bluuebunny/crossref_metadata_2025 huggingface dataset:
Human Golgi Antiapoptotic Protein Modulates Intracellular Calcium Fluxes
Authors: de Mattia Fabrizio , Gubser Caroline , van Dommelen Michiel M.T. , Visch Henk-Jan , Distelmaier Felix , Postigo Antonio , Luyten Tomas , Parys Jan B. , de Smedt Humbert , Smith Geoffey L. , Willems Peter H.G.M. , van Kuppeveld Frank J.M.
Published: June 2009

Golgi antiapoptotic protein (GAAP) is a novel regulator of cell death that is highly conserved in eukaryotes and present in some poxviruses, but its molecular mechanism is unknown. Given that alterations in intracellular Ca2+ homeostasis play an important role in determining cell sensitivity to apoptosis, we investigated if GAAP affected Ca2+ signaling. Overexpression of human (h)-GAAP suppressed staurosporine-induced, capacitative Ca2+ influx from the extracellular space. In addition, it reduced histamine-induced Ca2+ release from intracellular stores through inositol trisphosphate receptors. h-GAAP not only decreased the magnitude of the histamine-induced Ca2+ fluxes from stores to cytosol and mitochondrial matrices, but it also reduced the induction and frequency of oscillatory changes in cytosolic Ca2+. Overexpression of h-GAAP lowered the Ca2+ content of the intracellular stores and decreased the efficacy of IP3, providing possible explanations for the observed results. Opposite effects were obtained when h-GAAP was knocked down by siRNA. Thus, our data demonstrate that h-GAAP modulates intracellular Ca2+ fluxes induced by both physiological and apoptotic stimuli.
Ghrelin resistance occurs in severe heart failure and resolves after heart transplantation
Authors: Lund Lars H. , Williams Jill J. , Freda Pamela , LaManca John J. , LeJemtel Thierry H. , Mancini Donna M.
Published: June 2009

Aims

Severe heart failure (HF) is often associated with cachexia that reverses post‐heart transplantation (HTx) with frequent development of obesity. Ghrelin is a novel appetite‐stimulating hormone. The aim was to determine the role of ghrelin in regulating appetite, food intake, and body composition in HF and post‐HTx.

Methods and results

We measured serial ghrelin, hunger sensation, caloric intake, and body composition in 12 HF patients awaiting HTx, 12 patients 12.7 ± 8.6 months post‐HTx, and 7 controls. Seven of 12 HF patients were followed for longitudinal analysis post‐HTx. Body mass index was 23.1 ± 3.1 in HF and 31.5 ± 5.5 post‐HTx (P < 0.001). Heart transplantation patients had gained 18.0 ± 7.7 kg since HTx. Ghrelin area under the curve between controlled meals (control: 186 ± 39; HF: 264 ± 71; HTx: 194 ± 47 ng min/mL, P < 0.007) was higher in HF, but test meal caloric intake (control: 1185 ± 650; HF: 391 ± 103; HTx: 831 ± 309 kcal, P < 0.008) was lower in HF. The longitudinal analysis confirmed these findings.

Conclusion

Heart failure may be associated with resistance to the appetite‐stimulating effects of ghrelin, which may contribute to cachexia. Heart transplantation may be associated with resolution of ghrelin resistance, which may contribute to weight gain. These findings are preliminary and should be confirmed in larger trials.
A reanalysis of North Sea plaice spawning-stock biomass using the annual egg production method
Authors: van Damme Cindy J. G. , Bolle Loes J. , Fox Clive J. , Fossum Petter , Kraus Gerd , Munk Peter , Rohlf Norbert , Witthames Peter R. , Dickey-Collas Mark
Published: June 2009

van Damme, C. J. G., Bolle, L. J., Fox, C. J., Fossum, P., Kraus, G., Munk, P., Rohlf, N., Witthames, P. R., and Dickey-Collas, M. 2009. A reanalysis of North Sea plaice spawning-stock biomass using the annual egg production method. – ICES Journal of Marine Science, 66: 1999–2011. Uncertainty about the quality of current virtual population analysis-based stock assessment for North Sea plaice (Pleuronectes platessa) has led to various abundance indices. We compared biomass estimates from the annual egg production (AEP) method with current stock assessments based on catch-at-age to validate the current and historical perception of exploitation. The AEP method was also used to investigate the dynamics of the spatial components of plaice in the North Sea. We corrected for fecundity down-regulation and changes in sex ratio. Estimates from both methods were similar in trend and absolute biomass. On the Dogger Bank, there was a dramatic decline in biomass from 1948 and 1950 to 2004, and in the Southern Bight, the stock appeared to increase from 1987 and 1988 to 2004, although not reaching the historically high levels of 1948 or 1950. The timing of spawning of North Sea plaice does not appear to have changed throughout the period of high exploitation. We conclude that the AEP method is a useful way to hindcast the spatial dynamics of heavily exploited flatfish stocks.
The risk to fishery performance associated with spatially resolved management of Antarctic krill (Euphausia superba) harvesting
Authors: Hill Simeon L. , Trathan Philip N. , Agnew David J.
Published: June 2009

Hill, S. L., Trathan, P. N., and Agnew, D. J. 2009. The risk to fishery performance associated with spatially resolved management of Antarctic krill (Euphausia superba) harvesting. – ICES Journal of Marine Science, 66: 2148–2154. The ecosystem approach to fisheries attempts to define objectives for target species, the wider ecosystem, and critically, the fishery itself. Proposals for implementing the approach often include spatial restrictions on harvesting, so it is important to understand how these will affect fishery performance. One metric of potential performance is the probability of encountering exploitable densities of a target species at the scale of fishing operations. The probability of encountering exploitable densities of Antarctic krill, Euphausia superba, at the scale of 1 nautical mile during an acoustic survey was predicted by bathymetry and the mean krill density at the larger scale at which the fishery is managed. This suggests that the risk to fishery performance will increase if management actions relocate the fishery into deeper water. The results also suggest that ecosystem models resolved to the spatial scale of management units could usefully predict effects at the scale of fishing operations. However, correct parameterization of these models will require better characterization of threshold densities for efficient exploitation. Finally, the distribution of catch and fishing effort over an entire fishing season reflected the distribution of krill density observed during the survey.
Measuring Bias and Uncertainty in DW-NOMINATE Ideal Point Estimates via the Parametric Bootstrap
Authors: Carroll Royce , Lewis Jeffrey B. , Lo James , Poole Keith T. , Rosenthal Howard
Published: June 2009

