bluuebunny/crossref_metadata_2025 huggingface dataset:
Intravasal microdialysis is superior to intramyocardial microdialysis in detecting local ischaemia in experimental porcine myocardial infarction
Authors: Bäckström T. , Franco‐Cereceda A.
Published: January 2004

Objective:  A novel application of microdialysis was studied, where myocardial outflow of energy metabolites was monitored by intravasal microdialysis in the myocardial venous outflow during ischaemia and reperfusion. These levels where related to levels monitored by microdialysis catheters placed intramyocardially.

Methods:  Microdialysis catheters were introduced into the great cardiac vein (GCV), ischaemic myocardium and non‐ischaemic myocardium in 10 anaesthetized pigs. The left anterior descending coronary artery was occluded for 60 min in five pigs and five pigs served as controls. Ischaemia was followed by 120 min of reperfusion. Microdialysis samples were analysed for glucose, lactate, pyruvate and glycerol. Venous lactate and glucose levels were measured by blood samples from the femoral vein.

Results:  All animals subjected to ischaemia developed myocardial infarction. Lactate, lactate/pyruvate ratio and glycerol increased in the microdialysis samples from the GCV and the catheter placed in ischaemic myocardium while no changes were detected in samples from the catheter placed in the non‐ischaemic myocardium.

Conclusion:  In this study, we have demonstrated that intravasal microdialysis catheters rapidly and reliably detect local myocardial ischaemia, while intramyocardially placed microdialysis catheters will not show these changes if placed in a non‐ischaemic area.
Stimulation of HSP72 expression following ATP depletion and short‐term exercise training in fast‐twitch muscle
Authors: Ecochard L. , Roussel D. , Sempore B. , Favier R.
Published: January 2004

Aim:  Previous data have reported increases in HSP72 expression in skeletal muscles after endurance training but the physiological and biochemical signals that induce HSP72 accumulation remain largely unknown. In this study, we tested the hypothesis that energy status is a key regulatory event for HSP72 accumulation in skeletal muscles.

Methods:  Reduction of high‐energy phosphate levels was induced by supplementation with a creatine analogue, beta‐guanidinopropionic acid (GPA) for 3 weeks while control rats received distilled water in the same conditions. Half of the animals were kept sedentary while the others were submitted to a short‐term (2 weeks) training program on a treadmill (30 m min−1, 0% slope; 50–70 min day−1).

Results:  GPA supplementation resulted in a large drop (∼50%) in adenosine triphosphate (ATP) level in both fast and slow muscles whether the animals were trained or remained sedentary. HSP72 level did not change with GPA alone, but the training‐induced increase in HSP72 level was strongly enhanced by superimposition of GPA diet in fast but not in slow skeletal muscles. The changes in HSP72 level were not linked to changes in fibre typology and/or mitochondrial capacities.

Conclusions:  The results of the present investigation indicate that levels of high‐energy phosphate per se do not play a direct role in determining HSP72 level in skeletal muscles. However, during superimposition of training to GPA, then the adaptive strategy of fast‐twitch muscle (e.g. plantaris) seems to be directed towards appearance of some properties of red, oxidative fibres (increase in oxidative capacities and HSP72 level).
Effect of anaesthetic and rat strain on heart rate responses to simulated haemorrhage
Authors: Holobotovskyy V. V. , Arnolda L. F. , McKitrick D. J.
Published: January 2004

Aim and methods:  Haemorrhage is characterized by two distinct responses, sympathoexcitation that evokes tachycardia and supports blood pressure, followed by sympathoinhibition contributing to bradycardia and hypotension. It has been shown that anaesthetics alter the response to haemorrhage and we hypothesized that rat strain may also influence the response. We investigated the effect of simulated haemorrhage on heart rate (HR) responses in three strains of conscious rats, and the effect of three common anaesthetics, by comparing HR responses under anaesthesia to the conscious response. Haemorrhage was simulated by constricting the inferior vena cava. We demonstrate differential effects of anaesthetics, including both maintenance and elimination of HR responses to haemorrhage depending on anaesthetic.

Results:  We also show that both phases of the HR response differ in different conscious rat strains, and we have demonstrated a transient increase in HR during the decompensatory phase of haemorrhage, a novel ‘second HR peak’ with advanced hypotension.

Conclusion:  Both rat strain and anaesthetic influence HR responses to haemorrhage, and some anaesthetics appear less suitable than others for studies of haemodynamic responses in rats. There was evidence of an additional compensatory mechanism that operates at advanced levels of hypotension in the rat.
Sweating responses during activation of the muscle metaboreflex in humans is altered by time of day
Authors: Aoki K. , Kondo N. , Shimomura Y. , Yokoi M. , Iwanaga K. , Harada H. , Katsuura T.
Published: January 2004

Aim: The aim of the present study was to test for a time‐of‐day effect on sweating responses to activation of the muscle metaboreflex.

Methods: Eight male subjects each participated in two exercise sessions, one in the morning and one in the evening. Within each session there were two 60‐s bouts of isometric handgrip (IHG) exercise at 50% maximal voluntary contraction. Prior to IHG, whole body warming by a water‐perfused suit initiated mild sweating. The first bout of IHG exercise began at 06.00 hours (am) and 18.00 hours (pm). Blood circulation to the forearm was occluded for 120 s, beginning 5 s before the end of the second bout of IHG to activate the muscle metaboreflex.

