bluuebunny/crossref_metadata_2025 huggingface dataset:
Substrate specificity of Helicobacter pylori histone-like HU protein is determined by insufficient stabilization of DNA flexure points
Authors: CHEN Christina , GHOSH Sharmistha , GROVE Anne
Published: October 2004

The histone-like HU protein is ubiquitous in the eubacteria. A role for Escherichia coli HU in compaction of the bacterial genome has been reported, along with regulatory roles in DNA replication, transposition, repair and transcription. We show here that HU from the human pathogen Helicobacter pylori, which has been implicated in the development of ulcers and gastric cancer, exhibits enhanced thermal stability and distinct DNA substrate specificity. Thermal denaturation of HpyHU (H. pylori HU) measured by CD spectroscopy yields a melting temperature (Tm) of 56.4±0.1 °C. HpyHU binds linear duplex DNA with a site size of ∼19 bp and with low affinity, but in striking contrast to E. coli HU, HpyHU has only modest preference for DNA with mismatches, nicks or gaps. Instead, HpyHU binds stably to four-way DNA junctions with half-maximal saturation of 5 nM. Substitution of two residues adjacent to the DNA-intercalating prolines attenuates both the preference for flexible DNA and the ability to bend and supercoil DNA. These observations suggest that proline intercalation generates hinges that must be stabilized by adjacent residues; insufficient stabilization leads to reduced bending and a failure to bind preferably to DNA with flexure points, such as gaps and mismatches.
Hazelnut (Corylus avellana) vicilin Cor a 11: molecular characterization of a glycoprotein and its allergenic activity
Authors: LAUER Iris , FOETISCH Kay , KOLARICH Daniel , BALLMER-WEBER Barbara K. , CONTI Amedeo , ALTMANN Friedrich , VIETHS Stefan , SCHEURER Stephan
Published: October 2004

In Europe, hazelnuts (Corylus avellana) are a frequent cause of food allergies. Several important hazelnut allergens have been previously identified and characterized. Specific N-glycans are known to induce strong IgE responses of uncertain clinical relevance, but so far the allergenic potential of glycoproteins from hazelnut has not been investigated. The aim of the study was the molecular characterization of the glycosylated vicilin Cor a 11 from hazelnut and the analysis of its allergenic activity. Although MALDI–TOF (matrix-assisted laser-desorption ionization–time-of-flight) MS showed that one of two potential glycosylation sites of Cor a 11 was glycosylated, CD spectroscopy indicated that recombinant and natural Cor a 11 share similar secondary structures. Thus to analyse the impact of the glycan residues of Cor a 11 on IgE binding, the allergenic activity of natural glycosylated Cor a 11 and recombinant Cor a 11 was compared. In addition, the IgE sensitization pattern to recombinant Cor a 11, Cor a 1, Cor a 2 and Cor a 8 of 65 hazelnut allergic patients was determined in vitro. The prevalence of IgE reactivity to hazelnut vicilin Cor a 11 was below 50%. Basophil histamine-release assays were used to determine the allergenic activity of both natural and recombinant Cor a 11 in comparison with Cor a 1, a birch (Betula verrucosa) pollen-related major hazelnut allergen. Both forms of Cor a 11 induced mediator release from basophils to a similar extent, indicating that the hazelnut allergic patients had cross-linking IgE antibodies binding to the protein backbone and not to carbohydrate structures. In comparison to Cor a 1, a 10000-fold higher concentration of Cor a 11 was required to induce similar basophil mediator release. In conclusion, the hazelnut vicilin Cor a 11 is a minor allergen both in regard to prevalence and allergenic potency, whereas its glycan does not contribute to its allergenic activity.
Determination of electron temperature from spectral line intensity decay for radiation dominated plasmas
Authors: Michael C. A. , Howard J.
Published: October 2004

We describe a technique to absolutely estimate the electron temperature in radiation dominated plasmas from the temporal decay during the plasma afterglow of the intensity of a single spectral line. The model and underlying assumptions are described. We apply the model to data in both rf heated argon discharges and electron cyclotron heated He/H discharges in the H-1 heliac. The results agree well with probe measurements.
Design aspects of a MSE diagnostic for ITER
Authors: Casper T. A. , Jayakumar J. , Makowski M. A. , Ellis R.
Published: October 2004

The motional Stark effect (MSE) diagnostic is unique in its ability to measure the current profile and will be essential in the International Thermonuclear Experimental Reactor (ITER) for detailed analysis of Advanced Tokamak and other types of discharges. However, design of a MSE diagnostic for ITER presents many unique challenges. Among these is optical analysis for the convoluted optical path, required for effective neutron shielding, that employs several reflective optics arranged to form a labyrinth. The geometry of the diagnostic has been laid out and the expected Doppler shifts and channel resolution calculated. A model of the optical train has also been developed based on the Mueller matrix formalism. Unfolding the pitch angle for this complicated geometry is not straightforward and possible methods are evaluated. The CORSICA code is used to model a variety of ITER discharges including start-up, Ip-ramp, and reverse shear. The code also incorporates a synthetic MSE diagnostic that can be used to evaluate different viewing locations and optimize channel locations for the above discharges. Simulation of the optical emission spectrum is also under way.
Electric field and temperature effects on water in the narrow nonpolar pores of carbon nanotubes
Authors: Vaitheeswaran Subramanian , Rasaiah Jayendran C. , Hummer Gerhard
Published: October 2004

