bluuebunny/crossref_metadata_2025 huggingface dataset:
DrosophilaTwins regulates Armadillo levels in response to Wg/Wnt signal
Authors: Bajpai Ruchi , Makhijani Kalpana , Rao Prashanth Ramesh , Shashidhara L. S.
Published: February 2004

Protein Phosphatase 2A (PP2A) has a heterotrimeric-subunit structure,consisting of a core dimer of ∼36 kDa catalytic and ∼65 kDa scaffold subunits complexed to a third variable regulatory subunit. Several studies have implicated PP2A in Wg/Wnt signaling. However, reports on the precise nature of PP2A role in Wg/Wnt pathway in different organisms are conflicting. We show that twins (tws), which codes for the B/PR55 regulatory subunit of PP2A in Drosophila, is a positive regulator of Wg/Wnt signaling. In tws- wing discs both short- and long-range targets of Wingless morphogen are downregulated. Analyses of tws- mitotic clones suggest that requirement of Tws in Wingless pathway is cell-autonomous. Epistatic genetic studies indicate that Tws functions downstream of Dishevelled and upstream of Sgg and Armadillo. Our results suggest that Tws is required for the stabilization of Armadillo/β-catenin in response to Wg/Wnt signaling. Interestingly,overexpression of, otherwise normal, Tws protein induce dominant-negative phenotypes. The conflicting reports on the role of PP2A in Wg/Wnt signaling could be due to the dominant-negative effect caused by the overexpression of one of the subunits.
Segmental development of reticulospinal and branchiomotor neurons in lamprey: insights into the evolution of the vertebrate hindbrain
Authors: Murakami Yasunori , Pasqualetti Massimo , Takio Yoko , Hirano Shigeki , Rijli Filippo M. , Kuratani Shigeru
Published: February 2004

During development, the vertebrate hindbrain is subdivided along its anteroposterior axis into a series of segmental bulges called rhombomeres. These segments in turn generate a repeated pattern of rhombomere-specific neurons, including reticular and branchiomotor neurons. In amphioxus(Cephalochordata), the sister group of the vertebrates, a bona fide segmented hindbrain is lacking, although the embryonic brain vesicle shows molecular anteroposterior regionalization. Therefore, evaluation of the segmental patterning of the central nervous system of agnathan embryos is relevant to our understanding of the origin of the developmental plan of the vertebrate hindbrain. To investigate the neuronal organization of the hindbrain of the Japanese lamprey, Lethenteron japonicum, we retrogradely labeled the reticulospinal and branchial motoneurons. By combining this analysis with a study of the expression patterns of genes identifying specific rhombomeric territories such as LjKrox20, LjPax6, LjEphC and LjHox3, we found that the reticular neurons in the lamprey hindbrain, including isthmic,bulbar and Mauthner cells, develop in conserved rhombomere-specific positions,similar to those in the zebrafish. By contrast, lamprey trigeminal and facial motor nuclei are not in register with rhombomere boundaries, unlike those of gnathostomes. The trigeminal-facial boundary corresponds to the rostral border of LjHox3 expression in the middle of rhombomere 4. Exogenous application of retinoic acid (RA) induced a rostral shift of both the LjHox3 expression domain and branchiomotor nuclei with no obvious repatterning of rhombomeric segmentation and reticular neurons. Therefore,whereas subtype variations of motoneuron identity along the anteroposterior axis may rely on Hox-dependent positional values, as in gnathostomes, such variations in the lamprey are not constrained by hindbrain segmentation. We hypothesize that the registering of hindbrain segmentation and neuronal patterning may have been acquired through successive and independent stepwise patterning changes during evolution.
TheArabidopsis JAGGEDgene encodes a zinc finger protein that promotes leaf tissue development
Authors: Ohno Carolyn K. , Reddy G. Venugopala , Heisler Marcus G. B. , Meyerowitz Elliot M.
Published: February 2004

Important goals in understanding leaf development are to identify genes involved in pattern specification, and also genes that translate this information into cell types and tissue structure. Loss-of-function mutations at the JAGGED (JAG) locus result in Arabidopsisplants with abnormally shaped lateral organs including serrated leaves, narrow floral organs, and petals that contain fewer but more elongate cells. jag mutations also suppress bract formation in leafy,apetala1 and apetala2 mutant backgrounds. The JAG gene was identified by map-based cloning to be a member of the zinc finger family of plant transcription factors and encodes a protein similar in structure to SUPERMAN with a single C2H2-type zinc finger, a proline-rich motif and a short leucine-rich repressor motif. JAG mRNA is localized to lateral organ primordia throughout the plant but is not found in the shoot apical meristem. Misexpression of JAG results in leaf fusion and the development of ectopic leaf-like outgrowth from both vegetative and floral tissues. Thus, JAG is necessary for proper lateral organ shape and is sufficient to induce the proliferation of lateral organ tissue.
Gibberellin regulatesArabidopsisfloral development via suppression of DELLA protein function
Authors: Cheng Hui , Qin Lianju , Lee Sorcheng , Fu Xiangdong , Richards Donald E. , Cao Dongni , Luo Da , Harberd Nicholas P. , Peng Jinrong
Published: February 2004

