bluuebunny/crossref_metadata_2025 huggingface dataset:
Absence of the kdr mutation in the molecular ‘M’ form suggests different pyrethroid resistance mechanisms in the malaria vector mosquito Anopheles gambiae s.s.
Authors: Awolola T. S. , Brooke B. D. , Koekemoer L. L. , Coetzee M.
Published: December 2004

Field tests conducted on adult Anopheles mosquitoes using standard WHO procedures, diagnostic kits and test papers in south‐western Nigeria showed pyrethroid (deltamethrin and permethrin) resistance in adult populations of Anopheles gambiae sensu stricto. The knock‐down resistance (kdr) mutation involved in pyrethroid resistance was only found in the molecular S form of An. gambiae s.s. even in area where both molecular M and S forms occurred in sympatry. The absence of the kdr mutation in the M form suggests an additional pyrethroid resistance mechanism in An. gambiae s.s.
Présence d'Angiostrongylus cantonensis en Haïti
Authors: Raccurt Christian P. , Blaise Jacques , Durette‐Desset Marie‐Claude
Published: December 2004

Resumé

L'angiostrongylose nerveuse due àAngiostrongylus (Parastrongylus) cantonensis est une zoonose qui a été signalée dans la majorité des régions d'Asie méridionale et orientale, du Pacifique, de l'Océan indien, en Australie, dans quelques pays d'Afrique et de la Caraïbe. Sur 23 rats capturés en Haiti, 7 ont été parasités par des angiostrongyles dans les cavités droites du cœur ou dans les artères pulmonaires. Haïti est ainsi la dernière des grandes Antilles où la présence de A. cantonensis est confirmée. L'impact de ce parasite sur la santé humaine en Haïti n'est pas connu. Il demanderait àêtre évalué d'autant que 1,3% des décès recensés pendant l'année 1999 étaient dus à une méningite d'étiologie inconnue.
An estimation of the incidence of noma in north‐west Nigeria
Authors: Fieger Alexander , Marck Klaas W. , Busch Raymonde , Schmidt Andreas
Published: December 2004

Noma (cancrum oris, stomatitis gangrenosa) is a quickly spreading orofacial gangrene in children, caused by a combination of malnutrition, debilitation because of concomitant diseases (measles) and intraoral infections. The global incidence of noma in the world is uncertain. By comparing large numbers of noma patients and cleft lip patients in a large referral hospital for these disorders in Sokoto, Nigeria, we calculated the incidence of noma in north‐west Nigeria as 6.4 per 1000 children. Extrapolation of this incidence to the developing countries bordering the Sahara Desert (the noma belt of the world) gives an incidence of 25 600 for that region and a global incidence of 30 000–40 000. Noma is a good biological parameter of extreme poverty, and hence a global monitoring system for noma can be justified. Though economic progress is the most effective preventive measure against noma, medical prevention by vaccination programmes against measles should be enhanced as well.
Comparison of screening methods for anaemia in pregnant women in Awassa, Ethiopia
Authors: Gies S. , Brabin B. J. , Yassin M. A. , Cuevas L. E.
Published: December 2004

Screening for anaemia in pregnancy is essential for implementing and monitoring effective antenatal programmes. We compared the diagnostic accuracy of invasive and non‐invasive screening methods in a cross‐sectional survey of 403 pregnant women attending an urban health centre in Awassa, southern Ethiopia. Overall anaemia prevalence [haemoglobin (Hb): <11 g/dl] was 15.1% (95% CI: 12.1–19.9), mild anaemia (Hb: 10–10.9 g/dl) 10.4%, moderate anaemia (Hb: 7–9.9 g/dl) 4.2% and severe anaemia (Hb < 7 g/dl) 0.3%. Sensitivity, specificity and predictive values of conjunctival pallor and the WHO Hb colour scale were calculated for Hb cut‐off points <11, <10 and <9 g/dl. All methods in combination with the symptoms and complaints reported by the mothers were entered into a predictive scoring system. None of the methods tested or models predicted anaemia with suitable accuracy in this population. The diagnosis of anaemia based on clinical signs and symptoms remains unreliable despite attempts to develop predictive models.
Recognizing childhood illnesses and their traditional explanations: exploring options for care‐seeking interventions in the context of the IMCI strategy in rural Ghana
Authors: Hill Zelee , Kendall Carl , Arthur Paul , Kirkwood Betty , Adjei Eunice
Published: December 2004

Objectives Interventions that promote appropriate care‐seeking for severely ill children have the potential to substantially reduce child mortality in developing countries, but little is known about the best approach to address the issue. This paper explores the relative importance of illness recognition as a barrier to care‐seeking and the feasibility and potential impact of improving recognition.

Methods The study combined qualitative and quantitative methods including in‐depth interviews exploring the local illness classification system, a Rapid Anthropological Assessment (RAA) recording narratives of recent episodes of child illness and a survey designed to test the hypotheses that emerged from the RAA.

Results Several danger symptoms were not recognized by caregivers. There were recognition problems which may not be feasibly addressed in an intervention. Other significant care‐seeking barriers included classifying certain illnesses as ‘not‐for‐hospital’ and untreatable by modern medicine; problems of access; and frequent use of traditional medicines.