Water molecules in the narrow cylindrical pore of a (6,6) carbon nanotube form single-file chains with their dipoles collectively oriented either up or down along the tube axis. We study the interaction of such water chains with homogeneous electric fields for finite closed and infinite periodically replicated tubes. By evaluating the grand-canonical partition function term-by-term, we show that homogeneous electric fields favor the filling of previously empty nanotubes with water from the bulk phase. A two-state description of the collective water dipole orientation in the nanotube provides an excellent approximation for the dependence of the water-chain polarization and the filling equilibrium on the electric field. The energy and entropy contributions to the free energy of filling the nanotube were determined from the temperature dependence of the occupancy probabilities. We find that the energy of transfer depends sensitively on the water-tube interaction potential, and that the entropy of one-dimensionally ordered water chains is comparable to that of bulk water. We also discuss implications for proton transfer reactions in biology.
Deletion of the synaptic protein interaction site of the N-type (Ca
V
2.2) calcium channel inhibits secretion in mouse pheochromocytoma cells
Authors: Harkins Amy B. , Cahill Anne L. , Powers James F. , Tischler Arthur S. , Fox Aaron P.
Published: October 2004

Presynaptic N-type Ca
2+
channels (Ca
v
2.2, α
1B
) are thought to bind to SNARE (SNAP-25 receptor) complex proteins through a synaptic protein interaction (synprint) site on the intracellular loop between domains II and III of the α
1B
subunit. Whether binding of syntaxin to the N-type Ca
2+
channels is required for coupling Ca
2+
ion influx to rapid exocytosis has been the subject of considerable investigation. In this study, we deleted the synprint site from a recombinant α
1B
Ca
2+
channel subunit and transiently transfected either the wild-type α
1B
or the synprint deletion mutant into mouse pheochromocytoma (MPC) cell line 9/3L, a cell line that has the machinery required for rapid stimulated exocytosis but lacks endogenous voltage-dependent Ca
2+
channels. Secretion was elicited by activation of exogenously transfected Ca
2+
channel subunits. The current-voltage relationship was similar for the wild-type and mutant α
1B
-containing Ca
2+
channels. Although total Ca
2+
entry was slightly larger for the synprint deletion channel, compared with the wild-type channel, when Ca
2+
entry was normalized to cell size and limited to cells with similar Ca
2+
entry (≈150 × 10
6
Ca
2+
ions/pF cell size), total secretion and the rate of secretion, determined by capacitance measurements, were significantly reduced in cells expressing the synprint deletion mutant channels, compared with wild-type channels. Furthermore, the amount of endocytosis was significantly reduced in cells with the α
1B
synprint deletion mutant, compared with the wild-type subunit. These results suggest that the synprint site is necessary for efficient coupling of Ca
2+
influx through α
1B
-containing Ca
2+
channels to exocytosis.
Engineering high-level aluminum tolerance in barley with the
ALMT1
gene
Authors: Delhaize Emmanuel , Ryan Peter R. , Hebb Diane M. , Yamamoto Yoko , Sasaki Takayuki , Matsumoto Hideaki
Published: October 2004

Acidity is a serious limitation to plant production on many of the world's agricultural soils. Toxic aluminium (Al) cations solubilized by the acidity rapidly inhibit root growth and limit subsequent uptake of water and nutrients. Recent work has shown that the
ALMT1
gene of wheat (
Triticum aestivum
) encodes a malate transporter that is associated with malate efflux and Al tolerance. We generated transgenic barley (
Hordeum vulgare
) plants expressing
ALMT1
and assessed their ability to exude malate and withstand Al stress.
ALMT1
expression in barley conferred an Al-activated efflux of malate with properties similar to those of Al-tolerant wheat. The transgenic barley showed a high level of Al tolerance when grown in both hydroponic culture and on acid soils. These findings provide additional evidence that
ALMT1
is a major Al-tolerance gene and demonstrate its ability to confer effective tolerance to acid soils through a transgenic approach in an important crop species.
Regional crustal thickness and precipitation in young mountain chains
Authors: Ernst W. G.
Published: October 2004