Results: During both bouts of exercise, sweating rate (SR) on both the chest and right forearm significantly increased from the pre‐exercise period in bothamandpmsessions. SR rapidly decreased during first minute of recovery after the first bout of IHG exercise. However, during post‐exercise ischaemia (PEI) after the second bout of IHG exercise, SR was maintained significantly above the pre‐exercise level only in thepmsession. The increases in SR on the chest and right forearm during PEI were significantly greater in thepm, than in theam, session. However, SR of the palm was not maintained during PEI.

Conclusions: We conclude that under mild hyperthermic conditions, the sweating response in non‐glabrous skin to activation of the muscle metaboreflex exhibits a time‐of‐day effect.
Reopening of L‐type calcium channels in human ventricular myocytes during applied epicardial action potentials
Authors: Fülöp L. , Bányász T. , Magyar J. , Szentandrássy N. , Varró A. , Nánási P. P.
Published: January 2004

Aims:  Present study was performed to compare the dynamics of human L‐type calcium current (ICa,L) flowing during rectangular voltage pulses, voltage ramps, and action potentials (APs) recorded from epicardiac and endocardiac canine ventricular cells.

Methods:  I
Ca,L was recorded in single myocytes isolated from undiseased human hearts using the whole cell voltage clamp technique.

Results:  The decay of ICa,L was monotonic when using rectangular pulses or endocardial APs as voltage commands, whereas the current became double‐peaked (displaying a second rise and fall) during epicardial (EPI) APs or voltage ramps used to mimic EPI APs. These ICa,L profiles were associated with single‐hooked and double‐hooked phase‐plane trajectories, respectively. No sustained current was observed during the AP commands. Kinetics of deactivation and recovery from inactivation of human ICa,L were determined using twin‐pulse voltage protocols and voltage ramps, and the results were similar to those obtained previously in canine cells under identical experimental conditions.

Conclusions:  I
Ca,L can inactivate partially before and deactivate during the phase‐1 repolarization of the epicardiac AP, and reopening of these channels seems to be associated with formation of the dome.
Stretch reflex excitability of the anti‐gravity ankle extensor muscle in elderly humans
Authors: Kawashima N. , Nakazawa K. , Yamamoto S.‐I. , Nozaki D. , Akai M. , Yano H.
Published: January 2004

Aim:  To examine whether the stretch reflex excitability of the soleus muscle changes with age, stretch reflexes at rest (REST) and during weak voluntary contractions (ACT) were elicited in 18 older and 14 younger subjects.

Method:  The amplitude of the stretch reflex responses and gain, defined as the gradient of the regression line for the relation between stretch reflex responses against the angular velocity of the applied perturbation, were evaluated in each short‐latency (M1) and two long‐latency components (M2 and M3).

Results:  It was found that in the older group, both the amplitude and gain of the M1 component did not change from the REST to the ACT conditions, whereas in the younger group both variables significantly increased from the REST to ACT conditions. The latency of the M1 component was significantly shorter under the REST condition (older vs. younger: 51.8 ± 7.37 vs. 55.1 ± 8.69 ms), while no group differences were found in those variables under the ACT condition, suggesting that the muscle‐tendon complexes of SOL muscles of the older subjects were less elastic and had less slack, probably due to age‐related histochemical alterations. Further, the Hoffman reflex (H‐reflex), elicited during the REST condition in 10 older and 11 younger subjects showed no significant differences, suggesting that the soleus motoneuron response to the Ia input was comparable between the two subject groups.

Conclusion:  The histochemical alterations occurring with the ageing process might augment the short‐latency stretch reflex in the SOL muscle without enhancement of motoneuronal excitability, and this effect might be masked when the muscle is voluntarily activated.
Effective high‐dose chemotherapy combined with CD34+‐selected peripheral blood stem cell transplantation in a patient with cutaneous involvement of nasal NK/T‐cell lymphoma
Authors: Koizumi Kazuki , Fujimoto Katsuya , Haseyama Yoshihito , Endo Tomoyuki , Nishio Mitsufumi , Yokota Kouichi , Itoh Tomoo , Sawada Ken‐ichi , Koike Takao
Published: January 2004

The prognosis of nasal natural killer (NK)/T‐cell lymphoma with cutaneous involvement especially is morbid despite intensive chemotherapy and radiotherapy. We treated a 52‐yr‐old Japanese woman with cutaneous dissemination of nasal NK/T‐cell lymphoma. Six cycles of chemotherapy, irradiation to skin lesion were administered and complete remission (CR) was attained. High‐dose chemotherapy (HDC; etoposide 750 mg/m2 × 2 d, cyclophosphamide 60 mg/kg × 2 d, total body irradiation 12 Gy two daily fractions × 3 d) followed by CD34+‐selected autologous peripheral blood stem cell transplantation (CD34+‐APBSCT) was then prescribed. Complete remission (CR) was obtained and she has been free of disease for 34 months since CD34+‐APBSCT. We suggest that marrow‐ablative chemotherapy facilitated by autologous stem cell transplantation should be considered part of the primary therapy for subjects with a poor prognosis for nasal NK/T‐cell lymphoma with cutaneous involvement.
Quality of life and brain function following high‐dose recombinant human erythropoietin in low‐risk myelodysplastic syndromes: a preliminary report
Authors: Clavio Marino , Nobili Flavio , Balleari Enrico , Girtler Nicola , Ballerini Filippo , Vitali Paolo , Rosati Paola , Venturino Claudia , Varaldo Riccardo , Gobbi Marco , Ghio Riccardo , Rodriguez Guido
Published: January 2004

Objective:  In this prospective study we evaluate the effects of high‐dose recombinant human erythropoietin (rHuEPO) on quality of life (QOL) and brain function in patients with low‐risk myelodysplastic syndromes (MDS) (<10% marrow blasts). Preliminary data are reported.

