Ribosomal protein S14 is not responsible for the Minute phenotype associated with the M(1)7C locus in Drosophila melanogaster.

Summary A locus associated with a severe Minute effect has been mapped at 7C on the X chromosome of Drosophila melanogaster. Previous work has suggested that this Minute encodes ribosomal proteins S14A and S141B. We have made a chromosomal deficiency that removes the S14 ribosomal protein genes, yet does not display the Minute phenotype. These data suggest that the S14 genes do not actually correspond to the Minute locus.     

A region immediately adjacent to the origin of replication of bovine papilloma virus type 1 interacts in vitro with the nuclear matrix.

We have investigated the interaction of the 69% transforming fragment of the Bovine Papilloma Virus type 1 (BPV1) with the nuclear matrix from 1361.5 cells (NIH-3T3 cells transformed by a BPV chimeric construct). In vitro studies performed with end-labelled DNA fragments and nuclear matrices prepared using a high-salt extraction procedure demonstrate the binding of a 672 bp fragment adjacent to the viral origin of replication and containing the plasmid maintenance sequence (PMS-1). This fragment can be cleaved into two pieces (393 and 279 bp), both interacting equally well with the nuclear matrix. This indicates that a least two regions of the 672 bp DNA fragment are involved in the interaction.     

Eight decades of invasion by Chromolaena odorata (Asteraceae) and its biological control in West Africa: the story so far

ABSTRACT

Chromolaena odorata (L.) R.M. King and H. Robinson (Asteraceae) is a perennial weedy shrub of neotropical origin and a serious biotic threat in its invasive range. The Asian-West Africa (AWA) biotype of C. odorata present in West Africa is both morphologically and genetically different from the southern African (SA) biotype. The AWA biotype was first introduced into Nigeria in the late 1930s and rapidly spread across West Africa. Currently, 12 of the 16 countries in West Africa have been invaded, with significant negative effects on indigenous flora and fauna. However, locals in West Africa have found several uses for the weed. As chemical, physical and other conventional methods were unsustainable, costly and largely ineffective, three biological control agents, Apion brunneonigrum (Coleoptera: Brentidae), Pareuchaetes pseudoinsulata (Lepidoptera: Erebidae) and Cecidochares connexa (Diptera: Tephritidae), have been released in West Africa between the 1970s and the early 2000s. However, only C. connexa and P. pseudoinsulata established, contributing to the control of the weed, in six and four countries in West Africa respectively. Limited research funding, the absence of post-release evaluations of the established agents, and the ‘conflict of interest’ status of C. odorata (i.e. being beneficial for local use but damaging to ecosystem services and agriculture), are serious factors deterring the overall biological control effort. Here, using historical records and field surveys, we examine the invasion history, spread, impacts, and management of C. odorata in West Africa and make recommendations for the sustainable management of C. odorata in the region.     

A probable role of dihydropyridine receptors in repression of Ca2+ sparks demonstrated in cultured mammalian muscle

To activate skeletal muscle contraction, action potentials must be sensed by dihydropyridine receptors (DHPRs) in the T tubule, which signal the Ca²⁺ release channels or ryanodine receptors (RyRs) in the sarcoplasmic reticulum (SR) to open. We demonstrate here an inhibitory effect of the T tubule on the production of sparks of Ca²⁺ release. Murine primary cultures were confocally imaged for Ca²⁺ detection and T tubule visualization. After 72 h of differentiation, T tubules extended from the periphery for less than one-third of the myotube radius. Spontaneous Ca²⁺ sparks were found away from the region of cells where tubules were found. Immunostaining showed RyR1 and RyR3 isoforms in all areas, implying inhibition of both isoforms by a T tubule component. To test for a role of DHPRs in this inhibition, we imaged myotubes from dysgenic mice (mdg) that lack DHPRs. These exhibited T tubule development similar to that of normal myotubes, but produced few sparks, even in regions where tubules were absent. To increase spark frequency, a high-Ca²⁺ saline with 1 mM caffeine was used. Wild-type cells in this saline plus 50 µM nifedipine retained the topographic suppression pattern of sparks, but dysgenic cells in high-Ca²⁺ saline did not. Shifted excitation and emission ratios of indo-1 in the cytosol or mag-indo-1 in the SR were used to image [Ca²⁺] in these compartments. Under the conditions of interest, wild-type and mdg cells had similar levels of free [Ca²⁺] in cytosol and SR. These data suggest that DHPRs play a critical role in reducing the rate of spontaneous opening of Ca²⁺ release channels and/or their susceptibility to Ca²⁺-induced activation, thereby suppressing the production of Ca²⁺ sparks. excitation-contraction coupling; sarcoplasmic reticulum; ryanodine receptors; Ca²⁺ imaging     

Regional carbon footprint analysis of dairy feeds for milk production in the USA

Purpose

A greenhouse gas emissions analysis (carbon footprint) was conducted for cultivation, harvesting, and production of common dairy feeds used for the production of dairy milk in the USA. The goal was to determine the carbon footprint (grams CO₂ equivalents (gCO₂e)/kg of dry feed) in the USA on a regional basis, identify key inputs, and make recommendations for emissions reduction.

Methods

Commonly used dairy feeds in the USA, such as soybeans, alfalfa, corn, and others, were identified based on a recent literature review and information from dairy farm surveys. The following input data for the cultivation and harvesting of dairy feeds were collected for five US regions: crop production data, energy input, soil amendments, and crop protection chemicals. Life cycle inventory input data were mainly collected from the US Department of Agriculture National Agricultural Statistical Service on a state-by-state basis as well as from state extension services forage crop budget estimates. In addition to consulting other life cycle assessment studies and published articles and reports, this cradle-to-farm gate carbon footprint analysis was conducted using the Ecoinvent? unit processes in SimaPro version 7.1? (PRé Consultants 2009).

