Axle-less F1-ATPase rotates in the correct direction

F1-adenosine triphosphatase (ATPase) is an ATP-driven rotary molecular motor in which the central gamma subunit rotates inside a cylinder made of three alpha and three beta subunits alternately arranged. The rotor shaft, an antiparallel alpha-helical coiled coil of the amino and carboxyl termini of the gamma subunit, deeply penetrates the central cavity of the stator cylinder. We truncated the shaft step by step until the remaining rotor head would be outside the cavity and simply sat on the concave entrance of the stator orifice. All truncation mutants rotated in the correct direction, implying torque generation, although the average rotary speeds were low and short mutants exhibited moments of irregular motion. Neither a fixed pivot nor a rigid axle was needed for rotation of F1-ATPase.     

Growth and fatty acid composition of sesame (<Emphasis Type="Italic">Sesamum indicum</Emphasis> L.) genotypes as influence by planting dates and nitrogen fertilization in semiarid region of Northwest,Pakistan

A field experiment was conducted on silty clay loam soil in the years 2011–2012. Two sesame (Sesamum indicum L.) cultivars (Local Black and Local White) were evaluated using various 3 different sowing dates (20th June, 10th and 30th July) and four agrotechnical level (0, 40, 80 and 120 kg N ha^–1) at New Developmental Farm The University of Agriculture, Peshawar, Pakistan. The objective of this study was to evaluate the effects of sowing dates on growth, yield and oil fatty acid composition of two sesame cultivars grown under different nitrogen fertilization. Results showed that cv. Local Black was characterized by significantly higher content of oil (47%), seed yield (696 kg ha^–1) and oil yield (335 kg ha^–1) while cv. Local White had higher palmitic acid (8%) and linoleic acid (38.7%). Yield and its main components were positively affected by the earlier sowing date. With regard to fatty acid composition, a decrease in oleic and stearic acid and an increase in linoleic and palmitic acid were observed. At early sowing, oleic and palmitic acid decreased whereas linoleic and stearic acid increased. The decrease in the oleic/linoleic acid ratio observed at early sowing, suggests a possible role of temperature on the activity of oleate desaturase in the developing seeds. Intensive technology of cultivation (120 kg N ha^–1), compared to the economical technology (40 kg N ha^–1), significantly increased the seed yield of both sesame cultivars. This was due to higher number of branches, Capsules m^–2, capsules plant^–1, seeds capsule^–1 and 1000 seed weight (g). The intensive technology of cultivation had a beneficial effect on the content of palmitic acid, linolenic acid and oleic acid in sesame seed.     

Single-cell mass cytometry of differential immune and drug responses across a human hematopoietic continuum

Flow cytometry is an essential tool for dissecting the functional complexity of hematopoiesis. We used single-cell "mass cytometry" to examine healthy human bone marrow, measuring 34 parameters simultaneously in single cells (binding of 31 antibodies, viability, DNA content, and relative cell size). The signaling behavior of cell subsets spanning a defined hematopoietic hierarchy was monitored with 18 simultaneous markers of functional signaling states perturbed by a set of ex vivo stimuli and inhibitors. The data set allowed for an algorithmically driven assembly of related cell types defined by surface antigen expression, providing a superimposable map of cell signaling responses in combination with drug inhibition. Visualized in this manner, the analysis revealed previously unappreciated instances of both precise signaling responses that were bounded within conventionally defined cell subsets and more continuous phosphorylation responses that crossed cell population boundaries in unexpected manners yet tracked closely with cellular phenotype. Collectively, such single-cell analyses provide system-wide views of immune signaling in healthy human hematopoiesis, against which drug action and disease can be compared for mechanistic studies and pharmacologic intervention.     

Conserved eukaryotic fusogens can fuse viral envelopes to cells

Caenorhabditis elegans proteins AFF-1 and EFF-1 [C. elegans fusion family (CeFF) proteins] are essential for developmental cell-to-cell fusion and can merge insect cells. To study the structure and function of AFF-1, we constructed vesicular stomatitis virus (VSV) displaying AFF-1 on the viral envelope, substituting the native fusogen VSV glycoprotein. Electron microscopy and tomography revealed that AFF-1 formed distinct supercomplexes resembling pentameric and hexameric "flowers" on pseudoviruses. Viruses carrying AFF-1 infected mammalian cells only when CeFFs were on the target cell surface. Furthermore, we identified fusion family (FF) proteins within and beyond nematodes, and divergent members from the human parasitic nematode Trichinella spiralis and the chordate Branchiostoma floridae could also fuse mammalian cells. Thus, FF proteins are part of an ancient family of cellular fusogens that can promote fusion when expressed on a viral particle.     

