bluuebunny/crossref_metadata_2025 huggingface dataset:
Amyloid β‐peptide(1–42) and hydrogen peroxide‐induced toxicity are mediated by TRPM2 in rat primary striatal cultures
Authors: Fonfria E. , Marshall I. C. B. , Boyfield I. , Skaper S. D. , Hughes J. P. , Owen D. E. , Zhang W. , Miller B. A. , Benham C. D. , McNulty S.
Published: October 2005

Amyloid β‐peptide (Aβ) is the main component of senile plaques which characterize Alzheimer's disease and may induce neuronal death through mechanisms which include oxidative stress. To date, the signalling pathways linking oxidant stress, a component of several neurodegenerative diseases, to cell death in the CNS are poorly understood. Melastatin‐like transient receptor potential 2 (TRPM2) is a Ca2+‐permeant non‐selective cation channel, which responds to increases in oxidative stress levels in the cell and is activated by oxidants such as hydrogen peroxide. We demonstrate here that Aβ and hydrogen peroxide both induce death in cultured rat striatal cells which express TRPM2 endogenously. Transfection with a splice variant that acts as a dominant negative blocker of TRPM2 function (TRPM2‐S) inhibited both hydrogen peroxide‐ and Aβ‐induced increases in intracellular‐free Ca2+ and cell death. Functional inhibition of TRPM2 activation by the poly(ADP‐ribose)polymerase inhibitor SB‐750139, a modulator of intracellular pathways activating TRPM2, attenuated hydrogen peroxide‐ and Aβ‐induced cell death. Furthermore, a small interfering RNA which targets TRPM2, reduced TRPM2 mRNA levels and the toxicity induced by hydrogen peroxide and Aβ. These data demonstrate that activation of TRPM2, functionally expressed in primary cultures of rat striatum, contributes to Aβ‐ and oxidative stress‐induced striatal cell death.
Genomic profiling of the neuronal target genes of the plasticity‐related transcription factor – Zif268
Authors: Allan B.  , James  , Ann‐Marie  , Conway  , Brian J.  , Morris
Published: October 2005

The later phases of neuronal plasticity are invariably dependent on gene transcription. Induction of the transcription factor Zif268 (Egr‐1) in neurones is closely associated with many forms of functional plasticity, yet the neuronal target genes modulated by Zif268 have not been characterized. After transfection of a neuronal cell line with Zif268 we identified genes that show altered expression using high density microarrays. Although some of the genes identified have previously been associated with forms of neuronal plasticity, the majority have not been linked with neuronal plasticity or Zif268 action. Altered expression of a representative sample of the novel target genes was confirmed in Zif268‐transfected PC12 neurones, and in in vitro and in vivo models of Zif268‐associated neuronal plasticity. In particular, altered expression of the protease inhibitor Cystatin C and the chemokine Cxcl10 was observed in striatal tissue after haloperidol administration. Surprisingly, the group of identified genes is enriched for components of the proteasome and the major histocompatibility complex. Our findings suggest that altered expression of these genes following Zif268 induction may be a key component of long lasting plasticity in the CNS.
Polyglutamine tract‐binding protein‐1 dysfunction induces cell death of neurons through mitochondrial stress
Authors: Marubuchi Shigeki , Wada Yo‐ichi , Okuda Tomohiro , Hara Yukiko , Qi Mei‐ling , Hoshino Masataka , Nakagawa Masaya , Kanazawa Ichiro , Okazawa Hitoshi
Published: October 2005

Polyglutamine tract‐binding protein‐1 (PQBP‐1) is a nuclear protein that interacts and colocalizes with mutant polyglutamine proteins. We previously reported that PQBP‐1 transgenic mice show a late‐onset motor neuron disease‐like phenotype and cell death of motor neurons analogous to human neurodegeneration. To investigate the molecular mechanisms underlying the motor neuron death, we performed microarray analyses using the anterior horn tissues of the spinal cord and compared gene expression profiles between pre‐symptomatic transgenic and age‐matched control mice. Surprisingly, half of the spots changed more than 1.5‐fold turned out to be genes transcribed from the mitochondrial genome. Northern and western analyses confirmed up‐regulation of representative mitochondrial genes, cytochrome c oxidase (COX) subunit 1 and 2. Immunohistochemistry revealed that COX1 and COX2 proteins are increased in spinal motor neurons. Electron microscopic analyses revealed morphological abnormalities of mitochondria in the motor neurons. PQBP‐1 overexpression in primary neurons by adenovirus vector induced abnormalities of mitochondrial membrane potential from day 5, while cytochrome c release and caspase 3 activation were observed on day 9. An increase of cell death by PQBP‐1 was also confirmed on day 9. Collectively, these results indicate that dysfunction of PQBP‐1 induces mitochondrial stress, a key molecular pathomechanism that is shared among human neurodegenerative disorders.
Prevention of mitochondrial dysfunction in post‐traumatic mouse brain by superoxide dismutase
Authors: Xiong Ye , Shie Feng‐Shiun , Zhang Jing , Lee Chuan‐Pu , Ho Ye‐Shih
Published: October 2005