DW-NOMINATE scores for the U.S. Congress are widely used measures of legislators' ideological locations over time. These scores have been used in a large number of studies in political science and closely related fields. In this paper, we extend the work of Lewis and Poole (2004) on the parametric bootstrap to DW-NOMINATE and obtain standard errors for the legislator ideal points. These standard errors are in the range of 1%–4% of the range of DW-NOMINATE coordinates.
Mycelial carton galleries of
Azteca brevis
(Formicidae) as a multi-species network
Authors: Mayer Veronika E. , Voglmayr Hermann
Published: June 2009

Apart from growing fungi for nutrition, as seen in the New World Attini, ants cultivate fungi for reinforcement of the walls of their nests or tunnel-shaped runway galleries. These fungi are grown on organic material such as bark, epiphylls or trichomes, and form stable ‘carton structures’. In this study, the carton of the runway galleries built by
Azteca brevis
(Formicidae, Dolichoderinae) on branches of
Tetrathylacium macrophyllum
(Flacourtiaceae) is investigated. For the first time, molecular tools are used to address the biodiversity and phylogenetic affinities of fungi involved in tropical ant carton architecture, a previously neglected ant–fungus mutualism.

The
A. brevis
carton involves a complex association of several fungi. All the isolated fungi were unequivocally placed within the Chaetothyriales by DNA sequence data. Whereas five types of fungal hyphae were morphologically distinguishable, our DNA data showed that more species are involved, applying a phylogenetic species concept based on DNA phylogenies and hyphal morphology. In contrast to the New World Attini with their many-to-one (different ant species—one fungal cultivar) pattern, and temperate
Lasius
with a one-to-two (one ant species—two mutualists) or many-to-one (different ant species share the same mutualist) system, the
A. brevis
–fungi association is a one-to-many multi-species network. Vertical fungus transmission has not yet been found, indicating that the
A. brevis
–fungi interaction is rather generalized.
Analysis of the human E2 ubiquitin conjugating enzyme protein interaction network
Authors: Markson Gabriel , Kiel Christina , Hyde Russell , Brown Stephanie , Charalabous Panagoula , Bremm Anja , Semple Jennifer , Woodsmith Jonathan , Duley Simon , Salehi-Ashtiani Kourosh , Vidal Marc , Komander David , Serrano Luis , Lehner Paul , Sanderson Christopher M.
Published: June 2009

In eukaryotic cells the stability and function of many proteins are regulated by the addition of ubiquitin or ubiquitin-like peptides. This process is dependent upon the sequential action of an E1-activating enzyme, an E2-conjugating enzyme, and an E3 ligase. Different combinations of these proteins confer substrate specificity and the form of protein modification. However, combinatorial preferences within ubiquitination networks remain unclear. In this study, yeast two-hybrid (Y2H) screens were combined with true homology modeling methods to generate a high-density map of human E2/E3-RING interactions. These data include 535 experimentally defined novel E2/E3-RING interactions and >1300 E2/E3-RING pairs with more favorable predicted free-energy values than the canonical UBE2L3–CBL complex. The significance of Y2H predictions was assessed by both mutagenesis and functional assays. Significantly, 74/80 (>92%) of Y2H predicted complexes were disrupted by point mutations that inhibit verified E2/E3-RING interactions, and a ∼93% correlation was observed between Y2H data and the functional activity of E2/E3-RING complexes in vitro. Analysis of the high-density human E2/E3-RING network reveals complex combinatorial interactions and a strong potential for functional redundancy, especially within E2 families that have undergone evolutionary expansion. Finally, a one-step extended human E2/E3-RING network, containing 2644 proteins and 5087 edges, was assembled to provide a resource for future functional investigations.
Involvement of Pin1 induction in epithelial–mesenchymal transition of tamoxifen‐resistant breast cancer cells
Authors: Kim Mi Ra , Choi Hoo‐Kyun , Cho Kyoung Bin , Kim Hyung Sik , Kang Keon Wook
Published: June 2009

Acquisition of resistance to tamoxifen is a critical therapeutic problem in breast cancer patients. Epithelial–mesenchymal transition (EMT), where cells undergo a developmental switch from a polarized epithelial phenotype to a highly motile mesenchymal phenotype, is associated with invasion and motility of cancer cells. Here, we found that tamoxifen‐resistant (TAMR)‐MCF‐7 cells had undergone EMT, as evidenced by mesenchymal‐like cell shape, downregulation of basal E‐cadherin expression, and overexpression of N‐cadherin and vimentin, as well as increased Snail transcriptional activity and protein expression. Given the roles of glycogen synthase kinase (GSK)‐3β and nuclear factor (NF)‐κB in Snail‐mediated E‐cadherin deregulation during EMT, we examined the role of these signaling pathways in the EMT of TAMR‐MCF‐7 cells. Both Ser9‐phosphorylated GSK‐3β (inactive form) and NF‐κB reporter activity were increased in TAMR‐MCF‐7 cells, as was activation of the phosphatase and tensin homolog depleted on chromosome ten (PTEN)–phosphoinositide 3 (PI3)‐kinase–Akt pathway. Pin1, a peptidyl‐prolyl isomerase, was overexpressed in TAMR‐MCF‐7 cells, and Snail transcription and the expression of EMT markers could be decreased by Pin1 siRNA treatment. These results imply that Pin1 overexpression in TAMR‐MCF‐7 cells is involved in the EMT process via PTEN–PI3‐kinase–Akt–GSK‐3β and/or GSK‐3β–NF‐κB‐dependent Snail activation, and suggest the potential involvement of Pin1 in EMT during breast cancer development. (Cancer Sci 2009; 100: 1834–1841)
Insulin resistance is a major determinant of liver stiffness in nondiabetic patients with HCV genotype 1 chronic hepatitis
Authors: PETTA S. , CAMMÀ C. , DI MARCO V. , CALVARUSO V. , ENEA M. , BRONTE F. , BUTERA G. , CABIBI D. , CRAXÌ A.
Published: June 2009