Conclusion The recognition component of any care‐seeking intervention should identify the type of recognition problem present in the community. Many of the care‐seeking barriers identified in the study revolved around the local illness classification system, which should be explored and built on as part of any care‐seeking intervention.
Anopheles species of the mount Cameroon region: biting habits, feeding behaviour and entomological inoculation rates
Authors: Wanji Samuel , Tanke Theodore , Atanga Sali N. , Ajonina Caroline , Nicholas Tendongfor , Fontenille Didier
Published: December 2004

There is a lack of data on the Anopheles fauna, its biology and the roles played by different vector species in the transmission of malaria in the mount Cameroon region. The biting habits, feeding behaviour and entomological inoculation rates of different Anopheles species during the dry and rainy season were investigated. A total of 2165 Anopheles was collected, 805 in the rainy season and 1360 in the dry season. Five Anopheles species were identified: Anopheles gambiae s.l., An. funestus, An. hancocki, An. moucheti and An. nili. An. gambiae, An. funestus and An. hancocki, recorded during both seasons, were the main vectors of malaria in the region. An. gambiae s.s. was the only member of the An. gambiae (Giles) complex. These three species had their peak activity between 1 and 2 am. A human blood index (HBI) of 98.29% was recorded for fed Anopheles. The sporozoite rate, for all vectors together, was significantly higher in the rainy season (9.4%) than in the dry season (4.2%) with all the species infected by Plasmodium falciparum. The average inoculation rate was 0.44 infective bites per man per night, which adds up to 161 infective bites per year in this study area. Analyses of relative abundance and infection rate of malaria vectors at different sites situated along a transect of 20 km during the dry season showed high heterogeneity in biting and sporozoite rates. No malaria vector was caught at 1200 m a.s.l. The mount Cameroon region should be considered an area of high malaria transmission intensity.
Syndromic management of sexually transmitted diseases in Botswana's primary health care: quality of care aspects
Authors: Boonstra E. , Lindbæk M. , Klouman E. , Ngome E. , Romøren M. , Sundby J.
Published: December 2004

Objectives To evaluate the quality of care of the syndromic management of sexually transmitted diseases (STDs) in Botswana's primary health care.

Methods Participative observations of 224 consecutive consultations of patients with STDs (135 females and 89 males) by nurses. Twenty‐one cases were excluded because no STD checklist was filled in. Criteria for acceptable history taking, physical examination and correct treatment were agreed upon.

Results The quality of history taking and physical examination was acceptable for 25% and 23% of the women and for 54% and 57% of the men, respectively. Approximately, 65% of the women and 81% of the men received appropriate treatment. On average, consultations took 5.4 min for women and 4.6 min for men. STD contacts comprised 11% of STD cases. Advice on partner notification was provided to 66% of the women and 86% of men, and 75% and 89%, respectively, were counselled on the use of condoms. In half of the health facilities the lack of a fixed light source was the main constraint in carrying out a vaginal speculum examination. The availability of antibiotics and condoms was excellent. In 40% of the health facilities, all STD algorithms were displayed in the consultation room.

Conclusion One‐third of women and one‐fifth of men did not receive appropriate treatment for their STD, in spite of excellent provision of drugs. Although Botswana health workers perform relatively well on partner notification and counselling, there is considerable scope for improving the quality of medical history and clinical examination, especially in women. Emphasis should be given on training health workers in clinical examinations, in particular in pelvic examinations, and to supervision and in‐service training.
Incidence and risk factors of paediatric rotavirus diarrhoea in northern Ghana
Authors:   The Navrongo Rotavirus Research Group
Published: December 2004

We measured the type‐specific incidence of paediatric rotavirus diarrhoea in an area of northern Ghana. Over 1 year, diarrhoea 1717 episodes were identified, of which 677 (39%) were positive for rotavirus. Risk factors for rotavirus infection included old age, wasting, high Vesikari score and the episode occurring in the dry season. Rotavirus‐positive episodes tended to be more acute, causing vomiting and greater dehydration, and were more likely to require hospitalization. The incidence was 0.089 episodes per person‐year for all diarrhoea, and 0.035 for rotavirus diarrhoea. The observed incidence decreased markedly with distance from the nearest health centre, suggesting a large unobserved burden. G2P[6], G3P[4] and G9P[8] made up more than half the genotypes detected, but the remainder were diverse. There is a large burden of rotavirus diarrhoea, but the effectiveness of future vaccines could be diluted by the high polymorphism of the virus, and the difficulty of reaching remote populations.
Changing scenario of malaria in central India, the replacement of Plasmodium vivax by Plasmodium falciparum (1986–2000)
Authors: Singh Neeru , Nagpal A. C. , Saxena Ajay , Singh M. P.
Published: December 2004

Objectives  Since 1986, we have been studying the changing epidemiology of malaria in a forest belt of Mandla, which has the highest number of malaria cases in central India (Madhya Pradesh) to define the epidemiological characteristics of the infection with each Plasmodium species in different seasons of the year. Our long‐term objective was to determine the dynamics of Plasmodium vivaxvs.P. falciparum infections.

Methods  Five villages underwent fortnightly surveillance of fever cases. Drugs were distributed within 24 h after results of blood smears became available as per Indian National Anti‐Malaria Programme. Indoor resting mosquitoes were also collected fortnightly.

Results  The only two Plasmodium species encountered were P. vivax and P. falciparum in both children and adults. Relatively more malaria infections were recorded in children (≤14 years) than adults (>14 years) (χ2 = 89.94, P < 0.00001). However, there were significant falling trends in P. vivax from 1986 to 2000 in both age groups (≤14 years from 63 to 13, P < 0.0001 and >14 years from 84 to 7, P < 0.0001). The indoor resting density of Anopheles culicifacies, an efficient vector resistant to both dichlorodiphenyltrichloroethane (DDT) (4%) and hexachlorocyclohexane (HCH) (0.4%), was very high throughout this period in all villages (52.35 ± 31.8, range 5–200 per man hour). Anopheles fluviatilis was present in small numbers 0.78 ± 1.24 (range 0–7 per man hour).