Reactive oxygen species (ROS) are known to be involved in the pathogenesis of traumatic brain injury (TBI). Previous studies have shown that the susceptibility of mice to TBI‐induced formation of cortical lesion is determined by the expression levels of copper‐zinc and manganese superoxide dismutase (CuZnSOD and MnSOD, respectively). However, the underlying biochemical mechanisms are not understood. In this study, we measured the efficiency of mitochondrial respiration in mouse brains with altered expression of these two enzymes. While controlled cortical impact injury (CCII) with a deformation depth of 2 mm caused a drastic decrease in NAD‐linked bioenergetic capacity in brain mitochondria of wild‐type mice, the functional decrease was not observed in brains of littermate transgenic mice overexpressing CuZnSOD or MnSOD. In addition, a 1 mm CCII greatly compromised brain mitochondrial function in mice deficient in CuZnSOD or MnSOD, but not wild‐type mice. Inclusion of the calcium‐chelating agent, EGTA, in the assay solution could completely prevent dysfunction of oxidative phosphorylation in all mitochondrial samples, suggesting that the observed impairment of mitochondrial function was a result of calcium overloading. In conclusion, our results imply that mitochondrial dysfunction induced by superoxide anion radical contributes to lesion formation in mouse brain following physical trauma.
Carboxypeptidase E is required for normal synaptic transmission from photoreceptors to the inner retina
Authors: Zhu Xuemei , Wu Kebin , Rife Lawrence , Cawley Niamh X. , Brown Bruce , Adams Tiffany , Teofilo Karen , Lillo Concepcion , Williams David S. , Loh Y. Peng , Craft Cheryl M.
Published: October 2005

Defects in the gene encoding carboxypeptidase E (CPE) in either mouse or human lead to multiple endocrine disorders, including obesity and diabetes. Recent studies on Cpe–/– mice indicated neurological deficits in these animals. As a model system to study the potential role of CPE in neurophysiology, we carried out electroretinography (ERG) and retinal morphological studies on Cpe–/– and Cpefat/fat mutant mice. Normal retinal morphology was observed by light microscopy in both Cpe–/– and Cpefat/fat mice. However, with increasing age, abnormal retinal function was revealed by ERG. Both Cpe–/– and Cpefat/fat animals had progressively reduced ERG response sensitivity, decreased b‐wave amplitude and delayed implicit time with age, while maintaining a normal a‐wave amplitude. Immunohistochemical staining showed specific localization of CPE in photoreceptor synaptic terminals in wild‐type (WT) mice, but in both Cpe–/– and Cpefat/fat mice, CPE was absent in this layer. Bipolar cell morphology and distribution were normal in these mutant mice. Electron microscopy of retinas from Cpefat/fat mice revealed significantly reduced spherule size, but normal synaptic ribbons and synaptic vesicle density, implicating a reduction in total number of vesicles per synapse in the photoreceptors of these animals. These results suggest that CPE is required for normal‐sized photoreceptor synaptic terminal and normal signal transmission to the inner retina.
Transcriptional profiling of spinal cord injury‐induced central neuropathic pain
Authors: Nesic Olivera , Lee Julieann , Johnson Kathia M. , Ye Zaiming , Xu Guo‐Ying , Unabia Geda C. , Wood Thomas G. , McAdoo David J. , Westlund Karin N. , Hulsebosch Claire E. , Regino Perez‐Polo J.
Published: October 2005

Central neuropathic pain (CNP) is an important problem following spinal cord injury (SCI), because it severely affects the quality of life of SCI patients. As in the patient population, the majority of rats develop significant allodynia (CNP rats) after moderate SCI. However, about 10% of SCI rats do not develop allodynia, or develop significantly less allodynia than CNP rats (non‐CNP rats). To identify transcriptional changes underlying CNP development after SCI, we used Affymetrix DNA microarrays and RNAs extracted from the spinal cords of CNP and non‐CNP rats. DNA microarry analysis showed significantly increased expression of a number of genes associated with inflammation and astrocytic activation in the spinal cords of rats that developed CNP. For example, mRNA levels of glial fibrilary acidic protein (GFAP) and Aquaporin 4 (AQP4) significantly increased in CNP rats. We also found that GFAP, S100β and AQP4 protein elevation persisted for at least 9 months throughout contused spinal cords, consistent with the chronic nature of CNP. Thus, we hypothesize that CNP development results, in part, from dysfunctional, chronically “over‐activated” astrocytes. Although, it has been shown that activated astrocytes are associated with peripheral neuropathic pain, this has not previously been demonstrated in CNP after SCI.
The mitochondrial complex I inhibitor rotenone triggers a cerebral tauopathy
Authors: Höglinger Günter U. , Lannuzel Annie , Khondiker Myriam Escobar , Michel Patrick P. , Duyckaerts Charles , Féger Jean , Champy Pierre , Prigent Annick , Medja Fadia , Lombes Anne , Oertel Wolfgang H. , Ruberg Merle , Hirsch Etienne C.
Published: October 2005

Reduced activity of the mitochondrial respiratory chain – particularly complex I – may be implicated in the etiology of both Parkinson's disease and progressive supranuclear palsy, although these neurodegenerative diseases differ substantially as to their distinctive pattern of neuronal cell loss and the predominance of cerebral α‐synuclein or tau protein pathology. To determine experimentally whether chronic generalized complex I inhibition has an effect on the distribution of α‐synuclein or tau, we infused rats systemically with the plant‐derived isoflavonoid rotenone. Rotenone‐treated rats with a pronounced metabolic impairment had reduced locomotor activity, dystonic limb posture and postural instability. They lost neurons in the substantia nigra and in the striatum. Spherical deposits of α‐synuclein were observed in a few cells, but cells with abnormal cytoplasmic accumulations of tau immunoreactivity were significantly more numerous in the striatum of severely lesioned rats. Abnormally high levels of tau immunoreactivity were found in the cytoplasm of neurons, oligodendrocytes and astrocytes. Ultrastructurally, tau‐immunoreactive material consisted of straight 15‐nm filaments decorated by antibodies against phosphorylated tau. Many tau+ cell bodies also stained positive for thioflavin S, nitrotyrosine and ubiquitin. Some cells with abnormal tau immunoreactivity contained activated caspase 3. Our data suggest that chronic respiratory chain dysfunction might trigger a form of neurodegeneration in which accumulation of hyperphosphorylated tau protein predominates over deposits of α‐synuclein.
The Crow Corvus corone hybrid zone in southern Denmark and northern Germany
Authors: HAAS FREDRIK , BRODIN ANDERS
Published: October 2005