Methods:  Eleven consecutive patients were given rHuEPO (40 000 IU two times a week) for 12 wk. Responsive patients continued with 40 000 IU/wk for further 12 wk. Changes in QOL were assessed by the Functional Assessment of Cancer Therapy‐Anemia (FACT‐An) self‐report. Neurophysiological evaluation at the start of the therapy (t0) included duplex scanning of neck vessels, transcranial Doppler sonography (TCD), a complex neuropsychological evaluation, and quantitative electroencephalography (qEEG). Eight patients completed the neurophysiological evaluation after 24 wk (t1).

Results:  Six patients (55%) achieved an erythroid response after 12 wk, which was maintained after 24 wk of treatment. FACT‐An score showed a relevant improvement between t0 and t1 in these patients. At baseline, TCD showed a mean cerebral blood flow (CBF) velocity in the upper normal range. Abnormalities in brain function were observed in five patients. In the eight patients who were re‐evaluated at t1, improvement was observed in three responding patients, two of them with abnormal values at t0. A strict correlation between QOL and neurophysiological improvements was not observed.

Conclusions:  A high‐dose induction phase with rHuEPO followed by maintenance therapy may be an effective therapeutic schedule for low‐risk MDS patients. The erythroid response was associated with positive changes in the QOL. Neurophysiological improvements occurred only in a part (50%) of responding patients, mainly those who showed altered results at baseline.
Optimizing donor selection in a cord blood bank
Authors: Solves Pilar , Perales Alfredo , Mirabet Vicente , Blasco Ignacio , Blanquer Amando , Planelles D. , Larrea Luis , Monleon Javier , Carbonell‐Uberos F. , Angeles Soler M.
Published: January 2004

Objectives:  The main limitation factor for the wide use of umbilical cord blood (UCB) as a source of hematopoietic progenitor for transplantation is cell dose. One of the specific areas identified by some studies for improvement of UCB collection is donor selection.

Methods:  Over a 3–mth period, 391 consecutive maternal‐neonatal pairs were evaluated during the pre–partum period in the maternity ward at La Fe University Hospital (Valencia) by the Cord Blood Bank staff. Reasons for discarding umbilical cord blood donors and collected UCB units at the Cord Blood Bank in Valencia have been analysed. Obstetric factors influencing TNC content of 1300 collected UCB units have been determined, in order to establish obstetric criteria for cord blood donors selection in our geographic area.

Results:  Only 32.5% of potential cord blood donors were refused. Among 1300 UCB collected, 506 (38.9%) were discarded before cryopreservation, mainly due to low cell counts. Multivariate analyses showed that the main significant factors influencing nucleated cell count were the weight of the placenta, sex of newborn and mode of collection.

Conclusions:  Our study shows that maternal medical histories must be completely reviewed by medical staff before collection of the UCB. Obstetrical factors influence cell content of UCB and could be added to standard cord blood donor criteria in order to improve the bank efficiency.
Possible role of cytokine‐induced tryptophan degradation in anaemia of inflammation
Authors: Weiss Guenter , Schroecksnadel Katharina , Mattle Verena , Winkler Christiane , Konwalinka Guenther , Fuchs Dietmar
Published: January 2004

Anaemia of inflammation (AI) is a frequent complication in patients suffering from chronic inflammatory disorders including infections, autoimmune and malignant disease. Cytokine imbalance with a shift towards T‐helper (Th)1‐type immune response seems to be important in the pathogenesis of this type of anaemia. Interferon‐γ (IFN‐γ) and tumour necrosis factor‐α may affect the growth and differentiation of erythroid progenitor cells. In macrophages, IFN‐γ strongly induces indoleamine (2,3)‐dioxygenase, an enzyme which degrades tryptophan (trp) to kynurenine (kyn). Trp availability is rate limiting for protein biosynthesis and thus cell growth, including erythropoiesis. In this study, trp and kyn concentrations and their relationship to haemoglobin concentrations and to immune activation was examined in 22 patients with AI. Patients with AI presented with lower trp concentrations than healthy controls of similar age, and a significantly higher kyn to trp ratio, suggesting enhanced trp degradation and, because of a positive correlation with neopterin, immune activation. The kyn to trp ratio was inversely correlated to haemoglobin levels. Thus, the limitation of trp availability to erythroid progenitors may be a key mechanism in cytokine‐mediated inhibition of erythropoiesis, and the therapeutic modulation of indoleamine (2,3)‐dioxygenase and trp levels may be promising targets for AI therapy.
Successful treatment for patients with von Willebrand disease undergoing urgent surgery using factor VIII/VWF concentrate (Humate‐P®)
Authors: Thompson A. R. , Gill J. C. , Ewenstein B. M. , Mueller‐Velten G. , Schwartz B. A. ,   for the Humate‐P Study Group
Published: January 2004

