bluuebunny/crossref_metadata_2025 huggingface dataset:
Long PDE4 cAMP specific phosphodiesterases are activated by protein kinase A‐mediated phosphorylation of a single serine residue in Upstream Conserved Region 1 (UCR1)
Authors: MacKenzie Simon J , Baillie George S , McPhee Ian , MacKenzie Carolynn , Seamons Rachael , McSorley Theresa , Millen Jenni , Beard Matthew B , van Heeke Gino , Houslay Miles D
Published: January 2005

Challenge of COS1 cells with the adenylyl cyclase activator forskolin led to the activation of recombinant PDE4A8, PDE4B1, PDE4C2 and PDE4D5 cAMP‐specific phosphodiesterase long isoforms.

Forskolin challenge did not activate mutant long PDE4 isoforms where the serine target residue (STR) within the protein kinase A (PKA) consensus phosphorylation site in Upstream Conserved Region 1 (UCR1) was mutated to alanine.

The PKA inhibitor, H89, ablated forskolin activation of wild‐type long PDE4 isoforms.

Activated PKA caused the in vitro phosphorylation of recombinant wild‐type long PDE4 isoforms, but not those where the STR was mutated to alanine.

An antiserum specific for the phosphorylated form of the STR detected a single immunoreactive band for recombinant long PDE4 isoforms expressed in COS1 cells challenged with forskolin. This was not evident in forskolin‐challenged cells treated with H89. Neither was it evident in forskolin‐challenged cells expressing long isoforms where the STR had been mutated to alanine.

In transfected COS cells challenged with forskolin, only the phosphorylated PDE4D3 long form showed a decrease in mobility in Western blotting analysis. This decreased mobility of PDE4D3 was ablated upon mutation of either of the two serine targets for PKA phosphorylation in this isoform, namely Ser54 in UCR1 and Ser13 in the isoform‐specific N‐terminal region.

Activation by forskolin challenge did not markedly alter the sensitivity of PDE4A8, PDE4B1, PDE4C2 and PDE4D5 to inhibition by rolipram.

Long PDE4 isoforms from all four sub‐families can be phosphorylated by protein kinase A (PKA). This leads to an increase in their activity and may thus contribute to cellular desensitization processes in cells where these isoforms are selectively expressed.

British Journal of Pharmacology (2002) 136, 421–433; doi:10.1038/sj.bjp.0704743
Dual effect mediated by protease‐activated receptors on the mechanical activity of rat colon
Authors: Mulè Flavia , Baffi Maria Carmela , Cerra Maria Carmela
Published: January 2005

The present study examined the mechanical effects of agonist enzymes and receptor‐activating peptides for protease‐activated receptor (PAR)‐1 and PAR‐2 on longitudinal and circular muscle of rat isolated colonic segments in the attempt to clarify the PAR functional role in intestinal motility.

The responses to PAR‐1 and PAR‐2 activation were examined in vitro by recording simultaneously the changes of endoluminal pressure (index of circular muscle activity) and of isometric tension (index of longitudinal muscle activity).

Both PAR‐1 agonists, thrombin (0.1 nM – 3 μM) and SFLLRN‐NH2 (1 nM – 3 μM), and PAR‐2 agonists, trypsin (0.1 nM – 10 μM) and SLIGRL‐NH2 (1 nM – 10 μM), induced different effects in the two muscular layers: a reduction of the spontaneous contractions in the circular muscle and a contractile effect or biphasic, relaxation followed by contraction, depending on the concentration, in the longitudinal muscle.

The inhibitory effects were greatly reduced or abolished by apamin (0.1 μM) indicating that they mainly occur via activation of Ca2+‐dependent small conductance, K+‐channels.

The responses to PAR‐1 and PAR‐2 were unaffected by tetrodotoxin (1 μM) or indomethacin (1 μM) suggesting that are independent by products of cyclooxygenase or by neural action potentials.

These findings indicate that both PAR‐1 and PAR‐2 are functionally expressed in rat colon. PARs mediate changes of the mechanical activity of longitudinal and circular muscle which might explain the alterations of colonic motility observed during inflammatory conditions.

British Journal of Pharmacology (2002) 136, 367–374; doi:10.1038/sj.bjp.0704746
Sirolimus/cyclosporine/tacrolimus interactions on bile flow and biliary excretion of immunosuppressants in a subchronic bile fistula rat model
Authors: Deters Michael , Klabunde Til , Kirchner Gabriele , Resch Klaus , Kaever Volkhard
Published: January 2005

The new immunosuppressive agent sirolimus generally is combined in transplant patients with cyclosporine and tacrolimus which both exhibit cholestatic effects. Nothing is known about possible cholestatic effects of these combinations which might be important for biliary excretion of endogenous compounds as well as of immunosuppressants.

Rats were daily treated with sirolimus (1 mg kg−1 p.o.), cyclosporine (10 mg kg−1 i.p.), tacrolimus (1 mg kg−1 i.p.), or a combination of sirolimus with cyclosporine or tacrolimus. After 14 days a bile fistula was installed to investigate the effects of the immunosuppressants and their combinations on bile flow and on biliary excretion of bile salts, cholesterol, and immunosuppressants.

Cyclosporine as well as tacrolimus reduced bile flow (−22%; −18%), biliary excretion of bile salts (−15%;−36%) and cholesterol (−15%; −47%). Sirolimus decreased bile flow by 10%, but had no effect on cholesterol or bile salt excretion.

Combination of sirolimus/cyclosporine decreased bile flow and biliary bile salt excretion to the same extent as cyclosporine alone, but led to a 2 fold increase of biliary cholesterol excretion. Combination of sirolimus/tacrolimus reduced bile flow only by 7.5% and did not change biliary bile salt and cholesterol excretion.

Sirolimus enhanced blood concentrations of cyclosporine (+40%) and tacrolimus (+57%). Sirolimus blood concentration was increased by cyclosporine (+400%), but was not affected by tacrolimus.