In hybrid zones genetically differentiated populations meet and interbreed. As they result from ongoing divergence and potential speciation, such zones provide opportunities to study how different factors affect this process. One well‐known avian hybrid zone is that between the Hooded Crow Corvus corone cornix and Carrion Crow C. c. corone, which extends throughout much of Europe. We compare the current position and width of the zone in southern Denmark and northern Germany with that described in detail by Meise in 1928. A GPS technique was used to determine the geographical position of more than 1000 Crows in established pairs in which all individuals were classified as either Carrion, Hooded or hybrid phenotype. The data show that the Carrion Crow has expanded northwards and that this is most pronounced at the eastern side of the study area. Here the hybrid zone has moved around 19 km. However, the width of the zone seems to have remained stable over time. Possible reasons for why the zone may have moved are discussed.
Is nest‐site availability limiting Lesser Kestrel populations? A multiple scale approach
Authors: FRANCO ALDINA M. A. , MARQUES JOÃO T. , SUTHERLAND WILLIAM J.
Published: October 2005

The Lesser Kestrel, a colonial migratory falcon, is one of the most endangered birds in Europe and, due to a sharp population decline across much of the breeding range, is globally threatened. The reasons for this decline are unclear, but reduced nest‐site availability might be a major cause. To test this hypothesis we looked at nest‐site availability within Portuguese colonies in rural and urban buildings. Nest holes were larger, longer, higher and older than unoccupied cavities. A typical nest cavity was approximately 29–30 cm long, 300–340 cm high and had an inner chamber 16.5–18 cm wide. Large‐scale surveys of existing buildings in Portuguese villages suggested that 85% of sites lacked suitable nest cavities. The model for selection of buildings indicated that Lesser Kestrels prefer buildings with many roof and wall cavities, and that are surrounded by extensive cereal and fallow fields. The villages selected had many old buildings and monuments, were located in areas with few rivers, and a low percentage cover of cereal, olive groves and forest. The conservation implications of these results are discussed.
Ontogeny of sexual dimorphism in the Long‐tailed ManakinChiroxiphia linearis: long maturation of display trait morphology
Authors: ARÉVALO JOSÉ EDGARDO , HEEB PHILIPP
Published: October 2005

Males of dimorphic species often show ornaments that are thought to have evolved through female choice or/and male–male competition. The sexual differentiation of similar morphologies occurs during ontogeny, resulting in differential sex and age‐specific selection. The Long‐tailed Manakin is a dimorphic species with a highly skewed mating system, the males of which delay plumage maturation over 3 to 4 years. We describe ontogenetic changes in feather morphology in this species through sexual maturity. Males showed a significant increase in length of the central rectrices with age, hence their degree of sexual dimorphism increased from zero in 1‐year‐old males to 189.5% in adults. In contrast, male tail length decreased with age. Wing length did not vary significantly with age, but females had relatively longer wings than males. Wing loading was greater in females and decreased with age in males. In adults, rectrix length was positively correlated with testis volume, supporting the hypothesis that secondary sexual characters can signal the condition of primary sexual characters. Rectrix length showed positive allometry with body size in males less than 4 years old, whereas older males showed negative allometry and females showed isometry. Wing area and wing loading shifted from negative to positive allometry in males of 2 to 3 years of age. Changes in male morphology during ontogeny in the Long‐tailed Manakin appeared to be associated with their specific display behaviours. Age‐related changes in allometric growth of rectrices in the Long‐tailed Manakin suggested that young males invest disproportionately more in the length of this trait relative to their body size. This investment could act as a signal of competitive ability to move status position in their orderly queue.
A new species of rail (Aves: Rallidae) from the Upper Pleistocene and Holocene of Eivissa (Pityusic Islands, western Mediterranean)
Authors: MCMINN M. , PALMER M. , ALCOVER J. A.
Published: October 2005

A new species of rail is described from a Pleistocene and Holocene cave deposit on the island of Eivissa, Pityusic Islands (western Mediterranean Sea). Rallus eivissensis sp. nov. was an insular relative of the European Water Rail Rallus aquaticus. Compared with the extant Water Rail, the new species was smaller and stouter, had shorter and more robust hind limbs and shorter wings, with probably reduced flight ability. The Pityusics were the only Mediterranean islands with a vertebrate Quaternary fauna lacking terrestrial mammals, and this absence is no doubt related to the Eivissan rail evolution. The chronology of the Rallus eivissensis sp. nov. extinction overlaps broadly with a period of uncertainty for the arrival of humans at Eivissa, suggesting a relationship between the two events.
The winter fattening model: a test at low latitude using the Clamorous Reed Warbler
Authors: MEROM KOBY , QUADER SUHEL , YOM‐TOV YORAM
Published: October 2005