Summary.  von Willebrand disease (VWD) is characterized by insufficient von Willebrand factor (VWF) activity. It has been proposed that VWF:ristocetin cofactor (VWF:RCo) activity may be useful in evaluating the response to VWD treatment in patients who require replacement therapy. This prospective, open‐label, non‐randomized study evaluated the safety and efficacy of a factor VIII (FVIII)/VWF concentrate (Humate‐P®) used in treatment regimens based on VWF:RCo activity in subjects with VWD in situations requiring urgent and necessary surgery. This article summarizes the results for 39 subjects with 42 evaluable surgical treatment events, 100% of which were rated as excellent/good for overall efficacy (achievement of haemostasis). The median loading dose based upon VWF:RCo activity was 82.3 international units/kilogram (IU kg−1; range 32.5–216.8 IU kg−1), and the median maintenance dose per infusion was 52.8 IU kg−1 (range 24.2–196.5 IU kg−1) for a median of 3 days (range 1–50 days). The median number of infusions per event was 6 (range 1–67 infusions). Three unanticipated adverse events (peripheral oedema, extremity pain and pseudo‐thrombocytopenia) from two surgical treatment events were reported that were potentially treatment‐related. No serious drug‐related adverse events (AEs) were observed, and no thrombotic events were reported in this study. This study supports the safety and efficacy of the FVIII/VWF concentrate Humate‐P® for the prevention of surgical haemorrhage in patients with VWD when administered in doses calculated in VWF:RCo units.
Frequency and causes of hospitalization in HIV‐negative children and adolescents with haemophilia A or B and its effect on academic achievement
Authors: Wong W. Y. , Donfield S. M. , Rains E. , FitzGerald G. , Pearson S. K. , Gomperts E. D. ,   the Hemophilia Growth and Development Study
Published: January 2004

Summary.  The aim of the study is to determine the causes and frequency of hospitalization in HIV‐negative boys and adolescents with haemophilia and evaluate their impact on academic achievement. One hundred and twenty‐six HIV‐negative boys and adolescents were followed prospectively from 1989–96, at 14 comprehensive haemophilia treatment centres. One hundred and fifteen participants with haemophilia A or B were included in the investigation. These participants contributed an average of 57.8 months of follow‐up. There were 203 hospitalizations in 65 participants and 50 participants were never hospitalized. Haemarthroses and soft tissue bleeds accounted for 46 and 44 causes of hospitalization. Central line infection was the third most common cause. Participants with inhibitor had the majority of central line infections and hospitalizations. Intracranial haemorrhage resulted in five hospitalizations in two participants. Other causes of bleeding accounted for 22% of hospitalizations. The median number of hospitalizations per year was 0.18. Duration of hospital stay was significantly related to lower spelling scores. Acute and chronic joint problems and soft tissue bleeds still account for the majority of hospitalizations. Positive inhibitor status was associated with higher numbers of hospitalizations and central line infections. Academic achievement was affected, to some degree, by length of hospital stay.
HIV and HCV coinfection in haemophilia
Authors: Wilde J. T.
Published: January 2004

Summary.  A substantial number of haemophilic patients are infected with both human immunodeficiency virus (HIV) and hepatitis C (HCV). HIV has been shown to accelerate the course of HCV chronic liver disease and there is evidence that HCV infection may worsen the prognosis of HIV. As many HIV infected patients are stable on highly active antiretroviral therapy (HAART) HCV should be actively managed in coinfected individuals. Pegylated interferon (Peg‐IFN)/ribavirin combination therapy is the treatment of choice for HCV infection and should be considered in patients with stable HIV on or off HAART with CD4 counts >200 × 106/l. Results of on‐going trials of combination therapy in coinfected individuals are awaited. For coinfected patients with end stage liver disease who are stable on HAART liver transplantation should be considered.
Role of epsilon amino caproic acid in the management of haemophilic patients with inhibitors
Authors: Ghosh K. , Shetty S. , Jijina F. , Mohanty D.
Published: January 2004

Summary. We managed bleeding crisis in 10 consecutive severe haemophilic patients with inhibitors (eight had an inhibitor level of >5 BU mL−1) mainly with the antifibrinolytic agent, i.e. epsilon amino caproic acid (EACA). EACA was used by local, oral or intravenous routes either in combination or separately. Five patients developed inhibitors postoperatively and among the remaining five, four had recurrent haemarthrosis or soft tissue bleeds and one patient presented with severe gastrointestinal bleeding without demonstrable lesion. In all the patients, addition of EACA to their management protocol resulted in stoppage and/or reduced frequency of bleeding. In six of 10 patients, the results were excellent; of these six patients, five developed inhibitors postoperatively. Although a reduction in the frequency of bleeding was observed in patients with haemarthrosis and soft tissue bleed, it was not spectacular and the patients required additional therapy. Hence the results could be described as poor. No patient needed to stop the medicine because of the side‐effect of EACA. Symptoms like mild nausea and vertigo were seen as the side‐effects of this medicine when high intravenous dosage was administered. EACA thus appears to be an excellent adjuvant therapy for haemophilic patients with inhibitors. Besides its well‐recognized antifibrinolytic activity, EACA may have additional mechanisms of action in haemophilic patients with inhibitors . More extensive use of this cheap and safe product is warranted in haemophilic patients with inhibitors. If larger studies confirm this observation, then using antifibrinolytics will allow substantial reduction of FEIBA or activated prothrombin complex (APCC) usage in such patients without necessarily increasing the thrombotic complications or reduction of the clinical efficacy, when compared with higher dosage of FEIBA or APCC alone. This will lead to substantial financial savings in countries where up to 35% of severe haemophilia A patients develop inhibitors.
Prothrombin Shanghai: hypoprothrombinaemia caused by substitution of Gla29 by Gly
Authors: Wang W. , Fu Q. , Zhou R. , Wu W. , Ding Q. , Hu Y. , Wang X. , Wang H. , Wang Z.
Published: January 2004