Background  In patients with chronic hepatitis C (CHC), liver stiffness measurement (LSM) by transient elastography (TE), is closely related to the stage of fibrosis, but may be affected by necroinflammation. Other factors, such as insulin resistance (IR), might influence the performance of LSM.

Aims  To evaluate in a cohort of nondiabetic patients with genotype 1 CHC, whether IR and other anthropometric, biochemical, metabolic and histological factors contribute to LSM and to identify the best cut‐off values of LSM for predicting different stages of fibrosis.

Methods  Nondiabetic patients with genotype 1 CHC (n = 156) were evaluated by liver biopsy (Metavir score), anthropometric, biochemical and metabolic features including IR. Furthermore, all subjects underwent LSM by TE.

Results  Severe fibrosis (F3–F4) was associated with LSM (OR 1.291; 95%CI 1.106–1.508). LSM was also independently correlated with low platelets (P = 0.03), high γGT (P < 0.001) and high HOMA (P = 0.004) levels. A stiffness value ≥8 KPa was identified as the best cut‐off for predicting severe fibrosis (AUC 0.870); yet this cut‐off still failed to rule out F3–F4 fibrosis in 22.7% of patients (false‐negative rate) or rule in F3–F4 in 19.6% (false‐positive rate). Platelets <200 × 103/mmc and a HOMA of >2.7 were the major determinants of these diagnostic errors in predicting severe fibrosis.

Conclusions  In nondiabetic patients with genotype 1 CHC, insulin resistance, γGT and platelet levels contribute to LSM independently of liver fibrosis. The identification of these three factors contributes to a more correct interpretation of LSM.
The effects of urotensin II and urantide on forearm blood flow and systemic haemodynamics in humans
Authors: Cheriyan Joseph , Burton Timothy J. , Bradley Timothy J. , Wallace Sharon M. L. , Mäki‐Petäjä Kaisa M. , Mackenzie Isla S. , McEniery Carmel M. , Brown John , Wilkinson Ian B.
Published: June 2009

WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT

• In vitro studies have shown that urotensin II is a potent arterial vasoconstrictor.

• Previous studies investigating the in vivo cardiovascular actions of intra‐arterial administration of urotensin II in humans have provided conflicting results.

• The cardiovascular actions of intra‐arterial administration of the urotensin II receptor antagonist, urantide, are unknown.

WHAT THIS STUDY ADDS

• We have shown no in vivo effect of urotensin II or urantide on human forearm resistance vessels.

• Previous study discrepancies do not seem to relate to either the age or cardiovascular disease (CVD) status of subjects.

AIMS

(i) To compare the effects of intra‐arterial administration of urotensin II in patients with CVD with healthy volunteers, and (ii) to study the haemodynamic effects of intra‐arterial infusion of the urotensin II receptor antagonist, urantide.

METHODS

Ten healthy volunteers and 10 patients with CVD received a dose‐ramped brachial artery infusion of urotensin II. A further six healthy male volunteers received a prolonged urotensin II infusion and 11 healthy male volunteers received a dose‐ramped infusion of urantide. Forearm blood flow (FBF) was measured every 20 min and blood pressure and heart rate were assessed every 20 min.

RESULTS

In healthy volunteers and patients with CVD, intra‐arterial infusion of urotensin II had no effect on FBF ratio. A dose‐ramped infusion of urantide similarly had no effect on FBF ratio. During dose‐ramped infusions of urotensin II and urantide, systolic and mean arterial blood pressure increased significantly. In healthy volunteers, urotensin II and urantide, respectively, increased systolic blood pressure from 133 ± 6 to 137 ± 5 mmHg (P < 0.01) and from 113 ± 4 to 120 ± 4 mmHg (P < 0.01). In patients with CVD, heart rate also significantly increased during dose‐ramped infusion of urotensin II from 59 ± 3 to 62 ± 4 bpm (P < 0.05).

CONCLUSIONS

We have shown no in vivo effect of urotensin II or urantide on human forearm resistance vessels. Previous discrepancies do not seem to relate to either the age or CVD status of subjects. Changes in systemic cardiovascular haemodynamics during the dose‐ramped infusion studies are unlikely to be caused by urotensin II receptor modulation.
Trends in prescribing and self‐poisoning in relation to UK regulatory authority warnings against use of SSRI antidepressants in under‐18‐year‐olds
Authors: Bergen Helen , Hawton Keith , Murphy Elizabeth , Cooper Jayne , Kapur Navneet , Stalker Carol , Waters Keith
Published: June 2009

WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT

• Ecological studies have shown conflicting evidence in relation to associations between trends in selective serotonin reuptake inhibitor (SSRI) prescription rates and suicide rates in adolescents.

• After regulatory warnings in the UK against SSRI use in children and adolescents, prescribing of antidepressants in general declined in this group; there were no related changes in rates of suicide or hospital admissions for self‐harm.

WHAT THIS STUDY ADDS

• Based on all presentations to general hospitals, nonfatal self‐poisoning with SSRI antidepressants (but not fluoxetine) declined in 12–19‐year‐olds in three centres in England in line with UK prescribing trends.

