bluuebunny/crossref_metadata_2025 huggingface dataset:
Significance of Cryptosporidium as an aetiology of acute infectious diarrhoea in elderly Indians
Authors: Gambhir I. S. , Jaiswal J. P. , Nath G.
Published: December 2004

objective  To explore the significance of Cryptosporidium isolation in elderly Indian with acute infectious diarrhoea and its clinical correlates.

design  A hospital‐based clinico‐aetiological study of 120 patients aged 60 years or older, 25 adults younger than 60 years and 25 children up to age 14 with acute diarrhoea, and 57 apparently healthy elderly individuals.

results  Cryptosporidium was isolated in 22 older patients with diarrhoea (18.3%) and was highly significant (P < 0.01) compared with healthy age‐matched controls. Of these patients 66% had a history of close contact with animals. Most (68%) Cryptosporidium infections occurred during the rainy season. Among the elderly patients 17% suffered from vomiting and abdominal pain, 31% were febrile; none were severely dehydrated. Stools numbered three to nine per day with duration of 5–17 days. Stool leucocytes were <6/hpf and no RBCs were seen. Isolation of Cryptosporidium in older persons was associated with diabetes mellitus (22.7%), tuberculosis (9.0%), malignancy (4.5%) and coronary artery disease (4.5%).

conclusion  Cryptosporidium is an important cause of diarrhoea in elderly Indians, especially those with close contact with animals. The infection has a mild clinical course, is self‐limiting and does not cause dysenteric stool. Its effect on nutrition requires further study.
Infertility in Central Africa
Authors: Larsen Ulla
Published: December 2004

objectives To determine the prevalence and risk factors of primary and secondary infertility in the four Central African countries of Cameroon, Chad, Central African Republic and Gabon.

methods Primary infertility was approximated by the percentage of women childless after at least 5 years of marriage. The percentage with no birth at least 5 years subsequent to a previous birth was considered to have secondary infertility. Logistic regression and discrete logistic regression models were estimated to determine the risk factors of primary and secondary infertility, respectively. The relatively few women who were defined as infertile and reported using a traditional or modern method of contraception at survey date were considered fertile to reduce bias from falsely classifying effective contraceptive users as infertile.

results The prevalence of infertility was highest in Central African Republic and lowest in Chad: primary infertility ranged from 6.9% to 3.1% and secondary infertility for women aged 20–44 years ranged from 26.5% to 18.9%. Women married more than once vs. only once and formerly married women vs. women living with their husbands had higher odds of primary and secondary infertility. Also, younger cohorts had relatively higher risks of primary (born after 1970) and secondary infertility (born after 1960) compared with women born before 1960. In contrast, place of residence, religion, education of wife or husband and socio‐economic status were generally not associated with primary or secondary infertility.

conclusions Infertility is still prevalent in Central Africa and new interventions aimed at reducing the incidence and social implications of pathologic infertility are needed.
Attitude of women in western Uganda towards pre‐packed, unit‐dosed malaria treatment for children
Authors: Kilian A. H. D. , Tindyebwa D. , Gülck T. , Byamukama W. , Rubaale T. , Kabagambe G. , Korte R.
Published: December 2004

In the context of a larger study on malaria related knowledge, attitudes, practices and beliefs in western Uganda 813 women aged 15–49 years were shown a sample of a pre‐packed, unit‐dosed malaria treatment for children, its use was explained and attitudes of the women were investigated. Of all women, 90.5% (86% urban, 92% rural) said they would prefer the pre‐packed over the conventional type of treatment and 93.9% of these were willing to pay between 0.17 (rural) and 0.29 (urban) US$ more for this treatment. Two‐thirds (67.8%) thought that they would not have to ask their spouses before making a decision on the kind of treatment and 59.5% said they would rather stock the treatment at home than buy it when a child gets sick. The most mentioned reason for preferring pre‐packs was their safety and cleanliness, while ease of application, dosing and compliance were secondary. We conclude that pre‐packed, unit‐dosed malaria treatment is accepted by the caretakers of children in the area studied and that they readily understand and accept its concept. This indicates a high potential for this approach to improve the home management of malaria fevers and reduce malaria‐related morbidity and mortality if adequate coverage can be achieved and if the intervention is embedded into an appropriate programme of behavioural change communication and provider training.
Malaria and property accumulation in rice production systems in the savannah zone of Côte d'Ivoire
Authors: Audibert M. , Mathonnat J. , Henry M. C.
Published: December 2004

Irrigation stabilizes agricultural production and hence improves farmers’ living standards and conditions. The permanent presence of water may, however, increase the burden of water‐related parasitic diseases and counter the economic benefits of irrigation by reducing farmers’ health. The purpose of this study was to assess the impact of malaria on farm household property, beyond the health risk (studied elsewhere). The research question was: by weakening individuals, does malaria reduce productive capacities and income workers, and consequently limit their property accumulation? To test this hypothesis, we use data on property (farming equipment, livestock and durable consumer goods) and Plasmodium falciparum indicators generated by a study carried out in 1998 in the Ivorian savannah zone characterized by inland valley rice cultivation, with a sample of nearly 750 farming households. Property is influenced by many factors related to the size of the family, the area under cultivation and high parasite density infection rate of P. falciparum. A significant negative correlation between high‐density infection rate and the property values confirms that by reducing the living standards of households, malaria is a limiting factor for property accumulation.
Cardiovascular risk factor levels in a lower middle‐class community in Ankara, Turkey
Authors: Tezcan S. , Altıntaş H. , Sönmez R. , Akinci A. , Doğan B. , Çakır B. , Bilgin Y. , Klör H. U. , Razum Oliver
Published: December 2004

Summary Study objective  To assess the prevalence of risk factors for coronary heart disease (CHD) in a lower middle‐class urban community of Turkey.