Summary.  Prothrombin deficiency is a rare bleeding disorder inherited as an autosomal recessive trait. In this study, we reported a Chinese family with hereditary prothrombin deficiency. The proposita had a prolonged activated partial thromboplastin time (APTT, 71.6 s) and prothrombin time (PT, 28.0 s). The coagulation factors activities were normal except that prothrombin coagulation activity was markedly reduced, and the prothrombin antigen level was moderately decreased. Nucleotide sequencing of amplified DNA revealed a novel mutation, Glu (GAG) to Gly (GGG) at residue 29, which normally undergoes γ‐carboxylation within the Gla domain of prothrombin. The proposita was identified as homozygous, while her father, mother and maternal grandmother were heterozygous for the mutation. Gla29 has been demonstrated as one of the key residue for Ca2+‐binding, membrane interaction and biological activity of prothrombin.
External fixators in haemophilia
Authors: Lee V. , Srivastava A. , PalaniKumar C. , Daniel A. J. , Mathews V. , Babu N. , Chandy M. , Sundararaj G. D.
Published: January 2004

Summary.  External fixators (EF) are not commonly used for patients with haemophilia. We describe the use of EF (Ilizarov, AO‐ uni‐ and bi‐planar fixators and Charnley clamp) in nine patients (mean age: 19.2 years; range: 9–37) with haemophilia for the following indications – arthrodesis of infected joints, treatment of open fractures and osteoclasis. EF required an average of nine skin punctures [range: 4–17 were maintained for a period of 15 weeks (range: 8–29.5), without regular factor replacement, till bone healing was adequate and were removed with a single dose of factor infusion]. The mean preoperative factor level achieved was 85% (range: 64–102%). Much lower levels were subsequently maintained till wound healing. The average total factor consumption was 430 IU kg−1 (range: 240–870), administered over a period of 17 days (range: 9–44). There were no major complications related to EF except in a patient who developed inhibitors. In conclusion, EF can be used safely in haemophilic patients who do not have inhibitors and does not require prolonged factor replacement.
Hepatitis C virus quasispecies in the natural course of HCV‐related disease in patients with haemophilia
Authors: Tagariello G. , Gerotto M. , Pontisso P. , Belvini D. , Salviato R. , Radossi P. , Alberti A.
Published: January 2004

Summary.  Patients with haemophilia show high prevalence of hepatitis C infection but low rate of progressive liver disease when they are not co‐infected with HIV. The balance between host immune system and hepatitis C virus (HCV) variability seems to play a major role in the evolution of the HCV‐related disease. To address this point we have studied, in a group of selected patients with haemophilia, the composition and in some cases the evolution, of the highest variable envelope gene within the hypervariable region 1 (HVR1) of the HCV, which is the region more directly exposed to the host immune response. Five of 12 patients show a very high homogeneity of the HVR1 and four of those had severe progressive liver disease. These results seem to confirm the major role of the immunity in driving the variability of the HCV rather than the high degree of different HCV strains to which haemophiliacs have been in touch with, during their long‐term replacement therapy. Our results seem in keeping with other studies on different type of patients, where a low degree of quasispecies variability has been demonstrated in relationship with the progression and the severity of their liver disease.
West Nile Virus inactivation by the solvent/detergent steps of the second and third generation manufacturing processes for B‐domain deleted recombinant factor VIII
Authors: Jakubik J. J. , Vicik S. M. , Tannatt M. M. , Kelley B. D.
Published: January 2004

West Nile Virus (WNV) was immediately and completely inactivated to below assay detection limits upon addition of solvent/detergent (S/D) to intermediate process pools of second and third generation B‐domain deleted recombinant Factor VIII (BDDrFVIII; ReFacto, Wyeth, Cambridge, MA, USA). We verify that the S/D step provides robust enveloped virus inactivation (>5 log10) and functions as a WNV barrier.
The economic impact of factor VIII inhibitors in patients with haemophilia
Authors: Bohn R. L. , Aledort L. M. , Putnam K. G. , Ewenstein B. M. , Mogun H. , Avorn J.
Published: January 2004