• There was some evidence of a possible small substitution effect from use of other SSRIs for nonfatal self‐poisoning to use of fluoxetine

• Overall rates of nonfatal self‐harm in 12–19‐year‐olds in three centres in England were stable, indicating no major substitution of method to self‐injury or overall adverse impact of the Medicines and Healthcare products Regulatory Authority warning.

AIMS

To assess the impact of the UK Medicines and Healthcare products Regulatory Authority (MHRA) warning in December 2003 not to prescribe selective serotonin reuptake inhibitor (SSRI) antidepressants, except fluoxetine, to under‐18‐year‐olds.

METHODS

Interrupted time series analysis of prescriptions (UK) and general hospital presentations for nonfatal self‐poisoning (three centres in England) for 2000–2006.

RESULTS

Following the MHRA warning in December 2003 there were significant decreases in prescribing of SSRI antidepressants (conservative estimate 51%) to young people aged 12–19 years. Surprisingly, this decrease also affected fluoxetine (conservative estimate 20%) and tricyclics (conservative estimate 27%). Nonfatal self‐poisoning in this age group following the warning also declined significantly for SSRIs (conservative estimate 44%), but not for fluoxetine, tricyclic antidepressants, or all drugs and other substances. Rates of nonfatal self‐harm did not change significantly over the study period.

CONCLUSIONS

The reduction in both prescribing and self‐poisoning with SSRI antidepressants (except fluoxetine) following the MHRA warning is in keeping with reduced availability of these drugs. There was some evidence of substitution from other SSRIs to fluoxetine for use in self‐poisoning. Importantly, overall rates of nonfatal self‐harm and self‐poisoning did not change, indicating no substitution of method or increases in self‐injury.
Investigation of endothelial function by pulse contour analysis: a protocol for drug administration and timing of pulse contour assessment
Authors: Gordon Sarah E. , Cartwright Neil , Griffiths Mark J. D.
Published: June 2009

WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT

• Pulse contour analysis is a relatively new technique used for non‐invasive assessment of endothelial function.

• Endothelial dysfunction is a component of a number of common conditions including hypertension, diabetes mellitus and a variety of inflammatory conditions.

• A standardized method for assessing endothelial function using pulse contour analysis is required to allow comparison between studies.

WHAT THIS STUDY ADDS

• This study recommends the optimal dose of vasoactive agents and timing of measurements for the assessment of endothelial function using pulse contour analysis.

AIMS

Pulse contour analysis (PCA) obtained by finger photoplethysmography produces a digital volume pulse (DVP) including an inflection point in its down‐slope. The reflection index (RI: ratio of the inflection point height over the maximal DVP) is responsive to vasodilatation. We aimed to optimize the drug dose and time interval for assessing endothelial function using PCA in healthy volunteers and patients with severe coronary artery disease.

METHODS

Time and dose to RI response relationships were constructed in 16 volunteers and nine patients to inhaled salbutamol (100–400 µg) or sublingual nitroglycerin (NTG; 25–400 µg).

RESULTS

For the volunteers, the time to maximum RI response to inhaled salbutamol and sublingual NTG was 10.73 ± 0.41 and 3.66 ± 0.21 min, respectively. A plateau in the RI response to salbutamol occurred between 5 and 15 min after inhalation and results were averaged over this period. A dose‐dependent response was observed to inhaled salbutamol and sublingual NTG (P= 0.05 and P < 0.001 by repeated‐measures anova, respectively) in healthy volunteers. By contrast, in patients with severe coronary artery disease inhaled salbutamol (100–400 µg) did not cause a significant change in RI.

CONCLUSIONS

In healthy volunteers the RI response to inhaled salbutamol (100–200 µg) averaged over 5–15 min after administration may be used to investigate endothelial function by PCA. The response to sublingual NTG (50 µg) should be determined at 4 min. This technique may not be suitable for the assessment of endothelial function in subjects with extensive coronary artery disease owing to the small responses observed and potential confounding effect of vasoactive medication.
Short term effects of spironolactone on blood lipid profile: a 3‐month study on a cohort of young women with hirsutism
Authors: Nakhjavani Manouchehr , Hamidi Sepehr , Esteghamati Alireza , Abbasi Mehrshad , Nosratian‐Jahromi Simindokht , Pasalar Parvin
Published: June 2009

WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT

• Spironolactone is known to have antiandrogenic features and agonist activity at progesterone receptors, which are responsible for several of its hormonal side‐effects.

• The potential unfavourable influences of this medication on serum lipoproteins have long been a concern but literature lacks sufficient data on this issue.

WHAT THIS STUDY ADDS

• Spironolactone can adversely affect serum lipids by decreasing high‐density lipoprotein and increasing low‐density lipoprotein in women treated for hirsutism on a short‐term basis.

AIMS

To investigate the effects of spironolactone on serum lipids in women with hirsutism over a 3‐month period.

METHODS

In a prospective setting, 27 hirsute women (20 with polycystic ovary syndrome and seven with idiopathic hirsutism) with a mean age of 23.0 ± 5.1 years were studied at baseline and 3 months after receiving a daily dose of 100 mg of spironolactone. Patients did not receive any other medications and did not go through a specific diet during the study. Lipid profile, fasting blood glucose, testosterone, dehydroepiandrosterone sulphate (DHEAS) and prolactin (PRL) were measured at baseline and 3 months after therapy.

