Putting Soybeans into Permanent Farming

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C-terminal residues of skeletal muscle calsequestrin are essential for calcium binding and for skeletal ryanodine receptor inhibition

BACKGROUND Skeletal muscle function depends on calcium signaling proteins in the sarcoplasmic reticulum (SR), including the calcium-binding protein calsequestrin (CSQ), the ryanodine receptor (RyR) calcium release channel, and skeletal triadin 95 kDa (trisk95) and junctin, proteins that bind to calsequestrin type 1 (CSQ1) and ryanodine receptor type 1 (RyR1). CSQ1 inhibits RyR1 and communicates store calcium load to RyR1 channels via trisk95 and/or junctin. METHODS In this manuscript, we test predictions that CSQ1's acidic C-terminus contains binding sites for trisk95 and junctin, the major calcium binding domain, and that it determines CSQ1's ability to regulate RyR1 activity. RESULTS Progressive alanine substitution of C-terminal acidic residues of CSQ1 caused a parallel reduction in the calcium binding capacity but did not significantly alter CSQ1's association with trisk95/junctin or influence its inhibition of RyR1 activity. Deletion of the final seven residues in the C-terminus significantly hampered calcium binding, significantly reduced CSQ's association with trisk95/junctin and decreased its inhibition of RyR1. Deletion of the full C-terminus further reduced calcium binding to CSQ1 altered its association with trisk95 and junctin and abolished its inhibition of RyR1. CONCLUSIONS The correlation between the number of residues mutated/deleted and binding of calcium, trisk95, and junctin suggests that binding of each depends on diffuse ionic interactions with several C-terminal residues and that these interactions may be required for CSQ1 to maintain normal muscle function.This work was supported by the Australian Research Council (DP1094219 to AFD and NAB) and a NHMRC Career Development Award (NAB)

Rickettsial pathogens and their arthropod vectors.

Rickettsial diseases, important causes of illness and death worldwide, exist primarily in endemic and enzootic foci that occasionally give rise to sporadic or seasonal outbreaks. Rickettsial pathogens are highly specialized for obligate intracellular survival in both the vertebrate host and the invertebrate vector. While studies often focus primarily on the vertebrate host, the arthropod vector is often more important in the natural maintenance of the pathogen. Consequently, coevolution of rickettsiae with arthropods is responsible for many features of the host-pathogen relationship that are unique among arthropod-borne diseases, including efficient pathogen replication, long-term maintenance of infection, and transstadial and transovarial transmission. This article examines the common features of the host-pathogen relationship and of the arthropod vectors of the typhus and spotted fever group rickettsiae

Crocodiles as dinosaurs: Behavioural thermoregulation in very large ectotherms leads to high and stable body temperatures

Empirical field data describing daily and seasonal cycles in body temperature (Tb) of free-ranging Crocodylus porosus (32-1010 kg) can be predicted by a mathematical analysis. The analysis provides a mechanistic explanation for the decreased amplitude of daily cycles in Tb and the increase in 'average' Tb with increasing mass. Assessments of 'average' daily Tb were made by dividing the integral of the difference between measured values of Tb and minimum operative temperature by the period of integration, to yield a thermal index expressing relative 'warmth' of crocodiles. The average daily Tb of a 1010 kg crocodile was 3.7 degrees C warmer than that of a 42 kg individual in summer and 1.9 degreesC warmer than that of a 32 kg individual in winter. The success of this mathematical approach confirms that crocodiles are simple ectotherms and that there is unlikely to be a significant contribution to their thermal biology from physiological mechanisms. Behaviour, however, is very important even in large individuals. Crocodiles in the field typically move daily between land and water in cycles that vary seasonally. We predicted Tb for the reverse of these behavioural cycles, which more than doubled seasonal fluctuations in Tb compared with the observed fluctuations. We were also able to predict the Tb of very large, dinosaur-sized crocodiles in a similar climate to that at our study site. A 10 000 kg 'crocodile', for example, would be expected to have a Tb of 31 degreesC in winter, varying by less than 0.1 degrees C during a day when operative temperatures varied by nearly 20 degrees C, from 20 to 38 degrees C. The study confirms that, in low latitudes at least, large dinosaurs must have had an essentially high and stable value of Tb, without any need for endothermy. Also, access to shade or water must have been crucial for the survival of large dinosaurs at low latitudes. Furthermore, the finding of increasing 'average' Tb as ectotherms grow larger may have implications for the metabolic rates of very large reptiles, because the Q10 effect could counteract the downscaling of metabolic rate with mass, an effect that seems not to have been recognised previously

A Skeletal Muscle Ryanodine Receptor Interaction Domain in Triadin

Excitation-contraction coupling in skeletal muscle depends, in part, on a functional interaction between the ligand-gated ryanodine receptor (RyR1) and integral membrane protein Trisk 95, localized to the sarcoplasmic reticulum membrane. Various domains on Trisk 95 can associate with RyR1, yet the domain responsible for regulating RyR1 activity has remained elusive. We explored the hypothesis that a luminal Trisk 95 KEKE motif (residues 200-232), known to promote RyR1 binding, may also form the RyR1 activation domain. Peptides corresponding to Trisk 95 residues 200-232 or 200-231 bound to RyR1 and increased the single channel activity of RyR1 by 1.49 ± 0.11-fold and 1.8 ± 0.15-fold respectively, when added to its luminal side. A similar increase in [(3)H]ryanodine binding, which reflects open probability of the channels, was also observed. This RyR1 activation is similar to activation induced by full length Trisk 95. Circular dichroism showed that both peptides were intrinsically disordered, suggesting a defined secondary structure is not necessary to mediate RyR1 activation. These data for the first time demonstrate that Trisk 95's 200-231 region is responsible for RyR1 activation. Furthermore, it shows that no secondary structure is required to achieve this activation, the Trisk 95 residues themselves are critical for the Trisk 95-RyR1 interaction.This work was supported by the Australian Research Council (DP1094219 to A.F.D. and N.A.B.)

Air Conditions Close to the Ground and the Effect on Airplane Landings

This report presents the results of an investigation undertaken to determine the feasibility of making glide landings in gusty air. Wind velocities were measured at several stations between the ground and a height of 51 feet, and flight tests were made to determine the actual influence of gusts on an airplane gliding close to the ground

Cardiac cell modelling: Observations from the heart of the cardiac physiome project

In this manuscript we review the state of cardiac cell modelling in the context of international initiatives such as the IUPS Physiome and Virtual Physiological Human Projects, which aim to integrate computational models across scales and physics. In particular we focus on the relationship between experimental data and model parameterisation across a range of model types and cellular physiological systems. Finally, in the context of parameter identification and model reuse within the Cardiac Physiome, we suggest some future priority areas for this field