Design Cross‐sectional study in an age‐ and sex‐stratified random community sample with equal sample size per stratum. Direct age‐standardization using the standard world population to allow international comparison of findings. Logistic regression modelling to identify risk factors for obesity.

Setting Gülveren, a residential area in Ankara, total population 23 000 persons.

Participants A total of 1672 adults aged 25–64 years and resident in the study community were interviewed, 1272 (76.1%) of those came for physical examination.

Main results The age‐standardized prevalence of hypertension according to WHO MONICA criteria was 18.6% (95% confidence interval: 16.1–21.1%) among women and 12.3% (9.7–14.9%) among men; of obesity (body mass index, BMI ≥30 kg/m2) 51.0% (47.6–54.3%) among women and 15.1% (12.0–18.2%) among men; of current smoking 20.1% (17.5–22.6%) among women and 64.8% (61.4–68.2%) among men; of hypercholesterolaemia 20.1% (17.4–22.9%) among women and 13.8% (10.8–16.8%) among men; and of low high density lipoprotein (HDL) 48.4% (44.8–52.1%) among women and 40.6% (36.0–45.2%) among men. In the regression model, age, female sex, non‐ and ex‐smoking were associated with obesity.

Conclusions The prevalence of smoking, obesity and low HDL is high in this urban, lower middle‐class population, even in comparison with industrialized countries. Unexpectedly, women have less favourable CHD risk profiles than men, except for smoking. Preventive action should be community‐wide and address the common risk factors simultaneously to avoid replacement effects such as becoming obese after quitting smoking.
Short communication: Comparison of serology and isolates for the identification of infecting leptospiral serogroups in Hawaii, 1979–1998
Authors: Katz Alan R. , Effler Paul V. , Ansdell Vernon E.
Published: December 2004

Laboratory confirmation of leptospirosis is usually accomplished serologically, without isolates, using the microscopic agglutination test (MAT). However, optimal performance of the MAT is dependent on the knowledge of enzootic serogroups and serovars so that an appropriate MAT antigen testing battery can be established. Infecting leptospiral serogroups can be identified serologically without isolates, using the MAT, or by serogrouping of isolates, but little information is available regarding the correlation between these methods. The identification of infecting serogroups for 53 culture‐confirmed leptospirosis cases, diagnosed in Hawaii between 1979 and 1998, using serology and culture isolates were compared. The overall agreement between the two methods was good (κ = 0.71, 95% CI: 0.56, 0.86). However, the agreement varied between serogroups from 0 to 100%. In establishing the prevalence of serogroups, results obtained via MAT serology (in the absence of serogrouped isolates) should be considered presumptive rather than definitive.
Climatic and social risk factors for Aedes infestation in rural Thailand
Authors: Nagao Yoshiro , Thavara Usavadee , Chitnumsup Pensri , Tawatsin Apiwat , Chansang Chitti , Campbell‐Lendrum Diarmid
Published: December 2004

An intense epidemic of dengue haemorrhagic fever in 1998 prompted the Thai government to investigate the feasibility of focalized vector (Aedes aegypti) control programmes. We tested for correlations of three indices of Aedes larval abundance (housing index, container index and Breteau index) against 38 socio‐economic and four climatic variables. Availability of public water wells, existence of transport services and proportion of tin houses were positively associated with larval indices. Private water wells, health education, health insurance coverage, thatched houses and use of firewood for cooking were negatively associated. These probably represent both direct effects on breeding sites (private vs. public wells decrease necessity to store water, and health education may encourage breeding site removal), and more general effects of health‐related attitudes, housing quality and remoteness from urban areas. Indices were positively associated with daily minimum temperature, an increase in precipitation from the previous month (reflecting the onset of the rainy season) and daily maximum temperatures of approximately 33–34 °C. The associations were used to derive statistical models to predict the rank order of larval indices within the study area (Spearman's correlation coefficients = 0.525–0.554). The study provides a rational basis for identifying possible social interventions, and for prioritizing previously unsurveyed villages for further monitoring and focalized vector control.
Short communication: Towards saliva‐based screening for sleeping sickness?
Authors: Lejon V. , Kwete J. , Büscher P.
Published: December 2004

A pilot study was carried out on the detection of trypanosome‐specific antibodies in saliva for diagnosis of sleeping sickness. All twenty‐three saliva samples of parasitologically confirmed Trypanosoma brucei gambiense patients tested positive in an indirect enzyme‐linked immunosorbent assay, whereas all 14 saliva samples of a negative control group remained negative. Trypanosome‐specific antibody levels in patient saliva correlated with antibody levels in serum, but were about 250‐fold lower. Eight of 23 undiluted saliva samples of sleeping sickness patients tested positive in CATT/T. b. gambiense and two of 23 in LATEX/T. b. gambiense. All fourteen saliva samples of the negative control group were also positive in CATT/T. b. gambiense, as were four of 14 in LATEX/T. b. gambiense. CATT and LATEX were thus inappropriate for antibody detection in saliva. These results indicate that trypanosome‐specific antibody detection in saliva is possible. This could lead to the development of a simple, non‐invasive, reliable saliva field test for diagnosis of sleeping sickness.
DEET mosquito repellent sold through social marketing provides personal protection against malaria in an area of all‐night mosquito biting and partial coverage of insecticide‐treated nets: a case–control study of effectiveness
Authors: Rowland Mark , Freeman Tim , Downey Gerald , Hadi Abdul , Saeed Mohammed
Published: December 2004