RESULTS

Mean body mass index of patients was 26.1 ± 5.1 kg m−2 before treatment and 25.9 ± 5.7 kg m−2 after treatment (NS). The therapy was associated with a significant decline of mean high‐density lipoprotein (HDL), 39.5 mg dl−1[95% confidence interval (CI) 35.6, 43.4]vs. 32.2 mg dl−1 (95% CI 29.2, 35.2), and a significant increase in mean low‐density lipoprotein (LDL), 133.1 mg dl−1 (95% CI 120.2, 146) vs. 150.8 mg dl−1 (95% CI 139.1, 162.5), and cholesterol/HDL ratio, 5 (95% CI 4.4, 5.6) vs. 6.4 (95% CI 5.7, 7.1) (P < 0.05). No significant change was noted in total cholesterol, triglyceride or fasting blood glucose levels. Serum values of testosterone, DHEAS and PRL decreased significantly after 3 months of therapy (P < 0.05).

CONCLUSIONS

Spironolactone might have adverse effects on serum lipoprotein levels by increasing LDL and decreasing HDL over a short course of treatment. While treating hirsutism with spironolactone, special care should be given to women with metabolic disorders such as dyslipidaemia.
Calcitonin and carcinoembryonic antigen doubling times as prognostic factors in medullary thyroid carcinoma: a structured meta‐analysis
Authors: Meijer Johannes A. A. , Le Cessie Saskia , Van Den Hout Wilbert B. , Kievit Job , Schoones Johannes W. , Romijn Johannes A. , Smit Johannes W. A.
Published: June 2009

Context  In the management of patients with medullary thyroid carcinoma (MTC), calcitonin doubling time (dt) has gained interest as an independent predictor of recurrence and survival.

Objective  To perform a structured meta‐analysis of the diagnostic value of calcitonin dt, carcinoembryonic antigen (CEA) dt and the combination and to define dt strata with the highest predictive power.

Design  The study was a meta‐analysis using individual data.

Methods  Ten studies containing data on the post‐operative kinetics of tumour marker(s) and (recurrence free) survival were included.

Results  Calcitonin‐ and CEA‐dt are significant indicators for survival (hazard ratios (HR) 21·52 respectively infinite for dt 0–1 year compared to dt >1 year) and recurrence (HR 5·33 respectively 6·80 for dt 0–1 year compared to dt >1 year). The highest predictive power was found for the dt classification 0–1 year vs. >1 year. CEA dt has a higher predictive value than calcitonin dt in the subgroup of patients for which both parameters were available.

Conclusion  The dts of both calcitonin and CEA are strong prognostic indicators for MTC recurrence and death. CEA dt has a higher predictive value than calcitonin dt and therefore measuring both tumour markers is essential for proper risk stratification.
Despite their apparent integration, spur length but not perianth size affects reproductive success in the moth‐pollinated orchidPlatanthera bifolia
Authors: Boberg Elin , Ågren Jon
Published: June 2009

1. The evolution of many floral traits is thought to have been shaped by pollinator‐mediated selection for increased attractiveness or an improved mechanical fit of flowers to pollinators. Yet, few studies have examined experimentally the independent and interactive effects of different aspects of flower morphology on plant reproductive success.

2. In the orchidPlatanthera bifolia,perianth size and spur length are positively correlated within and among populations. To test the hypothesis that pollination success and seed output increases with increasing perianth size and spur length, we manipulated the two traits in a factorial design in a long‐spurred population ofP. bifoliapollinated by long‐tongued hawkmoths. Additionally, to determine whether differences in selfing rate can explain variation in fruit set and fruit size, we performed controlled self‐ and cross‐pollination.

3. Plants with long spurs had more flowers pollinated, more pollen removed and produced more and larger fruits compared to plants with short spurs. In contrast, perianth size did not affect the pollination success or fruit production ofP. bifolia.

4. Fruit production and fruit size did not differ among flowers pollinated with self‐ and cross pollen, respectively. This indicates that reduced pollen deposition rather than pollinator‐mediated self‐pollination caused the reduction in fruit set and fruit volume observed after shortening of the spur.

5. The results demonstrate that spur length, but not perianth size, is critical for reproductive success inP. bifolia, and suggest that among‐population differentiation in perianth size may reflect a correlated response to selection on spur length. The results are consistent with the hypothesis that visual display is less important than other cues for the reproductive success ofP. bifolia, and underscore the necessity to experimentally examine the functional significance of putatively adaptive traits.
Ecological and physiological determinants of dive duration in the freshwater crocodile
Authors: Campbell Hamish A. , Sullivan Scott , Read Mark A. , Gordos Mathew A. , Franklin Craig E.
Published: June 2009

1.
 Body mass is a key determinant of diving performance in endotherms. In air‐breathing ectotherms however, this paradigm occurs with considerably less force. Here, through remote recordings of dive behaviour over a wide size range (5–42 kg body mass, n = 17) of freshwater crocodiles (Crocodylus johnstoni), we demonstrate why body mass is such a poor determinant of dive duration for ectothermic divers.

2.
 Crocodiles were released into the wild with a time‐depth‐recorder attached to their dorsal scutes, and a movement activated radio‐tag attached to their tail. Over 15 days, 652·6 ± 58·4 (mean ± SE, n = 17) dives were recorded, with all individuals exhibiting two specific dive‐types. These were, a resting‐dive (62·7 ± 5·4% of total dive no.), where no activity occurred during the dive, and an active‐dive (37·1 ± 6·3% of total no.) associated with swimming.

3.
 The durations of resting‐dives (∼12 min) were similar for all crocodiles. Smaller crocodiles (6·3 ± 0·7 kg, mean ± SE, n = 9) exhibited a significant correlation between dive duration and post‐dive surface‐interval, whilst larger crocodiles (17·9 ± 3·75 kg, mean ± SE, n = 8) did not. This demonstrated that aerobic dive duration was mass‐specific during resting‐dives, but other mass specific factors, presumably ecological, determined dive duration.

4.  The durations of active‐dives were never >1 min, showed no relationship with body mass and no correlation with the post‐dive‐surface interval. In crocodiles, aerobic metabolic scope is independent of body mass but anaerobic capacity is mass dependent, suggesting that active‐dive duration was determined by sustained activity and dives were terminated before anaerobic metabolism became significant.