The phytohormone gibberellin (GA) regulates the development and fertility of Arabidopsis flowers. The mature flowers of GA-deficient mutant plants typically exhibit reduced elongation growth of petals and stamens. In addition, GA-deficiency blocks anther development, resulting in male sterility. Previous analyses have shown that GA promotes the elongation of plant organs by opposing the function of the DELLA proteins, a family of nuclear growth repressors. However, it was not clear that the DELLA proteins are involved in the GA-regulation of stamen and anther development. We show that GA regulates cell elongation rather than cell division during Arabidopsis stamen filament elongation. In addition, GA regulates the cellular developmental pathway of anthers leading from microspore to mature pollen grain. Genetic analysis shows that the Arabidopsis DELLA proteins RGA and RGL2 jointly repress petal, stamen and anther development in GA-deficient plants, and that this function is enhanced by RGL1 activity. GA thus promotes Arabidopsis petal, stamen and anther development by opposing the function of the DELLA proteins RGA, RGL1 and RGL2.
Distinct activities of Msx1 and Msx3 in dorsal neural tube development
Authors: Liu Ying , Helms Amy W. , Johnson Jane E.
Published: February 2004

Patterning of the dorsal neural tube involves Bmp signaling, which results in activation of multiple pathways leading to the formation of neural crest,roof plate and dorsal interneuron cell types. We show that constitutive activation of Bmp signaling at early stages (HH10-12) of chick neural tube development induces roof-plate cell fate, accompanied by an increase of programmed cell death and a repression of neuronal differentiation. These activities are mimicked by the overexpression of the homeodomain transcription factor Msx1, a factor known to be induced by Bmp signaling. By contrast, the closely related factor, Msx3, does not have these activities. At later stages of neural tube development (HH14-16), dorsal progenitor cells lose their competence to generate roof-plate cells in response to Bmp signaling and instead generate dorsal interneurons. This aspect of Bmp signaling is phenocopied by the overexpression of Msx3 but not Msx1. Taken together, these results suggest that these two different Msx family members can mediate distinct aspects of Bmp signaling during neural tube development.
De novo DNA methylation is dispensable for the initiation and propagation of X chromosome inactivation
Authors: Sado Takashi , Okano Masaki , Li En , Sasaki Hiroyuki
Published: February 2004

Xist (X-inactive specific transcript) plays a crucial role in X-inactivation. This non-coding RNA becomes upregulated on the X chromosome that is to be inactivated upon differentiation. Previous studies have revealed that although maintenance-type DNA methylation is not essential for X-inactivation to occur, it is required for the stable repression of Xist in differentiated cells. However, it is unknown whether differential de novo methylation at the Xist promoter, which is mediated by Dnmt3a and/or Dnmt3b, is a cause or a consequence of monoallelic expression of Xist. We show that Xist expression is appropriately regulated in the absence of Dnmt3a and Dnmt3b and that a single X chromosome undergoes proper inactivation in mutant females. Our results indicate that a mechanism(s) other than DNA methylation plays a principal role in initiating X-inactivation. We also demonstrate that delayed upregulation of Xistdoes not induce X-inactivation, consistent with a crucial developmental window for the chromosomal silencing.
Metalloproteinase pregnancy-associated plasma protein A is a critical growth regulatory factor during fetal development
Authors: Conover Cheryl A. , Bale Laurie K. , Overgaard Michael T. , Johnstone Edward W. , Laursen Ulla H. , Füchtbauer Ernst-Martin , Oxvig Claus , van Deursen Jan
Published: February 2004

Pregnancy-associated plasma protein A (PAPPA) is a metzincin superfamily metalloproteinase in the insulin-like growth factor (IGF) system. PAPPA increases IGF bioavailability and mitogenic effectiveness in vitro through regulated cleavage of IGF-binding protein 4 (IGFBP4). To determine its function in vivo, we generated PAPPA-null mice by gene targeting. Mice homozygous for targeted disruption of the PAPPA gene were viable but 60% the size of wild-type littermates at birth. The impact of the mutation was exerted during the early embryonic period prior to organogenesis, resulting in proportional dwarfism. PAPPA, IGF2 and IGFBP4 transcripts co-localized in wild-type embryos, and expression of IGF2 and IGFBP4 mRNA was not altered in PAPPA-deficient embryos. However,IGFBP4 proteolytic activity was completely lacking in fibroblasts derived from PAPPA-deficient embryos, and IGFBP4 effectively inhibited IGF-stimulated mitogenesis in these cells. These results provide the first direct evidence that PAPPA is an essential growth regulatory factor in vivo, and suggest a novel mechanism for regulated IGF bioavailability during early fetal development.
HtrA1 serine protease inhibits signaling mediated by Tgfβ family proteins
Authors: Oka Chio , Tsujimoto Rumi , Kajikawa Miwa , Koshiba-Takeuchi Kazuko , Ina Junko , Yano Masato , Tsuchiya Akiho , Ueta Yoshihumi , Soma Akinobu , Kanda Hidenobu , Matsumoto Michio , Kawaichi Masashi
Published: February 2004