DEET (Diethyl‐3‐methylbenzamide), the widely used mosquito repellent, has the potential to prevent malarial infection but hitherto there has been no study demonstrating this possibility during normal everyday use. MosbarTM, a repellent soap containing DEET, was promoted through social marketing in villages in eastern Afghanistan. This was followed up with a case–control study of effectiveness against malarial infection conducted through local clinics. Mosbar was purchased by 43% of households. Reported use of insecticide‐treated nets (ITNs) was 65% among the control group. There was a strong association between Mosbar use and ITN use, as 81% of Mosbar users also possessed ITN. The use of Mosbar was associated with a 45% reduction in the odds of malaria (95% CI: −11% to 72%, P = 0.08) after adjusting for ITN and other unmatched factors. Ownership of ITNs was associated with a 46% reduction in the odds of malaria (95% CI: 12% to 67%, P = 0.013) after adjusting for Mosbar and other unmatched factors. The greatest reduction in the odds of malaria was associated with combined use of Mosbar and ITN (69% reduction, 95% CI: 28% to 87%, P = 0.007). The association between recalled use of Mosbar 10 days ago (nearer the time of infection) and reduction in malarial infections (adjusted odds ratio 0.08, 95% CI: 0.01–0.61, P = 0.001) was significantly stronger than that shown by current use of Mosbar. Most purchasers of Mosbar were satisfied with the product (74%), although a minority said they preferred to use only ITN (8%). The local mosquito vectors, Anopheles stephensi and A. nigerrimus, started biting shortly after dusk and continued biting until early morning. It was shown that Mosbar prevented biting throughout this period. In regions where mosquito vectors bite during evening and night, repellents could have a useful supplementary role to ITN and their use should be more widely encouraged.
Immunohistochemical evaluation of mononuclear infiltrates in canine lupoid onychodystrophy
Authors: Mueller R. S. , West K. , Bettenay S. V.
Published: December 2004

The purpose of this study was to characterize the immunophenotype of the inflammatory infiltrate in canine lupoid onychodystrophy and to determine if decalcification interferes with immunoreactivity. Claw biopsies of 14 dogs with lupoid onychodystrophy were stained with CD3, BLA36 and HM57 (CD97α), MAC 387, lysozyme and MHC class II using an immunoperoxidase and avidin/biotin technique. Cells infiltrating the claw matrix were counted in two high‐power fields; numbers were expressed as a percentage of the cellular infiltrate. The inflammatory infiltrate consisted predominantly of B and T cells; macrophages were typically only present in small numbers. There was no correlation between the predominant inflammatory cell and clinical parameters. CD3, BLA36, lysozyme and MHC II preserved significant antigenicity during formalin fixation and short decalcification, while staining for CD79A and particularly MAC 387 was less reliable. Prolonged decalcification of 7–14 days abolished immunostaining with all antibodies. Short decalcification periods allow immunostaining of samples with some antibodies (CD3, BLA36, lysozyme, MHC II), but not others (CD79A, MAC387), while prolonged decalcification prohibits immunostaining. The nature of the mononuclear infiltrate in lupoid onychodystrophy is not helpful clinically.

 This study was funded by the American Academy of Veterinary Dermatology.
Feline cutaneous neuroendocrine carcinoma (Merkel cell tumour): clinical and pathological findings
Authors: Bagnasco Giorgio , Properzi Roberto , Porto Roberto , Nardini Vincenzo , Poli Alessandro , Abramo Francesca
Published: December 2004

Abstract A case of a feline Merkel cell tumour is described. An 8‐year‐old, female cat developed a round, alopecic, reddish mass on the nose. Wide excisional surgery was performed with cartilage resection. Histologically the mass was composed of solid islands of mostly basophilic densely packed cells with a scant cytoplasm, which was suggestive of a neuroendocrine origin. Results of immunohistochemical studies using antibodies against neurone‐specific enolase, chromogranin, synaptophysin and pan‐cytokeratin allowed classification of the lesion as a Merkel cell tumour. Ultrastructurally, dense core granules were identified in the cytoplasm. In a 2‐year follow‐up no relapses or metastases were observed. The clinical course recorded is in contrast with the malignant nature of a Merkel cell tumour recently described in a cat and of the human Merkel cell tumour, but is similar to the course of the canine Merkel cell tumour which is often benign. Early diagnosis along with the use of wide surgical excision might be considered an important factor in preventing relapse of this tumour.
Ecological aspects of seed dormancy‐break and germination in Heteranthera limosa (Pontederiaceae), a summer annual weed of rice fields
Authors: Baskin C C , Baskin J M , Chester E W
Published: December 2004

Summary Heteranthera limosa seeds were buried in flooded and in non‐flooded soil and exposed to natural seasonal temperature changes in Lexington, Kentucky, USA. Seeds exhumed after various periods of burial ranging from 2 to 36 months were tested for germination under both flooded and non‐flooded conditions. Seeds were dormant at maturity in September and became non‐dormant during winter. Seeds buried in non‐flooded soil during winter germinated to higher percentages and over a wider range of temperatures when tested under flooded conditions (in light) during spring and summer, than did those buried in flooded soil during winter. Thus, the water regime associated with rice culture (non‐flooded in winter and flooded in summer) is optimal for dormancy‐break and germination of H. limosa seeds. A portion of the buried seeds exhibited an annual dormancy/non‐dormancy cycle, whereas others had a conditional dormancy/non‐dormancy cycle. Regardless of the type of cycle, seeds buried in non‐flooded soil retained the ability to germinate in light at high temperatures under flooded conditions throughout the summer. Thus, seeds potentially can germinate at any time during the growing season, whenever rice fields are flooded. Flooding fields during winter and/or sowing rice relatively early in the growing season may help in establishing rice before seeds of H. limosa germinate.
The role of weeds in supporting biological diversity within crop fields*
Authors: Marshall E J P , Brown V K , Boatman N D , Lutman P J W , Squire G R , Ward L K
Published: December 2004