Conclusion  Major contributors of the changing epidemiology of malaria in this area are changing drug sensitivity along with insecticide sensitivity.
Four examples of metastatic canine cutaneous nodules
Authors: Hubert B. , Magnol J. P.
Published: December 2004

The causes of single or multiple cutaneous nodules are many. It is important to perform fine‐needle aspirations for cytological examination and to perform skin biopsies. In certain cases a surgical excision of the entire nodule, with a subsequent histological examination, is a simpler approach. When the nodule represents a cutaneous metastasis of a primary neoplasia and is the only sign of the tumour, the aetiology of the nodule can sometimes be rather uncommon. The diagnosis may be made more difficult when the lesion is isolated, appears benign, is in an unusual location (cranial aspect of foreleg, dorsal trunk or scapular region) or is difficult to excise due to haemostasis and local tissue infiltration. The histological interpretation can sometimes prove to be a complicated matter, requiring the use of specific markers to identify anapaestic tumour, but nonetheless is necessary in order to be able to characterize the primary tumour. Using these four examples, a description is provided of the distant localization of an ovarian dysgerminoma in an Afghan hound bitch, a mammary adenocarcinoma in a Labrador bitch, and an extraskeletal mammary osteosarcoma in a doberman bitch and a vesicular adenocarcinoma in a Briard dog. In the last two cases cited, the Alamartine–Ball–Cadiot syndrome was in its final stage of development.
The biology of melanocytes
Authors: Sulaimon Shola S. , Kitchell Barbara E.
Published: December 2004

Abstract In veterinary medicine, our understanding of the biology and regulation of melanocytic function is mostly based on information realized from human and murine studies. Improved understanding of the biology of melanocytes is needed to develop more effective treatment regimens for malignant melanoma and other melanocytic disorders. In vertebrates, melanocytes are well known for their role in skin pigmentation, hair and feather coloration, and for their ability to produce and distribute melanin to surrounding keratinocytes. Enzymes involved in melanin synthesis are present exclusively in melanosomes. The type of melanin synthesized by melanocytes in mammals is regulated at a genetic, biochemical and environmental level. These regulatory factors affect not only the phenotypic appearance, but also the photoprotective properties of melanin. This review addresses the biology of melanocytes, melanin synthesis and the photoprotective properties of melanin.
Comparison of serum dexamethasone concentrations in cats after oral or transdermal administration using Pluronic Lecithin Organogel (PLO): A pilot study
Authors: Willis‐Goulet Heather S. , Schmidt Bill A. , Nicklin Constance F. , Marsella Rosanna , Kunkle Gail A. , Tebbett Ian r.
Published: December 2004

Abstract The objective of this study was to measure and compare the serum concentrations of dexamethasone after oral and transdermal administration using pluronic lecithin organogel in six healthy cats. The study was designed as a crossover, in which the cats were randomly assigned to two groups. The cats received a single dose (0.05 mg kg−1) of dexamethasone either orally or transdermally on the inner pinna. Blood samples were taken at 0, 5, 15, 30, 60, 90 and 120 min, and 3, 4, 6, 8, 12 and 24, 48 and 72 h post dexamethasone administration. A mean peak serum concentration of 30.1 ng mL−1was detected 15 min after oral administration. Serum concentrations were below detection limits by 24 h. In contrast, there was no significant increase in serum concentrations of dexamethasone after transdermal administration. In cats, transdermal administration of a single dose of dexamethasone did not result in significant serum concentrations compared to oral administration.
Gene polymorphisms in glyphosate‐resistant and ‐susceptible biotypes of Eleusine indica from Malaysia
Authors: Ng C H , Wickneswari R , Salmijah S , Teng Y T , Ismail B S
Published: December 2004

Summary Resistant (R) and susceptible (S) biotypes of Eleusine indica were collected from four areas, namely Chaah, Lenggeng, Bidor and Temerloh, in Malaysia. Restriction fragment length polymorphism (RFLP) and polymerase chain reaction (PCR)‐RFLP analyses using SphI restriction enzyme were able to differentiate the R biotype from the S biotype by showing R‐specific and S‐specific polymorphisms in E. indica from three of the areas, with the exception of Temerloh where no polymorphisms were detected. The different DNA profiles for the R biotypes obtained indicate that SphI is not a useful diagnostic marker. The DNA polymorphisms detected in the 5‐enolpyruvylshikimate 3‐phosphate (EPSP) synthase gene suggest that there are different mutation events leading to development of resistance to glyphosate. Partial sequencing of the EPSP synthase gene confirmed different mutations occurring with substitution of proline with serine or threonine at amino acid 106 for the R biotype in Chaah, Bidor and Temerloh.
Xenognosin production and tolerance to Striga asiatica infection of high‐yielding maize cultivars
Authors: Pierce S , Mbwaga A M , Press M C , Scholes J D
Published: December 2004

Summary Ten cultivars of maize, a number of which are high yielding in the presence of the weed Striga asiatica in sub‐Saharan Africa, were tested for resistance and tolerance to this root hemiparasite. Observed in rhizotrons, Striga asiatica attached and developed on roots of all cultivars, indicating that none exhibited a true resistance mechanism. However, root exudates from two cultivars, IWD STR Co and Zea diploperennis BC4C2, stimulated relatively little germination of S. asiatica, indicating low xenognosin production, with few parasites attaching and no significant decrease in host height or biomass apparent after 55 days of infection. Cultivar 98 Syn WEC also exhibited no significant differences in height and biomass on infection despite high xenognosin production and a lack of resistance to parasite attachment and development, indicating the operation of a tolerance mechanism. Cross‐breeding of these low xenognosin‐producing maize cultivars and Striga‐tolerant lines could produce cultivars that perform exceptionally well on Striga‐infested soils. This study highlights the need for cereal breeders to distinguish between the different mechanisms underpinning host performance in the presence of Striga.
Amaranthus hybridus populations resistant to triazine and acetolactate synthase‐inhibiting herbicides
Authors: Maertens K D , Sprague C L , Tranel P J , Hines R A
Published: December 2004