HtrA1, a member of the mammalian HtrA serine protease family, has a highly conserved protease domain followed by a PDZ domain. Because HtrA1 is a secretory protein and has another functional domain with homology to follistatin, we examined whether HtrA1 functions as an antagonist of Tgfβfamily proteins. During embryo development, mouse HtrA1 was expressed in specific areas where signaling by Tgfβ family proteins plays important regulatory roles. The GST-pulldown assay showed that HtrA1 binds to a broad range of Tgfβ family proteins, including Bmp4, Gdf5, Tgfβs and activin. HtrA1 inhibited signaling by Bmp4, Bmp2, and Tgfβ1 in C2C12 cells, presumably by preventing receptor activation. Experiments using a series of deletion mutants indicated that the binding activity of HtrA1 required the protease domain and a small linker region preceding it, and that inhibition of Tgfβ signaling is dependent on the proteolytic activity of HtrA1. Misexpression of HtrA1 near the developing chick eye led to suppression of eye development that was indistinguishable from the effects of noggin. Taken together, these data indicate that HtrA1 protease is a novel inhibitor of Tgfβ family members.
A Raf/MEK/ERK signaling pathway is required for development of the sea urchin embryo micromere lineage through phosphorylation of the transcription factor Ets
Authors: Röttinger Eric , Besnardeau Lydia , Lepage Thierry
Published: February 2004

In the sea urchin embryo, the skeleton of the larva is built from a population of mesenchymal cells known as the primary mesenchyme cells (PMCs). These derive from the large micromeres that originate from the vegetal pole at fourth cleavage. At the blastula stage, the 32 cells of this lineage detach from the epithelium and ingress into the blastocoel by a process of epithelial-mesenchymal transition. We report that shortly before ingression,there is a transient and highly localized activation of the MAP-kinase ERK in the micromere lineage. We show that ingression of the PMCs requires the activity of ERK, MEK and Raf, and depends on the maternal Wnt/β-catenin pathway. Dissociation experiments and injection of mRNA encoding a dominant-negative form of Ras indicated that this activation is probably cell autonomous. We identified the transcription factors Ets1 and Alx1 as putative targets of the phosphorylation by ERK. Both proteins contain a single consensus site for phosphorylation by the MAP kinase ERK. In addition, the Ets1 protein sequence contains a putative ERK docking site. Overexpression of ets1 by injection of synthetic mRNA in the egg caused a dramatic increase in the number of cells becoming mesenchymal at the blastula stage. This effect could be largely inhibited by treating embryos with the MEK inhibitor U0126. Moreover, mutations in the consensus phosphorylation motif substituting threonine 107 by an aspartic or an alanine residue resulted respectively in a constitutively active form of Ets1 that could not be inhibited by U0126 or in an inactive form of Ets1. These results show that the MAP kinase pathway, working through phosphorylation of Ets1, is required for full specification of the PMCs and their subsequent transition from epithelial to mesenchymal state.
missing oocyteencodes a highly conserved nuclear protein required for the maintenance of the meiotic cycle and oocyte identity inDrosophila
Authors: Iida Takako , Lilly Mary A.
Published: February 2004

In Drosophila, a single oocyte develops within a 16-cell germline cyst. Although all 16 cells initiate meiosis and undergo premeiotic S phase,only the oocyte retains its meiotic chromosome configuration and remains in the meiotic cycle. The other 15 cells in the cyst enter the endocycle and develop as polyploid nurse cells. A longstanding goal in the field has been to identify factors that are concentrated or activated in the oocyte, that promote meiotic progression and/or the establishment of the oocyte identity. We present the characterization of the missing oocyte gene, an excellent candidate for a gene directly involved in the differentiation of the oocyte nucleus. The missing oocyte gene encodes a highly conserved protein that preferentially accumulates in pro-oocyte nuclei in early prophase of meiosis I. In missing oocyte mutants, the oocyte enters the endocycle and develops as a polyploid nurse cell. Genetic interaction studies indicate that missing oocyte influences meiotic progression prior to pachytene and may interact with pathways that control DNA metabolism. Our data strongly suggest that the product of the missing oocyte gene acts in the oocyte nucleus to facilitate the execution of the unique cell cycle and developmental programs that produce the mature haploid gamete.
Active cell migration drives the unilateral movements of the anterior visceral endoderm
Authors: Srinivas Shankar , Rodriguez Tristan , Clements Melanie , Smith James C. , Beddington Rosa S. P.
Published: February 2004

The anterior visceral endoderm (AVE) of the mouse embryo is a specialised extra-embryonic tissue that is essential for anterior patterning of the embryo. It is characterised by the expression of anterior markers such as Hex, Cerberus-like and Lhx1. At pre-gastrula stages, cells of the AVE are initially located at the distal tip of the embryo, but they then move unilaterally to the future anterior. This movement is essential for converting the existing proximodistal axis into an anteroposterior axis. To investigate this process, we developed a culture system capable of imaging embryos in real time with single cell resolution. Our results show that AVE cells continuously change shape and project filopodial processes in their direction of motion, suggesting that they are actively migrating. Their proximal movement stops abruptly at the junction of the epiblast and extra-embryonic ectoderm, whereupon they move laterally. Confocal microscope images show that AVE cells migrate as a single layer in direct contact with the epiblast, suggesting that this tissue might provide directional cues. Together, these results show that the anteroposterior axis is correctly positioned by the active movement of cells of the AVE in response to cues from their environment, and by a `barrier' to their movement that provides an endpoint for this migration.
Target-derived BMP signaling limits sensory neuron number and the extent of peripheral innervation in vivo
Authors: Guha Udayan , Gomes William A. , Samanta Jayshree , Gupta Meenakshi , Rice Frank L. , Kessler John A.
Published: February 2004