Summary Weeds are major constraints on crop production, yet as part of the primary producers within farming systems, they may be important components of the agroecosystem. Using published literature, the role of weeds in arable systems for other above‐ground trophic levels are examined. In the UK, there is evidence that weed flora have changed over the past century, with some species declining in abundance, whereas others have increased. There is also some evidence for a decline in the size of arable weed seedbanks. Some of these changes reflect improved agricultural efficiency, changes to more winter‐sown crops in arable rotations and the use of more broad‐spectrum herbicide combinations. Interrogation of a database of records of phytophagous insects associated with plant species in the UK reveals that many arable weed species support a high diversity of insect species. Reductions in abundances of host plants may affect associated insects and other taxa. A number of insect groups and farmland birds have shown marked population declines over the past 30 years. Correlational studies indicate that many of these declines are associated with changes in agricultural practices. Certainly reductions in food availability in winter and for nestling birds in spring are implicated in the declines of several bird species, notably the grey partridge, Perdix perdix. Thus weeds have a role within agroecosystems in supporting biodiversity more generally. An understanding of weed competitivity and the importance of weeds for insects and birds may allow the identification of the most important weed species. This may form the first step in balancing the needs for weed control with the requirements for biodiversity and more sustainable production methods.
A Karyopherin α2 Nuclear Transport Pathway is Regulated by Glucose in Hepatic and Pancreatic Cells
Authors: Cassany Aurélia , Guillemain Ghislaine , Klein Christophe , Dalet Véronique , Brot‐Laroche Edith , Leturque Armelle
Published: December 2004

We studied the role of the karyopherin α2 nuclear import carrier (also known as importin α2) in glucose signaling. In mhAT3F hepatoma cells, GFP‐karyopherin α2 accumulated massively in the cytoplasm within minutes of glucose extracellular addition and returned to the nucleus after glucose removal. In contrast, GFP‐karyopherin α1 distribution was unaffected regardless of glucose concentration. Glucose increased GFP‐karyopherin α2 nuclear efflux by a factor 80 and its shuttling by a factor 4. These glucose‐induced movements were not due to glycolytic ATP production. The mechanism involved was leptomycin B‐insensitive, but phosphatase‐ and energy‐dependent. HepG2 and COS‐7 cells displayed no glucose‐induced GFP‐karyopherin α2 movements. In pancreatic MIN‐6 cells, the glucose‐induced movements of karyopherin α2 and the stimulation of glucose‐induced gene transcription were simultaneously lost between passages 28 and 33. Thus, extracellular glucose regulates a nuclear transport pathway by increasing the nuclear efflux and shuttling of karyopherin α2 in cells in which glucose can stimulate the transcription of sugar‐responsive genes.
Reduction of anti‐Galα1,3Gal antibodies by infusion of types 2 and 6 gal trisaccharides conjugated to poly‐l‐lysine
Authors: Kuwaki K. , Knosalla C. , Moran K. , Alt A. , Katopodis A.G. , Duthaler R.O. , Schuurman H.‐J. , Awwad M. , Cooper D.K.C.
Published: December 2004

Abstract:  To investigate the specificity of anti‐Galα1,3Gal (Gal) antibodies (Abs) with respect to Gal oligosaccharides of types 2 and 6, eight baboons received an intravenous infusion of either a poly‐l‐lysine conjugate of Gal type 2 (n = 5) or type 6 (n = 3), followed 48 h later by the alternative Gal type 6 or 2 conjugate, respectively. IgM Abs reactive to Gal type 2 were depleted by 80 to 89% by either Gal conjugate. IgM reactive to Gal type 6 was less efficiently depleted by the Gal type 2 conjugate (57% depletion) than the Gal type 6 (82% depletion). Gal‐reactive IgG was depleted more slowly and less efficiently by either glycoconjugate (initially by only 28 to 54%). Our results indicate that the Gal type 6 conjugate depletes most anti‐Gal IgM, but the Gal type 2 conjugate is less efficient in depleting anti‐Gal IgM reactive with type 6. There remain small fractions of antibody that are unadsorbed, particularly of IgG, probably due to their low affinity and distribution in both the intra‐ and extra‐vascular compartments.
Intraportal pig islet xenotransplantation into athymic mice as an in vivo model for the study of the instant blood‐mediated inflammatory reaction
Authors: Goto Masafumi , Groth Carl G. , Nilsson Bo , Korsgren Olle
Published: December 2004

Abstract:  One of the main obstacles to successful intraportal islet transplantation is the instant blood‐mediated inflammatory reaction (IBMIR) elicited by the isolated islets when exposed to fresh human blood. In the present study, we investigated whether intraportal transplantation of pig islets into diabetic athymic mice could be used as a small animal model to study xenogeneic IBMIR in vivo.