Amaranthus hybridus L. populations (A, B and C) obtained from escapes in Massac County and Pope County fields in southern Illinois, USA were subjected to greenhouse and laboratory experiments to measure multiple resistance to triazine and acetolactate synthase (ALS)‐inhibiting herbicides and cross‐resistance between sulfonylurea and imidazolinone herbicides. Phytotoxicity responses of the three populations revealed that only population B exhibited multiple resistances to triazine and ALS‐inhibiting herbicides. This population was >167‐, >152‐ and >189‐fold resistant to atrazine, imazamox and thifensulfuron, respectively, at the whole plant level compared with the susceptible population. Population A was only resistant to triazines and population C was only resistant to ALS‐inhibiting herbicides. Results from in vivo ALS enzyme and chlorophyll fluorescence assays confirmed these findings and indicated that an altered site‐of‐action mediated resistance to both triazine and ALS‐inhibiting herbicides. Gene sequencing revealed that a glycine for serine substitution at residue 264 of the D1 protein, and a leucine for tryptophan substitution at residue 574 of ALS were the causes of resistance for the three populations.
Fibrin Sealant: Scientific Rationale, Production Methods, Properties, and Current Clinical Use
Authors: Radosevich M. , Goubran H. A. , Burnouf T.
Published: December 2004

Fibrin sealant is a complex plasma‐derived product which is increasingly used as a biodegradable tissue adhesive or sealant to stop or control bleeding or provide air and fluid tightness in many surgical situations. This review describes the historical development of current fibrin sealant preparations and the scientific rationale behind the alleged physiological benefits of its major plasma‐derived components. A comparison in the extraction methods and viral reduction treatments applied to current commercial products and autologous preparations, and their respective advantages and limits, are discussed. Application devices used for surgical applications are described. A survey of the major clinical applications in various surgical areas is presented. Current issues in terms of viral safety, definition of optimal fibrin sealant composition, and regulatory concerns, especially to demonstrate clinical efficacy, are also included.
Heating Lyophilised Factor VIII Does Not Alter Its Recognition by Specific Antibodies
Authors: Gilles J. G. , di Giambattista M. , Laub R. , Saint‐Remy J. M. R.
Published: December 2004

Background and objectives: Alterations of factor VIII (FVIII) during preparation procedures can potentially affect its immunogenicity. One method evaluating such alterations could be by determining the reactivity of FVIII with specific antibodies. Materials and methods: Since heat treatment is currently used to reduce the risk of viral transmission, we evaluated the immunoreactivity of plasma‐derived FVIII before and after heating at different temperatures and for different periods. Freeze‐dried FVIII was used for these experiments as part of the validation procedure of a novel FVIII preparation. Results: Heating FVIII for up to 72 h at 80 °C does not alter its reactivity with specific rabbit antibodies or mouse monoclonal antibodies, although some loss of FVIII activity occurred after 72 h. After heating for 2 h at 100 °C, a procedure that reduced FVIII activity by about 50%, there were still no significant effects on FVIII reactivity with monoclonal antibodies. Conclusions: Freeze‐dried preparations of plasma‐derived FVIII seem to be resistant to heat‐induced structural denaturation.
In vitro Evaluation of the Hemostatic Effectiveness of Non Viable Platelet Preparations: Studies with Frozen‐Thawed, Sonicated or Lyophilized Platelets
Authors: Alemany Monica , Hernandez Maria Rosa , Bozzo Jordi , Galan Ana Maria , Reverter Joan Carles , Mazzara Roberto , Ordinas Antonio , Escolar Gines
Published: December 2004

Objectives:Because of the limited life span of platelets in regular storage systems, we were interested in investigating the effects on hemostasis of nonliving platelet derivatives.Methods:We evaluated the effects of different platelet preparations on primary hemostasis in a well‐established perfusion model. Studies were carried out with blood anticoagulated with low molecular weight heparin. Similar amounts of frozen‐thawed, sonicated or lyophilized platelets were added to normal blood or to blood which had been experimentally depleted of platelets. Platelet interaction with the subendothelium and fibrin deposition were morphometrically evaluated.Results:Addition of nonviable platelet preparations to thrombocytopenic blood always promoted a statistically significant increase in the deposition of fibrin on the subendothelium, but only lyophilized platelets retained some ability to interact with the subendothelium. Flow cytometry studies demonstrated the presence of GPIb, GPIIIa and P‐selectin on lyophilized platelets.Conclusions:Preparations containing nonviable platelets may still retain some hemostatic properties.
Thrombocytopenia Induced by Vancomycin‐Dependent Platelet Antibody
Authors: Mizon P. , Kiefel V. , Mannessier L. , Mueller‐Eckhardt C. , Goudemand J.
Published: December 2004