Results

The final carbon footprint results (gCO₂e/kg of dry dairy feed) varied regionally depending on a number of factors such as lime and fertilizer application rates. The average national US carbon footprint results of the main feeds were: corn grain (390), corn silage (200), dried distillers grains with solubles (910 dry mill, 670 wet mill), oats (850), soybeans (390), soybean meal (410), winter wheat (430), alfalfa hay (170), and forage mix (160).

Conclusions and recommendations

The southeast dairy region generally showed a relatively high level of carbon footprint for most feeds, and this is attributable to the higher application rates of both synthetic fertilizers and lime. The highest contributor to carbon footprint for most regions (apart from soybeans and soybean meal) was due to the application of inorganic nitrogen fertilizer. Efficient transfer of knowledge to farmers with regards to fertilizer best management practices such as precision application of farm nutrients may contribute significantly to reducing regional crop carbon footprints.     

Intracellular Ca(2+) release as irreversible Markov process

In striated muscles, intracellular Ca(2+) release is tightly controlled by the membrane voltage sensor. Ca(2+) ions are necessary mediators of this control in cardiac but not in skeletal muscle, where their role is ill-understood. An intrinsic gating oscillation of Ca(2+) release-not involving the voltage sensor-is demonstrated in frog skeletal muscle fibers under voltage clamp. A Markov model of the Ca(2+) release units is shown to reproduce the oscillations, and it is demonstrated that for Markov processes to have oscillatory transients, its transition rates must violate thermodynamic reversibility. Such irreversibility results in permanent cycling of the units through a ring of states, which requires a source of free energy. Inhibition of the oscillation by 20 to 40 mM EGTA or partial depletion of Ca(2+) in the sarcoplasmic reticulum (SR) identifies the SR [Ca(2+)] gradient as the energy source, and indicates a location of the critical Ca(2+)-sensing site at distances greater than 35 nm from the open channel. These results, which are consistent with a recent demonstration of irreversibility in gating of cardiac Ca(2+) sparks, (Wang, S.-Q., L.-S. Song, L. Xu, G. Meissner, E. G. Lakatta, E. Ríos, M. D. Stern, and H. Cheng. 2002. Biophys. J. 83:242-251) exemplify a cell-wide oscillation caused by coupling between ion permeation and channel gating.     

Unitary Ca2+ current through mammalian cardiac and amphibian skeletal muscle ryanodine receptor Channels under near-physiological ionic conditions

Ryanodine receptor (RyR) channels from mammalian cardiac and amphibian skeletal muscle were incorporated into planar lipid bilayers. Unitary Ca2+ currents in the SR lumen-to-cytosol direction were recorded at 0 mV in the presence of caffeine (to minimize gating fluctuations). Currents measured with 20 mM lumenal Ca2+ as exclusive charge carrier were 4.00 and 4.07 pA, respectively, and not significantly different. Currents recorded at 1-30 mM lumenal Ca2+ concentrations were attenuated by physiological [K+] (150 mM) and [Mg2+] (1 mM), in the same proportion (approximately 55%) in mammalian and amphibian channels. Two amplitudes, differing by approximately 35%, were found in amphibian channel studies, probably corresponding to alpha and beta RyR isoforms. In physiological [Mg2+], [K+], and lumenal [Ca2+] (1 mM), the Ca2+ current was just less than 0.5 pA. Comparison of this value with the Ca2+ flux underlying Ca2+ sparks suggests that sparks in mammalian cardiac and amphibian skeletal muscles are generated by opening of multiple RyR channels. Further, symmetric high concentrations of Mg2+ substantially reduced the current carried by 10 mM Ca2+ (approximately 40% at 10 mM Mg2+), suggesting that high Mg2+ may make sparks smaller by both inhibiting RyR gating and reducing unitary current.     

Concerted vs. sequential. Two activation patterns of vast arrays of intracellular Ca2+ channels in muscle

To signal cell responses, Ca(2+) is released from storage through intracellular Ca(2+) channels. Unlike most plasmalemmal channels, these are clustered in quasi-crystalline arrays, which should endow them with unique properties. Two distinct patterns of local activation of Ca(2+) release were revealed in images of Ca(2+) sparks in permeabilized cells of amphibian muscle. In the presence of sulfate, an anion that enters the SR and precipitates Ca(2+), sparks became wider than in the conventional, glutamate-based solution. Some of these were "protoplatykurtic" (had a flat top from early on), suggesting an extensive array of channels that activate simultaneously. Under these conditions the rate of production of signal mass was roughly constant during the rise time of the spark and could be as high as 5 microm(3) ms(-1), consistent with a release current >50 pA since the beginning of the event. This pattern, called "concerted activation," was observed also in rat muscle fibers. When sulfate was combined with a reduced cytosolic [Ca(2+)] (50 nM) these sparks coexisted (and interfered) with a sequential progression of channel opening, probably mediated by Ca(2+)-induced Ca(2+) release (CICR). Sequential propagation, observed only in frogs, may require parajunctional channels, of RyR isoform beta, which are absent in the rat. Concerted opening instead appears to be a property of RyR alpha in the amphibian and the homologous isoform 1 in the mammal.     

First record of a specialist folivore of Chromolaena odorata (Asteraceae) in Togo,and indices of its range expansion in Nigeria: implications for biological control

Pareuchaetes pseudoinsulata is reported for the first time in Togo and widespread in south-western Nigeria, albeit in low densities and difficult to detect. Country-wide surveys are warranted in both countries, with the intention of renewing efforts in the biological control of Chromolaena odorata in western Africa, where the socio-ecological impact of the weed is significant.