Adult porcine islets (APIs) or rat islets were implanted into the portal vein or under the renal subcapsular space of diabetic athymic mice. Graft survival and morphology were evaluated by measuring blood glucose levels and by performing immunohistochemical staining, respectively. Transplantation of rat islets, irrespective of implantation site, cured all diabetic athymic mice. APIs transplanted subcapsularly also cured all diabetic athymic mice, while none of the animals transplanted with an equivalent amount of APIs via the portal vein remained normoglycemic for more than 10 days after transplantation. Immunohistochemical staining on day 7 showed that most of intraportally transplanted APIs were entrapped in clots and infiltrated with CD11b+ leukocytes. Intraportal transplantation of APIs into athymic mice induced IBMIR, thus providing a small animal model for studying xenogeneic IBMIR.
Comparison between the Conventional Tube Technique and the Gel Technique in Direct Antiglobulin Tests
Authors: Nathalang Oytip , Chuansumrit Ampaiwan , Prayoonwiwat Wichai , Siripoonya Praput , Sriphaisal Thip
Published: December 2004

Objectives: The direct antiglobulin test (DAT) is a diagnostic procedure demonstrating in‐vivo antibody or complement coating on red cells. The gel technique (GT) for this test is sensitive and easier to do than the conventional tube technique (CTT). Methods: We tested 52 newborns with hyperbilirubinemia and 6 children and 17 adults with autoimmune hemolytic anemia (AIHA) using the (DAT) in the form of the conventional tube technique (CTT) and the gel technique (GT) simultaneously. The gradings of the agglutination reactions of both techniques were scored as 12, 10, 8, 5, 3, 0 for 4+, 3+, 2+, 1+ and w+ and negative, respectively. Results: The GT yielded higher scores than the CTT (p < 0.01). The overall sensitivity and specificity of the GT were 93.5 and 88.6%, respectively. The specificity of the DAT‐positive results in newborns was determined by IgG only, while in AIHA, it was determined by IgG and/or C3d and, in only one sample, by IgM. Conclusion: The GT is equal to or better than the CTT. Since the GT is simple, the exposure of blood bank personnel to the blood sample is low. We highly recommend the GT, especially in areas where HIV infection is prevalent.
Prolonged Holding of Whole Blood at 22 °C Does Not Increase Activation in Platelet Concentrates
Authors: Sanz C. , Pereira A. , Faúndez A.I. , Ordinas A.
Published: December 2004

Objectives: Platelets prepared after holding of whole blood overnight at 22 °C have a well‐preserved metabolism. However, the possibility that such prolonged incubation with active granulocytes may increase platelet activation has not been fully tested. Methods: We investigated this possibility by flow cytometric analysis of membrane glycoproteins (GPs) Ib and IIb/IIIa and the activation markers CD62P and CD63 in platelet concentrates (PCs) prepared from whole blood that was held for either 6 h without cooling plates (n = 20) or for 24 h on cooling plates of 1,4‐butanediol (n = 20). PCs were prepared by the platelet‐rich plasma method and analyzed on the second storage day. Results: Platelet yield and aggregation response to ristocetin, collagen and epinephrine + ADP were similar in both types of PCs, as was the mean fluorescence intensity for GPs Ib and IIb/IIIa. PCs prepared by the overnight‐hold method did not differ from those obtained 6 h after collection in the percentage of platelets expressing CD62P (12.3±6.2% vs. 14.1±4.0%; p > 0.1) or CD63 (9.8±6.4% vs. 8.8±3.6%; p > 0.1). Conclusion: Prolonged holding of whole blood at 22 °C prior to component preparation does not increase the level of platelet activation.
Platelet Alloimmunization after Transfusion
Authors: Taaning Ellen , Simonsen Anne Catrine , Hjelms Erik , Svejgaard Arne , Morling Niels
Published: December 2004

Background and objectives: The frequency of platelet‐specific antibodies after one series of blood transfusions has not been reported, and in multiply transfused patients is controversial. Materials and methods: We studied the frequency of alloimmunization against platelet antigens in 117 patients who received a single series of blood transfusions. They received mostly saline‐adenine‐glucose + mannitol red blood cell components (poor in leukocytes and platelets) in connection with cardiac surgery. Platelet‐specific antibodies were detected with the platelet ELISA and the monoclonal‐antibody‐specific immobilization of platelet antigen assay. HLA antibodies were detected by the standard lymphocyte cytotoxicity techniques. Results: We found platelet‐specific anti‐HPA‐5b (anti‐Bra) in 2 cases (1.7%). One antibody was the result of de novo immunization. We detected lymphocytotoxic HLA antibodies in 21 patients (17.9%), of whom 18 (15.4%) had had no detectable antibodies before transfusion. There was a positive correlation between the transfused load of immunogenic materials and the frequency of alloimmunization against HLA antigens. In one third of the immunized patients, there was no history of previous immunization. Conclusion: There was a low incidence of platelet‐specific antibodies after one series of blood transfusions in this group of patients. This is similar to the results of some previous studies in multiply transfused patients, but not with those of others who found a higher incidence.
Gene Frequencies of the HPA‐1 and HPA‐2 Platelet Antigen Alleles among the Amerindians
Authors: Covas D. T. , Delgado M. , Zeitune M. M. , Guerreiro J. F. , Santos S. E. B. , Zago M. A.
Published: December 2004