Crustal thickness is related to climate through precipitation-induced erosion. Along the Andes, the highest mountains and thickest crust (≈70 km) occur at 25° south, a region of low precipitation. Westerly winds warm passing over the Atacama Desert; precipitation is modest in the High Andes and eastward over the Altiplano. Severe aridity, hence low erosion rates, helps to account for the elevated volcanogenic contractional arc and high, internally draining plateau in its rain shadow. Weak erosion along the north-central arc provides scant amounts of sediment to the Chile–Peru Trench, starving the subduction channel. Subcrustal removal might be expected to reduce the crustal thickness, but is not a factor at 25° south. The thickness of the gravitationally compensated continental crust cannot reflect underplating and/or partial fusion of sediments, but must be caused chiefly by volcanism-plutonism and contraction. Contrasting climate typifies the terrane at 45° south where moisture-laden westerly winds encounter a cool margin, bringing abundant precipitation. The alpine landscape is of lower average elevation compared with the north-central Andes and is supported by thinner continental crust (≈35 km). Intense erosion supplies voluminous clastic debris to the offshore trench, and vast quantities are subducted. However, the southern Andean crust is only about half as thick as that at 25° south, suggesting that erosion, not subcrustal sediment accretion or anatexis, is partly responsible for the thickness of the mountain belt. The Himalayas plus Tibetan Plateau, the Sierra Nevada plus Colorado Plateau, and the Japanese Islands exhibit analogous relationships between crustal thickness and climate.
Visualization of a functionally enhanced GFP-tagged galanin R2 receptor in PC12 cells: Constitutive and ligand-induced internalization
Authors: Xia Sheng , Kjær Svend , Zheng Kang , Hu Ping-Sheng , Bai Li , Jia Jun-Yong , Rigler Rudolf , Pramanik Aladdin , Xu Tao , Hökfelt Tomas , Xu Zhi-Qing David
Published: October 2004

Trafficking of the galanin R2 receptor (GALR2) fused with enhanced GFP (EGFP) was studied by using confocal fluorescence microscopy. The fusion protein was predominantly localized on the plasma membrane with some intracellular fluorescent structures (vesicles), mainly in the perinuclear region. Incubation with galanin resulted in a concentration-dependent increase in intracellular Ca
2+
concentration levels, suggesting that the GALR2-EGFP conjugate is functional. After blocking endocytosis with methyl-β-cyclodextrin GALR2-EGFP expression was increased on the surface and decreased in the cytoplasm. Blocking endocytic recycling with monensin caused an increase of intracellular GALR2-EGFP accumulation and a decrease of fluorescence on the plasma membrane. GALR2-EGFP on the plasma membrane was internalized within 5-10 min after treatment with galanin or AR-M1896, a selective GALR2 agonist, with a dramatic reduction in plasma membrane localization and appearance in intracellular vesicles. Neither M35 nor M40, two galanin analogues with putative antagonistic action, prevented GALR2 agonist-induced internalization of GALR2-EGFP, suggesting that they are not antagonists at this receptor under the present circumstances. Galanin stimulation at low temperature caused GALR2-EGFP aggregation and clustering on the surface but no translocation to cytoplasm. After coincubation with galanin the GALR2-EGFP was colocalized with internalized Texas red-transferrin, a marker of the clathrin endocytic pathway. Hyperosmotic sucrose inhibited internalization of GALR2-EGFP. Taken together these findings indicate that GALR2 undergoes constitutive endocytosis and recycling and that both ligand-independent and ligand-dependent internalization use the clathrin-dependent endocytic recycling pathway.
RNA polymerase I remains intact without subunit exchange through multiple rounds of transcription in
Saccharomyces cerevisiae
Authors: Schneider David A. , Nomura Masayasu
Published: October 2004

Previous experiments using mammalian cells suggested that after each round of transcription, RNA polymerase I (Pol I) dissociates into subunits that leave and reenter the nucleolus as individual subunits, before formation of a new initiation complex. In this study, we show that the size and subunit composition of Pol I did not change significantly when Pol I was not engaged in rRNA transcription, brought about by either the absence of Pol I-specific rDNA template or specific inhibition of the transcription initiation step that requires Rrn3p. In fact, Pol I purified from cells completely lacking rDNA repeats was more active than when purified from wild-type cells in an
in vitro
transcription system designed to assay active Pol I–Rrn3p complexes. Furthermore, measurements of the exchange of A135 and A190 subunits between preexistent Pol I and newly synthesized Pol I showed that these two largest subunits of Pol I do not disassociate through many rounds of transcription
in vivo
. Thus, Pol I is not a dynamic protein complex but rather a stable enzyme.
The Maturing of Hormesis as a Credible Dose-Response Model
Authors: Calabrese Edward J.
Published: October 2004

Hormesis is a dose-response phenomenon that has received little recognition, credibility and acceptance as evidenced by its absence from major toxicological/risk assessment texts, governmental regulatory dose-response modeling for risk assessment, and non-visibility in major professional toxicological society national meetings. This paper traces the historical evolution of the hormetic dose-response hypothesis, why this model is not only credible but also more common than the widely accepted threshold model in direct comparative evaluation, and how the toxicological community made a critical error in rejecting hormesis, a rejection sustained over 70 years.
Spectroscopic Analyses of Oscillations in ECTO-NOX-Catalyzed Oxidation of NADH
Authors: Morré D. James , Morré Dorothy M.
Published: October 2004