The role of target-derived BMP signaling in development of sensory ganglia and the sensory innervation of the skin was examined in transgenic animals that overexpress either the BMP inhibitor noggin or BMP4 under the control of a keratin 14 (K14) promoter. Overexpression of noggin resulted in a significant increase in the number of neurons in the trigeminal and dorsal root ganglia. Conversely, overexpression of BMP4 resulted in a significant decrease in the number of dorsal root ganglion neurons. There was no significant change in proliferation of trigeminal ganglion neurons in the noggin transgenic animals, and neuron numbers did not undergo the normal developmental decrease between E12.5 and the adult, suggesting that programmed cell death was decreased in these animals. The increase in neuron numbers in the K14-noggin animals was followed by an extraordinary increase in the density of innervation in the skin and a marked change in the pattern of innervation by different types of fibers. Conversely, the density of innervation of the skin was decreased in the BMP4 overexpressing animals. Further Merkel cells and their innervation were increased in the K14-noggin mice and decreased in the K14-BMP4 mice. The changes in neuron numbers and the density of innervation were not accompanied by a change in the levels of neurotrophins in the skin. These findings indicate that the normal developmental decrease in neuron numbers in sensory ganglia depends upon BMP signaling, and that BMPs may limit both the final neuron number in sensory ganglia as well as the extent of innervation of targets. Coupled with prior observations, this suggests that BMP signaling may regulate the acquisition of dependence of neurons on neurotrophins for survival, as well as their dependence on target-derived neurotrophins for determining the density of innervation of the target.
Upregulation of Mitimere and Nubbin acts through Cyclin E to confer self-renewing asymmetric division potential to neural precursor cells
Authors: Bhat Krishna Moorthi , Apsel Nora
Published: February 2004

In the Drosophila CNS, neuroblasts undergo self-renewing asymmetric divisions, whereas their progeny, ganglion mother cells (GMCs),divide asymmetrically to generate terminal postmitotic neurons. It is not known whether GMCs have the potential to undergo self-renewing asymmetric divisions. It is also not known how precursor cells undergo self-renewing asymmetric divisions. Here, we report that maintaining high levels of Mitimere or Nubbin, two POU proteins, in a GMC causes it to undergo self-renewing asymmetric divisions. These asymmetric divisions are due to upregulation of Cyclin E in late GMC and its unequal distribution between two daughter cells. GMCs in an embryo overexpressing Cyclin E, or in an embryo mutant for archipelago, also undergo self-renewing asymmetric divisions. Although the GMC self-renewal is independent of inscuteable and numb, the fate of the differentiating daughter is inscuteable and numb-dependent. Our results reveal that regulation of Cyclin E levels, and asymmetric distribution of Cyclin E and other determinants, confer self-renewing asymmetric division potential to precursor cells, and thus define a pathway that regulates such divisions. These results add to our understanding of maintenance and loss of pluripotential stem cell identity.
Cytoplasmic and molecular reconstruction ofXenopusembryos:synergy of dorsalizing and endo-mesodermalizing determinants drives early axial patterning
Authors: Katsumoto Keiichi , Arikawa Tomohiro , Doi Jun-ya , Fujii Hidefumi , Nishimatsu Shin-ichiro , Sakai Masao
Published: February 2004

Ablation of vegetal cytoplasm from newly fertilized Xenopus eggs results in the development of permanent blastula-type embryos (PBEs). PBEs cleave normally and develop into a very simple tissue consisting only of atypical epidermis. We tried to restore complete embryonic development in PBEs by cytoplasmic transplantation or by mRNA injection. We show a two-step reconstruction of the body plan. In the first step, PBEs injected with either marginal cytoplasm or synthetic VegT RNA restored gastrulation and mesoderm formation, but not axial patterning. Injection of Xwnt8 mRNA (acting upstream of β-catenin and thus substitutes for the dorsal determinant)did not restore axial development in PBEs. Simultaneous injections of Xwnt8 and VegT into PBEs resulted in dorsal axis development, showing the synergy of these molecules in axial development. These results suggest that the mixing of two cytoplasmic determinants, i.e. the dorsal determinant in the vegetal pole and the endo-mesodermal determinant in the whole vegetal half, triggers the early axial developmental process in Xenopus embryos.
Early developmental failure of substantia nigra dopamine neurons in mice lacking the homeodomain genePitx3
Authors: Smidt Marten P. , Smits Simone M. , Bouwmeester Hans , Hamers Frank P. T. , van der Linden Annemarie J. A. , Hellemons Anita J. C. G. M. , Graw Jochen , Burbach J. Peter H.
Published: February 2004