5.  All individuals showed similar diel phase shifts in dive duration, type and depth, illustrating the overwhelming influence of the external environment on dive behaviour. Dive durations which resulted in significant anaerobic debt occurred rarely, but were undertaken in response to a potential threat.

6.
 Body mass was a poor predictor of diving in C. johnstoni because the external environmental and ecological factors exerted a greater influence on dive duration than oxygen reserves.
The return of the vole cycle in southern Finland refutes the generality of the loss of cycles through ‘climatic forcing’
Authors: BROMMER JON E. , PIETIÄINEN HANNU , AHOLA KARI , KARELL PATRIK , KARSTINEN TEUVO , KOLUNEN HEIKKI
Published: June 2009

Multiannual cycles in the abundance of voles and other animals have been collapsing in the last decades. It has been proposed that this phenomenon is ‘climatically forced’ by milder winters. We here consider the dynamics of bank and field voles during more than two decades in two localities (170 km apart) in southern Finland. Using wavelet analysis, we show that a clear 3‐year cycle disappeared in the mid 1990s. However, the vole cycle returned in both localities after about 5 years despite winters becoming increasingly milder. In both localities, vole cycles were mainly determined by bank voles after the period of noncyclic dynamics, whereas field voles were dominant before this irregularity. Wavelet coherency analysis shows that spatial synchrony temporarily broke down during the period of noncyclic dynamics, but was fully restored afterwards. The return of the cycle despite ongoing rapid climate change argues against ‘climatic forcing’ as a general explanation for loss of cycles. Rather, the population‐dynamical consequences of climate change may be dependent on the local species composition and mechanism of delayed density dependence.
Soil organic carbon stocks in Laos: spatial variations and controlling factors
Authors: CHAPLOT VINCENT , BOUAHOM BOUNTHONG , VALENTIN CHRISTIAN
Published: June 2009

Surface soils, which contain the largest pool of terrestrial organic carbon (C), may be able to sequester atmospheric C and thus mitigate climate change. However, this remains controversial, largely due to insufficient data and knowledge gaps in respect of organic C contents and stocks in soils and the main factors of their control. Up to now and despite numerous evaluations of soil organic carbon (SOC) stocks worldwide, the sloping lands of southeast Asia, one of the most biogeochemically active regions of the world, remain uninvestigated. Our main objective was to quantify SOC stocks and to evaluate the impact of various environmental factors. We, therefore, selected Laos with 230 566 km2of mostly forested steep slopes, and where cultivation is still mainly traditional, i.e. a system of shifting cultivation without fertilization or mechanical tillage. Analytical data from 3471 soil profiles demonstrated that the top 1 m of soil depth holds an estimated 4.64 billion tons of SOC, 65% of which is in the first 0.3 m. SOC stocks to 0.3 m exhibit a high coefficient of variation (CV=62%) with values from 1.8 to 771 Mg C ha−1and a mean at 129 Mg C ha−1. Furthermore, these stocks are significantly (atP<0.05 level) affected by land use as shown by principal components analysis andt‐tests with the largest amount being found under forest, less under shifting cultivation and the smallest under continuous cultivation. Moreover, SOC stocks correlated regionally to total annual rainfalls and latitude, and locally at the hill‐slope level to the distance to the stream network and the slope angle. It is hypothesized that this correlation is through actions on mineral weathering, soil clay content, soil fertility and SOC redistributions in landscapes. These relationships between SOC stocks and environmental factors may be of further use in (1) predicting the impact of global changes on future SOC stocks; and (2) identifying optimal strategies for land use planning so as to minimize soil C emissions to the atmosphere while maximizing carbon sequestration in soils.
Risk factors for inhibitor formation in haemophilia: a prevalent case–control study
Authors: RAGNI M. V. , OJEIFO O. , FENG J. , YAN J. , HILL K. A. , SOMMER S. S. , TRUCCO M. N. , BRAMBILLA D. J. ,   AND THE HEMOPHILIA INHIBITOR STUDY
Published: June 2009

Summary.  Inhibitor formation is a major complication of haemophilia treatment. In a prevalent case–control study, we evaluated blood product exposure, genotype and HLA type on haemophilia A inhibitor formation. Product exposure was extracted from medical records. Genotype was determined on stored DNA samples by detection of virtually all mutations‐SSCP (DOVAM‐S) and subcycling PCR. HLA typing was performed by PCR amplification and exonuclease‐released fluorescence. Cases experienced higher intensity factor, 455 vs. 200 U per exposure, P < 0.005, more frequent central nervous system (CNS) bleeding, seven of 20 (35.0%) vs. one of 57 (1.7%), P = 0.001 and more commonly from inhibitor families, seven of 20 (35.0%) vs. zero of 57 (0%), P < 0.001, and African‐American, 12 of 63 (19.0%) vs. six of 117 (5.1%), P = 0.015. Among the latter, CNS bleeding was more commonly the initial bleed, 60% vs. 0%, P < 0.001, and survival was shorter, 14 vs. 38 yr, P = 0.025. Inhibitor formation was uncommon in those with missense mutations, two of 65 (3.1%) vs. 31 of 119 (26.0%), P = 0.008, and unrelated to factor VIII immunogenic epitope, P = 0.388, or HLA type, P > 0.100. Genotype was not associated with race. Time to immune tolerance was shorter for titres <120 vs. ≥120 BU/mL, six vs. 16 months, P < 0.01, but unaffected by tolerizing dose regimen, P > 0.50. Inhibitor formation is associated with high intensity product exposure, CNS bleeding, African‐American race and low frequency of missense mutations. The ideal time to initiate prophylaxis to reduce CNS bleeding and inhibitor formation will require prospective studies.
Immunological aspects and therapeutic significance of an autoantibody against histone H1 in a rat model of concanavalin A‐induced hepatitis
Authors: Nakano Toshiaki , Goto Shigeru , Lai Chia‐Yun , Hsu Li‐Wen , Takaoka Yuki , Kawamoto Seiji , Chiang Kuei‐Chen , Shimada Yayoi , Ohmori Naoya , Goto Takeshi , Sato Shuji , Ono Kazuhisa , Cheng Yu‐Fan , Chen Chao‐Long
Published: June 2009