Small birds at high latitudes accumulate fat during the day so that they can survive long and cold winter nights. The winter fattening model suggests that birds increase their minimum (morning) mass in cold weather, build up their mass during the day, and then rely on the energy reserves so accumulated until the morning. While data from mid and high latitudes support this model, little is known about the strategies of birds inhabiting lower latitudes (< 40°N). We use an 18‐year data set to investigate whether the winter fattening model holds in a mid‐latitude (32°N) population of the Clamorous Reed Warbler Acrocephalus stentoreus. We show that morning mass and fat score vary as predicted by the winter fattening model. In addition, adults appear to be better adapted to long and cold winter nights than first‐year birds. A long‐term trend of increased body mass may be attributable to character release following reduced interspecific competition.
European flyway permeability and abmigration in Teal Anas crecca, an analysis based on ringing recoveries
Authors: GUILLEMAIN MATTHIEU , SADOUL NICOLAS , SIMON GÉRALDINE
Published: October 2005

Abmigration, the switch between flyways, is thought to be frequent in dabbling ducks, and supposedly results from high female philopatry combined with winter pairing: a male pairing in winter with a female from another flyway may follow her to her breeding area and thus abmigrate. On this basis, the frequency of abmigration should be much higher amongst males than amongst females, should increase with time since ringing, and be higher if environmental conditions force birds from different flyways into the same geographical area in winter (in this case the Iberian peninsula in the case of cold spells). Analysing more than 9000 recoveries of Teal ringed in the Camargue, southern France, showed that even with the most conservative flyway definition, 15% of Camargue (Mediterranean flyway) individuals (both males and females) were subsequently recovered in the North‐west European flyway. Moving a flyway boundary closer to the Camargue (i.e. to the south‐east) increased this percentage to c. 32%. None of the predicted patterns were supported by the data: there was no systematic sex difference in abmigration rate; abmigration showed no consistent increase with time from ringing, and was not significantly more frequent after cold spells. These results suggest that there is permeability between these two flyways for all classes of individuals, which might better be considered as a continuum. This should be taken into account in the development of management and conservation policies for Teal in Europe.
The use of mitochondrial and nuclear sequence data in assessing the taxonomic status of the endangered Uluguru Bush Shrike Malaconotus alius
Authors: FUCHS JÉRÔME , FJELDSÅ JON , PASQUET ERIC
Published: October 2005

The endangered Uluguru Bush Shrike Malaconotus alius is a large, black‐headed bush shrike, strictly endemic to the Uluguru Mountains in Tanzania. It has recently been suggested that this species has been misplaced within the genus Malaconotus and might instead be related to Malagasy Vangidae. To assess its systematic affinities, we analysed 1518 bp of sequence data obtained from a nuclear intron (myoglobin intron‐2) and a mitochondrial protein‐coding gene (ND2) using parsimony, maximum likelihood and Bayesian inference. Both genes strongly support the traditional placement within the genus Malaconotus, suggesting that similarities between the Uluguru Bush Shrike and the Vangidae are due to convergence. These results caution that taxonomic changes should not be made without a proper character analysis (i.e. assignment of character homology).
Unravelling the migration and moult strategies of a long‐distance migrant using stable isotopes: Red Knot Calidris canutus movements in the Americas
Authors: ATKINSON PHILIP W. , BAKER ALLAN J. , BEVAN RICHARD M. , CLARK NIGEL A. , COLE KIMBERLY B. , GONZALEZ PATRICIA M. , NEWTON JASON , NILES LAWRENCE J. , ROBINSON ROBERT A.
Published: October 2005

For long‐distance migrants, such as many of the shorebirds, understanding the demographic implications of behavioural strategies adopted by individuals is key to understanding how environmental change will affect populations. Stable isotopes have been used in the terrestrial environment to infer migratory strategies of birds but rarely in marine or estuarine systems. Here, we show that the stable isotope ratios of carbon and nitrogen in flight feathers can be used to identify at least three discrete wintering areas of the Red Knot Calidris canutus on the eastern seaboard of the Americas, ranging from southeastern USA to Patagonia and Tierra del Fuego. In spring, birds migrate northwards via Delaware Bay, in the northeastern USA, the last stopping point before arrival in Arctic breeding areas, where they fatten up on eggs of spawning Horseshoe Crabs Limulus polyphemus. The isotope ratios of feather samples taken from birds caught in the Bay during May 2003 were compared with feathers obtained from known wintering areas in Florida (USA), Bahia Lomas (Chile) and Rio Grande (Argentina). In May 2003, 30% of birds passing through the Bay had Florida‐type ‘signatures’, 58% were Bahia Lomas‐type, 6% were Rio Grande‐type and 7% were unclassified. Some of the southern wintering birds had started moulting flight feathers in northern areas, suspended this, and then finished their moult in the wintering areas, whereas others flew straight to the wintering areas before commencing moult. This study shows that stable isotopes can be used to infer migratory strategies of coastal‐feeding shorebirds and provides the basis for identifying the moult strategy and wintering areas of birds passing through Delaware Bay. Coupled with banding and marking birds as individuals, stable isotopes provide a powerful tool for estimating population‐specific demographic parameters and, in this case, further our understanding of the migration systems of the declining Nearctic populations of Red Knot.
Flight speeds of two seabirds: a test of Norberg's hypothesis
Authors: ELLIOTT KYLE HAMISH , GASTON ANTHONY J.
Published: October 2005