Background and objectives: Many drugs are associated with thrombocytopenic purpura through immune‐mediated platelet destruction. The case of a woman who suffered lifethreatening thrombocytopenia during vancomycin treatment for Staphylococcus aureus septicemia is reported. Materials and methods: Conventional clinical and laboratory methods, including flow cytometry. Results: After treatment of septicemia with vancomycin, severe thrombocytopenia and bleeding occurred, without detection of drug‐dependent platelet antibodies (DDPA). This was followed by vegetative endocarditis, whereupon antibiotics were withdrawn so as to isolate the organism. The thrombocytopenia was corrected. On day 34, antibiotics including vancomycin were reinstituted, and three days later thrombocytopenia recurred. With a change in antibiotics, the platelet count corrected itself within four days. Conclusions: Vancomycin may induce potentially severe immunological thrombocytopenia.
Storage of Buffy Coat Preparations at 22°C in Plastic Containers with Different Gas Permeability
Authors: Eriksson L. , Eriksson G. , Högman C.F.
Published: December 2004

Background: Buffy coats (BCs) are used as an alternative to platelet‐rich plasma in the preparation of platelet concentrates (PCs). For this purpose the BCs have to be stored for some time at 20–24°C which implies cellular metabolic activity. However, little information is available concerning the effects of a number of factors which may influence the suitability of the preparation as the source of PC. Study design and methods:We studied the effects on BCs of a high and low gas permeability of the wall of the plastic containers, PL2209 and PL146, respectively, mixing versus non‐mixing during storage for 48 h at 22°C, and two types of anticoagulant solutions, CPD and half strength citrate CPD (0.5CPD). The buffy coats were prepared by the bottom and top technique. The median values of volume and haematrocrit were 58–64 ml and 39–45%, respectively. A total of 48 BCs were tested. Blood gases, pH, bicarbonate concentration and haemolysis were determined in the blood mixtures and β‐thromboglobulin (β‐TG), lactate dehydrogenase (LDH), complement factor 3a, and elastase in the extracellular fluid. Results: The pH decreased in all units but to a lesser extent in PL2209 containers than in PL146 units. In the former the pCO2 decreased slowly in contrast to the latter where it increased by about 50%. Mixing during storage increased the pH and decreased the pCO2 in 0.5CPD‐PL146 and CPD‐PL2209 units, as compared to resting, while no effects of mixing were observed in the other groups. The pO2 decreased to low levels in PL146 units. The haemolysis and LDH release were higher in mixed than in unmixed units. The initial β‐TG levels were lowest in 0.5CPD‐PL146 units which also had the lowest 24‐hour levels. The release of β‐TG during storage was smallest in CPD resting units. The elastase release was significantly higher in 0.5CPD than in CPD units already from the beginning of storage and increased during storage at about the same rate irrespective of mixing. The C3a levels were higher in 0.5CPD‐PL2209 units than in the other units at 2 h. Storage for 24 h caused an increase by 2–3 times of the original level without any clear relation to storage conditions. Conclusions: In BC units accumulation of CO2 occurs in containers with low gas permeability. These also show the most rapid pH decrease during storage. Prolonged holding of BCs puts extra emphasis on the need of satisfactory gas permeability of the container for platelet storage in BC‐derived PCs. Continuous mixing causes red cell damage and does not seem to have any clear benefit. The release of granulocyte elastase was higher in 0.5CPD than in CPD units but there was no indication of an associated increase in platelet activation. Summary: Study of buffy coats stored in various media and containers at 22°C suggests that it is better to restrict storage to 24 h or less to avoid activation or other deleterious effects on the platelets.
Removal of Methylene Blue from Plasma via an Adsorbent Filter
Authors: AuBuchon James P. , Pickard Constance , Herschel Louise , O'Connor James L. , Lee Eric
Published: December 2004

Objective: To evaluate adsorbent filtration of methylene blue (MB) and leukocytes from plasma. Methods: Plasma (750 ml) from apheresis of 10 normal subjects was split into three aliquots: control (A), filtration (B) and MB addition (to 1 μM), phototreatment and filtration (C). Biochemical and coagulation tests were performed; units A and B were reinfused. Results: Filtration reduced MB to undetectable (<0.05 μM) levels and leukocytes by 3 log10. Biochemical analytes were unchanged. The partial thromboplastin time was prolonged with MB addition (11±13%) or filtration (26±12%, p < 0.05), but the effects were not additive. Autologous transfusion was well tolerated. Conclusion: Adsorbent filtration can reduce residual MB to undetectable levels and yields a component suitable for transfusion.
Hypotensive Reactions Associated with Platelet Transfusions and Angiotensin‐Converting Enzyme Inhibitors
Authors: Mair B. , Leparc G. F.
Published: December 2004

Background and Objectives: To determine the cause of hypotensive reactions associated with platelet transfusions in coronary artery bypass surgery patients. Materials and Methods: Platelet transfusion reactions that occurred during a 3‐month period were retrospectively reviewed. Results: Eighteen transfusion reactions occurred in 16 patients. Sixteen of the reactions were hypotensive, and occurred in 14 patients transfused with platelets through negatively charged bedside leukocyte reduction filters. All 14 patients had received angiotensin‐converting enzyme (ACE) inhibitors prior to transfusion. Conclusion: Preliminary findings suggest a new type of transfusion reaction associated with the use of negatively charged leukocyte reduction filters during platelet transfusions to patients on ACE inhibitors.
Fas and Fas Ligand Expression on Human Peripheral Blood Leukocytes
Authors: Mincheff Milcho , Loukinov Dmitri , Zoubak Serguei , Hammett Michael , Meryman Harold
Published: December 2004