We conclude that a combination of sirolimus/tacrolimus could be the better alternative to the cotreatment of sirolimus/cyclosporine in cholestatic patients and in those facing difficulties in reaching therapeutic ranges of sirolimus blood concentration.

British Journal of Pharmacology (2002) 136, 604–612; doi:10.1038/sj.bjp.0704756
Airway function, oedema, cell infiltration and nitric oxide generation in conscious ozone‐exposed guinea‐pigs: effects of dexamethasone and rolipram
Authors: Toward Toby J , Broadley Kenneth J
Published: January 2005

The effects of ozone inhalation (90 min, 2.15±0.05 p.p.m.) and their modification by dexamethasone (20 mg kg−1) or the phosphodiesterase‐4 inhibitor, rolipram (1 mg kg−1), administered (i.p.) 24 and 0.5 h before and 24 h after ozone exposure were examined in conscious guinea‐pigs.

Ozone caused an early‐phase bronchoconstriction (EPB) as a fall in specific airways conductance (sGaw) measured by whole body plethysmography, followed at 5 h by a late‐phase bronchoconstriction (LPB) and increased respiratory rate. Rolipram did not alter this profile but dexamethasone inhibited the EPB.

Airway hyperreactivity to inhaled histamine (1 mM, 20 s) occurred at 0.5, 2, 12, 24 and 48 h after ozone inhalation, the 2 h change being abolished by rolipram and dexamethasone.

Bronchoalveolar lavage fluid (BALF) macrophages, eosinophils and neutrophils were significantly (P<0.05) elevated at 12, 24 and 48 h after ozone exposure, the 48 h influx being significantly attenuated (P<0.05) by rolipram and dexamethasone.

BALF nitric oxide (NO) metabolites decreased 0.5 h after ozone exposure by 52%, recovered at 2 h and significantly increased at 12 (101%) and 24 h (127%). The elevated NO was unaffected by rolipram or dexamethasone.

Lung oedema, measured from wet/dry weight differences, was significant 12, 24 and 48 h after ozone exposure, the latter being significantly attenuated (P<0.05) by rolipram and dexamethasone.

Ozone exposure of guinea‐pigs produced features common to COPD. Although rolipram and dexamethasone did not affect the airway function changes, they inhibited the inflammation, airway hyperreactivity and oedema.

British Journal of Pharmacology (2002) 136, 735–745; doi:10.1038/sj.bjp.0704764
Endocannabinoid levels in rat limbic forebrain and hypothalamus in relation to fasting, feeding and satiation: stimulation of eating by 2‐arachidonoyl glycerol
Authors: Kirkham Tim C , Williams Claire M , Fezza Filomena , Marzo Vincenzo Di
Published: January 2005

Endocannabinoids are implicated in appetite and body weight regulation. In rodents, anandamide stimulates eating by actions at central CB1 receptors, and hypothalamic endocannabinoids may be under the negative control of leptin. However, changes to brain endocannabinoid levels in direct relation to feeding or changing nutritional status have not been investigated.

We measured anandamide and 2‐arachidonoyl glycerol (2‐AG) levels in feeding‐associated brain regions of rats, during fasting, feeding of a palatable food, or after satiation. Endocannabinoid levels were compared to those in rats fed ad libitum, at a point in their daily cycle when motivation to eat was absent. Fasting increased levels of anandamide and 2‐AG in the limbic forebrain and, to a lesser extent, of 2‐AG in the hypothalamus. By contrast, hypothalamic 2‐AG declined as animals ate. No changes were detected in satiated rats. Endocannabinoid levels in the cerebellum, a control region not directly involved in the control of food intake, were unaffected by any manipulation.

As 2‐AG was most sensitive to variation during feeding, and to leptin regulation in a previous study, we examined the behavioural effects of 2‐AG when injected into the nucleus accumbens shell, a limbic forebrain area strongly linked to eating motivation. 2‐AG potently, and dose‐dependently, stimulated feeding. This effect was attenuated by the CB1 receptor antagonist SR141716.

These findings provide the first direct evidence of altered brain levels of endocannabinoids, and of 2‐AG in particular, during fasting and feeding. The nature of these effects supports a role for endocannabinoids in the control of appetitive motivation.

British Journal of Pharmacology (2002) 136, 550–557; doi:10.1038/sj.bjp.0704767
Effects of sildenafil on erectile activity in mice lacking neuronal or endothelial nitric oxide synthase
Authors: Cashen Doreen E , MacIntyre D Euan , Martin William J
Published: January 2005

Using an in vivo model of erectile activity, the effects of sildenafil were studied in mice lacking neuronal or endothelial nitric oxide synthase (nNOS and eNOS, respectively).

Under pentobarbitone anaesthesia, intracavernous pressure (ICP) and mean arterial pressure (MAP) were monitored continuously in wild‐type, nNOS−/− and eNOS−/− mice. The magnitude of erectile activity was quantified as the ratio of ICP to MAP.

No differences in basal ICP or MAP were observed amongst wild‐type, eNOS−/− and nNOS−/− mice. Electrical stimulation of the cavernous nerve (ESCN; 4.0 V, 16 Hz, 1 ms, 30 s) evoked increases in ICP and ICP/MAP as well as penile tumescence. Responses to ESCN were reduced in nNOS−/−, but not in eNOS−/− mice.

L‐NAME (50 mg kg−1, i.v.) significantly increased MAP and attenuated erectile responses in both wild‐type and eNOS−/− mice.

Sildenafil (1 mg kg−1, i.v.) augmented electrically‐evoked erectile activity in a voltage‐dependent manner in wild‐type mice and facilitated erectile responses in eNOS−/− mice. By contrast, sildenafil failed to augment the diminished erectile responses in mice lacking the nNOS isoform.