Background and objectives: Platelet‐specific alloantigens are important in neonatal alloimmune thrombocytopenia, posttransfusion purpura, refractoriness to platelet transfusions, and population genetics. Data are scarce on allele frequencies in ethnic groups other than whites and Asians. Materials and methods: Using allele‐specific restriction enzyme analysis, we studied the distribution of HPA‐1 and HPA‐2 alleles in six Brazilian Amazon tribes of Amerindians, belonging to five different language stocks. We compared these with the values obtained for blacks and whites. Results: Only the HPA‐la allele was found among 132 Amerindian chromosomes, compared with a gene frequency of HPA‐lb of 0.115 and 0.133, respectively, among blacks and whites. The frequency of HPA‐2b among the Amerindians (0.042) is lower than that obtained for blacks and whites (0.148 and 0.100, respectively), and the lowest thus far observed in a population of Asian origin. Conclusion: Differences in DNA polymorphisms in Amerindian populations have not only anthropological and genetic interest, but also practical applications when they involve coding regions that may change the functional or immunologic features of the protein.
Complement‐Binding Anti‐Jka Not Detectable by DiaMed Gels
Authors: O'Brien P. , Hopkins L. , McCarthy D. , Murphy S.
Published: December 2004

Background: An anti‐Jka was not detectable by the DiaMed‐ID Micro Typing System but was strongly reactive when tested using other indirect antiglobulin test (IAT) methods. However, it was detectable in the DiaMed‐ID system when the recommended diluent ID‐Diluent 2, was replaced by other low ionic Strength saline (LISS) solutions. This led us to believe that the problem lay with the ID‐Diluent 2. Methods: The anti‐Jka was investigated using various low ionic strength diluents in the IAT against both polyspecific and monospecific antiglobulin reagents. We tested the ID‐Diluent 2 for its ability to inhibit complement activation, and bind calcium. Results: The anti‐Jka was shown to be a ‘complement‐only’ binding antibody that reacted in all IATs performed except when tested with ID‐Diluent 2. The ID‐Diluent was shown to prevent haemolysis of ABO lytic antibodies, and further testing shows that it binds calcium. Conclusion: The use of a diluent which prevents haemolysis and binds calcium in an IAT system that contains anti‐complement as well as anti‐IgG must be questioned.
IVIG Adverse Reactions: Potential Role of Cytokines and Vasoactive Substances
Authors: Bagdasarian A. , Tonetta S. , Harel W. , Mamidi R. , Uemura Y.
Published: December 2004

Background and Objectives: A clinical study was conducted to determine the effect of IVIG infusion rates on adverse experiences (AE) and on serum levels of cytokines and vasoactive substances. Materials and Methods: Forty‐two healthy volunteers were randomized into 3 groups with maximum IVIG infusion rates of 0.04, 0.06, and 0.08 ml/kg/min, and a final dose of 0.5 g IgG/kg body weight. Results: Adverse reactions were noted only at the highest infusion rate of 0.08 ml/kg/min, except in 1 subject infused at 0.06 ml/kg/min. There were significant increases in IL‐6 (p = 0.011) and thromboxane B2 (p = 0.007) in AE subjects as compared to non‐AE subjects. Conclusion: IVIG‐induced adverse reactions occur more often with rapid infusion rates and may be mediated by elevated levels of inflammatory cytokines and vasoactive substances.
Bacterial Contamination of Autologous Bone Marrow: Reinfusion of Culture‐Positive Grafts Does Not Result in Clinical Sequelae during the Posttransplantation Course
Authors: Schwella N. , Rick O. , Heuft H. G. , Miksits K. , Zimmermann R. , Zingsem J. , Eckstein R. , Huhn D.
Published: December 2004

Objectives: Microbiological cultures and posttransplantation course were analyzed in order to investigate the incidence and clinical significance of bacterial contamination of autologous bone marrow (BM) grafts. Methods: Cultures were obtained from BM after collection, BM concentrate after processing, contaminated/cryopreserved BM at thawing, and from peripheral blood (PB) following autologous BM transplantation (ABMT). The posttransplantation course of patients grafted with culture‐positive BM was recorded and compared with patients who underwent ABMT with noncontaminated BM grafts. Results: In 239 BM grafts processed, the incidence of microbiological contamination was 26.4% (n = 63). Fifty marrow grafts were contaminated by bacteria from the skin flora: coagulase‐negative Staphylococcus (CNSC), Propionibacterium, and Coryne‐bacterium species (79%). Thirty‐eight patients underwent ABMT (day 0) with cryopreserved culture‐positive BM, and 32 patients were evaluable for microbiological cultures at thawing: in 10 of 32 BM grafts CNSC was found prior to reinfusion. Following ABMT, PB cultures revealed CNSC in 5 of 38 patients between days +4 and +12. However, the late occurrence of positive PB cultures after BM reinfusion made a relationship between BM CNSC and PB CNSC unlikely. In 33 of 38 patients, no graft‐contaminating bacteria were detected in PB. Comparison of the posttransplantation course of patients who received contaminated BM with that of patients grafted with noncontaminated BM showed no significant differences concerning time to engraftment, febrile days, and days on antibiotics. Conclusion: (1) Collection and/or ex vivo processing can result in microbiological contamination of BM grafts predominantly with bacteria from the skin flora, and (2) only CNSC can be cultured at thawing from previously contaminated/cryopreserved BM. Since patients undergoing ABMT usually receive oral antibiotics from beginning of the conditioning regimen which are active against CNSC, no further administration of antibiotics is recommended for the reinfusion of bacterially contaminated BM grafts.
The Complement‐Activating Capacity of Maternal IgG Antibodies to Blood Group A in Paired Mother/Child Serum Samples
Authors: Hari Yvonne , von Allmen Edith C. , Boss Gabriela M. , Naiem Abdelbaky , Gittermann Matthias , Nydegger Urs E.
Published: December 2004