The mesencephalic dopamine (mesDA) system is involved in the control of movement and behavior. The expression of Pitx3 in the brain is restricted to the mesDA system and the gene is induced relatively late, at E11.5, a time when tyrosine hydroxylase (Th) gene expression is initiated. We show here that, in the Pitx3-deficient aphakia(ak) mouse mutant, the mesDA system is malformed. Owing to the developmental failure of mesDA neurons in the lateral field of the midbrain,mesDA neurons are not found in the SNc and the projections to the caudate putamen are selectively lost. However, Pitx3 is expressed in all mesDA neurons in control animals. Therefore, mesDA neurons react specifically to the loss of Pitx3. Defects of motor control where not seen in the ak mice,suggesting that other neuronal systems compensate for the absence of the nigrostriatal pathway. However, an overall lower activity was observed. The results suggest that Pitx3 is specifically required for the formation of the SNc subfield at the onset of dopaminergic neuron differentiation.
Gata2 specifies serotonergic neurons downstream of sonic hedgehog
Authors: Craven Sarah E. , Lim Kim-Chew , Ye Weilan , Engel James Douglas , de Sauvage Frederic , Rosenthal Arnon
Published: February 2004

Distinct classes of serotonergic (5-HT) neurons develop along the ventral midline of the vertebrate hindbrain. Here, we identify a Sonic hedgehog(Shh)-regulated cascade of transcription factors that acts to generate a specific subset of 5-HT neurons. This transcriptional cascade is sufficient for the induction of rostral 5-HT neurons within rhombomere 1 (r1), which project to the forebrain, but not for the induction of caudal 5-HT neurons,which largely terminate in the spinal cord. Within the rostral hindbrain, the Shh-activated homeodomain proteins Nkx2.2 and Nkx6.1 cooperate to induce the closely related zinc-finger transcription factors Gata2 and Gata3. Gata2 in turn is necessary and sufficient to activate the transcription factors Lmx1b and Pet1, and to induce 5-HT neurons within r1. In contrast to Gata2, Gata3 is not required for the specification of rostral 5-HT neurons and appears unable to substitute for the loss of Gata2. Our findings reveal that the identity of closely related 5-HT subclasses occurs through distinct responses of adjacent rostrocaudal progenitor domains to broad ventral inducers.
Notch signaling: control of cell communication and cell fate
Authors: Lai Eric C.
Published: February 2004

Notch is a transmembrane receptor that mediates local cell-cell communication and coordinates a signaling cascade present in all animal species studied to date. Notch signaling is used widely to determine cell fates and to regulate pattern formation; its dysfunction results in a tremendous variety of developmental defects and adult pathologies. This primer describes the mechanism of Notch signal transduction and how it is used to control the formation of biological patterns.
Overexpression ofbroad: a new insight into its role in theDrosophilaprothoracic gland cells
Authors: Zhou Xiaofeng , Zhou Baohua , Truman James W. , Riddiford Lynn M.
Published: February 2004

Insect molting is triggered by ecdysteroids, which are produced in the prothoracic glands (PG). The broad (br) gene is one of the`early genes' directly regulated by ecdysteroids. Ectopic expression of the BR-Z3 isoform in early second instar Drosophila larvae (L2) before the rise of the ecdysteroid titer prevented molting to the third instar, but the larvae subsequently formed L2 prepupae after prolonged feeding. When these larvae were fed on diet containing 20-hydroxyecdysone (20E), they formed pharate third instar larvae. The critical weight for normal L3 pupariation of w1118 larvae was found to be 0.8 mg and that for L2 pupariation was 0.45 mg. We also defined a threshold weight for metamorphosis of 0.3 mg, above which L2 larvae will metamorphose when provided with 20E. BR-Z3 apparently works through the PG cells of the ring gland but not the putative neurosecretory cells that drive ecdysone secretion, because ectopic expression of BR-Z3 specifically in the ring gland caused 53% of the larvae to become permanent first instar larvae. Driving other BR isoforms in the ring gland prevented larval molting or pupariation to varying degrees. These molting defects were rescued by feeding 20E. Overexpression of each of the BR isoforms caused degeneration of the PG cells but on different time courses,indicating that BR is a signal for the degeneration of the PG cells that normally occurs during the pupal–adult transition.
The anterior cardiac plexus: an intrinsic neurosecretory site within the stomatogastric nervous system of the crabCancer productus
Authors: Christie Andrew E. , Cain Shaun D. , Edwards John M. , Clason Todd A. , Cherny Elena , Lin Minhui , Manhas Amitoz S. , Sellereit Kirsten L. , Cowan Nicholas G. , Nold Kellen A. , Strassburg Hans-Peter , Graubard Katherine
Published: February 2004