These data reveal the relative importance of nNOS, compared to eNOS, as the critical NOS isoform in the control of erectile function and illustrate that the nNOS isoform is required for sildenafil‐induced facilitation of erectile responses in vivo in mice.

British Journal of Pharmacology (2002) 136, 693–700; doi:10.1038/sj.bjp.0704772
Enhancement of apomorphine‐induced penile erection in the rat by a selective α1D‐adrenoceptor antagonist
Authors: Mizusawa Hiroya , Hedlund Petter , Sjunnesson Johan , Brioni Jorge D , Sullivan James P , Andersson Karl‐Erik
Published: January 2005

Effects of A‐322312 (α1B‐adrenoceptor (AR) antagonist), A‐119637 (α1D‐AR antagonist), prazosin (non‐selective α1‐AR antagonist), and yohimbine (α2‐AR antagonist) were studied in rat corpus cavernosum (CC) and cavernous artery (Acc) preparations. Effects of intracavernous (i.c.) or intraperitoneal (i.p.) administration of α1‐AR antagonists on apomorphine‐induced erections were investigated.

A‐119637 attenuated electrically induced contractions in isolated CC (−logIC50; 8.12±0.15), and relaxed noradrenaline (NA)‐contracted preparations by more than 90% at 10−7 M. At the same concentration, the −logEC50 value for NA in Acc was altered from 6.79±0.07 to 4.86±0.13. In the CC and Acc, prazosin similarily inhibited contractile responses.

Inhibitory effects of A‐322312 (10−7 M) in electrically activated CC were 32.3±5.1%, whereas no effect on concentration‐response curves for NA was observed in the Acc. Yohimbine (10−8 M and 10−7 M), enhanced electrically‐induced contractions in isolated CC by 20 to 50%. At 10−6 M, inhibitory effects of yohimbine were obtained.

A‐119637 (0.3 μmol kg−1, i.p.) tripled the number of erections, and produced a 6 fold increase in the duration of apomorphine‐induced erectile responses. A‐322312, prazosin, or yohimbine did not enhance erections induced by apomorphine. None of the α1‐AR antagonists significantly increased ICP upon i.c. administration. Decreases in blood pressure were seen with A‐119637 and prazosin.

The present findings show that there is a functional predominance of the α1D‐AR subtype in the rat erectile tissue, and that blockade of this receptor facilitates rat penile erection induced by a suboptimal dose of apomorphine.

British Journal of Pharmacology (2002) 136, 701–708; doi:10.1038/sj.bjp.0704773
The extracellular regulated kinases (ERK) 1/2 mediate cannabinoid‐induced inhibition of gap junctional communication in endothelial cells
Authors: Brandes R P , Popp R , Ott G , Bredenkötter D , Wallner C , Busse R , Fleming I
Published: January 2005

Cannabinoids are potent inhibitors of endothelium‐derived hyperpolarizing factor (EDHF)‐mediated relaxations. We set out to study the mechanism underlying this effect and the possible role of cannabinoid‐induced changes in intercellular gap junction communication.

In cultured endothelial cells, Δ9‐tetrahydrocannabinol (Δ9‐THC) and the cannabinoid receptor agonist HU210, increased the phosphorylation of extracellular regulated kinases 1/2 (ERK1/2) and inhibited gap junctional communication, as determined by Lucifer Yellow dye transfer and electrical capacity measurements.

Δ9‐THC elicited a pronounced increase in the phosphorylation of connexin 43, which was sensitive to PD98059 and U0126, two inhibitors of ERK1/2 activation. Inhibition of ERK1/2 also prevented the Δ9‐THC‐induced inhibition of gap junctional communication.

Δ9‐THC prevented both the bradykinin‐induced hyperpolarization and the nitric oxide and prostacyclin‐independent relaxation of pre‐contracted rings of porcine coronary artery. These effects were prevented by PD98059 as well as U0126.

In the absence of Δ9‐THC, neither PD98059 nor U0126 affected the NO‐mediated relaxation of coronary artery rings but both substances induced a leftward shift in the concentration – relaxation curve to bradykinin when diclofenac and Nωnitro‐L‐arginine were present. Moreover, PD98059 and U0126 prolonged the bradykinin‐induced hyperpolarization of porcine coronary arteries, without affecting the magnitude of the response.

These results indicate that the cannabinoid‐induced activation of ERK1/2, which leads to the phosphorylation of connexin 43 and inhibition of gap junctional communication, may partially account for the Δ9‐THC‐induced inhibition of EDHF‐mediated relaxation. Moreover, the activation of ERK1/2 by endothelial cell agonists such as bradykinin, appears to exert a negative feedback inhibition on EDHF‐mediated responses.

British Journal of Pharmacology (2002) 136, 709–716; doi:10.1038/sj.bjp.0704776
YC‐1 increases cyclo‐oxygenase‐2 expression through protein kinase G‐ and p44/42 mitogen‐activated protein kinase‐dependent pathways in A549 cells
Authors: Chang Ming‐Shyan , Lee Wen‐Sen , Teng Che‐Ming , Lee Horng‐Mo , Sheu Joen‐Rong , Hsiao George , Lin Chien‐Huang
Published: January 2005

YC‐1, an activator of soluble guanylate cyclase (sGC), has been shown to increase the intracellular cGMP concentration. This study was designed to investigate the signaling pathway involved in the YC‐1‐induced COX‐2 expression in A549 cells.

YC‐1 caused a concentration‐ and time‐dependent increase in COX activity and COX‐2 expression in A549 cells. Pretreatment of the cells with the sGC inhibitor (ODQ), the protein kinase G (PKG) inhibitor (KT‐5823), and the PKC inhibitors (Go 6976 and GF10923X), attenuated the YC‐1‐induced increase in COX activity and COX‐2 expression.