Insecticidal activity of Arum maculatum tuber lectin and its binding to the glycosylated insect gut receptors

Mannose binding approximately 50 kDa homotetrameric lectin, purified from edible Arum maculatum tuber, was analyzed through SDS-PAGE and studied for its agglutination property using rabbit erythrocytes. Cross reactivity of the purified lectin was verified through western blot using Colocasia esculantum(Family, Araceae) tuber lectin antibody. The insecticidal activity of Arum maculatum tuber lectin (ATL) was tested against two economically important sucking pests, Lipaphis erysimi and Aphis craccivora, in an artificial diet. The LC(50) values for L. erysimi and A. craccivora were determined to be 21 microg/mL and 16 microg/mL, respectively. Addition of alpha-d-mannose in ATL-supplemented diet reduced the aphid mortality. Two major receptor proteins of ATL (approximately 40 kDa and approximately 35 kDa) were detected from the brush border membrane vesicle (BBMV) protein of L. erysimi and A. craccivora guts, respectively, using ligand-binding assay. Alpha-d-Mannose was found to be a deterrent to such binding of ATL to the BBMV receptors.     

Effect of 3-O-octanoyl-(+)-catechin on the responses of GABA(A) receptors and Na+/glucose cotransporters expressed in xenopus oocytes and on the oocyte membrane potential

Recently, 3-O-octanoyl-(+)-catechin (OC) was synthesized from (+)-catechin (C) by incorporation of an octanoyl chain into C in the light of (-)-epicatechin gallate (ECg) and (-)-epigallocatechin gallate (EGCg), which are the major polyphenols found in green tea and have strong physiological activities. OC was found to inhibit the response of ionotropic gamma-aminobutyric acid (GABA) receptors (GABA(A) receptors) and Na+/glucose cotransporters expressed in Xenopus oocytes in a noncompetitive manner more strongly than does C. OC also induced a nonspecific membrane current and decreased the membrane potential of the oocyte, and thus death of the oocyte occurred even at lower concentrations than that induced by C or EGCg. Although EGCg produced H2O2 in aqueous solution, OC did not. This newly synthesized catechin derivative OC possibly binds to the lipid membrane more strongly than does C, Ecg, or EGCg and as a result perturbs the membrane structure.     

Boron-Calcium Synergically Alleviates Aluminum Toxicity in Wheat Plants (Triticum aestivum L.)

The effects of B and Ca treatments on root growth, nutrient localization and cell wall properties in wheat ( Triticum aestivum L.) plants with and without Al stress were investigated. Seedlings were grown hydroponically in a complete nutrient solution for 7 d and then treated with B (0, 40 μM), Ca (0, 2,500 μM), and Al (0, 100 μM) in a 500 μM CaCl₂ solution for 8 d. The cell wall materials (CWM) were extracted with a phenol: acetic acid: water (2:1:1 w/v/v) solution and used for subsequent pectin extraction with trans -1,2-diami-nocyclohexane- N,N,N,N -tetraacetic acid (CDTA) and Na₂CO₃ solutions. Boron, Ca, and B + Ca treatments enhanced root growth by 19.5, 15.2, and 27.2%, respectively, compared to the control (pH 4.5). Calcium and B+Ca treatments enhanced root growth with Al stress by 43 and 54%, respectively, while B did not exert any effect. The amounts of CWM and pectin per unit of root fresh weight increased by Al treatment, whereas the Ca and B+Ca treatments slightly reduced the contents of these components. Seventy-four percent of total B, 69% of total Ca, and 85% of total Al were located in the cell wall in the B, Ca, and Al treatments, respectively and 32% of total B, 33% of total Ca, and 33% of total Al were located in the CDTA-soluble and Na₂CO₃-soluble pectin fractions. A more conspicuous localization of B was observed in the presence of Al. Aluminum treatment markedly decreased the Ca content in the cell wall as well as pectin fractions, mainly in the case of the CDTA-soluble pectin fraction. Boron + Ca treatment decreased the Al content in the cell wall and pectin fractions compared to the Ca treatment alone in the presence of Al. It is concluded that the B+Ca treatment enhanced root growth and, B and Ca uptake, and helped to maintain a normal B and Ca metabolism in the cell walls even in the presence of Al.     

Efficiency and Flow Regime of a Highway Stormwater Detention Pond in Washington, USA

Wet detention ponds are a preferable alternative in treating stormwater runoff. Literature suggests that a detention pond’s efficiency in removing principal pollutants of concern, TSS and metals, is highly variable and is affected by a complex array of factors including its geographic location. The objective of this paper was to investigate the TSS and metal removal efficiency of a highway stormwater detention pond in Spokane, Washington along with its flow regime. Pond influent and effluent data for TSS and metal were collected for approximately two years. TSS removal by the pond was found to be 68.1–99.4% with an average of 83.9%. Average metal removal efficiency was 54.7–64.6% which is 72.5–86.9% of the TSS removal. The pond’s flow regime was found to vary with its changing surface topography, a result of sedimentation of suspended solids.     

Maceration of plant tissue by fungi is inhibited by recombinant antipectinase antibodies

Polyclonal antiserum from mice immunized with extracellular proteins from Rhizoctonia solani inhibited pectinase and cellulase activities in cell free culture supernatants of Rhizoctonia solani. Spleen mRNA from these mice was used to construct a cDNA library from which antipectinase ScFv antibodies were isolated using phage display techniques. Soluble ScFv antibodies produced by individual clones in Escherichia coli inhibited polygalacturonase in the culture supernatants of a range of fungal pathogens, including ascomycetes, basidiomycetes, and oomycetes. The soluble antibodies also inhibited maceration of potato tissue by these pathogens.