Objectives: Study of Fas and Fas ligand(Fas‐L) expression, as well as sFas–L release, by fresh human peripheral blood leukocytes. Methods: Flow cytometry, cytotoxicity, immunofluorescence staining of fresh smears, Western blotting, Results: Granulocytes and monocytes express a low level of Fas receptor, but no Fas–L. These cells, as well as NK cells, contain presynthesized depots of Fas–L which they express following activation by brief storage (60 min) at room temperature or during separation from whole blood. Such activation also leads to Fas receptor upregulation. NK cells do not express Fas receptor. Once expressed on blood leukocytes, fully functional Fas–L can be released from the membrane and can be detected in plasma–free cell supernatants. Conclusion: Human peripheral blood granulocytes, monocytes and NK cells contain intracellular presynthesized Fas–L which they readily express following blood anticoagulation, blood storage or cell separation. Soluble Fas–L is released from those cells and can be detected in protein–free supernatants by immunoblotting.
Outcome of Transfusion of K:11 Erythrocytes in a Patient with Anti‐K11 Antibody
Authors: Kelley Carolyn M. , Karwal Mark W. , Schlueter Annette J. , Olson John D.
Published: December 2004

Background and objectives: The clinical significance of anti‐K11 in red cell transfusion therapy is unknown. We report the outcome of transfusion of K:11 erythrocytes into a patient with a known anti‐K11 antibody. Materials and methods: The patient was monitored clinically following transfusion of 11 units of K: 11 erythrocytes. A red cell survival study with K: 11 erythrocytes and a monocyte monolayer assay (MMA) were performed. Results: No adverse clinical outcome was detected. The red cell survival study showed normal survival of K: 11 erythrocytes, and the MMA showed no increase in reactive monocytes. Conclusion: These findings suggest that K:11 red cells can safely be transfused to individuals with anti‐K11 antibody.
Estimated Risk of Transmission of Hepatitis C Virus by Blood Transfusion
Authors: Koerner K. , Cardoso M. , Dengler Th. , Kerowgan M. , Kubanek B.
Published: December 2004

Objective: The risk of transmitting hepatitis C (HCV) by transfusion of anti‐HCV‐negative screened blood was estimated for the blood donor population of Baden‐Württemberg (southwestern Germany). Methods: The data from the blood donors screened for anti‐HCV and for HBsAg during 1990‐1995 were analyzed. Results: The prevalence of confirmed anti‐HCV‐positive blood donations decreased continuously during the last 5 years, reaching 121 per 100,000 blood donations. A higher anti‐HCV prevalence rate was found in female than in male blood donors (p<0.05). The estimated risk of transmitting HCV during the window period is 1:200,000 (1:97,000–1:1,400,000) for repeat donors. In 1995, the calculated risk for first‐time donors was 1:20,000 (1:15,000–28,000). The incidence for HCV was 1.2 per 100,000 blood donations. Conclusion: The risk of transmitting hepatitis C by blood transfusion is low. Additional tests to shorten the window period to detect antibodies to HCV might increase the safety of blood transfusion.
Monocyte Activation in Platelet Concentrates
Authors: Grey D. , Erber W. N. , Saunders K. M. , Lown J. A. G.
Published: December 2004

Background and Objectives: Cytokines in platelet concentrates contribute to febrile transfusion reactions. Activated monocytes are a major source of inflammatory cytokines, however the role of monocytes in cytokine production in platelet concentrates has not been clarified. This study undertook to quantitate monocytes, determine whether monocyte activation occurs and identify an association with IL‐6 and IL‐β concentrations in platelet concentrates. Materials and Methods: 17 platelet concentrates were analysed for total leucocyte and monocyte counts, CD14 and CD16 monocyte‐associated antigen expression and IL‐1β and IL‐6 measurements on days 1, 2, 3, 4 and 5. Results: Monocytes in all platelet concentrates expressed increased levels of CD14 and CD16 from day 1 of storage. 10/17 platelet concentrates had elevated IL‐6 levels by day 3. Platelet concentrates with IL‐6 levels above 15 pg/ml on day 5 had monocyte counts between 0.14 and 15.6×106/unit on day 1, while those with IL‐6 levels below 15 pg/ml had low monocyte counts of <0.01 to 1.2×106/unit on day 1. Conclusion: Monocytes present in platelet concentrates exhibit features of activation. Monocyte activation is present following the preparation of platelet concentrates, implicating the manufacturing process in its development. Increased IL‐6 and IL‐1β levels during platelet concentrate storage are commonly associated with a higher monocyte count. However, no direct association could be identified between the extent of monocyte activation and the level of cytokine release.
Erythropoiesis in Patients Stimulated with Erythropoietin: The Relevance of Storage Iron
Authors: Goodnough Lawrence Tim , Marcus Randall E.
Published: December 2004

Background and Objectives: The clinical importance of iron‐restricted erythropoiesis in erythropoietin (EPO)‐stimulated patients is controversial. Materials and Methods: We therefore reviewed 70 patients randomized into clinical trials of aggressive autologous donation and oral iron supplementation, with or without recombinant human EPO therapy. Results: Nineteen (27%) iron‐depleted patients produced 5.4±2.8 ml RBC/kg compared to 4.8±2.3 ml RBC/kg (nonsignificant) in iron‐replete patients due to endogenous EPO (placebo group) stimulation. EPO‐treated iron‐depleted patients produced 20% less RBC than iron‐replete patients (8.23±3.3 vs. 10.2±4.0, p=0.066). RBC volume expansion correlated with initial storage iron only in iron‐replete patients who received EPO therapy. Conclusion: Initial storage iron status is a marginally important limitation to EPO‐mediated erythropoiesis in the setting of oral iron supplementation. Strategies to maintain plasma transferrin saturation with intravenous iron therapy may be desirable to improve the erythropoietic response to EPO in this setting.
Increased Lipid Peroxidation of Erythrocytes in Blood Stored in Polyvinyl Chloride Blood Storage Bags Plasticized with Di‐[2‐Ethyl Hexyl] Phthalate and Effect of Antioxidants
Authors: Deepa Devi K. V. , Kumar V. Manoj , Arun P. , Santhosh Anitha , Padmakumaran Nair K. G. , Lakshmi L. R. , Kurup P. A.
Published: December 2004