Background and objectives: The aim of this study was to investigate whether IgG antibodies to blood group A bind or activate complement after crossing the placenta. Materials and methods: IgG anti‐A concentrations as well as C1‐q‐binding and the capacity of anti‐A to form the C5b‐9 membrane attack complex were measured in sera of 61 mother/child pairs. The enzyme‐linked immunosorbent assay was used to quantitate anti‐A or anti‐B in maternal and neonatal serum samples. Results: The concentrations of IgG anti‐A in ABO‐identical and ABO‐compatible children born to type O or B mothers correlated significantly with maternal concentrations (p < 0.05), whereas type A or AB children had strikingly diminished concentrations of IgG anti‐A. The IgG anti‐A concentrations were analyzed in two different groups: first, 44 mother/child pairs with mothers and children group O or B (group I), and second, 17 pairs with mothers group O or B and children group A or AB (group II). In group I, identical or compatible pairs with regard to IgG anti‐A, a significant correlation was found by Spearman's rank analysis, whereas the same analysis revealed no significant correlation in group II. The capacity of anti‐A to activate complement was also more marked in newborns who were ABO‐identical or ABO‐compatible with their mothers, and this capacity was virtually absent in cord blood samples from incompatible babies. Conclusions: Tests for complement‐dependent or complement‐independent ABO blood group antibodies must be interpreted in the light of the blood group of the child.
Application of the Human Hepatitis B Virus Core Antigen from Transgenic Tobacco Plants for Serological Diagnosis
Authors: Tsuda Shinya , Yoshioka Keiko , Tanaka Takeshi , Iwata Akiko , Yoshikawa Akira , Watanabe Yuichiro , Okada Yoshimi
Published: December 2004

Background and objectives: The aim was to produce HBcAg from plants more cheaply than can be done by other currently available means, and to apply such antigen to immunoassay procedures for pretransfusion testing of donor blood. Materials and methods: Transgenic Nicotiana tabacum cv. SR‐1 plants expressing the human hepatitis B virus (HBV) core antigen (HBcAg) gene were generated by Agrobacterium‐mediated transformation. The recombinant product, called tHBcAg, can assemble itself into a spherical particle with a diameter of 25 to 30 nm, and can maintain two antigenic determinants of HBcAg, namely HBc/α and HBc/β. Partly purified tHBcAg was used in the hemagglutinationinhibition (HI) test, as routinely used by the Japanese Blood Center, to test a panel of 524 blood units taken from HBV‐positive donors. Results: In the HI test, tHBcAg showed serologic properties comparable to that from Escherichia coli, the standard antigen used in the Japanese Blood Center. Conclusions: Transgenic plants can produce reagents for serologic testing and perhaps even such medical materials as oral vaccines.
Alternative Therapies for Neonatal Autoimmune Thrombocytopenia
Authors: Ovali Fahri , Samanci Nedim , Ermiş Bahri , Akdoǧan Zeliha , Daǧoǧlu Türkan
Published: December 2004

Objective: This study was performed to test whether corticosteroids were superior to intravenous immunoglobulins (IVIG) in the treatment of neonatal autoimmune thrombocytopenia (NAT). Methods: All cases received IVIG, and unresponsive cases received corticosteroids. Results: Of 7 babies who received IVIG, only 1 responded. The 6 remaining cases received corticosteroids therafter, and all of them responded well to this therapy. Conclusions: Corticosteroids may be more effective than IVIG in NAT.
Severe Neonatal Alloimmune Thrombocytopenia due to Anti‐HPA‐3a
Authors: Boehlen F. , Kaplan C. , de Moerloose P.
Published: December 2004

Background and Objectives: Neonatal alloimmune thrombocytopenia is usually attributable to HPA‐la antibodies. We report a case of parental platelet antigen incompatibility associated with a severe neonatal thrombocytopenia secondary to alloimmunization to HPA‐3a. Materials and Methods: Platelet antibodies were detected by the monoclonal antibody‐specific immobilization of platelet antigens, genotyping of the platelets by PCR, and HLA typing by serologic procedures and PCR. Results: Genotyping of maternal and paternal platelets confirmed the incompatibility in the HPA‐3a system. It is noteworthy that the mother is of the HLA type DRB3*0101, is ABO‐incompatible with her husband, and also has HLA class I antibodies. Conclusion: Severe neonatal thrombocytopenia associated with HPA‐3a alloimmunization is infrequent and all the factors mentioned above could have played a role in the severity of the disease.
Kinetics of Bradykinin Levels during and after Leucocyte Filtration of Platelet Concentrates
Authors: Hild M. , Söderström T. , Egberg N , Lundahl J.
Published: December 2004

Objective: To measure beadykinin levels in platelet concentrates during and after leucocyte filtration. Methods: Platelet concentrates were leukocyte depleted using three different leucocyte‐depletion filters selected to represent filters with negative, positive or neutral charge, respectively. Bradykinin levels were analysed before, during and up to 90 min post‐filtration. Results: Significant levels of bradykinin were generated when negatively charged leucocyte depletion filters were used. However, we found a high variation between platelet concentrates prepared from different donors as well as within the same concentrate when this was divided into three aliquots. Generated bradykinin was rapidly degraded in the collection bag and no bradykinin was detectable 60 min post‐filtration. Conclusion: Bradykinin generation is more pronounced when negatively charged leucocyte removal filters are used. Due to a rapid degradation of bradykinin in the storage bag, pre‐storage filtration should minimise the risk of bradykinin related adverse reactions during transfusion with some filters.
Use of Overlapping Synthetic Peptides to Characterize Samples from Blood Donors with Indeterminate Results to Hepatitis C Virus Core Antigen
Authors: León Pilar , López José A. , Elola Consuelo , Lee Stephen R. , Calmann Mark , Echevarría José M.
Published: December 2004