Exposure of A549 cells to YC‐1 caused an increase in PKC activity; this effect was inhibited by ODQ, KT‐5823 or Go 6976. Western blot analyses showed that PKC‐α, ‐ι, ‐λ, ‐ζ and ‐μ isoforms were detected in A549 cells. Treatment of A549 cells with YC‐1 or PMA caused a translocation of PKC‐α, but not other isoforms, from the cytosol to the membrane fraction. Long‐term (24 h) treatment of A549 cells with PMA down‐regulated the PKC‐α.

The MEK inhibitor, PD 98059 (10–50 μM), concentration‐dependently attenuated the YC‐1‐induced increases in COX activity and COX‐2 expression. Treatment of A549 cells with YC‐1 caused an activation of p44/42 MAPK; this effect was inhibited by KT‐5823, Go 6976, long‐term (24 h) PMA treatment or PD98059, but not the p38 MAPK inhibitor, SB 203580.

These results indicate that in human pulmonary epithelial cells, YC‐1 might activate PKG through an upstream sGC/cGMP pathway to elicit PKC‐α activation, which in turn, initiates p44/42 MAPK activation, and finally induces COX‐2 expression.

British Journal of Pharmacology (2002) 136, 558–567; doi:10.1038/sj.bjp.0704777
NO‐ and haem‐independent activation of soluble guanylyl cyclase: molecular basis and cardiovascular implications of a new pharmacological principle
Authors: Stasch Johannes‐Peter , Schmidt Peter , Alonso‐Alija Cristina , Apeler Heiner , Dembowsky Klaus , Haerter Michael , Heil Markus , Minuth Torsten , Perzborn Elisabeth , Pleiss Ulrich , Schramm Matthias , Schroeder Werner , Schröder Henning , Stahl Elke , Steinke Wolfram , Wunder Frank
Published: January 2005

Soluble guanylyl cyclase (sGC) is the only proven receptor for the ubiquitous biological messenger nitric oxide (NO) and is intimately involved in many signal transduction pathways, most notably in regulating vascular tone and platelet function. sGC is a heterodimeric (α/ß) protein that converts GTP to cyclic GMP; NO binds to its prosthetic haem group. Here, we report the discovery of a novel sGC activating compound, its interaction with a previously unrecognized regulatory site and its therapeutic implications.

Through a high‐throughput screen we identified BAY 58‐2667, an amino dicarboxylic acid which potently activates sGC in an NO‐independent manner. In contrast to NO, YC‐1 and BAY 41‐2272, the sGC stimulators described recently, BAY 58‐2667 activates the enzyme even after it has been oxidized by the sGC inhibitor ODQ or rendered haem deficient.

Binding studies with radiolabelled BAY 58‐2667 show a high affinity site on the enzyme.

Using photoaffinity labelling studies we identified the amino acids 371 (α‐subunit) and 231 – 310 (ß‐subunit) as target regions for BAY 58‐2667.

sGC activation by BAY 58‐2667 results in an antiplatelet activity both in vitro and in vivo and a potent vasorelaxation which is not influenced by nitrate tolerance.

BAY 58‐2667 shows a potent antihypertensive effect in conscious spontaneously hypertensive rats. In anaesthetized dogs the hemodynamic effects of BAY 58‐2667 and GTN are very similar on the arterial and venous system.

This novel type of sGC activator is a valuable research tool and may offer a new approach for treating cardiovascular diseases.

British Journal of Pharmacology (2002) 136, 773–783; doi:10.1038/sj.bjp.0704778
Mg2+ sensitizes KATP channels to inhibition by DIDS: dependence on the sulphonylurea receptor subunit
Authors: Gojkovic‐Bukarica Ljiljana , Hambrock Annette , Löffler‐Walz Cornelia , Quast Ulrich , Russ Ulrich
Published: January 2005

ATP‐sensitive potassium channels (KATP channels) consist of pore‐forming Kir6.x subunits and of sulphonylurea receptors (SURs). In the absence of Mg2+, the stilbene disulphonate, DIDS, irreversibly inhibits KATP channels by binding to the Kir subunit. Here, the effects of Mg2+ on the interaction of DIDS with recombinant KATP channels were studied in electrophysiological and [3H]‐glibenclamide binding experiments.

In inside‐out macropatches, Mg2+ (0.7 mM) increased the sensitivity of KATP channels towards DIDS up to 70 fold (IC50=2.7 μM for Kir6.2/SUR2B). Inhibition of current at DIDS concentrations 10 μM was irreversible.

Mg2+ sensitized the truncated Kir6.2Δ26 channel towards inhibition by DIDS only upon coexpression with a SUR subunit (SUR2B). The effect of Mg2+ did not require the presence of nucleotides.

[3H]‐glibenclamide binding to SUR2B(Y1206S), a mutant with improved affinity for glibenclamide, was inhibited by DIDS. The potency of inhibition was increased by Mg2+ and by coexpression with Kir6.2.

In the presence of Mg2+, DIDS inhibited binding of [3H]‐glibenclamide to Kir6.2/SUR2B(Y1206S) with IC50=7.9 μM by a non‐competitive mechanism. Inhibition was fully reversible.

It is concluded that the binding site of DIDS on SUR that is sensed by glibenclamide does not mediate channel inhibition. Instead, Mg2+ binding to SUR may allosterically increase the accessibility and/or reactivity of the DIDS site on Kir6.2. The fact that the Mg2+ effect does not require the presence of nucleotides underlines the importance of this ion in modulating the properties of the KATP channel.

