find that when more referenda are held on the same day, constituents are more likely to refer to parliamentary recommendations when making their decisions. This finding holds true even when we narrow our focus to referenda with a relatively lower voter turnout on days on which more than one referendum is held. We also demonstrate that when constituents face a higher level of complexity, they follow the parliamentary recommendations rather than those of interest groups. “Viewed as a geometric figure, the ant’s path is irregular, complex, hard to describe. But its complexity is really a complexity in the surface of the beach, not a complexity in the ant.” ( [1] p. 51)     

Fluorescence techniques in biotechnology

The high specificity and sensitivity of fluorescence techniques have made them important analytical tools in medicine and biotechnology. Besides monitoring and quantitative detection of biomolecules these methods can be used for controlling bacterial activities or for measuring physiological states of cells or tissues. Three topics of importance in biotechnology — immunoassays, photosynthesis and fermentation — are treated in detail.     

Stem-like cells in human hepatoblastoma.

Hepatoblastoma is a pediatric liver tumor with epithelial components resembling embryonal and fetal liver cells. The existence of teratoid hepatoblastoma suggests the presence of stem cells in hepatoblastoma. The aim of this study was to analyze the expression of stem cell markers in hepatoblastomas. We studied specimens from 10 hepatoblastomas. Five of the hepatoblastomas were of epithelial and five of mixed type. Immunohistochemistry (IHC) for the stem cell markers CD34, Thy1, c-kit, and the hepatic or biliary lineage markers CK-18, OCH, CK-7, and CD56 was performed. Double IHC for stem cell and lineage markers was used to identify putative liver stem cells. The different markers showed distinct distributions on the tumor cells. Cells in atypical ducts were found to express simultaneously stem cell markers and hepatocytic or biliary lineage markers. Other cells in connective tissue showed c-kit expression, but not hepatic or biliary marker expression. The data show the presence of different cell populations bearing stem cell markers in human hepatoblastoma. Ductal cells co-expressing stem cell markers and hepatic lineage markers phenotypically resemble hepatic stem-like cells. These findings support the thesis that stem cells play a role in the histogenesis of hepatoblastoma.     

Not Just an empty cavity: the inter-rhabdomeral space in the Jamaican cavefly Neoditomyia farri (Diptera, Mycetophilidae)

Contrary to most other Diptera, the inter-rhabdomeral spaces of the retina of the Jamaican cavefly Neoditomya farri are filled neither by extracellular matrix nor dense cytoplasmic material. Instead, a foamy organization of loose vacuoles, measuring approximately 0.7 microm in diameter, appears to keep the rhabdomeres of retinula cells 7 and 8 in place. The vacuoles are bounded by membranes and traces of actin, determined immunocytochemically, are present. The origin of the vacuoles is unclear, but evidence in support of a retinula cell rather than cone cell origin is advanced.     

Muscle activity reduces soft-tissue resonance at heel-strike during walking

Muscle activity has previously been suggested to minimize soft-tissue resonance which occurs at heel-strike during walking and running. If this concept were true then the greatest vibration damping would occur when the input force was closest to the resonant frequency of the soft-tissues at heel-strike. However, this idea has not been tested. The purpose of this study was to test whether muscle activity in the lower extremity is used to damp soft-tissue resonance which occurs at heel-strike during walking. Hard and soft shoe conditions were tested in a randomized block design. Ground reaction forces, soft-tissue accelerations and myoelectric activity were measured during walking for 40 subjects. Soft-tissue mass was estimated from anthropologic measurements, allowing inertial forces in the soft-tissues to be calculated. The force transfer from the ground to the tissues was compared with changes in the muscle activity. The soft condition resulted in relative frequencies (input/tissue) to be closer to resonance for the main soft-tissue groups. However, no increase in force transmission was observed. Therefore, the vibration damping in the tissues must have increased. This increase concurred with increases in the muscle activity for the biceps femoris and lateral gastrocnemius. The evidence supports the proposal that muscle activity damps soft-tissue resonance at heel-strike. Muscles generate forces which act across the joints and, therefore, shoe design may be used to modify muscle activity and thus joint loading during walking and running.