Spectroscopic strategies that substantiate periodic oscillations in low rates of NADH oxidation exhibited by ECTO-NOX proteins at the animal and plant cell surface are described. Both continuous display and discontinuous rate determinations exhibit the oscillations but continuous displays lack sufficient resolution to discern details. A procedure is documented where rates are determined by least squares analyses of traces recorded over 1 min at intervals of 1.5 min. These traces recapitulate the continuous displays but offer an opportunity to reliably estimate changes in reaction rates over short time intervals not afforded by the continuous traces. Results from previously used rate determination over 5 min intervals are included for comparison. Turbidity is identified as the major contributor to losses in resolution. Even highly purified NOX preparations tend to aggregate to form turbid suspensions. With turbid suspensions, double beam or dual wavelength instrumentation where the sample is placed immediately adjacent to the photomultiplier tube are required to reduce losses in resolution from turbidity. Also required are high levels of synchronous ECTO-NOX function. Blue or red (plants) light, small molecules (i.e., melatonin) and autosynchrony alone or in combination were used to synchronize the oscillations. Special problems posed by preparations containing more than one ECTO-NOX form and where the different ECTO-NOX forms do not cross entrain are discussed.
Optimal Design for Estimating Parameters of the 4-Parameter Hill Model
Authors: Khinkis Leonid A. , Levasseur Laurence , Faessel Hélène , Greco William R.
Published: October 2004

Many drug concentration-effect relationships are described by nonlinear sigmoid models. The 4-parameter Hill model, which belongs to this class, is commonly used. An experimental design is essential to accurately estimate the parameters of the model. In this report we investigate properties of D-optimal designs. D-optimal designs minimize the volume of the confidence region for the parameter estimates or, equivalently, minimize the determinant of the variance-covariance matrix of the estimated parameters. It is assumed that the variance of the random error is proportional to some power of the response. To generate D-optimal designs one needs to assume the values of the parameters. Even when these preliminary guesses about the parameter values are appreciably different from the true values of the parameters, the D-optimal designs produce satisfactory results. This property of D-optimal designs is called robustness. It can be quantified by using D-efficiency. A five-point design consisting of four D-optimal points and an extra fifth point is introduced with the goals to increase robustness and to better characterize the middle part of the Hill curve. Four-point D-optimal designs are then compared to five-point designs and to log-spread designs, both theoretically and practically with laboratory experiments.

D-optimal designs proved themselves to be practical and useful when the true underlying model is known, when good prior knowledge of parameters is available, and when experimental units are dear. The goal of this report is to give the practitioner a better understanding for D-optimal designs as a useful tool for the routine planning of laboratory experiments.
A single-field consistency relation for the three-point function
Authors: Paolo Creminelli  , Matias Zaldarriaga
Published: October 2004

We point out the existence of a consistency relation involving the three-point function of
scalar perturbations which is valid in any inflationary model, independently of the inflaton
Lagrangian under the assumption that the inflaton is the only dynamical field. The
three-point function in the limit in which one of the momenta is much smaller than the
other two is fixed in terms of the power spectrum and its tilt. This relation, although very
hard to verify experimentally, could be easily proved wrong by forthcoming data, thus
ruling out any scenario with a single dynamical field in a model independent way.
Economic Partnership Agreements Between Sub‐Saharan Africa and the EU: A Development Perspective
Authors: Hinkle Lawrence E. , Schiff Maurice
Published: October 2004

This paper draws on Hinkle and Schiff (2003). It analyses the planned Economic Partnership Agreements (EPAs) between the EU and Sub‐Saharan Africa (SSA) from a development perspective. It does not take a position on whether SSA should enter into EPAs with the EU. Rather, it starts from the notion that the process of forming EPAs is unlikely to be reversed and examines the conditions that will maximise SSA's benefits from the EPAs. If this notion is correct, then the analysis presented in the paper applies. On the other hand, Pascal Lamy, the EU Trade Commissioner, made a proposal at the May 2004 G‐90 summit in Dakar that might lead to a change in the EPA process. He proposed that the G‐90, a group consisting of ACP and non‐ACP LDC countries, should not have to make concessions at the WTO Doha Round of multilateral trade negotiations, i.e., he proposed a ‘free round’ for the G‐90. This proposal opens the door to the possibility that the same might apply to the ACP countries in the EU‐ACP negotiations and that the EPA process might be reversed. The paper considers the key issues raised by the planned EPAs, their relationship to the WTO's Doha Round and the EU's Everything‐but‐Arms Initiative, the changes needed to make the EPAs internally consistent, the domestic reforms in SSA that would need to accompany trade liberalisation in both goods and services, and the potential effects of the EPAs on regional integration in SSA. The EPAs will pose a number of policy challenges for SSA countries, including: restructuring of indirect tax systems, reduction of MFN tariffs, liberalisation of service imports on an MFN basis and related regulatory reforms in the services sector, and liberalisation of trade in both goods and services within the regional trading blocs in SSA. The paper also finds that the EPAs provide an opportunity to accelerate regional and global trade integration in SSA. To realise the potential development benefits of the planned EPAs, two steps are essential. First, the EU must, as it has stated, truly treat the EPAs as instruments of development, subordinating its commercial interests in the agreements to the development needs of SSA. Second, the SSA countries need to implement a number of EPA‐related trade policy reforms. However, the latter is far from certain, given the lack of reform momentum in SSA.
Trade Policy in Burundi: Reform Without Political Stability
Authors: Milner Chris
Published: October 2004