British Journal of Pharmacology (2002) 137, 429–440. doi:10.1038/sj.bjp.0704905
Speech recognition with amplitude and frequency modulations
Authors: Zeng Fan-Gang , Nie Kaibao , Stickney Ginger S. , Kong Ying-Yee , Vongphoe Michael , Bhargave Ashish , Wei Chaogang , Cao Keli
Published: January 2005

Amplitude modulation (AM) and frequency modulation (FM) are commonly used in communication, but their relative contributions to speech recognition have not been fully explored. To bridge this gap, we derived slowly varying AM and FM from speech sounds and conducted listening tests using stimuli with different modulations in normal-hearing and cochlear-implant subjects. We found that although AM from a limited number of spectral bands may be sufficient for speech recognition in quiet, FM significantly enhances speech recognition in noise, as well as speaker and tone recognition. Additional speech reception threshold measures revealed that FM is particularly critical for speech recognition with a competing voice and is independent of spectral resolution and similarity. These results suggest that AM and FM provide independent yet complementary contributions to support robust speech recognition under realistic listening situations. Encoding FM may improve auditory scene analysis, cochlear-implant, and audiocoding performance.
NMR screening and crystal quality of bacterially expressed prokaryotic and eukaryotic proteins in a structural genomics pipeline
Authors: Page Rebecca , Peti Wolfgang , Wilson Ian A. , Stevens Raymond C. , Wüthrich Kurt
Published: January 2005

In the Joint Center for Structural Genomics, one-dimensional (1D)
1
H NMR spectroscopy is routinely used to characterize the folded state of protein targets and, thus, serves to guide subsequent crystallization efforts and to identify proteins for NMR structure determination. Here, we describe 1D
1
H NMR screening of a group of 79 mouse homologue proteins, which correlates the NMR data with the outcome of subsequent crystallization experiments and crystallographic structure determination. Based on the 1D
1
H NMR spectra, the proteins are classified into four groups, “A” to “D.” A-type proteins are candidates for structure determination by NMR or crystallography; “B”-type are earmarked for crystallography; “C” indicates folded globular proteins with broadened line shapes; and “D” are nonglobular, “unfolded” polypeptides. The results obtained from coarse- and fine-screen crystallization trials imply that only A- and B-type proteins should be used for extensive crystallization trials in the future, with C and D proteins subjected only to coarse-screen crystallization trials. Of the presently studied 79 soluble protein targets, 63% yielded A- or B-quality 1D
1
H NMR spectra. Although similar yields of crystallization hits were obtained for all four groups, A to D, crystals from A- and B-type proteins diffracted on average to significantly higher resolution than crystals produced from C- or D-type proteins. Furthermore, the output of refined crystal structures from this test set of proteins was 4-fold higher for A- and B-type than for C- and D-type proteins.
Three GnRH receptor types in laser-captured single cells of the cichlid pituitary display cellular and functional heterogeneity
Authors: Parhar Ishwar S. , Ogawa Satoshi , Sakuma Yasuo
Published: January 2005

The role of multiple gonadotropin-releasing hormone receptor (GnRH-R) types in the regulation of gonadotropic and nongonadotropic cells remains speculative. To address this issue, we developed a technology integrating laser-captured microdissection of single digoxigenin-labeled pituitary cells coupled with real-time quantitative PCR to examine the expression profiles of three endogenous GnRH-R types (R1, R2, and R3) in immature and mature males of tilapia
Oreochromis niloticus.
Here, in addition to gonadotropes (luteinizing and folicle-stimulating hormone, FSH), we show GnRH-Rs are also present in lactotropes, somatotropes, thyrotropes, melanotropes (melanocyte-stimulating hormone, MSH), corticotropes and somatolactin cells. Subpopulations of pituitary cells express single (42.9%), multiple (32.4%) or lack (24.7%) GnRH-Rs. In immature males, the percentage of FSH cells containing combinations of GnRH-Rs was significantly higher (R1+R2: 24%,
P
< 0.05; R1+R2+R3: 25%,
P
< 0.01) than in mature males, whereas the percentage showing only R1 and R1 and R3 transcripts (
P
< 0.05) was higher in mature males. Significantly greater copies of R1 and R3 transcripts were found in MSH cells of immature and mature males, respectively (
P
< 0.05). GnRH-R transcripts in other pituitary cells (lactotropes, R1 and R2; somatolactin cells/thyrotropes/corticotropes, R1, R2, and R3) were significantly higher in mature males (
P
< 0.05) but were unaltered in somatotropes and luteinizing hormone cells. Thus, FSH and MSH cells are required for both reproductive states, whereas other pituitary cells are recruited only during testicular maturation. The differential expression of GnRH-Rs in gonadotropic and nongonadotropic cells demonstrates cellular and functional heterogeneity of mechanisms controlling normal sexual development.
Population dynamics and potential of fisheries stock enhancement: practical theory for assessment and policy analysis
Authors: Lorenzen Kai
Published: January 2005

The population dynamics of fisheries stock enhancement, and its potential for generating benefits over and above those obtainable from optimal exploitation of wild stocks alone are poorly understood and highly controversial. I review pertinent knowledge of fish population biology, and extend the dynamic pool theory of fishing to stock enhancement by unpacking recruitment, incorporating regulation in the recruited stock, and accounting for biological differences between wild and hatchery fish. I then analyse the dynamics of stock enhancement and its potential role in fisheries management, using the candidate stock of North Sea sole as an example and considering economic as well as biological criteria. Enhancement through release of recruits or advanced juveniles is predicted to increase total yield and stock abundance, but reduce abundance of the naturally recruited stock component through compensatory responses or overfishing. Economic feasibility of enhancement is subject to strong constraints, including trade–offs between the costs of fishing and hatchery releases. Costs of hatchery fish strongly influence optimal policy, which may range from no enhancement at high cost to high levels of stocking and fishing effort at low cost. Release of genetically maladapted fish reduces the effectiveness of enhancement, and is most detrimental overall if fitness of hatchery fish is only moderately compromised. As a temporary measure for the rebuilding of depleted stocks, enhancement cannot substitute for effort limitation, and is advantageous as an auxiliary measure only if the population has been reduced to a very low proportion of its unexploited biomass. Quantitative analysis of population dynamics is central to the responsible use of stock enhancement in fisheries management, and the necessary tools are available.
Ultrastructure of spermiogenesis and the spermatozoon in Castrada cristatispina Papi, 1951 (Platyhelminthes, Rhabdocoela, Typhloplanida)
Authors: Culioli Julia‐Laurence , Foata Joséphine , Mori Christophe , Orsini Antoine , Marchand Bernard
Published: January 2005