The stomatogastric nervous system (STNS) of decapod crustaceans is modulated by both locally released and circulating substances. In some species, including chelate lobsters and freshwater crayfish, the release zones for hormones are located both intrinsically to and at some distance from the STNS. In other crustaceans, including Brachyuran crabs, the existence of extrinsic sites is well documented. Little, however, is known about the presence of intrinsic neuroendocrine structures in these animals. Putative intrinsic sites have been identified within the STNS of several crab species,though ultrastructural confirmation that these structures are in fact neuroendocrine in nature remains lacking. Using a combination of anatomical techniques, we demonstrate the existence of a pair of neurosecretory sites within the STNS of the crab Cancer productus. These structures, which we have named the anterior cardiac plexi (ACPs), are located on the anterior cardiac nerves (acns), which overlie the cardiac sac region of the foregut. Each ACP starts several hundred μm from the origin of the acn and extends distally for up to several mm. Transmission electron microscopy done on these structures shows that nerve terminals are present in the peripheral portion of each acn, just below a well defined epineurium. These terminals contain dense-core and, occasionally,electron-lucent vesicles. In many terminals, morphological correlates of hormone secretion are evident. Immunocytochemistry shows that the ACPs are immunopositive for FLRFamide-related peptide. All FLRFamide labeling in the ACPs originates from four axons, which descend to these sites through the superior oesophageal and stomatogastric nerves. Moreover, these FLRFamide-immunopositive axons are the sole source of innervation to the ACPs. Collectively, our results suggest that the STNS of C. productus is not only a potential target site for circulating hormones, but also serves as a neuroendocrine release center itself.
Does she smell like a queen? Chemoreception of a cuticular hydrocarbon signal in the ant Pachycondyla inversa
Authors: D'Ettorre Patrizia , Heinze Jürgen , Schulz Claudia , Francke Wittko , Ayasse Manfred
Published: February 2004

Primitive ant societies, with their relatively simple social structure,provide an opportunity to explore the evolution of chemical communication, in particular of mechanisms underlying within-colony discrimination. In the same colony, slight differences in individual odours can be the basis for discrimination between different castes, classes of age and social status. There is some evidence from correlative studies that such inter-individual variation is associated with differences in reproductive status, but direct proof that certain chemical compounds are detected and recognized by ants is still lacking. In the ponerine ant Pachycondyla inversa, fertile queens and, in orphaned colonies, dominant egg-laying workers are characterized by the predominance of a branched hydrocarbon,3,11-dimethylheptacosane (3,11-diMeC27) on the cuticle. Using electroanntennography and gas chromatography with electroantennographic detection, we show that the antennae of P. inversa workers react to this key compound. 3,11-diMeC27 is correlated with ovarian activity and, because it is detected, is likely to assume the role of a fertility signal reflecting the quality of the sender.
Mechanisms and significance of reduced activity and responsiveness in resting frog tadpoles
Authors: Lambert Thomas D. , Howard Jenny , Plant Andy , Soffe Steve , Roberts Alan
Published: February 2004

Hatchling Xenopus laevis tadpoles spend most of their time attached to objects or the water surface by mucus secreted by a gland on the head. While attached, swimming activity and responsiveness to swim-initiating stimuli are reduced over long periods of time. We have investigated the mechanisms and significance of this apparent long-term inhibition. In behavioural experiments we show, firstly, that innervation of the cement gland and GABAA-mediated inhibition are necessary for attachment to reduce responsiveness, and secondly, that denervation of the cement gland increases tadpole activity and increases their predation by damselfly nymphs(Zygoptera). To investigate the neuronal pathway from the cement gland to GABAA inhibition, we have devised an immobilized, inverted tadpole preparation where a weight attached to the mucus simulates the force as it hangs. Simulated attachment reduces responsiveness and spontaneous fictive swimming activity. We have recorded the activity and responses of trigeminal neurons innervating the cement gland. They are spontaneously active and simulating attachment results in a sustained increase in this activity. We propose that hanging from a mucus strand increases firing in cement gland afferents. This leads to tonic GABA inhibition that reduces tadpole activity and responses, and leads to fewer attacks by predators.
Isolation of a novel aquaglyceroporin from a marine teleost (Sparus auratus): function and tissue distribution
Authors: Santos C. R. A. , Estêvão M. D. , Fuentes J. , Cardoso J. C. R. , Fabra M. , Passos A. L. , Detmers F. J. , Deen P. M. T. , Cerdà J. , Power D. M.
Published: February 2004

The aquaporins (formerly called the major intrinsic protein family) are transmembrane channel proteins. The family includes the CHIP group, which are functionally characterised as water channels and the GLP group, which are specialised for glycerol transport. The present study reports the identification and characterisation of a novel GLP family member in a teleost fish, the sea bream Sparus auratus. A sea bream aquaporin (sbAQP)cDNA of 1047 bp and encoding a protein of 298 amino acids was isolated from a kidney cDNA library. Functional characterization of the sbAQP using a Xenopus oocyte assay revealed that the isolated cDNA stimulated osmotic water permeability in a mercury-sensitive manner and also stimulated urea and glycerol uptake. Northern blotting demonstrated that sbAQP was expressed at high levels in the posterior region of the gut, where two transcripts were identified (1.6 kb and 2 kb), and in kidney, where a single transcript was present (2 kb). In situ hybridisation studies with a sbAQP riboprobe revealed its presence in the lamina propria and smooth muscle layer of the posterior region of the gut and in epithelial cells of some kidney tubules. sbAQP was also present in putative chloride cells of the gill. Phylogenetic analysis of sbAQP, including putative GLP genes from Fugu rubripes, revealed that it did not group with any of the previously isolated vertebrate GLPs and instead formed a separate group, suggesting that it may be a novel GLP member.
Fracture characteristics of acrylic bone cement-bone composites
Authors: Buckley P J , Orr J F , Revie I C , Breusch S J , Dunne N J
Published: February 2004