Norberg suggested that birds should increase their flight speed when rearing chicks in order to maximize chick energy intake by reducing commuting time. We measured the incubation and chick‐rearing flight speeds of a medium‐range (Brünnich's Guillemot Uria lomvia) and long‐range (Northern Fulmar Fulmarus glacialis) forager near the Prince Leopold Island colony, Nunavut, Canada. The mean flight speed for the long‐range forager was significantly higher during chick‐rearing than during incubation. The medium‐range forager showed no difference in mean flight speed during the two periods. We suggest that because petrels fly close to their minimum power velocity and have a low wing‐loading, whereas alcids fly close to their maximum range velocity and have a high wing‐loading, petrels have a greater ability than alcids to alter their flight speed according to changes in the demands of different breeding stages. Consequently, whereas Northern Fulmars adapt to the additional cost of chick‐rearing partially by altering flight speed, Brünnich's Guillemots can only do so by reducing mass.
The incidence of the parasitic disease trichomoniasis and its treatment in reintroduced and wild Pink PigeonsColumba mayeri
Authors: SWINNERTON K. J. , GREENWOOD A. G. , CHAPMAN R. E. , JONES C. G.
Published: October 2005

The recovery programme for the endangered Pink PigeonColumba mayeriPrévost, 1843 of Mauritius has involved intensive management to promote rapid population growth. As the Pink Pigeon population has increased in size and distribution, disease problems have emerged that have affected wild and reintroduced birds. Trichomoniasis first appeared in the remnant wild population in 1992, and became established in a reintroduced subpopulation on the offshore island of Ile aux Aigrettes. We aimed to control the disease by treating individuals and, on Ile aux Aigrettes, by routinely treating the population with medicated drinking water. On Ile aux Aigrettes, 49% of squabs were clinically diseased and only 27% of all squabs survived without treatment. Treatment of squabs that were not clinically diseased improved survival from 36 to 82%, suggesting that subclinical infections also affected survival. Subclinical infections were demonstrated by microscopy in 22% of Pink Pigeons and 17% of exotic doves on Ile aux Aigrettes. Clinical disease was observed in 4.5% of adults and 3.1% of juveniles, but more juveniles than adults died from the disease. It is likely that Trichomoniasis and other diseases will continue to affect the Pink Pigeon's recovery and the species may require long‐term management to counteract the effects of disease and other limiting factors.
Accidental extravascular insertion of a subclavian hemodialysis catheter is signaled by nonvisualization of catheter tip
Authors: Florescu Marius C , Moussa Hatem , Salifu Moro , Friedman Eli A
Published: October 2005

Subclavian hemodialysis (HD) catheter placement under fluoroscopy with perforation of the superior vena cava (SVC) is a rare complication that needs to be recognized and treated appropriately. We report the case of a 47‐year‐old black woman under treatment for end‐stage renal disease secondary to HIV‐associated nephropathy who sustained an extravascular insertion of fluoroscopy‐guided subclavian catheterization for HD. Subsequent successful removal of the extravascularly placed catheter along with repair of the lacerated SVC were effected by open thoracic surgery.
Middle molecule removal in low‐flux polysulfone dialyzers: Impact of flows and surface area on whole‐body and dialyzer clearances
Authors: Eloot Sunny , De Vos Jean‐Yves , De Vos Filip , Hombrouckx Remi , Verdonck Pascal
Published: October 2005

Some studies found that the removal of middle molecules has a long‐term effect on mortality and, even more, is enhanced by high‐flux dialysis. In order to enhance middle molecule removal in a low‐flux dialyzer, the present study aimed at investigating the combined impact of dialyzer flows and membrane surface area. Blood and dialysate flows were varied within the clinical range 300–500 and 500–800 mL/min, respectively, while the ultrafiltration rate was kept constant at 0.1 L/hr. Single‐pass tests were performed in vitro in a single Fresenius F6HPS dialyzer® (3 tests) and serially (5 tests) and parallel (3 tests) connected dialyzers. The blood substitution fluid consisted of dialysis fluid in which radioactive‐labeled vitamin B12 (molecular weight 1355 Da) was dissolved. Dialyzer clearance as well as whole‐body clearance was calculated from radioactivity concentrations of samples taken from the inlet and outlet bloodline. Adding a second dialyzer in series or parallel ameliorated the overall dialyzer and whole‐body clearance significantly, except for the highest applied blood flows of 500 mL/min. Better solute removal was also obtained with higher dialysate flows, while the use of higher blood flows seemed advantageous only when using a single dialyzer. Analysis of the ultrafiltration profiles in the different configurations illustrated that enhancing the internal filtration rate ameliorates convective transport of middle molecules. Adequate solute removal results from a number of interactions, as there are blood and dialysate flows, membrane surface area, filtration profile and concentration profiles in the blood and dialysate compartment.
Hyperhemolysis syndrome in anemia of chronic disease
Authors: Darabi Kamran , Dzik Sunny
Published: October 2005

BACKGROUND: Occasional cases of delayed hemolytic transfusion reaction (DHTR) demonstrate severe and persistent hemolysis and are referred to as hyperhemolysis syndrome. This syndrome usually occurs in patients with sickle cell disease and possibly thalassemia who receive multiple transfusions. There are few such clinical reports in patients without hemoglobinopathies.

CASE REPORT: A 67‐year‐old woman with anemia and a history of four previous transfusions was admitted with shortness of breath and a hematocrit (Hct) level of 27 percent. The patient was group O with a negative antibody screen. She received 1 unit of electronically cross‐matched red blood cells (RBCs) and was discharged. Thirteen days later she returned to hospital with weakness and a Hct level of 23 percent. The antibody screen now demonstrated anti‐K alloantibody. The direct antigloblulin test (DAT) was positive with both anti‐immunoglobulin G and anti‐complement (C3). Anti‐K was recovered in the eluate. The previously transfused RBC unit was positive for presence of the K antigen. The patient's RBCs were negative for the presence of K antigen. Other laboratory data confirmed ongoing hemolysis, and a diagnosis of DHTR was made. She continued to display findings of active hemolysis for 9 more weeks requiring 19 units of RBCs. Thirty‐four days after the original transfusion, her DAT remained positive and both the plasma sample and a RBC eluate demonstrated anti‐K.