Spermiogenesis in Castrada cristatispina begins with the formation of a zone of differentiation containing two centrioles with associated striated rootlets and an intercentriolar body between them. The centrioles give rise to two parallel, free flagella of the Trepaxonemata 9 + ‘1’ pattern, growing out in opposite directions. Spermatids undergo a latero‐ventral rotation of the flagella and a subsequent disto‐proximal rotation of centrioles, and a distal cytoplasmic projection appears. The former rotation involves the compression of a row of microtubules and allows the recognition of a ventral side and a dorsal side. At the end of the differentiation, the centrioles and cortical microtubules lie parallel to the sperm axis. The modifications of the intercentriolar body and the migration of the nucleus and the centrioles toward the distal projection are described. The mature spermatozoon of C. cristatispina is filiform, tapered at both ends and shares several features with the other Rhabdocoela gametes. Nevertheless, the posterior extremity is capped by an electron‐dense material. A gradient between mitochondria and dense bodies exists along the sperm axis. This study has enable us a phylogenetic approach of the Rhabdocoela through a comparison of the ultrastructural features of C. cristatispina with the other Rhabdocoela taxa. We propose the disto‐proximal rotation of centrioles as a synapomorphy of the Rhabdocoela.
Comparative analysis of the sperm ultrastructure of three species of Phyllomedusa (Anura, Hylidae)
Authors: Costa G. C. , Garda A. A. , Teixeira R. D. , Colli G. R. , Báo S. N.
Published: January 2005

We describe the sperm ultrastructure of three species of frogs in the genus Phyllomedusa. According to micrographs, total size of the spermatozoon of Phyllomedusa hypochondrialis is significantly smaller than those of Phyllomedusa bicolor and Phyllomedusa tarsius. The acrosome complex consists of two conical structures covering the nucleus, the acrosome vesicle and the subacrosomal cone. The subacrosomal cone of P. bicolor and P. tarsius is less electron‐dense and appears more granular in transverse section than in P. hypochondrialis. In P. bicolor and P. tarsius, the nuclear space is reduced and the subacrosomal cone fills most of the space between the acrosome vesicle and nucleus. The anterior region of the nucleus in the spermatozoa of P. bicolor and P. tarsius ends abruptly, while in P. hypochondrialis it is sharp‐ended. In P. bicolor and P. tarsius, the axial fibre is much larger than in P. hypochondrialis. The sperm ultrastructure of Phyllomedusa appears conservative at the intrageneric level. Future studies on the sperm ultrastructure of hylids can provide new insights on the systematics of the group and a larger database for a cladistic analysis.
A conversation with Dr Michael Fordham
Authors: Astor James
Published: January 2005

Abstract:  In this interview1 in Dr Fordham's 83rd year he describes how he started to work with children, and how Mrs Jung was supportive. He talks about the initial suspicion this interest generated in the wider Jungian community. He refers to his acceptance of and interest in the psychotic elements in child analysis and his transference‐based approach to working with these elements. He reflects on his own birth, his work with evacuee children in hostels during the war years and the politics of supervision. He describes the core Jungian concepts which underpinned his work and the theoretical differences from the Kleinian and Anna Freudian positions.
An interview with Michael Fordham
Authors: Roazen Paul
Published: January 2005

Abstract:  In the course of early interviews on the history of psychoanalysis, I saw Michael Fordham in the late summer of 1965. We concentrated primarily on the differences between Freud and Jung, as well as the characteristic distinctions between the two schools that they founded. Fordham also talked about some of his personal contacts with Jung.
‘Do be my enemy for friendship's sake’ (Blake)
Authors: Gordon Montagnon Rosemary
Published: January 2005

Abstract:  The author explores our psychological need for enemies and the contribution this makes to overt conflicts in the external world. Enemies serve as an opposite from which we can differentiate ourselves, either as an individual or as a group; they help us to define our physical and psychological boundaries. Enemies provide a target and an outlet for our aggression and also for the projection of the shadow. They also provide the stimulus to individuation, through the heroic encounter with the enemy in the unfamiliar world outside the home, particularly in adolescence. The psychic integration of ‘self’ and ‘enemy’ is explored as the outcome of individuation.
2. Freud's ‘id’ and Jung's ‘self’ as aids in self‐analysis
Authors: Plaut Alfred B. J.
Published: January 2005

Abstract:  The id and the self are described as constructs of our unconscious which can be deployed in describing the analytic process. They can be used like the x in mathematics or the joker in a pack of cards as fitting in almost anywhere. But we can call on them as important aids to concentration, when additional room is made for meditating on the past which then comes to life again. They are useful for analysing ourselves and others. In this way the id as well as the self become aide‐mémoires. The analysis of the repressed by means of the transference is not the only road to hitherto unconscious memories.