This paper evaluates Burundi's progress with trade policy reform, by comparing earlier analyses of Burundi's trade policies undertaken in the 1980s with that of the WTO's recent Trade Policy Review. Since the mid‐1980s Burundi has been trying to reform its trade and macroeconomic policies against the background of continuous socio‐political tensions and periodic outbreaks of violent tribal conflict. A ‘then and now’ comparison allows us to assess both the extent of the trade reforms and of the economic return to those reforms. It is evident that there has been a significant rationalisation and simplification of trade policy. Burundi has eliminated most quantitative import restrictions and reduced the average level and range of its tariffs. The scope for allocative distortions, undesirable redistributive effects and for impediments to investment and growth has been substantially reduced. However, a return to reform in terms of export growth or diversification and of overall economic growth is not discernible yet. This is unsurprising, given the scale of the economic disruption. Sustained socio‐political stability, among other things, will be required to induce the investment in human and physical capital needed for a positive return to trade policy reform.
Regionalism Within Multilateralism: The WTO Trade Policy Review of Canada
Authors: Head Keith , Ries John
Published: October 2004

The World Trade Organisation published a Trade Policy Review of Canada in 2003. In this paper, we discuss the WTO Review and augment the discussion by presenting original data and reviewing the empirical literature. The WTO concludes that Canada's trade regime is open and transparent but maintains barriers in a few important sectors. We subject this claim to empirical scrutiny, comparing Canada's actual imports to a multilateral benchmark based on the gravity equation. We show that Canada imports about what should be expected given the size of its economy and its location. In a second benchmarking exercise, we show that Canada's anti‐dumping initiations are in proportion to its imports and that Canada's exports are targeted less by other countries’ anti‐dumping investigations than what might be expected based on Canadian export levels. Like many other countries, Canada has pursued trade liberalisation through the World Trade Organisation while simultaneously signing multiple regional trade agreements. Our summary of the recent literature indicates that Canada's regional trade agreements have generated more trade creation than trade diversion. Canada has also spurred imports from the least developed countries by unilaterally eliminating tariffs and quota barriers on 48 of the world's poorest countries in January 2003. We also discuss Canadian progress in opening its agriculture and clothing industries. Overall, we conclude that Canada appears committed to advancing globalisation through multilateral trade liberalisation supplemented by unilateral and bilateral initiatives.
Does the Quality of Institutions Limit the MENA's Integration in the World Economy?
Authors: Méon Pierre‐Guillaume , Sekkat Khalid
Published: October 2004

Using a large sample of countries over the 1990s, this paper examines the extent to which institutions’ functioning disables a greater participation of the Middle East and North Africa (MENA) in the world economy. It focuses on the impact on manufactured exports and FDI attractiveness and considers a broad index of political risk as well as indices targeted toward specific aspects of governance (corruption, government effectiveness and the rule of law). The results are robust to different econometric approaches and lend strong support to the hypothesis that the functioning of institutions may disable the participation of MENA countries in the world economy. They suggest that the impact of an improvement in the quality of institutions may result in a sensitive increase of FDI inflows and manufactured exports. That increase is comparable to the one resulting from liberalisation policies. Hence, although institutional reforms can take time, they deserve the necessary efforts given their outcomes as compared to other reforms.
Rules of Origin and the EU‐Med Partnership: The Case of Textiles
Authors: Augier Patricia , Gasiorek Michael , Lai‐Tong Charles
Published: October 2004

This paper analyses the impact of rules of origin on patterns of trade in the context of the pan‐European system of diagonal cumulation. The paper first highlights the importance of rules of origin in all preferential trading arrangements while arguing that those rules can easily lead to trade suppression and/or trade diversion. We then focus on the introduction of the pan‐European system in 1997 and show evidence to suggest that the introduction of the system materially impacted on trade between the EU, and its CEFTA, EFTA and Baltic states partner countries. The main body of the paper then empirically explores the impact of the lack of cumulation in the textile industry on the countries of the Southern Mediterranean. The results suggest that rules of origin may indeed substantially constrain trade between non‐cumulating countries, possibly by as much as 70–80 per cent in aggregate. While preferential trading agreements thus serve to increase intra‐PTA trade through the liberalisation of trade barriers, they may also be doing so by effectively raising external barriers to trade through the use of constraining rules of origin. To the extent that they do so increases the likelihood of trade diversion and trade suppresion.
Nicotine enemas for treatment of ulcerative colitis: a study of the pharmacokinetics and adverse events associated with three doses of nicotine
Authors: Ingram J. R. , Routledge P. , Rhodes J. , Marshall R. W. , Buss D. C. , Evans B. K. , Feyerabend C. , Thomas G. A. O.
Published: October 2004

Background : Transdermal nicotine is of value in active ulcerative colitis but causes adverse events because of systemic absorption. Nicotine enemas may give rise to fewer adverse events.

Aim : To assess the pharmacokinetics of nicotine enemas in three doses.

Methods : Thirteen volunteers, all non‐smokers but three ex‐smokers, were given enemas on separate occasions containing 3, 6 and 9 mg of nicotine, in ascending dose order. Adverse events were recorded and blood samples taken over 8 h for measurement of serum nicotine and cotinine.