We previously demonstrated the immunosuppressive activity of anti‐histone H1 autoantibody induced in experimental and clinical liver allograft tolerance. This study aimed to explore the immunological aspects of anti‐histone H1 autoantibody in liver injury induced by concanavalin A (Con A). To establish a Con A‐hepatitis model, 20 mg/kg Con A was intravenously injected into rats, after which liver function and histopathological analyses were performed. In this model, anti‐histone H1 autoantibody was transiently induced in the sera during the natural recovery stage, 3–7 days after Con A injection. To evaluate the therapeutic significance of anti‐histone H1 autoantibody, a polyclonal antibody against histone H1 was intraperitoneally injected immediately after Con A injection. We found that injection of anti‐histone H1 antibody could reduce Con A‐induced liver damage. Further mechanical analyses revealed that anti‐histone H1 antibody altered the intracellular activation of mitogen‐activated protein kinase, nuclear factor‐κB and calcineurin via T‐cell receptor signalling, suggesting that anti‐histone H1 antibody may protect the liver from Con A‐induced injury by inhibiting activation of effector T cells. These findings suggest that anti‐histone H1 autoantibody may be a natural immune regulatory factor that protects inflamed livers suffering from autoimmune hepatitis and may lead to T‐cell unresponsiveness through the selective regulation of mitogen‐activated protein kinase/nuclear factor‐κB and calcineurin signalling.
Cooperative action of CD8 T lymphocytes and natural killer cells controls tumour growth under conditions of restricted T‐cell receptor diversity
Authors: Shanker Anil , Buferne Michel , Schmitt‐Verhulst Anne‐Marie
Published: June 2009

In mice expressing a transgenic T‐cell receptor (TCR; TCRP1A) of DBA/2 origin with reactivity towards a cancer‐germline antigen P1A, the number of TCRP1A CD8+ T cells in lymphoid organs is lower in DBA/2 than in B10.D2 or B10.D2(× DBA/2)F1 mice. This reduction results from haemopoietic cell autonomous differences in the differentiation of the major histocompatibility complex class I‐restricted TCRP1A thymocytes controlled by DBA/2 versus B10.D2‐encoded genes. We report here that the lower number of TCRP1A CD8+ T cells in DBA/2 mice correlated with their poor resistance to P1A‐expressing mastocytoma solid tumours. Functional potency of CD8+ cytolytic T lymphocytes (CTL) from the above strains was not compromised, but their number after expansion appeared to be influenced by their genetic background. Intriguingly, non‐transgenic DBA/2 mice resisted P1A+ tumours more efficiently despite poor representation of P1A‐specific CTL. This was partly the result of their more heterogeneous TCR repertoire, including reactivity to non‐P1A tumour antigens because mice that had rejected a P1A+ tumour became resistant to a P1A− variant of the tumour. Such ‘cross‐resistance’ did not develop in the TCRP1A transgenic mice. Nonetheless, reconstitution of RAGº/º mice with TCRP1A CD8+ T cells, with or without CD4+ T cells, or exclusive representation of TCRP1A CD8+ T cells in RAGº/º TCRP1A transgenic mice efficiently resisted the growth of P1A‐expressing tumours. Natural killer cells present at a higher number in RAGº/º mice also contributed to tumour resistance, in part through an NKG2D‐dependent mechanism. Hence, in the absence of a polyclonal T‐cell repertoire, precursor frequencies of natural killer cells and tumour‐specific CTL affect tumour resistance.
Tumor necrosis factor‐α augments lipopolysaccharide‐induced suppressor of cytokine signalling 3 (SOCS‐3) protein expression by preventing the degradation
Authors: Dagvadorj Jargalsaikhan , Naiki Yoshikazu , Tumurkhuu Gantsetseg , Shadat Mohammod Noman Abu , Iftakhar‐E‐Khuda Imtiaz , Komatsu Takayuki , Koide Naoki , Yoshida Tomoaki , Yokochi Takashi
Published: June 2009

The regulatory role of tumour necrosis factor‐α (TNF‐α) on the expression of suppressor of cytokine signalling 3 (SOCS‐3) in response to lipopolysaccharide (LPS) was examined using peritoneal macrophages from TNF‐α‐deficient mice. The LPS‐induced SOCS‐3 expression was markedly augmented in macrophages from wild‐type mice whereas such augmentation was not seen in the cells from TNF‐α‐deficient mice. However, there was no significant difference in the level of SOCS‐3 messenger RNA expression between macrophages from wild‐type mice and those from TNF‐α‐deficient mice. The addition of exogenous TNF‐α augmented the LPS‐induced SOCS‐3 expression in macrophages from TNF‐α‐deficient mice. The pulse chase analysis suggested augmented degradation of LPS‐induced SOCS‐3 protein in macrophages from TNF‐α‐deficient mice. Moreover, MG 132, a 26S proteasome inhibitor, sustained the LPS‐induced SOCS‐3 expression in those cells. The tyrosine phosphorylation of SOCS‐3 was definitely induced in LPS‐stimulated macrophages from TNF‐α‐deficient mice but not wild‐type mice. A tyrosine phosphatase inhibitor enhanced the tyrosine phosphorylation of SOCS‐3 in wild‐type mice and accelerated the degradation. Therefore, it was suggested that TNF‐α prevented the degradation of SOCS‐3 protein via inhibition of the tyrosine phosphorylation in LPS‐stimulated macrophages.
Dissecting the essentiality of the bifunctional trypanothione synthetase‐amidase in Trypanosoma brucei using chemical and genetic methods
Authors: Wyllie Susan , Oza Sandra L. , Patterson Stephen , Spinks Daniel , Thompson Stephen , Fairlamb Alan H.
Published: June 2009