This kind of meditation has helped me despite all reservations to undertake self‐analysis and to write an autobiography. I maintain that one can have a transference to oneself. It is, of course, narcissistic. But then, no one can write an autobiography without a healthy dose of narcissism.
Finding our way in the dark
Authors: Beebe John
Published: January 2005

Abstract:  Accepting the common lay definition of nightmare as any form of upsetting, dreamlike cognition occurring in the midst of sleep, the author argues for a classification of nightmares that would be based, not on physiological findings, but on what the upsetting nocturnal occurrence turns out to mean to the dreamer. Three types of nightmare identified on this basis are illustrated through dream sequences from classic Hollywood movies and amplified by clinical examples: (1) the dream that symbolizes the next stage of life as unusually daunting, (2) the dream that exposes the shadow of another person in a shocking way, and (3) the ‘empathy dream’, in which the dreamer experiences directly the anxieties of another subject. It is suggested that the accurate interpretation of a particular upsetting dream depends upon which type of nightmare the dream turns out to be.
Development of the pelvis and posterior part of the vertebral column in the Anura
Authors: Ročková Hana , Roček Zbyněk
Published: January 2005

The anuran pelvic girdle is unique among all amphibians in that its acetabular portion is located far posterior to the sacrum, lateral to the postsacral (= caudal) vertebral column, which is reduced to a single rod‐like element called the urostyle. This situation in the adult is strikingly different not only from that in ancestral temnospondyls but also in other modern amphibians. Because there is no fossil that would document this evolutionary anatomical modification except for Triadobatrachus, the only data may be inferred from development in modern anurans. We chose seven anuran species (belonging to the genera Discoglossus, Bombina, Pelobates, Bufo, Rana and Xenopus), representing the principal locomotory types (saltation, swimming, crawling and burrowing). Development of the pelvic girdle was studied on cleared and stained whole mounts and partly on serial histological sections. The basic developmental pattern was similar in all species: the pelvis on both sides develops from two centres (puboischiadic and iliac, respectively). The ilium then extends vertically towards the sacral vertebra and later rotates posteriorly so that ultimately the acetabulum is lateral to the tail (= urostyle). Only minor deviations from this pattern were found, mainly associated with differences in water and terrestrial dwelling.
Traversing the intact/fibrillated joint surface: a biomechanical interpretation
Authors: Broom Neil D. , Ngo Thuy , Tham Evelyn
Published: January 2005

Cartilage taken from the osteoarthritic bovine patellae was used to investigate the progression of change in the collagenous architecture associated with the development of fibrillated lesions. Differential interference contrast optical microscopy using fully hydrated radial sections revealed a continuity in the alteration of the fibrillar architecture in the general matrix consistent with the progressive destructuring of a native radial arrangement of fibrils repeatedly interconnected in the transverse direction via a non‐entwinement‐based linking mechanism. This destructuring is shown to occur in the still intact regions adjacent to the disrupted lesion thus rendering them more vulnerable to radial rupture. Two contrasting modes of surface rupture were observed and these are explained in terms of the absence or presence of a skewed structural weakening of the intermediate zone. A mechanism of surface rupture initiation based on simple bi‐layer theory is proposed to account for the intensification of surface ruptures observed in the intact regions on advancing towards the fibrillation front. Focusing specifically on the primary collagen architecture in the cartilage matrix, this study proposes a pathway of change from intact to overt disruption within a unified structural framework.
Mosquito transmission modulates the immune response in mice infected with the L3 of Brugia pahangi
Authors: Gillan Victoria , Devaney Eileen
Published: January 2005

Mice infected by syringe inoculation with the L3 of the filarial nematode Brugia pahangi generate a strong Th2 response. In this study we compared immune responses in mice infected via syringe with those infected by mosquito transmission of L3. Levels of antigen‐specific IL‐4, IL‐5 and IL‐10 were significantly reduced in mice infected via mosquito. A possible explanation of these results was that mice infected via mosquito received fewer L3 than those infected via syringe. To investigate this possibility, mice were infected with different numbers of L3 (50, 25 or 10). Howere there was no difference in responses in these animals, suggesting that the reduced immune reactivity in mice infected by mosquito cannot be solely ascribed to exposure to lower numbers of parasites. These results also demonstrate that the L3 is an extremely potent stimulus for Th2 differentiation, with 10 L3 sufficient to drive a strong Th2 response. The differences in immune reactivity between syringe and mosquito infected mice may relate to the presence of immuno‐suppressive factors in mosquito saliva inoculated at the time of transmission or may reflect the interaction of L3 with different populations of antigen presenting cells in the two groups of mice. Further studies will be required to differentiate between these possibilities.
Schistosomiasis co‐infection in humans influences inflammatory markers in uncomplicated Plasmodium falciparum malaria
Authors: Diallo T. O. , Remoue F. , Schacht A. M. , Charrier N. , Dompnier J.‐P. , Pillet S. , Garraud O. , N'Diaye A. A. , Capron A. , Capron M. , Riveau G.
Published: January 2005

Malaria and schistosomiasis are the two major parasite diseases present in developing countries. The epidemiological co‐infection with schistosomiasis could influence the development of the physiological reaction associated with Plasmodium falciparum infection in human. Most studies have demonstrated the association of circulating levels of interferon‐γ (IFN‐γ), tumour necrosis factor‐a (TNF‐α), interleukin‐10 (IL‐10), transforming growth factor (TGF‐β) and soluble Tumour Necrosis Factor Receptors (sTNF‐RI and sTNF‐RII) with the morbidity of malaria. In the present study, we showed that Schistosoma haematobium co‐infection influences, in an age‐dependent manner, the unbalance between pro‐ and anti‐inflammatory circulating cytokines that play a key role during malaria infection. Indeed, children co‐infected by S. haematobium have higher levels of IFN‐γ and sTNF‐RII than children infected only by P. falciparum. In contrast, co‐infected adults presented a significant increase of IFN‐γ, IL‐10, TGF‐β, sTNF‐RI and sTNF‐RII rates and IL‐10/TNF‐α ratio. Taken together, this study indicates that schistosomiasis co‐infection can unbalance the regulation of inflammatory factors in uncomplicated P. falciparum malaria. The possible consequences of the schistosomiasis co‐infection for age‐dependent malaria morbidity are discussed.
Gene cloning, expression and vaccine testing of Schistosoma japonicum SjFABP
Authors: Liu J. M. , Cai X. Z. , Lin J. J. , Fu Z. Q. , Yang G. Z. , Shi F. H. , Cai Y. M. , Shen W. , Taylor M. G. , Wu X. F.
Published: January 2005