Summary.  The impact on the cost of care for haemophilia patients with inhibitors is not well defined. To quantify the effect on health care expenditures associated with inhibitors to factor VIII (FVIII) or FIX, we conducted a retrospective cohort study examining product use and outcomes in adult and paediatric haemophilia patients with and without inhibitors. Twelve patients with inhibitors to FVIII or FIX (cases) identified in the haemophilia surveillance system (HSS) at two centres were matched on age, severity of haemophilia, and treatment centre to haemophilia patients without inhibitors. Patients with HIV or significant liver disease were excluded from the study. All eligible non‐inhibitor control patients were selected for inclusion in the study, resulting in a total of 28 controls. We then tracked product usage and hospitalizations from programme entry until 1998 or loss to follow‐up, producing a total database of 184 person‐years of experience. A descriptive matched analysis was conducted to examine annual differences in the cost of product used and hospitalizations. We found that the median cost for factor products among haemophilia patients with inhibitors was $55 853/year, $2760 less than comparable haemophilia patients without inhibitors. The median number of hospitalizations per year was 1.0 for both inhibitor and non‐inhibitor patients and the median number of days hospitalized was virtually the same. Although these findings do not appear to support the belief that there is a substantial increase in the cost of care for haemophilia patients with inhibitors, it does document that a few outlier patients can drive the cost of treatment for this disease. As the largest component of the cost of care is that of factor concentrate, it becomes imperative in the current health care environment to better define the true costs and benefits of treatments designed to eradicate or manage inhibitors. A careful cost accounting of immune tolerance induction (ITI) and other therapeutic strategies, taking into account successes and failures and duration and intensity of therapy, should help to better define the costs and benefits of such approaches. Methods to identify high cost inhibitor patients should be developed so that these strategies may be targeted to appropriate candidates.
Development of H1e histone linker‐specific antibodies by means of synthetic peptides
Authors: Foulon K. , Baltora‐Rosset S. , Fuentes V. , Mesnard F. , Da Nascimento S. , Bouhassira E. , Rochette J.
Published: January 2004

Abstract:  A large body of data suggests that the linker histones family (H1) affects gene expression. Investigation of the linker histones role is then of a major interest in cell cycle studies with implications in gene therapy. Indeed, it has been shown that in most tissues a switch of histone subtypes occurs when the cells cease to divide. To investigate linker histone role in gene or transgene expression, an antibody against subtypes of H1 would be useful for immunoprecipitation experiments and further assays measuring H1subtypes–DNA interactions in living cells. In order to produce an antibody against the H1e subtype of linker histones, two synthetic peptides derived from two regions of the H1e mouse histone protein were examined for their potential, [as keyhole limpet hemocyanin (KLH) conjugates] to elicit polyclonal anti‐H1e antibodies in New Zealand white rabbits. Selection of the peptide sequences was based on amino acid differences within the different classes of histones and between mice and rabbit histones as well. The evaluation of their potential immunogenic properties was based on examination of peptide hydropathy using predicting algorithms. Immunoglobulins (IgG) obtained from immunized and nonimmunized rabbits were tested using enzyme‐linked immunosorbent assay (ELISA) procedures, Western immunoblot, and immunofluorescence experiments. Results showed that the selected synthetic peptides gave rise to a high‐titer polyclonal antibody able to recognize the H1e histone under various conditions. This polyclonal antibody did not cross‐react with other histones. To our knowledge, this is the first antibody produced against the mouse H1e linker histone.
Potent bradykinin antagonists containing N‐benzylglycine or N‐benzyl‐l‐alanine in position 8
Authors: Dawidowska O. , Wierzba T.H. , Prahl A. , Kowalczyk W. , Derdowska I. , Neubert K. , Zabrocki J. , Olejniczak B. , Juzwa W. , Lammek B.
Published: January 2004

Abstract:  Two new analogues of a previously designed bradykinin (BK) antagonist, d‐Arg‐Arg‐Pro‐Hyp‐Gly‐Thi‐Ser‐d‐Phe‐Thi‐Arg, substituted in position 8 by N‐benzylglycine and N‐benzyl‐l‐alanine were designed, synthesized and bioassayed. The results show an impressive enhancement of B2 antagonistic potencies of both peptides in comparison with the model. In two further analogues these modifications were combined with acylation of the N‐terminus with 1‐adamantanacarboxylic acid. Acylated analogues exhibited higher antagonistic potency in comparison with the parent compounds, however, the range of effect was not as high as in previously described cases. The activity of analogues was assessed by their ability to inhibit vasodepressor response to exogenous BK (rat blood pressure test). Our results may be of value in the design of more potent BK antagonists.
Functional analysis of chemically synthesized derivatives of the human CC chemokine CCL15/HCC‐2, a high affinity CCR1 ligand
Authors: Escher S.E. , Vakili J. , Forssmann U. , Frimpong‐Boateng A. , Sticht H. , Detheux M. , Adermann K.
Published: January 2004

Abstract:  The CCL15 is a human CC chemokine that activates the receptors, CCR1 and CCR3. Unlike other chemokines, it contains an unusually long N‐terminal domain of 31 amino acids preceding the first cysteine residue and a third disulfide bond. To elucidate the functional role of distinct structural determinants, a series of sequential amino‐terminal truncated and point‐mutated CCL15 derivatives as well as mutants lacking the third disulfide bond and the carboxy‐terminal α‐helix were synthesized using 9‐fluorenylmethoxycarbonyl (Fmoc) chemistry. We demonstrate that a truncation of 24 amino acid residues (Δ24‐CCL15) converts the slightly active 92‐residue Δ0‐CCL15 into a potent agonist of CC chemokine receptor 1 (CCR1) and a weak agonist of CCR3 in cell‐based assays. The biological activity decreases from Δ24‐CCL15 to Δ29‐CCL15, and re‐increases from Δ29‐CCL15 to Δ30‐CCL15. Thus, an exocyclic N‐terminal region of only one amino acid residue is sufficient for efficient CCR1 activation. As none of the peptides investigated except for Δ24‐CCL15 activates CCR3, we suggest that CCR1 is the major receptor for CCL15 in vivo. Further we demonstrate that the third disulfide bond of CCL15 and an exchange of tyrosine in position 70 by a leucine residue, which is conserved in CXC chemokines, do not alter the interaction with CCR1. In contrast, a CCL15 derivative lacking the carboxy‐terminal α‐helix exhibits a complete loss of tertiary structure and hence loss of CCR1 agonistic and binding activity. This study demonstrates that specific protein residues in chemokines, which contribute to receptor–ligand interaction, vary significantly between chemokines and cannot be extrapolated using data from functionally related chemokines.
Ecological, morphological and phylogenetic correlates of interspecific variation in plasma carotenoid concentration in birds
Authors: Tella J. L. , Figuerola J. , Negro J. J. , Blanco G. , Rodríguez-Estrella R. , Forero M. G. , Blázquez M. C. , Green A. J. , Hiraldo F.
Published: January 2004