Results : Enemas were retained by most subjects. Eleven of 14 adverse events were ‘early’– 30–105 min after the enema, corresponding to maximum plasma nicotine concentrations; three events were later, 4–8 h after the enema and unrelated to the tmax. ‘Early’ adverse events occurred in eight subjects – six with 9 mg. The three highest plasma nicotine concentrations were with 9 mg and associated with headache, nausea and sweating. Only one had adverse events with 3 mg and withdrew from the study. Nicotine Cmax with 6 and 9 mg doses were respectively two and three times the value with 3 mg. Peak nicotine concentrations occurred 44–50 min after the enema.

Conclusion : The 6 mg dose of nicotine probably represents the dose to use in clinical practice – for the highest therapeutic dose with a low risk of adverse events.
Effect of xenon anaesthesia on accuracy of cardiac output measurement using partial CO2 rebreathing
Authors: Bein B. , Hanne P. , Hanss R. , Renner J. , Weber B. , Steinfath M. , Scholz J. , Tonner P. H.
Published: October 2004

Cardiac output (CO) determination based on partial CO2 rebreathing has recently been introduced into clinical practice. The determination of flow is crucial for exact CO readings and the physical properties of xenon, i.e. high density and viscosity, may influence flow readings. This study compared echocardiography‐derived CO measurements with the partial rebreathing method during total intravenous (TIVA) vs. xenon‐based anaesthesia. Thirty‐nine patients ASA physical status III undergoing aortic reconstruction were randomly assigned to receive either xenon (Xe, n = 20) or TIVA (T, n = 19) based general anaesthetic. Paired measurements were taken before xenon administration, after xenon administration, before and after clamping of the abdominal aorta and after declamping and at corresponding time points in the TIVA group. Data were analysed with a Bland‐Altmann plot. Bias and precision were acceptable and comparable before xenon administration (T 0.54 ± 0.92 l.min−1 vs. Xe 0.11 ± 1.1 l.min−1), but after xenon administration CO was largely overestimated by partial CO2 rebreathing (T 0.04 ± 0.91 l.min−1 vs. Xe −4.0 ± 2.1 l.min−1). In the TIVA group, bias and precision after declamping increased significantly (P < 0.01) compared to all time points except baseline. In its current application, the NICO cardiac output monitor appears to be inappropriate for determination of CO during xenon based anaesthesia.
The target joint
Authors: Mulder K. , Llinás A.
Published: October 2004

Summary.   Bleeding into the joints is the most common manifestation of severe haemophiliacs. Although it may resolve spontaneously or with treatment, some patients find that one particular joint has recurrent bleeding; this is termed a target joint. Recurrent bleeding prevents the joint from regaining its range of motion, muscle strength and normal appearance. These changes become permanent, leading eventually to osteoarthritis. A target joint requires urgent and comprehensive treatment, especially in young patients, if permanent damage is to be prevented. Treatment with factor concentrate prophylaxis and physiotherapy can help to prevent new bleeds and allow the synovitis to resolve, but for persistent synovitis, synovectomy is recommended. The target ankle joint is a special challenge as it often develops in very young children when the articular cartilage is susceptible and compliance with conservative treatment is difficult.
von Willebrand disease: laboratory aspects of diagnosis and treatment
Authors: Favaloro E. J. , Lillicrap D. , Lazzari M. A. , Cattaneo M. , Mazurier C. , Woods A. , Meschengieser S. , Blanco A. , Kempfer A. C. , Hubbard A. , Chang A.
Published: October 2004

Summary. 
von Willebrand disease is the most common inherited bleeding disorder in humans. VWD can be classified into three major types, designated Types 1, 2 and 3; Type 2 can be further separated into subtypes 2A, 2B, 2M and 2N. The diagnosis of VWD requires a personal and family history of bleeding and confirmation by laboratory analysis. Although Types 2 and 3 are relatively straightforward to diagnose, there may be a risk of overdiagnosis of Type 1 because of an overlap within the normal range. We also report on the clinical profile and diagnosis of VWD in a South American cohort of patients and on the in vitro characteristics of some factor concentrates available for treatment of VWD.
Hepatitis C in haemophilia: lights and shadows
Authors: Rumi M. G. , De Filippi F. , Santagostino E. , Colombo M.
Published: October 2004

Summary.  Hepatitis C is a major cause of morbidity and mortality in haemophiliacs who received clotting factor concentrates before the availability of virus‐inactivated factors in the mid‐1980s. Early studies gave conflicting indications as to the severity of hepatitis C (originally termed non‐A non‐B hepatitis), as mild, slowly progressive hepatitis was documented in several infants and young adults with haemophilia who were examined with repeat liver biopsies, whereas more progressive hepatitis and cirrhosis was documented in others. One major point of dispute was whether these discrepancies could in part be accounted for by epidemiological differences among studies, as hepatitis C acquired early in life may initially run a benign course and later worsen owing to spontaneous recrudescence of hepatitis or interference with such comorbidity factors as alcohol abuse or infection with the human immunodeficiency virus (HIV). In the mid 1990s, the latter infection overshadowed hepatitis C as a cause of death in this patient population. Because hepatocellular carcinoma is emerging as an important complication in haemophiliacs with long‐standing hepatitis C virus (HCV) infection who survived HIV infection, and because of recent advances in treating HIV, morbidity and mortality associated with chronic hepatitis C have regained emphasis amongst haemophiliacs. The development of newer interferon‐based therapies provides an opportunity for modifying the natural history of HCV infection in a substantial number of haemophilic patients.
Variant Creutzfeldt–Jakob disease: risk of transmission by blood and blood products
Authors: Ironside J. W. , Head M. W.
Published: October 2004