A 600 bp DNA fragment was amplified by PCR from an adult Schistosoma japonicum cDNA library. Sequence analysis confirmed that this fragment contained an S. japonicum Chinese mainland strain fatty acid binding protein (Sj14FABP) gene. This gene was subsequently expressed in Escherichia coli (E. coli) and in Baculovirus/silkworm systems. The recombinant protein from E. coli was a 41 kDa GST fusion protein (rSj14/GST), which could be purified by glutathione agarose affinity chromatography, with a yield of 25 mg/L E. coli culture. The recombinant protein from the Baculovirus/silkworm system was an 18 kDa fusion protein (rSj14/His), which could be purified by Ni‐NTA resin chromatography column with a yield of 3·5 mg per silkworm larva. Both rSj14/GST and rSj14/His could be recognized by S. japonicum‐infected mouse sera and anti‐rSj14/GST mouse sera in Western blotting. The purified recombinant protein was immunogenic in mice, rats and sheep, and 34·3%, 31·9% and 59·2% worm reductions, respectively, were obtained in vaccinated Kunming mice, Wistar rats and sheep vaccinated with Sj14/GST, compared to non‐vaccinated control groups. Worm reductions of 48·8% and 49·0% were recorded in Balb/c mice immunized with Sj14/His, compared to non‐vaccinated and BCG‐vaccinated groups, respectively. These results indicate that rSj14FABP is a promising candidate vaccine for schistosomiasis japonica, particularly as in the rat and sheep vaccination experiments, no adjuvant was used.
Increased Concentration in Banking: Megabanks and Their Implications for Deposit Insurance
Authors: Jones Kenneth D. , Nguyen Chau
Published: January 2005

During the past two decades, the U.S. banking industry has experienced an unprecedented wave of consolidation, marked by a substantial decline in the number of insured depository institutions and the emergence of banking behemoths with assets totaling in the hundreds of billions of dollars. This unparalleled concentration of assets and deposits among a handful of “megabanks” has important implications for deposit insurance. Most importantly, the Federal Deposit Insurance Corporation (FDIC) now faces a situation in which the failure of even a single megabank could overwhelm the resources immediately available to the deposit insurance system and expose both the banking industry and the government (i.e., taxpayers) to huge potential liabilities. This article highlights the current structure of the banking industry, examines the threat that this structure poses to the deposit insurance funds, and suggests possible approaches for dealing with megabanks and the increasing concentration of insured deposits.
Unintentional methadone‐related overdose death in New Mexico (USA) and implications for surveillance, 1998–2002
Authors: Shah Nina , Lathrop Sarah L. , Landen Michael G.
Published: January 2005

Aims  To determine death rates from methadone over time, to characterize methadone‐related death and to discuss public health surveillance of methadone‐related death.

Design  We analyzed medical examiner data for all unintentional drug overdose deaths in New Mexico, USA, between 1998 and 2002.

Measurements  Age‐adjusted death rates for methadone‐related death, logistic regression models for likelihood of methadone‐related death among all unintentional drug overdose deaths and bivariate comparisons within methadone‐related death.

Findings  Of 1120 drug overdose deaths during this period, there were 143 (12.8%) methadone‐related deaths; the death rate decreased over the time period, averaging 1.6 per 100 000. Of 143 methadone‐related deaths, 22.4% were due to methadone alone, 23.8% were due to methadone/prescription drugs (no illicit drugs), 50.3% were due to methadone/illicit drugs and 3.5% were due to methadone/alcohol. These groups were significantly different in demographics, health history and circumstances of death. Of 79 decedents (55.2%) with a known source of methadone, 68 obtained methadone through a physician prescription (31 for methadone maintenance treatment (MMT), 27 for managing pain and 10 had unknown reason for prescription).

Conclusions  Methadone‐related death rates and the proportion of methadone‐related death among all drug overdose deaths decreased in New Mexico from 1998 to 2002. It is important for surveillance of methadone‐related death to assess multiple drug causes, not just underlying cause. Also, methadone for pain management must be examined alongside MMT and when possible, methadone co‐intoxication should be described in the context of other drugs causing death.
Cut‐off levels for breath carbon monoxide as a marker for cigarette smoking
Authors: Javors Martin A. , Hatch John P. , Lamb Richard J.
Published: January 2005

Aims  Current clinical studies often use a breath carbon monoxide (BCO) cut‐off level of 8 parts per million (p.p.m.) or higher to identify smoking. In this study, the cut‐off level of BCO as an indicator of smoking over the past 24 hours was re‐examined.

Design  BCO and self‐reported smoking were obtained each weekday for up to 14 weeks in 213 subjects paid to deliver reduced BCO values. Analysis of 12 386 paired values for reported smoking and BCO were analyzed.

Findings  The 25% quartile, median and 75% quartile values for BCO were 1, 1 and 2 p.p.m. on non‐smoking days and 2, 5 and 12 p.p.m. on smoking days, respectively. Receiver‐operating characteristic (ROC) analysis indicated that BCO provided high diagnostic accuracy to distinguish between smoking and non‐smoking days [area under the curve (AUC) = 0.853, P < 0.0001]. The highest combined sensitivity and specificity was observed at a BCO cut‐off level of 3 p.p.m. (sensitivity = 71.5%; specificity = 84.8%). At a BCO cut‐off of 8 p.p.m. sensitivity and specificity were 40.6% and 98.2%, respectively, indicating that many smokers would be falsely classified as abstinent. Finally, the percentage of true tests (positive and negative) was highest at a BCO cut‐off of 2 p.p.m. (80.2%).

Conclusions  BCO cut‐off levels well below 8 p.p.m and as low as 2–3 p.p.m. may be more useful when it is important to maximize identification of smoking abstinence with a high degree of certainty.