Carotenoids are important as pigments for bright coloration of animals, and as physiologically active compounds with a wide array of health-related benefits. However, the causes of variation in carotenoid acquisition and physiology among species are poorly known. We measured the concentration of carotenoids in the blood of 80 wild bird species differing in diet, body size and the extent of carotenoid-based traits. Preliminary analyses showed that diet significantly explains interspecific variability in plasma carotenoids. However, dietary influences were apparently overridden by phylogenetic relationships among species, which explained most (65%) of this variability. This phylogenetic effect could be due partly to its covariation with diet, but may also be caused by interspecific differences in carotenoid absorption from food to the blood stream, mediated, for example by endothelial carriers or gut parasites. Carotenoid concentrations also decreased with body size (which may be explained by the allometric relationship between ingestion rate and body mass), and correlated positively with the extent of carotenoid-dependent coloration of plumage and bare parts. Therefore, the acquisition of carotenoids from the diet and their use for both health and display functions seem to be constrained by ecological and physiological aspects linked to the phylogeny and size of the species.
Estimation of relative fitnesses from relative risk data and the predicted future of haemoglobin alleles S and C
Authors: Hedrick P.
Published: January 2004

Epidemiological studies of genetic differences in disease susceptibility often estimate the relative risks (RR) of different genotypes. Here I provide an approach to calculate the relative fitnesses of different genotypes based on RR data so that population genetic approaches may be utilized with these data. Using recent RR data on human haemoglobin β genotypes from Burkina Faso, this approach is used to predict changes in the frequency of the haemoglobin sickle-cell S and C alleles. Overall, it generally appears that allele C will quickly replace the S allele in malarial environments. Explicit population genetic predictions suggest that this replacement may occur within the next 50 generations in Burkina Faso.
Genetic and morphological differentiation in Tephritis bardanae (Diptera: Tephritidae): evidence for host-race formation
Authors: Diegisser T. , Johannesen J. , Lehr C. , Seitz A.
Published: January 2004

The fruit fly Tephritis bardanae infests flower heads of two burdock hosts, Arctium tomentosum and A. minus. Observations suggest host-associated mating and behavioural differences at oviposition indicating host-race status. Previously, flies from each host plant were found to differ slightly in allozyme allele frequencies, but these differences could as well be explained by geographical separation of host plants. In the present study, we explicitly test whether genetic and morphological variance among T. bardanae are explained best by host-plant association or by geographical location, and if this pattern is stable over a 10-year period. Populations of A. tomentosum flies differed significantly from those of A. minus flies in (i) allozyme allele frequencies at the loci Pep-A and Pgd, (ii) mtDNA haplotype frequencies and (iii) wing size. In contrast, geographical location had no significant influence on the variance estimates. While it remains uncertain whether morphometric differentiation reflects genotypic variability or phenotypic plasticity, allozyme and mtDNA differentiation is genetically determined. This provides strong evidence for host-race formation in T. bardanae. However, the levels of differentiation are relatively low indicating that the system is in an early stage of divergence. This might be due to a lack of time (i.e. the host shift occurred recently) or due to relatively high gene flow preventing much differentiation at loci not experiencing selection.
The utility of AFLPs for supporting mitochondrial DNA phylogeographical analyses in the Taiwanese bamboo viper, Trimeresurus stejnegeri
Authors: Creer S. , Thorpe R. S. , Malhotra A. , Chou W.-H. , Stenson A. G.
Published: January 2004

An amplified fragment length polymorphism (AFLP) assay was performed on individuals representing discrete haplotypes from two genetically distinct mtDNA lineages of the bamboo viper, Trimeresurus stejnegeri (Schmidt), within Taiwan. AFLP (525 polymorphic markers from five primer pairs) and mtDNA genetic distances were highly correlated and an analysis of molecular variance, and a Bayesian approach similarly partitioned estimates of genetic similarity according to the mtDNA phylogeographical pattern. These results are discussed in relation to biogeographical hypotheses, comparative rates of mtDNA molecular evolution, and in the identification of evolutionary significant units of Taiwanese T. stejnegeri. In spite of the high degree of congruence between the genetic datasets, the AFLP phylogenetic analysis did not support the mtDNA tree, suggesting that no contemporary barriers to gene flow exist between individuals from the two mtDNA lineages.
The burgeoning field of statistical phylogeography
Authors: Knowles L. L.
Published: January 2004