Background and Objectives: Previous work in this laboratory has shown significant decrease in vitamin E in erythrocytes in blood stored in polyvinyl chloride (PVC) bags plasticized with di‐[2‐ethyl hexyl] phthalate (DEHP), and in erythrocytes incubated in vitro with DEHP. Since vitamin E is a major antioxidant, a study was carried out to find out whether this decrease observed in vitamin E has an effect on lipid peroxidation in blood stored in DEHP‐plasticized PVC blood bags. Materials and Methods: Blood was collected in Penpol blood storage bags (which is a DEHP‐plasticized PVC bag) and parameters of lipid peroxidation, i.e. activity of superoxide dismutase (SOD), catalase, glutathione peroxidase, glutathione reductase, concentration of malondialdehyde (MDA), conjugated dienes, hydroperoxides, glutathione and vitamin E studied in erythrocytes after various periods of storage as compared to glass bottles. Erythrocytes were also incubated in vitro with DEHP with and without vitamin E, and changes in lipid peroxidation studied. Results:Blood stored in Penpol bags showed increased lipid peroxidation in erythrocytes as compared to that stored in glass bottles, as is evident from a greater increase in MDA and a greater decrease in glutathione and a significant decrease in vitamin E. The addition of vitamin E decreased the formation of MDA and conjugated dienes and prevented the decrease in vitamin E. However in spite of increased lipid peroxidation in the presence of DEHP, the release of K+ and hemoglobin from erythrocytes was lower. When there was an increase in DEHP taken up by erythrocytes, there was a corresponding decrease in vitamin E. More important, whenever there was an increase in vitamin E in erythrocytes (when RBCs in the presence of DEHP were incubated with vitamin E), there was a progressive decrease in DEHP. Conclusion: DEHP caused increased lipid peroxidation in erythrocytes. At the same time, it decreased the release of K+ and hemoglobin from erythrocytes. It is possible that the stabilizing effect of DEHP on the erythrocyte membrane may offset the detrimental effects of the increased lipid peroxidation it causes.
Platelets Stored in a New‐Generation Container
Authors: de Wildt‐Eggen J. , Schrijver J. G. , Smid W. M. , Joie M. , Bollinne V. , Bins M.
Published: December 2004

Background and Objectives: In order to preserve platelet concentrates (PCs) with high yields, a new polyolefin platelet storage container (PL 2410, 1.3L, Baxter, La Châtre, France) with increased gas permeability in combination with a larger surface area has been developed. The storage capacity was studied with platelets in plasma and platelets additive solution. Materials and Methods: Platelet concentrate pools (PCs) of different yields suspended in either plasma (PCs‐PL; n = 30) or PAS II (PCs‐PAS; n = 37) were prepared. For preparation of PCs with a low, intermediate and high number of platelets 3, 5 and 6 buffy coat (BCs) were pooled with different volumes of plasma and 5 and 6 BCs were pooled with different volumes of PAS II, in order to obtain PCs of equal volumes comparable with routine conditions. All PCs were stored on a flatbed shaker at 22±2°C and evaluated on days 1, 2, 5 and 7 by determining platelet and white cell counts, pH (37°C), pO2, pCO2 and swirling score. Results: Platelet yields ranged from 1.5 up to 5.5 × 1011 platelets/U. On day 7 all PCs‐PL (n = 4) with platelet yields above 4.5×1011 had a pH value below 6.8 (range 5.91–6.79). While 7 of 8 PCs‐PAS units with platelet yields above 4.0×1011/U showed a pH value below 6.8 (range 6.31–6.70). Conclusion: During 7 days of storage in a new 1.3‐liter platelet container, the pH was maintained above 6.8 in PCs in plasma with a yield between 1.5‐and 4.5×1011/U; when PAS II was used, the maximum platelet yield allowed for comparable pH maintenance was somewhat lower (4.0x1011/U).
Autologous Platelet Collection and Storage to Support Thrombocytopenia in Patients Undergoing High‐Dose Chemotherapy and Circulating Progenitor Cell Transplantation for High‐Risk Breast Cancer
Authors: Torretta Lorella , Perotti Cesare , Pedrazzoli Paolo , Dornini Gianluca , Viarengo Gianluca , Livraghi Alessandra , Noris Patrizia , Prada Gian Antonio Da , Balduini Carlo L. , Cuna Gioacchino Robustelli della , Salvaneschi Laura
Published: December 2004

Objectives: The use of circulating progenitor cell support following high‐dose chemotherapy for malignancies decreases but does not entirely abolish platelet transfusion requirement. We investigated the feasibility of supporting the posttransplant thrombocytopenic phase exclusively with autologous platelets collected by apheresis and cryopreserved. Methods: 25 patients underwent plateletpheresis during the platelet rebound occurring after high‐dose cyclophosphamide. Autologous platelets were cryopreserved in 5% dimethylsulfoxide, thawed and transfused during the aplastic phase after the myeloablative regimen whenever clinically required. Results: A single plateletpheresis was carried out in all patients, allowing the harvest of a platelet concentrate with a mean value of 7.7 × 10 11 platelets. No significant procedure‐ or transfusion‐related side effects were recorded. Mean platelet recovery after freezing and thawing was 63% and the mean number of platelet reinfused was 4.8 × 1011; 23 of 25 patients were fully supported with autologous platelets. Conclusion: Plateletpheresis performed in our selected group of patients was found to be a safe and effective procedure to collect large amounts of autologous platelets; the numbers obtained proved to be sufficient for the transfusion demand of almost all patients.
Two Examples of an Inseparable Antibody That Reacts Equally Well with DW+ and Rh32+ Red Blood Cells
Authors: Reid Marion E. , Sausais Laima , Zaroulis Charles G. , Mohandas Kala , Coghlan Gail , Lomas‐Francis Christine
Published: December 2004