The bifunctional trypanothione synthetase‐amidase (TRYS) comprises two structurally distinct catalytic domains for synthesis and hydrolysis of trypanothione (N1,N8‐bis(glutathionyl)spermidine). This unique dithiol plays a pivotal role in thiol‐redox homeostasis and in defence against chemical and oxidative stress in trypanosomatids. A tetracycline‐dependent conditional double knockout of TRYS (cDKO) was generated in bloodstream Trypanosoma brucei. Culture of cDKO parasites without tetracycline induction resulted in loss of trypanothione and accumulation of glutathione, followed by growth inhibition and cell lysis after 6 days. In the absence of inducer, cDKO cells were unable to infect mice, confirming that this enzyme is essential for virulence in vivo as well as in vitro. To establish whether both enzymatic functions were essential, an amidase‐dead mutant cDKO line was generated. In the presence of inducer, this line showed decreased growth in vitro and decreased virulence in vivo, indicating that the amidase function is not absolutely required for viability. The druggability of TRYS was assessed using a potent small molecule inhibitor developed in our laboratory. Growth inhibition correlated in rank order cDKO, single KO, wild‐type and overexpressing lines and produced the predicted biochemical phenotype. The synthetase function of TRYS is thus unequivocally validated as a drug target by both chemical and genetic methods.
Viral vectors in malaria vaccine development
Authors: LIMBACH K. J. , RICHIE T. L.
Published: June 2009

Traditional vaccine technologies have resulted in an impressive array of efficacious vaccines against a variety of infectious agents. However, several potentially deadly pathogens, including retroviruses and parasites, have proven less amenable to the application of traditional vaccine platforms, indicating the need for new approaches. Viral vectors represent an attractive way to deliver and present vaccine antigens that may offer advantages over traditional platforms. Due to their ability to induce strong cell‐mediated immunity (CMI) in addition to antibodies, viral vectors may be suitable for infectious agents, such as malaria parasites, where potent CMI is required for protection. Poxvirus‐vectored malaria vaccines have been the most extensively studied in the clinic, achieving significant reductions in liver‐stage parasite burden. More recently, adenovirus‐vectored malaria vaccines have entered clinical testing. The most promising approach – heterologous prime‐boost regimens, in which different viral vectors are sequentially paired with each other or with DNA or recombinant protein vaccines – is now being explored, and could provide high‐grade protection, if findings in animal models are translatable to humans. Significant barriers remain, however, such as pre‐existing immunity to the vector particle and an unexplained safety signal observed in one trial suggesting an increased risk of HIV acquisition in volunteers with pre‐existing immunity to the vector.
Molecular analysis of knock down resistance (kdr) mutation and distribution of kdr genotypes in a wild population of Culex quinquefasciatus from India
Authors: Sarkar Manas , Borkotoki Aparajita , Baruah Indra , Bhattacharyya Indra Kumar , Srivastava Ravi Bihari
Published: June 2009

Objective  To investigate the presence of knock down resistance (kdr) mutation, its frequency distribution in the principal vector of bancroftian filariasis, Culex quinquefasciatus from northeastern India, and to relate kdr genotypes with susceptibility and/or resistance to DDT and deltamethrin in this vectors.

Methods  Adult female mosquitoes were collected by aspiration from human dwellings in two villages, Benganajuli and Rikamari, and two military establishments, Field Units I and II. Insecticide susceptibility tests were performed following WHO methods with 4% DDT and 0.05% deltamethrin. Molecular identification of kdr mutation and genotyping of kdr locus was performed by allele‐specific PCR (AS‐PCR) and direct sequencing in a subset of samples.

Results  Mosquitoes were resistant to DDT and showed 11.9–41.2% mortality, whereas the knock down bioassay for deltamethrin suggests complete susceptibility to this insecticide in all study sites except Benganajuli. The result of AS‐PCR confirmed the presence of three genotypes: susceptible (SS), resistant (RR) and heterozygous (SR) in the population. Genotype frequencies at kdr locus for DDT‐resistant individuals conformed with the Hardy–Weinberg proportion, whereas DDT and deltamethrin susceptible individuals differed significantly (P < 0.05). The efficacy of AS‐PCR in detecting the correct genotype was not encouraging.

Conclusions  This is the first report from India on kdr genotyping in C. quinquefasciatus, and it confirms the occurrence of kdr allele in this vector in northeastern India. This finding has serious implications for the filariasis control programmes in India.
Rivanol treatment of cystic fibrosis serum: effect of supernatant upon ciliary action
Authors: McCombs Michael L. , Bowman Barbara H.
Published: June 2009

Serum from cystic fibrosis patients and their parents will inhibit ciliary movement on oyster gill tissue. Previous studies have indicated the factor responsible for ciliary inhibition is a cationic protein with a molecular weight between 125,000 to 200,000, and has properties similar to γG‐globulin. To support this evidence, sera from CF patients, heterozygotes, and control individuals were subjected to rivanol precipitation and the supernatants tested on oyster cilia. In addition other sources of cilia and flagella were examined as possible assay systems for the CF factor.
Lesch‐Nyhan syndrome: Rapid detection of heterozygotes
Authors: Wood Stephen , Pinsky Leonard
Published: June 2009

A simple, rapid test has been developed for heterozygous carriers of the mutant gene responsible for the Lesch‐Nyhan syndrome. The test is based upon the ability of cultured skin fibroblasts to incorporate 14C‐adenine following exposure to the purine analogue 8‐azaguanine (8AG). Heterozygous cultures had an intermediate level of resistance to the inhibitory action of 8AG.