In the newly emerging field of statistical phylogeography, consideration of the stochastic nature of genetic processes and explicit reference to theoretical expectations under various models has dramatically transformed how historical processes are studied. Rather than being restricted to ad hoc explanations for observed patterns of genetic variation, assessments about the underlying evolutionary processes are now based on statistical tests of various hypotheses, as well as estimates of the parameters specified by the models. A wide range of demographical and biogeographical processes can be accommodated by these new analytical approaches, providing biologically more realistic models. Because of these advances, statistical phylogeography can provide unprecedented insights about a species’ history, including decisive information about the factors that shape patterns of genetic variation, species distributions, and speciation. However, to improve our understanding of such processes, a critical examination and appreciation of the inherent difficulties of historical inference and challenges specific to testing phylogeographical hypotheses are essential. As the field of statistical phylogeography continues to take shape many difficulties have been resolved. Nonetheless, careful attention to the complexities of testing historical hypotheses and further theoretical developments are essential to improving the accuracy of our conclusions about a species’ history.
On the origin of termite workers: weighing up the phylogenetic evidence
Authors: Thompson G. J. , Kitade O. , Lo N. , Crozier R. H.
Published: January 2004

Resolving the phylogenetic history of a ‘true’ worker caste in termites is essential to our understanding of termite eusocial evolution. Whether this caste is ancient and monophyletic or derived and polyphyletic will have a tremendous impact on our interpretation of termite eusocial history and remains an outstanding question in termite biology. Recent work has begun to re-examine this question in light of new phylogenetic information, but new questions have now arisen about how best to model character state changes in termite caste systems. In the present paper, we compare the models of Grandcolas and D'Haese [J. Evol. Biol. 15 (2002) 885] and Thompson et al. [J. Evol. Biol. 13 (2000) 869] and attempt to make explicit how these proposals differ with respect to the number of, and homology between, character states. We highlight the support each model has for the two principal, but competing, evolutionary hypotheses outlined above.
Premating barriers to gene exchange and their implications for the structure of a mosaic hybrid zone between Chorthippus brunneus and C. jacobsi (Orthoptera: Acrididae)
Authors: Bailey R. I. , Thomas C. D. , Butlin R. K.
Published: January 2004

Many hybrid zones contain a deficit of hybrid genotypes relative to expectations from tension zone models. This is often associated with separation of parental genotypes into distinct habitats (mosaicism), but sometimes parentals can be found co-occuring in the same local population (bimodality). In both cases, prezygotic isolation may play an important role in determining the genotypic composition of the zone. Chorthippus brunneus and C. jacobsi (Orthoptera: Acrididae) meet and form a complex hybrid zone in northern Spain. Analysis of stridulatory peg numbers reveals partial spatial and seasonal isolation in a 25 km2 area of the zone: C. jacobsi phenotypes predominate in June and July and are present in both valley and mountain habitats; C. brunneus phenotypes predominate in August and are restricted to valley habitats, always in sympatry with C. jacobsi. Strong assortative mating was observed in laboratory mating experiments. Spatial, seasonal and behavioural isolation combine to produce strong premating isolation in the study area. These results suggest a role for premating isolation in maintaining both the mosaic structure and bimodality of this hybrid zone.
Cranial evolution in sakis (Pithecia, Platyrrhini) II: evolutionary processes and morphological integration
Authors: Marroig G. , de Vivo M. , Cheverud J. M.
Published: January 2004

Patterns of interspecific differentiation in saki monkeys (Pithecia) are quantitatively described and possible evolutionary processes producing them are examined. The comparison of species correlation matrices to expected patterns of morphological integration reveal significant and similar patterns of development-based cranial integration among species. Aspects of the facial region are more heavily influenced by general size variation than features of the neural region. The comparison of pooled within- and between-groups V/CV matrices suggests that genetic drift might be a sufficient explanation for saki cranial evolution. Differential natural selection gradients are also reconstructed because selection may also have caused population differentiation through evolutionary time. These gradients illustrate the inherent multivariate nature of selection, being a consequence of the interaction between existing morphological integration (correlation) among traits and the action of natural selection. Yet, our attempt to interpret selection gradients in terms of their functional significance did not result in any clear association between selection and function. Perhaps this is also an indication that morphological evolution in sakis was mostly neutral.
Genetic variation in infestation with a directly transmitted ectoparasite
Authors: Møller A. P. , Martinelli R. , Saino N.
Published: January 2004

Genetic variation in levels of parasitism of hosts is an underlying assumption of studies of coevolution, but few such estimates are available from the field. We studied genetic variation in the abundance of the chewing louse Hirundoecus malleus on its barn swallow host Hirundo rustica. These parasites are directly transmitted and a test of genetic variation of parasite abundance would thus provide a particularly strong test. The prevalence and the abundance of the chewing lice did not differ significantly between adult male and female hosts. The resemblance in parasite intensity of H. malleus of offspring and their parents was positive and highly significant, and an analysis of extra-pair paternity in the host allowed partitioning of this resemblance between genetic and common environment effects. There was no significant resemblance in parasite intensity between extra-pair offspring and their foster parents, although the resemblance remained for within-pair offspring. This provides evidence for the abundance of directly transmitted parasites having an additive genetic component. We found no evidence of common environment effects as parents did not resemble each other with respect to lice abundance.