CONCLUSION: The delayed hemolytic transfusion reaction with hyperhemolysis can occur among patients without hemoglobinopathies.
Prevalence of viral markers among first‐time Arab blood donors in Kuwait
Authors: Ameen Reem , Sanad Nabil , Al‐Shemmari Salem , Siddique Iqbal , Chowdhury Rafiq I. , Al‐Hamdan Samia , Al‐Bashir Abdulaziz
Published: October 2005

BACKGROUND: The aim of this study was to assess the effect of blood donation modes on the prevalence of viral markers among Arab first‐time blood donors in Kuwait.

STUDY DESIGN AND METHODS: Donor ethnic background was classified as Kuwaiti nationals and non‐Kuwaiti Arabs. A total of 26,874 donors were screened in 2002 for the following viral markers: hepatitis C virus antibody (anti‐HCV), hepatitis B surface antigen (HBsAg), anti‐hepatitis B core antigen (HBc), human immunodeficiency virus‐1 and ‐2 antibody (anti‐HIV‐1 and ‐2), HIV p24, and human T lymphotropic virus‐I and ‐II antibody (anti‐HTLVI/II). All samples positive for the presence of anti‐HBc were tested for anti‐HBs. Among these donors, 12,798 were first‐time donors of which 74 percent were replacement and 26 percent were volunteers.

RESULTS: The prevalence of HCV among replacement donors was significantly higher than the volunteer group. The difference between the two modes of blood donations, however, was not significant for HBsAg. The prevalence of anti‐HCV among Kuwaiti national and non‐Kuwaiti Arab first‐time donors was 0.8 and 5.4 percent, respectively, whereas the prevalence of HBsAg was 1.1 and 3.5 percent, respectively, with the difference being significant at a p level of <0.0001. The difference observed for prevalence of anti‐HBc among Kuwaiti national and non‐Kuwaiti Arab donors (17 and 33.3%, respectively) was significant (p < 0.0001). Among first‐time donors, 13.7 percent were positive for the presence of anti‐HBs, indicating that 13.7 percent of the total Arab donor population might have had a previous infection and possible immunity to hepatitis B virus (HBV).

CONCLUSION: A high prevalence of HBV and HCV was found among non‐Kuwaiti Arab donors. The prevalence of anti‐HCV was only significantly higher among replacement versus volunteer first‐time donors. Therefore, there is a need to develop a strategic plan that incorporates the diverse background of the blood donors living in Kuwait.
Effective storage of granulocytes collected by centrifugation leukapheresis from donors stimulated with granulocyte–colony‐stimulating factor
Authors: Hubel Kai , Rodger Elin , Gaviria J. Milton , Price Thomas H. , Dale David C. , Liles W. Conrad
Published: October 2005

BACKGROUND: Donor stimulation with granulocyte–colony‐stimulating factor (G‐CSF) has increased the number of neutrophils (PMNs) that can be collected for granulocyte transfusion therapy. Clinical utility, however, has been limited by the inability to store functional PMNs ex vivo. This study was conducted to determine whether granulocyte products from G‐CSF–stimulated donors could be effectively stored at reduced temperature (22°C vs. 10°C) with maintenance of functional properties in vitro and in vivo.

STUDY DESIGN AND METHODS: Nine normal subjects received G‐CSF (600 µg subcutaneously) 12 hours before centrifugation leukapheresis. Granulocyte products were divided and stored for 24 and 48 hours under four conditions: 1) 22°C; 2) 22°C, with supplemental G‐CSF (100 ng/mL); 3) 10°C; and 4) 10°C, with supplemental G‐CSF. Functional PMN activity during ex vivo storage was assessed in vitro and in vivo by the skin‐window technique for granulocytes stored at 10°C for 24 hours.

RESULTS: Surface expression of CD11b/CD18, CD14, CD16, CD32, and CD64 was maintained during 48‐hour storage at reduced temperature. Inducible respiratory burst activity, bactericidal activity, and fungicidal activity were preserved during storage for 48‐hour storage at 10°C. Proinflammatory cytokine production was decreased in product stored at 10°C. Supplemental G‐CSF ex vivo did not substantially improve functional activity during storage. After storage at 10°C for 24 hours, in vitro chemotactic potential was maintained, and transfused granulocytes retained capacity to circulate and migrate appropriately in vivo.

CONCLUSIONS: Granulocyte product collected by centrifugation leukapheresis from G‐CSF–stimulated donors can be effectively stored at subphysiologic temperature for 24 hours with preservation of functional activity. Storage at 10°C appears to be slightly superior to storage at 22°C.
Cell loss and recovery in umbilical cord blood processing: a comparison of postthaw and postwash samples
Authors: Laroche Vincent , McKenna David H. , Moroff Gary , Schierman Therese , Kadidlo Diane , McCullough Jeffrey
Published: October 2005

BACKGROUND: Engraftment after umbilical cord blood (UCB) transplantation is highly dependent on nucleated cell (NC) and CD34+ cell content. Current standard postthaw (PT) processing includes a wash step to remove dimethyl sulfoxide (DMSO), lysed red cells, and stroma. The contribution of the wash step to cell loss and ultimately the dose of cells available for transplant have yet to be systematically reported. This study examines the effect of the wash step as well as that of PT storage on various quality control variables of UCB units.