Background and Objectives: Despite recent improvements in supplemental assays, isolated reactivity to the hepatitis C virus (HCV) core antigen continues as one of the main problems in the confirmation of anti‐HCV in blood donors. Reactivity against individual peptides from the c22‐3 HCV recombinant antigen has been described as a useful tool for anti‐HCV confirmation and donor counseling in such cases. Materials and Methods: We used a previously described set of overlapping peptides spanning the entire sequence of the c22‐3 antigen to study 87 single serum samples from blood donors with reactivity for c22‐3 antigen alone in second generation recombinant immunoblot assay (RIBA‐2). All of them had been previously studied by RIBA‐3, anti‐HCV E2 EIA and HCV PCR. Results: 66 of the 87 samples studied could be classified as positive or negative for anti‐HCV core using the multipeptide assay and such classification correlated well with the results obtained with RIBA‐3 and the anti‐E2 EIA. However, some discrepancies were found. The epitopes located along the N‐terminal half of the molecule were mainly responsible for the specific antibody recognition but those enclosed within amino acids 1–15 were frequently involved in nonspecific reactivity. Some 38% of samples were considered to have specific antibody to the c22‐3 antigen and a further 9% reacted for both anti‐E2 and single core peptides that were often involved in specific antibody recognision. Conclusion: Testing of blood donor samples indeterminate to the HCV c22‐3 antigen for reactivity against individual core peptides can confirm the presence of specific antibody and recognize nonspecific reactivity with certain cross‐reacting epitopes. Third generation supplemental tests have reduced such false reactivity, but confirmation of samples with anticore alone is still necessary. Single reactivity to the HCV core antigen is likely to reflect prior exposure to the virus, but rarely active infection, either acute or chronic.
Hepatitis E Virus: Relevance in Blood Donors and Other Risk Groups
Authors: Mateos María Luisa , Camarero Cristina , Lasa Elena , Teruel José Luis , Mir Nuria , Baquero Fernando
Published: December 2004

Background and Objectives: Hepatitis E virus (HEV) infection usually causes an acute self‐limited disease. HEV is associated with feces‐contaminated drinking water, but other vectors, such as blood, are possible. The aim of this study was to investigate the prevalence of HEV in blood donors and in two groups at high risk of parenteral infections, namely, hemodialysis patients and children infected with hepatitis C virus (HCV) via blood transfusion. Materials and Methods: We investigated the prevalence of anti‐HEV in 863 blood donors, 63 hemodialysis patients, and 42 children infected with HCV posttransfusion. Results: The prevalence rates were 2.8, 6.3%, and 0 respectively. Conclusions: (1) The incidence of HEV in Spain is similar to that in other western European countries, and (2) HEV is probably not transmitted parenterally to children.
Prevention and Management of Platelet Transfusion Refractoriness
Authors: Novotny V.M.J.
Published: December 2004

Platelet transfusion refractoriness is a major complication of long‐term platelet supportive care. Refractoriness may lead to fatal bleeding complications in thrombocytopenic patients. Major factors involved are factors related to the clinical condition of the patient as well as HLA alloimmunisation. Non‐alloimmune factors may occur in up to 80% of the patients. However, platelet transfusion outcome is impaired in only 50% of the patients having these conditions. HLA alloimmunisation has been convincingly reduced by the use of leucocytedepleted transfusions. UV‐B irradiation of platelet transfusions may be alternatively used to reduce HLA alloimmunisation. Despite these measures, patients with a history of pregnancy or non‐leucocyte‐depleted transfusions form HLA antibodies in a high proportion (up to 50%). HPA antibodies play a minor but relatively important role in patients with HLA antibodies. ABO antibodies may play a role in refractoriness, which can be abolished by transfusion of ABO‐identical platelets. Screening for the presence of HLA and/or HPA antibodies is indicated in case of transfusion failure after ABO‐identical or HLA‐matched platelets. If no alloantibodies are detected, further analysis to define a role of drugrelated or autoantibodies is required. In case of HLA and/or HPA alloimmunisation associated with refractoriness, matched platelet transfusions are indicated. In case of non‐alloimmune factors associated with increased platelet consumption, increasing the transfusion frequency can be considered. Additional investigations are still necessary to define risk factors for secondary HLA alloimmunisation and refractoriness due to non‐immune factors to further decrease the incidence of refractoriness.
Use of Bovine Viral Diarrhoea Virus as an Internal Control for Amplification of Hepatitis C Virus
Authors: Cleland A. , Nettleton P. , Jarvis L.M. , Simmonds P.
Published: December 2004

Background and Objectives:
Screening for hepatitis C virus (HCV) by polymerase chain reaction (PCR) will be mandatory for screening blood and plasma donors in Europe and elsewhere. This study describes an internally controlled, highly sensitive PCR method designed for screening blood donations in pools. Material and Methods: RNA extracted from bovine viral diarrhoea virus (BVDV) was used as an internal control to monitor the efficiency of extraction, reverse transcription and amplification steps in HCV PCR. Results: Sensitivity of PCR for single molecules of HCV in the presence of 33 genome equivalents of BVDV RNA was achieved by reducing the efficiency of BVDV amplification. BVDV could be recovered at high efficiency from large volume pools (2–5 ml) by ultracentrifugation and by the NucliSens extraction method. Conclusion: Detection of BVDV validates the extraction, reverse transcription and amplification methods used for HCV detection in plasma pools and provides valuable quality assurance for negative results.