In this study, the fracture properties of Perspex®, acrylic bone cement prepared using a commercially available reduced pressure mixing system and a bone cement-bone composite were compared under different test conditions. The method used was the double-torsion (DT) test. The observations made from this investigation are as follows. The fracture toughness and critical crack length for Perspex® significantly increased (ANOVA, p = 0.001) when tested in water compared to air. An increase in test temperature from 19 to 37°C resulted in a decrease in the fracture properties in water, this reduction being also statistically significant (ANOVA, p = 0.02). The mean fracture toughness and standard deviation of CMW3TM bone cement when mixed under reduced pressure was 2.19 ± 0.11 MN m−3/2 compared to 3.89 ± 0.10 MN m−3/2 for the cement-bone composite (ANOVA, p = 0.004). The crack length determined for CMW3TM bone cement and the cement-bone composite were 0.323 ± 0.031 and 1.1434 ± 0.61 mm respectively. The plateau loads of the composite material were higher than measured for the monolithic acrylic bone cement, 249.66 ± 67.75 N compared with 140.83 ± 6.82 N. The high level of variation recorded for the plateau loads of the bone cement-bone composite is due to the orientation and volume fraction of the cancellous bone. It can be concluded from this investigation that acrylic bone cement interdigitation into the cancellous bone results in a superior material with respect to crack resistance in comparison with the bone cement as a lone entity. Therefore it is an advantage if there is sufficient cancellous bone stock available within the intermedullary canal to allow bone cement penetration to occur, for the transfer of loads during daily activity. Additionally, it is paramount that the clinician ensures that adequate pressure is applied and maintained for an appropriate time during cement injection and prosthesis insertion in order to ensure optimum cement penetration into the pore openings of the cancellous bone, thus improving the resistance of the cement mantle to fracture and ultimately improving the longevity of the joint replacement.
Microbubble ultrasound contrast agents: A review
Authors: Stride E , Saffari N
Published: February 2004

The superior scattering properties of gas bubbles compared with blood cells have made microbubble ultrasound contrast agents important tools in ultrasound diagnosis. Over the past 2 years they have become the focus of a wide and rapidly expanding field of research, with their benefits being repeatedly demonstrated, both in ultrasound image enhancement, and more recently in drug and gene delivery applications. However, despite considerable investigation, their behaviour is by no means fully understood and, while no definite evidence of harmful effects has been obtained, there remain some concerns as to their safety. In this review the existing theoretical and experimental evidence is examined in order to clarify the extent to which contrast agents are currently understood and to identify areas for future research. In particular the disparity between the conditions considered in theoretical models and those encountered both in vitro, and more importantly in vivo is discussed, together with the controversy regarding the risk of harmful bio-effects.
A technique for the measurement of patellar tracking during weight-bearing activities using ultrasound
Authors: Shih Yi-Fen , Bull A M J , McGregor A H , Humphries K , Amis A A
Published: February 2004

A new fixation device, the femoral clamp, was developed in this study for the ultrasound measurement of patellar medio-lateral motion during sitting and squatting knee flexion/extension. Seventeen subjects, 6 males, 11 females, aged between 18 and 40 years were recruited for the test. Results showed that the patella moved medially then laterally from extension to flexion when sitting. Weight-bearing knee motion produced a more laterally tracked patella without the presence of the initial medial patellar translation. The tracking patterns of the patellae were similar regardless of knee movement direction. The patellar lateral position was greatly affected by the movement task ( p < 0.0005), and was also influenced by gender for maximum medial position ( p < 0.05). The reproducibility of the measurement was between 0.29 and 0.90 for the intra-rater and 0.34-0.75 for the inter-rater reliability. The accuracy of the ultrasound measurement was validated by interventional magnetic resonance (iMR) imaging of the patella and the mean error of the measurement was 1.4 ± 3.2 mm. Although further research is needed to improve the accuracy and reliability of this method, it has demonstrated the feasibility of obtaining patellar tracking data during load-bearing activities.
Medical device regulation for manufacturers
Authors: McAllister P , Jeswiet J
Published: February 2004

Manufacturers of medical devices are held to a higher standard than manufacturers of many other products due to the potential severity of the consequences of introducing inferior or unsafe products to the market-place. In Canada, the medical device industry is regulated by Health Canada under the Medical Device Regulations of the Food and Drug Act. The Medical Device Regulations define requirements of medical device design, development and manufacture to ensure that products reaching the public are safe and effective. Health Canada also requires that medical device manufacturers maintain distribution records to ensure that devices can be traced to the source and consumers can be contacted successfully in the event that a device is recalled.

Medical devices exported from Canada must be compliant with the regulations of the country of import. The Canadian Medical Device Regulations were based on the Medical Device Directives of the European Union thus facilitating approval of Canadian devices for the European market. The United States Food and Drug Administration has separate and distinct requirements for safety and quality of medical devices. While effort has been made to facilitate approval and trade of Canadian medical devices in the United States and the European Union, obtaining approval from multiple regulatory bodies can result in increased device development time and cost.