STUDY DESIGN AND METHODS: Ten units were thawed and washed based on the New York Blood Center method. Samples were removed from each unit at six time points: prefreeze (PF), immediately PT, immediately postwash (PW), and 1, 2, and 5 hours PW. On each sample, total nucleated cell (TNC) count, CD34+ cell enumeration, colony‐forming unit (CFU)–granulocyte‐macrophage, and viability assays (fluorescence microscopy [acridine orange/propidium iodide, or AO/PI] and flow cytometry [7‐aminoactinomycin]) were obtained.

RESULTS: TNC counts decreased PT and at subsequent time points; the PT TNC recovery was 89 percent compared to 82 percent PW (p < 0.01). TNC recovery decreased to 90 percent of PW (82% of PT) values (p < 0.01) and 83 percent of PW (76% of PT) values (p < 0.001), at 2 and 5 hours PW, respectively. CD34+ cell loss PT was not significant. Viability by AO/PI decreased PT and plateaued over time. In contrast, viability by flow cytometry remained higher and increased slightly over time. CFUs were significantly lower PT, recovering PW.

CONCLUSIONS: Our data indicate that the thawing and washing results in a substantial loss of cells, with TNC loss approaching 20 percent when compared with PF counts; the wash step was responsible for nearly half of the cell loss. The reduced PT viability was expected. Elapse of time PW resulted in further loss of NCs but no detectable significant changes in CD34+ cell content and viability and/or CFU.
Clinical safety of platelets photochemically treated with amotosalen HCl and ultraviolet A light for pathogen inactivation: the SPRINT trial
Authors: Snyder Edward , McCullough Jeffrey , Slichter Sherrill J. , Strauss Ronald G. , Lopez‐Plaza Ileana , Lin Jin‐Sying , Corash Laurence , Conlan Maureen G. ,   SPRINT Study Group
Published: October 2005

BACKGROUND:  A photochemical treatment (PCT) method utilizing a novel psoralen, amotosalen HCl, with ultraviolet A illumination has been developed to inactivate viruses, bacteria, protozoa, and white blood cells in platelet (PLT) concentrates. A randomized, controlled, double‐blind, Phase III trial (SPRINT) evaluated hemostatic efficacy and safety of PCT apheresis PLTs compared to untreated conventional (control) apheresis PLTs in 645 thrombocytopenic oncology patients requiring PLT transfusion support. Hemostatic equivalency was demonstrated. The proportion of patients with Grade 2 bleeding was not inferior for PCT PLTs.

STUDY DESIGN AND METHODS:  To further assess the safety of PCT PLTs, the adverse event (AE) profile of PCT PLTs transfused in the SPRINT trial is reported. Safety assessments included transfusion reactions, AEs, and deaths in patients treated with PCT or control PLTs in the SPRINT trial.

RESULTS:  A total of 4719 study PLT transfusions were given (2678 PCT and 2041 control). Transfusion reactions were significantly fewer following transfusion of PCT than control PLTs (3.0% vs. 4.1%; p = 0.02). Overall AEs (99.7% PCT vs. 98.2% control), Grade 3 or 4 AEs (79% PCT vs. 79% control), thrombotic AEs (3.8% PCT vs. 3.7% control), and deaths (3.5% PCT vs. 5.2% control) were comparable between treatment groups. Minor hemorrhagic AEs (petechiae [39% PCT vs. 29% control; p < 0.01] and fecal occult blood [33% PCT vs. 25% control; p = 0.03]) and skin rashes (56% PCT vs. 42% control; p < 0.001) were significantly more frequent in the PCT group.

CONCLUSION:  The overall safety profile of PCT PLTs was comparable to untreated PLTs.
Removal of exogenous (spiked) and endogenous prion infectivity from red cells with a new prototype of leukoreduction filter
Authors: Sowemimo‐Coker Samuel , Kascsak Regina , Kim Anzi , Andrade Fabiola , Pesci Susan , Kascsak Richard , Meeker Clifford , Carp Richard , Brown Paul
Published: October 2005

BACKGROUND: Two recent probable cases of transmission of a variant of human Creutzfeldt‐Jakob disease (vCJD) through blood transfusion suggest that the disease can be transmitted through transfusion of blood components from presymptomatic blood donors. In the absence of a preclinical screening test, removal of the infectious agent by processing is the only means by which risk to recipients of blood from donors with inapparent vCJD infections can be eliminated.

STUDY DESIGN AND METHODS: In the endogenous infectivity study, a pool of 500 mL of whole blood was collected into CP2D anticoagulant from 263K‐strain scrapie‐infected hamsters, processed into 300 mL of red cells (RBCs), and then passed through a prion removal filter. Pre‐ and postfiltration samples were tested for PrPsc by Western blot and for infectivity by inoculation of healthy hamsters. In the exogenous (spiking) infectivity study, 30 mL of 10 percent (wt/vol) scrapie‐infected brain homogenates was added to 270 mL of human RBCs and then filtered. Levels of PrPsc and infectivity were determined by Western blot and bioassay.

RESULTS: In the endogenous infectivity study, the prefiltered RBCs transmitted disease to 6 of 43 animals, whereas the postfiltered RBCs did not transmit disease to any of 35 animals, and a barely visible prefiltration PrPsc Western blot signal was reduced below the level of detection in the postfiltration sample. In the exogenous (spike) study, infectivity was reduced by 3.7 log LD50 per mL, from 9.2 to 5.5 log LD50 per mL.