Summary.  Variant Creutzfeldt–Jakob disease (CJD) is a novel acquired human prion disease apparently resulting from exposure to the bovine spongiform encephalopathy (BSE) agent. Variant CJD differs from other human prion diseases in that the disease‐associated form of the prion protein and infectivity are readily detectable in lymphoid tissues throughout the body. Lymphoid tissues and lymphocytes are implicated in the peripheral pathogenesis of prion diseases (where infectivity may be detected during the preclinical phase of the illness), giving rise to concerns that blood and blood products may also contain infectious particles, representing a possible source of iatrogenic spread of variant CJD. This concern has been reinforced following the experimental transmission of BSE in a sheep model by transfusion of blood and buffy coat from animals in the preclinical phase of the illness, and the recent identification of a UK case of variant CJD in a patient who had received packed red blood cells that had been donated by an individual who subsequently died from variant CJD. Studies in animal models suggest that most prion infectivity in blood may be cell‐associated, with lower levels in the plasma, and there is evidence to suggest that any infectivity present may be reduced during the process of plasma fractionation. However, the possibility that plasma or blood products could transmit the disease cannot be excluded. Further studies are required to develop more sensitive means to detect disease‐associated prion protein in blood; such techniques could be employed for screening purposes to reduce exposure to contaminated products and to assist with risk management in potentially exposed individuals.
Haemophilic factors produced by transgenic livestock: abundance that can enable alternative therapies worldwide
Authors: Van Cott K. E. , Monahan P. E. , Nichols T. C. , Velander W. H.
Published: October 2004

Summary.  Haemophilia replacement factors, both plasma‐derived and recombinant, are in relatively short supply and are high‐cost products. This has stymied the study and development of alternative methods of administration of haemophilia therapy even in the most economically advanced countries, owing to the large amounts of material needed because bioabsorption and bioavailability of haemophilic factors can be less than 10% when using non‐intravenous routes of delivery. There is therefore a need to increase access to therapy worldwide by decreasing the cost and increasing the abundance so that therapy can be achieved through simplified, alternative delivery methods. Transgenic livestock have been used to produce haemophilic factors in milk. Only the pig mammary gland has been shown to carry out the post‐translational processing necessary to enable both the biological activity and long circulation half‐life needed for therapeutic glycoproteins. Furthermore, the large amounts of recombinant protein that can be produced from pig milk make feasible the use of alternative delivery methods such as oral, intratracheal, subcutaneous, and intramuscular administration.
Monitoring haemophilia severity and treatment: new or old laboratory tests?
Authors: Barrowcliffe T. W.
Published: October 2004

Summary. Testing of factor levels is desirable for a number of reasons, including diagnosis of the level of severity of the disease, and the efficacy of factor‐replacement therapy. The older methods comprised the one‐stage, two‐stage, and chromogenic methods, and newer methods, including the thrombin generation test, are now available. This paper outlines the use of these methods and problems associated with them.
Haemophilia in the developing world: successes, frustrations and opportunities
Authors: Tezanos Pinto M. , Ortiz Z.
Published: October 2004

Summary.  There is a need to bridge key gaps between high‐ and low‐income countries and individuals; between health policy and clinical practice; and between producers and users of healthcare technology and scientific evidence. The objective of this report was to perform a diagnosis of the situation in the developing world through a survey. This survey was conducted to gather specific information on various aspects related to haemophilia. Countries were chosen by their comparability in infant and adult mortality rates according to the regionalization proposed by the World Health Organization. These indicators are very sensitive to socioeconomic conditions, and have been widely used to study health inequalities. All regions, except Africa and the East Mediterranean, were represented. Africa was excluded because its indicators were not comparable. The East Mediterranean was not represented because of difficulties in contacting investigators. Twenty‐one country representatives were contacted, and 11 answered the questionnaire. Successes obtained by developing countries are based more on the skills and creativity of the local professionals than on the availability of state‐of‐the‐art technology. Frustrations were related to disease underregistration and the limited availability of treatment products. Haemophilia care in the developing world is not as fair as we would like it to be. Governments do not always cover treatment costs, and a very small percentage of the national health budgets is alotted to haemophilia care. The role of the World Federation of Hemophilia was considered crucial by all the investigators surveyed. Training programmes and supply of factor concentrates were the main contributions identified by the respondents.
Towards the goal of prophylaxis: experience and treatment strategies from Sweden, France and Hungary
Authors: Petrini P. , Chambost H. , Nemes L.
Published: October 2004

Summary.  The only form of haemophilia treatment that is able to prevent arthropathy and other consequences of bleeding symptoms in patients with severe haemophilia is prophylaxis started at an early age (primary prophylaxis). It is also highly beneficial for the psychological and social wellbeing of patients and their families. Scientific institutions and international organizations such as WHO, the World Federation of Hemophilia (WFH) and the National Haemophilia Foundation (NHF) have recommended that prophylaxis be considered optimum therapy. This paper discusses the barriers to prophylaxis, such as the perceived need, costs and availability, and difficulty of venous access, and describes the authors' experiences with the therapy.
Essential issues of laboratory investigation for patients with haemophilia and bleeding disorders
Authors: Chuansumrit A. , McCraw A. , Preston E. F.
Published: October 2004