The Global Harmonization Task Force is an organization composed of members from Japanese, Australian, European, Canadian and American medical device regulatory bodies. This organization was formed with the objective of harmonizing medical device regulations in an effort to facilitate international trade and standardize the quality of medical devices available to all countries. This paper discusses the requirements that must be met by manufacturers when designing and manufacturing medical devices.
Microscopic asperity contact and deformation of ultrahigh molecular weight polyethylene bearing surfaces
Authors: Wang F C , Jin Z M , McEwen H M J , Fisher J
Published: February 2004

The effect of the roughness and topography of ultrahigh molecular weight polyethylene (UHMWPE) bearing surfaces on the microscopic contact mechanics with a metallic counterface was investigated in the present study. Both simple sinusoidal roughness forms, with a wide range of amplitudes and wavelengths, and real surface topographies, measured before and after wear testing in a simple pin-on-plate machine, were considered in the theoretical analysis. The finite difference method was used to solve the microscopic contact between the rough UHMWPE bearing surface and a smooth hard counterface. The fast Fourier transform (FFT) was used to cope with the large number of mesh points required to represent the surface topography of the UHMWPE bearing surface. It was found that only isolated asperity contacts occurred under physiological loading, and the real contact area was only a small fraction of the nominal contact area. Consequently, the average contact pressure experienced at the articulating surfaces was significantly higher than the nominal contact pressure. Furthermore, it was shown that the majority of asperities on the worn UHMWPE pin were deformed in the elastic region, and consideration of the plastic deformation only resulted in a negligible increase in the predicted asperity contact area. Microscopic asperity contact and deformation mechanisms may play an important role in the understanding of the wear mechanisms of UHMWPE bearing surfaces.
Computer simulation of the human leg subjected to impact loading
Authors: Xishi Wang , Turgut T S , Nuri A
Published: February 2004

One of the most important tasks in clinical practice is to determine accurately the forces and displacements of the various anatomical structural components of the human leg, especially when subjected to impact loading. This paper therefore develops a computer simulation which describes the biodynamic response of the human lower extremity for realistic activities such as kicking by foot etc. Firstly, the paper derives the nonlinear equations of motion and nonlinear constraint equation governing the sagittal plane response of the human leg subjected to impact loading. The equations are then solved using numerical techniques and the results are presented. Special attention is given to the ligament, contact and muscle group forces, the location of contact points, anterior-posterior displacements and the motion trajectories of the femur and the tibia.
Apparatus to test insertion and removal torque of bone screws
Authors: Koistinen A , Santavirta S , Lappalainen R
Published: February 2004

This paper describes affordable equipment for testing bone screw torque, corresponding to ASTM standard F543-00 for testing metallic medical bone screws. Correct testing of thin and long bone screws is essential due to screw failures during insertion and removal of the screws. Furthermore, insertion torque is an important factor in predicting fixation strength, screw pull-out force and effects of surface treatment of screws. The capability of the custom-built tester was determined using polytetrafluoroethylene and wood disc samples and bone screws. Bovine cortical bones allowed testing to the failure limit, i.e. the torque increased in long screws to the fracture limit. For 2.7 and 3.5 mm thick self-tapping cortical bone screws, the failure torques were 30–50 per cent higher than the minimum values required by the standard (1.0 and 2.3 N m respectively). The equipment provided reproducible results and fulfilled the ASTM standard very well. Preliminary testing with amorphous diamond coated bone screws showed good durability of the coating and on average 10–15 per cent lower torque values compared with uncoated screws. The equipment can be used to measure insertion and removal torques as described in the standard. Furthermore, it also allows testing of normal screws and bolts.
Corrections to the Mann et. al. (1998) Proxy Data Base and Northern Hemispheric Average Temperature Series
Authors: McIntyre Stephen , McKitrick Ross
Published: February 2004

The data set of proxies of past climate used in Mann, Bradley and Hughes (1998, “MBH98” hereafter) for the estimation of temperatures from 1400 to 1980 contains collation errors, unjustifiable truncation or extrapolation of source data, obsolete data, geographical location errors, incorrect calculation of principal components and other quality control defects. We detail these errors and defects. We then apply MBH98 methodology to the construction of a Northern Hemisphere average temperature index for the 1400–1980 period, using corrected and updated source data. The major finding is that the values in the early 15th century exceed any values in the 20th century. The particular “hockey stick” shape derived in the MBH98 proxy construction – a temperature index that decreases slightly between the early 15th century and early 20th century and then increases dramatically up to 1980 — is primarily an artefact of poor data handling, obsolete data and incorrect calculation of principal components.
On the Coherence between Dynamics of the World Fuel Consumption and Global Temperature Anomaly
Authors: Klyashtorin L.B. , Lyubushin A.A.
Published: February 2004

Analysis of the long-term dynamics of World Fuel Consumption (WFC) and the Global Temperature anomaly (dT) for the last 140 years (1961–2000) shows that unlike the monotonously and exponentially increasing WFC, the dynamics of global dT against the background of a linear, age-long trend, undergo quasi-cyclic fluctuations with about 60 a year period. No true linear correlation has taken place between the dT and WFC dynamics in the last century.

Spectral analysis of reconstructed temperature for the last 1420 years and instrumentally measured for the last 140 years global dT shows that dominant period for its variations for the last 1000 years lies in the 5060 years interval.

Modelling of roughly 60–years cyclic dT changes suggest that the observed rise of dT will flatten in the next 5–10 years, and that we might expect a lowering of dT by nearly 1–0.15°C to the end of the 2020s.