Background and Objectives: Anti‐DW is a rare specificity that detects an antigen on DVa red blood cells (RBCs). Some anti‐DW contain an inseparable component that cross‐reacts weakly with RBCs expressing the low‐incidence Rh antigen Rh32. RH32 is expressed by RBCs with either the RN or the DBT phenotype. Case Report: We describe here an antibody found in the serum of 2 patients that reacts equally well with RBCs possessing either DVa, RN, or DBT phenotypes. The reactivity for DW and Rh32 antigens could not be separated by adsorption onto and elution from DW+Rh32‐or from DW ‐Rh32+RBCs. Conclusions: We suggest that amino acids encoded by nucleotides at the junction of exon 4 of RHD to exon 5 of RHCE may induce a conformation that is recognized by these equally reactive inseparable antibodies. Until such time that the epitope recognized by these antibodies is defined, we recommend use of the descriptive name anti‐DW/Rh 32.
NA1/NA2 Antisera Inhibit FcγRI‐but Not FcγRII‐Mediated Phagocytosis
Authors: Flesch Brigitte K. , Maass Wiebke , Neppert Jürgen
Published: December 2004

Background and Objectives: Alloantibodies against the granulocyte‐specific NA antigens play an important role in alloimmune neonatal neutropenia. As the NA system is located on the FcγRIIIb, the influence of NA‐specific antibodies on granulocyte function is of special interest. Materials and Methods: We tested alloantisera specific for NA1 and NA2 for their ability to influence FcγR‐mediated phagocytosis of polymorphonuclear neutrophils by use of different FcγR specific targets. Red blood cells coated with human IgG anti‐D served as specific targets for FcγRI‐mediated phagocytosis while mouse IgG1 anti‐glycophorin A was used to coat red blood cells (RBCs) to obtain FcγRII specific targets. To test for a hypothetical induction of phagocytosis by FcγRIIIb we used D– RBCs coated with human monoclonal anti‐D as target cells for unprimed neutrophils. Results: Granulocyte phagocytosis was directly induced by FcγRI and FcγRII but not by FcγRIIIb. NA1 alloantisera significantly inhibited FcγRI‐mediated phagocytosis of IFN‐γ‐stimulated neutrophils if the corresponding antigen was expressed. Conversely, NA2 alloantisera inhibited FcγRI‐mediated phagocytosis in NA2‐positive individuals. There was no effect of NA1‐ and NA2‐specific alloantibodies on FcγRII‐mediated phagocytosis. Conclusion: NA‐specific alloantisera inhibit the FcγRI‐induced phagocytosis in primed neutrophils, but they do not significantly inhibit their FcγRIIa‐specific phagocytosis of mIgG1‐coated RBCs.
Progression to AIDS in Relation to Clinical Factors and Clotting Product Consumption
Authors: Roosendaal G. , van der Schouw Y. T. , Mauser‐Bunschoten E. P. , Borleffs J. C. , van den Berg H. M.
Published: December 2004

Objective: In order to describe the progression of HIV disease in a cohort of Dutch HIV‐1‐infected patients with haemophilia and to study the influence of clinical and clotting factors, we performed a 15‐year longitudinal clinical and laboratory study. Methods: The progression to AIDS in relation to type of haemophilia, antibodies against factor VII, age, cytomegalovirus (CMV) infection, and type and total amount of clotting factor consumption was studied in 52 haemophilic patients infected with HIV‐1 between 1981 and July 1985. Results: The progression to AIDS was faster in the group of patients with antibodies against factor VIII (inhibitors) and in the group of patients with haemophilia B than in the group of patients with haemophilia A without inhibitors. Conclusion: We concluded that HIV‐infected patients with haemophilia A with inhibitors and patients with haemophilia A without inhibitors. These differences are independent of the total amount of clotting factor used and other known risk factors such as age at seroconversion and CMV in fection and may be due to the type of clotting product used.
Sequence and Specificity Analysis of Recombinant Human Fab Anti‐Rh D Isolated by Phage Display
Authors: Miescher Sylvia , Vogel Monique , Biaggi Christine , Ramseyer Vreni , Hustinx Hein , Eicher Nicole , Imboden Martin A. , Spycher Martin O. , Amstutz Hanspeter , Stadler Beda M.
Published: December 2004

Background and Objectives: Hyperimmune anti‐Rh D serum is worldwide in short supply. As a first step to develop an alternative source of Rh D antibodies, we describe in this study the isolation and characterization of recombinant antiRh D Fab fragments. Materials and Methods: Peripheral blood mononuclear cells harvested from a hyperimmunized donor were used to construct two combinatorial Fab libraries. Phages expressing these Fab fragments were selected on whole red blood cells followed by testing of positive clones in an indirect hemagglutination assay. Results: Individual Fab clones are of high affinity and competitively inhibit the binding of a registered anti‐D immunoglobulin. The Fab clones are also specific against the partial D phenotypes, Rh33, DIII, DIVa, DIVb DVa, and DVII. The 13 different but highly homologous clones express preferentially VH3 segments. Conclusion: These Fab fragments show potential for the development of a new generation of therapeutic anti‐Rh D reagents.