Summary.  Apart from history‐taking and physical examination, laboratory investigation is one of the essential issues for the definite diagnosis of haemophilia and bleeding disorders. The limited resources of medical personnel, equipment and reagents should be shared among several departments in the hospital, especially for serving patients with common genetic diseases such as thalassemia and haemoglobinopathies. Medical personnel require appropriate training to expand their skills in laboratory techniques. Laboratory procedures can be created, modified and simplified using locally produced and shared equipment. Molecular genetic studies can also be set up at different levels of hospital service using simple, rapid and low‐cost methods. Finally, a system of periodic external quality control will guarantee the accuracy of laboratory results.
Clinical diagnosis of von Willebrand disease
Authors: Federici A. B.
Published: October 2004

Summary.  von Willebrand disease (VWD) is the most frequent inherited bleeding disorder and is caused by quantitative (Types 1 and 3) or qualitative (Type 2) defects of von Willebrand factor (VWF). VWD is inherited by autosomal dominant or recessive pattern, but women with milder VWD forms seem to be more symptomatic than men. Mild VWD forms are both under‐ and misdiagnosed. The clinical expression of VWD is usually mild in Type 1, increasing in severity in Types 2 and 3. Mucocutaneous bleeding (epistaxis, menorrhagia) is a typical manifestation of the disease, and bleeding after dental extraction is the most frequent postoperative bleeding type. Because FVIII levels are usually only slightly reduced in most VWD types, spontaneous haemarthroses or haematomas are rare in VWD Types 1, 2A and 2B, whereas in Type 3 the severity of bleeding may resemble haemophilia. In Type 1 VWD, bleeding after delivery is rare because FVIII/VWF levels become normal at the end of pregnancy. Post‐operative bleeding may not occur in Type 1 VWD patients, but in Type 3 VWD, prophylaxis is always required. Only a few retrospective studies on clinical diagnosis of VWD are available. In the 1234 cases enrolled by an Italian retrospective study, diagnosis of Types 1, 2 and 3 VWD occurred in young adults (83%), mainly in women (57%). Mucosal bleeding (64%) was more frequent than haematomas or haemarthrosis (15%), and 63% of patients did not require transfusions. In a more recent Italian prospective study (815/1234 cases observed for 1 year in 6/16 Italian centres), only 147 (18%) VWD patients showed bleeding episodes (n = 318) and minor or major surgeries (n = 87).
Inhibitors in haemophilia: clinical aspects
Authors: Dimichele D. , Rivard G. , Hay C. , Antunes S.
Published: October 2004

Summary.  Inhibitor development continues to be a severe complication worldwide of haemophilia therapy. Given the difficulties associated with the treatment of inhibitors in both the developed and the developing world, prediction and prevention of inhibitors following exposure to factor VIII or IX in the young child has become a management priority. Two different clinical approaches to minimizing or delaying inhibitor development are highlighted in this manuscript. Challenges in the therapy of existing inhibitors persist. However, the nature of the issues faced by physicians in the developed and developing worlds vary considerably. Both are discussed in this manuscript by representatives of both worlds, who face these challenges on a daily basis.
The impact of information technology on haemophilia care
Authors: Baker R. I. , Laurenson L. , Winter M. , Pritchard A. M.
Published: October 2004

Summary.  The past decade has seen an explosion in the use of computers, data programs, hand‐held electronic devices and the Internet. How these advances impact on haemophilia management both now and in the future are discussed from the perspective of haemophilia registries, Internet‐based electronic haemophilia treatment records, and the potential for haemophilia telehealth.
Models for psychosocial services in the developed and developing world
Authors: Spilsbury M.
Published: October 2004

Summary.  This paper is based on the assumption that psychosocial services can add important mental and social dimensions to the traditional concepts of help and medicine. Peer support which has been established world‐wide for people with haemophilia and their families is invaluable and irreplaceable. However the role of the professional psychosocial worker is underutilized in most countries, mostly because of lack of financial resources. This paper will argue that skilled workers can assist individuals and groups to reach their full potential in a variety of creative and non‐threatening ways. Psychosocial workers have strict codes of conduct and requirements relating to accreditation to ensure accountability in their work. They can work in a variety of individual roles and have a wide range of individual mandates within work settings.
Disease‐specific quality‐of‐life measurement tools for haemophilia patients
Authors: Remor E. , Young N. L. , Von Mackensen S. , Lopatina E. G.
Published: October 2004

Summary.  The purpose of this paper is to summarize the state of the art in measuring quality of life in haemophila populations. The paper reviews the measures recently included in haemophila trials in the published literature. It also summarizes the development of four new disease‐specific measures of health‐related quality of life. Two of these were developed for children (the Haemo‐QoL and the CHO‐KLAT), and two for adults (the Hemofilia‐QoL and the Hemolatin‐QoL). These new measures show promise for use in clinical trials. Further research is in progress to complete the psychometric testing and cross‐cultural validation.