CONCLUSION: The new filter was effective in removing both infectivity and PrPsc from RBCs. The use of this type of filter should reduce the risk of vCJD transmission through blood transfusion.
The impact of e‐mail in acquiring and retaining whole‐blood donors: a comparative analysis of the Puget Sound Blood Center donor e‐mail communication program
Authors: Geyer Maria Elena
Published: October 2005

BACKGROUND: With the emergence of e‐mail as a common form of communication, it is important to understand the role and impact e‐mail can have on acquiring and retaining whole‐blood donors.

STUDY DESIGN AND METHODS: Whole‐blood donors who opted in to receive e‐mail communications (n = 43,232) from the Puget Sound Blood Center (PSBC) and as a result participated in one or more of five e‐mail marketing campaigns between July 2002 and March 2003 were studied.

RESULTS: New donors with e‐mail addresses grew by 74 percent over the measurement period compared with 2 percent growth of those without e‐mail addresses. Nearly 15,000 prospective donors were reached as a result of donors passing along e‐mails they received from PSBC to friends and family. Nearly 5 percent of PSBC e‐mail campaign recipients registered to donate online. PSBC e‐mail campaigns have outperformed e‐mail marketing industry mean open rates by 44 percent and click‐through rates by 36 percent.

CONCLUSION: E‐mail is now a mainstream communication medium. It has proven to be an effective method of reaching new donors and generating new donation registrations. Donors respond more favorably to e‐mail communications from PSBC compared with e‐mail they receive from other organizations. Deeper understanding of how different segments of blood donors use e‐mail may help blood centers adjust their donor acquisition and retention programs. New recruitment methods are needed to grow the overall donor base, and e‐mail appears to have the potential to make a significant contribution to meeting this objective.
Cryopreservation of hematopoietic progenitor cells from apheresis at high cell concentrations does not impair the hematopoietic recovery after transplantation
Authors: Martin‐Henao Gregorio Angel , Torrico Carlos , Azqueta Carmen , Amill Begoña , Querol Sergio , Garcia Juan
Published: October 2005

BACKGROUND: The high number of nuclear cells (NCs) from hematopoietic progenitor cells‐apheresis (HPC‐A) requires cryopreservation in large volumes or at high NC concentrations. The effect of NC concentration during cryopreservation has yet to be examined.

STUDY DESIGN AND METHODS: In the experimental arm (n = 610, Protocol B), the first HPC‐A sample from the patient was cryopreserved in two cryobags and subsequent collections in one cryobag, resulting in high NC concentrations (>100 × 106 NCs/mL) in most cases. The effect of NC concentrations at freezing in NC recovery after thawing and engraftment kinetics was analyzed and compared with a group of HPC‐A cryopreserved at standard NC concentrations (n = 455, Protocol A).

RESULTS: The mean (SD) NC concentration at freezing was 78 (28) × 106 per mL (median, 82 × 106/mL; range, 12 × 106‐156 × 106/mL) and 183 (108) × 106 per mL (median, 156 × 106/mL; range, 16 × 106‐678 × 106/mL), for HPC‐A cryopreserved according to Protocols A and B, respectively. The NC viabilities of the test vials and HPC‐A components after thawing were 88 percent versus 85 percent and 85 percent versus 82 percent, and the cloning efficiency was 49 percent versus 33 percent for Protocols A and B, respectively (p < 0.001). Significant differences were not observed in the recovery of NCs. Days to neutrophil and platelet engraftment were not different between patients transplanted in the standard‐ (n = 143) or high‐cell‐concentration group (n = 238).

CONCLUSION: The cryopreservation of HPC‐A at higher than standard NC concentrations has no adverse impact on hematopoietic reconstitution after transplantation.
Freezing human platelets with 6 percent dimethyl sulfoxide with removal of the supernatant solution before freezing and storage at −80°C without postthaw processing
Authors: Valeri C. Robert , Ragno Gina , Khuri Shukri
Published: October 2005

BACKGROUND: Platelets (PLTs) can be frozen with 6 percent dimethyl sulfoxide (DMSO) at −80°C for up to 2 years. This method has been modified by concentrating the PLTs and removing the supernatant before freezing.

STUDY DESIGN AND METHODS: High‐yield leukoreduced PLTs stored at 22°C for up to 5 days were divided into three equal volumes: one was frozen with 6 percent DMSO at −80°C, thawed, washed, and resuspended in plasma (old method with DMSO); the second was treated with 6 percent DMSO, concentrated to remove the supernatant DMSO, frozen at −80°C, thawed, and diluted with 0.9 percent NaCl (new method with DMSO); and the third was treated with 0.9 percent NaCl without DMSO, concentrated to remove the supernatant solution, frozen at −80°C, thawed, and diluted with 0.9 percent NaCl (new method without DMSO).

RESULTS: Freeze‐thaw‐wash recovery of PLTs frozen by the old method with DMSO was 74 ± 2 percent with 5 percent PLT microparticles. Freeze‐thaw recovery was 94 ± 2 percent with 7 percent PLT microparticles (new method with DMSO) and 69 ± 9 percent with 15 percent PLT microparticles (new method without DMSO). Total DMSO in washed PLTs was 400 and 600 mg in PLTs concentrated before freezing. In vivo recovery of PLTs frozen by the new method with DMSO and transfused into normal volunteers was 30 percent and the life span was 7 days.

CONCLUSION: Concentrating PLTs before freezing simplified the procedure by eliminating postthaw washing. PLTs frozen by this method had more PLTs with reduced GPIb and increased annexin V binding than those frozen by the old method.