Interleukin-1beta-mediated induction of Cox-2 in the CNS contributes to inflammatory pain hypersensitivity

Inflammation causes the induction of cyclooxygenase-2 (Cox-2), leading to the release of prostanoids, which sensitize peripheral nociceptor terminals and produce localized pain hypersensitivity. Peripheral inflammation also generates pain hypersensitivity in neighbouring uninjured tissue (secondary hyperalgesia), because of increased neuronal excitability in the spinal cord (central sensitization), and a syndrome comprising diffuse muscle and joint pain, fever, lethargy and anorexia. Here we show that Cox-2 may be involved in these central nervous system (CNS) responses, by finding a widespread induction of Cox-2 expression in spinal cord neurons and in other regions of the CNS, elevating prostaglandin E2 (PGE2) levels in the cerebrospinal fluid. The major inducer of central Cox-2 upregulation is interleukin-1beta in the CNS, and as basal phospholipase A2 activity in the CNS does not change with peripheral inflammation, Cox-2 levels must regulate central prostanoid production. Intraspinal administration of an interleukin-converting enzyme or Cox-2 inhibitor decreases inflammation-induced central PGE2 levels and mechanical hyperalgesia. Thus, preventing central prostanoid production by inhibiting the interleukin-1beta-mediated induction of Cox-2 in neurons or by inhibiting central Cox-2 activity reduces centrally generated inflammatory pain hypersensitivity.     

Developing a genome-wide selection model for genetic improvement of residual feed intake and carcass merit in a beef cattle breeding program

Residual feed intake (RFI) and carcass merit (CM) are both complex traits emerging as critical targets for beef genetic improvement.RFI and CM traits are difficult and expensive to measure and genetic improvement for these traits through traditional selection methods is not very effective.Therefore,genome-wide selection using DNA markers may be a potential alternative for genetic improvement of these traits.In this study,the efficiency of a genome-wide selection model for genetic improvement of RFI and CM was assessed.The Illumina Bovine50K bead chip was used to genotype 922 beef cattle from the Kinsella Beef Research Ranch of the University of Alberta.A Bayes model and multiple marker regression using a stepwise method were used to conduct the association test.The number of significant SNP markers for carcass weight (CWT),carcass back fat (BF),carcass rib eye area (REA),carcass grade fat (GDF),lean meat yield (LMY),and residual feed intake (RFI) were 75,54,67,57,44 and 50,respectively.Bi-variate analysis of marker scores and phenotypes for all traits were made using DMU Software.The genetic parameter for each trait was estimated.The genetic correlations of marker score and phenotype for CWT,BF,REA,GDF,LMY and RFI were 0.75,0.69,0.87,0.77,0.78,and 0.85,respectively.The average prediction accuracies of phenotypic EBV for the six traits were increased by 0.05,0.16,0.24,0.23,0.17 and 0.19,respectively.The results of this study indicated that the two-trait marker-assisted evaluation model used was a suitable alternative of genetic evaluation for these traits in beef cattle.     

Variation in the bark call of the red squirrel ( Tamiasciurus hudsonicus )

Calls of the red squirrel ( Tamiasciurus hudsonicus; n = 122) were recorded in wild populations from 15 localities in Arizona, New Mexico, Colorado, Utah, Wyoming, Montana, Idaho, and Washington. Computer-generated audiospectrograms of 20- or 30-second samples from a calling bout of each individual were analyzed. Eighteen bark types (distinct forms of the bark call) were identified plus a 19th category that included rarely used, longer bark calls. The frequency of use of each bark type within the sample was recorded for each squirrel. Differences in frequency of use of the various bark types were found among subspecies, within subspecies, and within populations; additionally, the southern subspecies utilized a reduced number of bark types. The large number of different bark types and the variation in bark type usage within populations suggest the potential for communication of such information as individual identification, behavioral states, or gender identification.     

A common sequence of calcium and pH signals in the mitogenic stimulation of eukaryotic cells

When normal quiescent (G0) cells are stimulated by mitogens to enter the cell cycle, the metabolic derepression which occurs is similar in a variety of cells. The mechanisms initiating these responses and their relationship to subsequent progression through G1 to DNA synthesis in S phase, however, are generally undefined. The clearest evidence has been obtained in sea urchin eggs, where fertilization by sperm causes a rapid, transient increase in the concentration of free cytoplasmic Ca2+ [(Ca]i), followed by a sustained increase in cytoplasmic pH (pHi). It has been demonstrated clearly that these ionic responses are obligatory for progression to DNA synthesis by the normal pathway after fertilization, although the Ca2+ signal can be bypassed by parthenogenetic agents which elevate directly pHi (for example, NH+4 ions). These observations raise the questions of whether other eukaryotic cells show the same sequence of ionic responses when stimulated by mitogens and whether such signals are an obligatory component of their mitogenic pathways. We show here that a common sequence of [Ca]i and pHi responses occurs in both quiescent mouse thymocytes and Swiss 3T3 fibroblasts stimulated by appropriate mitogens. Furthermore, 'opportunistic' mitogens (those that do not act on the cells in vivo, such as concanavalin A (Con A), the Ca2+ ionophore A23187 and 12-o-tetradecanoyl phorbol 13-acetate CTPA] that are mitogenic for both mouse thymocytes and 3T3 fibroblast, each produce characteristic ionic responses that are the same in both types of cell.     

Enhanced myogenesis in NCAM-transfected mouse myoblasts

The fusion of mononucleate precursor myoblasts to form the multinucleated skeletal muscle fibre is proceeded by a series of complex cell-cell interactions but the cell-surface molecules involved in these events have not been characterized. During myogenesis in vivo and in vitro, expression of the neural cell adhesion molecule (NCAM) undergoes an isoform transition that precisely correlates with terminal myoblast differentiation and myotube formation. Altered processing of RNA results in the replacement of the transmembrane NCAM (relative molecular mass, 145,000 (145K) in proliferating myoblasts by a predominant 125K NCAM form linked to glycosyl phosphatidylinositol in myotubes. We now report that mouse myoblasts transfected to constitutively express the human muscle-specific 125K glycosylphosphatidylinositol-linked NCAM isoform more readily fuse to form myotubes. This suggests that NCAM plays a part in myoblast fusion and that the isoform switch may promote this function.     

Bis-methionine axial ligation of haem in bacterioferritin from Pseudomonas aeruginosa

The iron-containing bacterioferritins contain the protoporphyrin IX haem group. It has been established that Escherichia coli cytochrome b1, cytochrome b557 and bacterioferritin are identical. The optical spectra at room temperature of the haem group show it to be predominantly low-spin in both the ferrous and ferric states. The nature of the axial ligands binding the haem group to the polypeptide has, however, remained unknown. Low-spin, bis-coordinate haem centres in proteins typically have a role in rapid electron transfer as redox changes at the metal ion lead to little structural rearrangement. There are only four amino acids with side-chains that have ligand field strengths sufficient to generate the low-spin state of haem, namely, histidine, lysine, methionine and cysteine. Hence there are, potentially, ten different pairs of these four ligands which could be discovered in electron transfer haemoproteins. To date only three have been established with certainty. They are bis-histidine, as in mammalian cytochrome b5, methionine-histidine, typified by cytochrome c and lysine-histidine, recently recognized by spectroscopic methods in cytochrome f. Here we report the electron paramagnetic resonance and near infrared magnetic circular dichroism spectra of the oxidized state of Ps. aeruginosa bacterioferritin which enable the axial ligands to be identified as the thioether side chains of two methionine residues, a ligation scheme not previously reported for haem in any protein.