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Biomedical subjects

R Warwick

Publications and source records attributed to R Warwick.

At least 19 recordsLinked to original sources

Cord blood banking in London: the first 1000 collections.

The London Cord Blood Bank was established with the aim of collecting, processing and storing 10000 unrelated stem cell donations for the significant number of children in the UK requiring transplantation, for whom a matched unrelated bone marrow donor cannot be found. Collection is performed at two hospitals by dedicated cord blood bank staff after delivery of the placenta. Mothers are interviewed regarding medical, ethnic and behavioural history by nurse counsellors and sign a detailed consent form. Donations are returned to the bank for processing. Volume reduction is undertaken by a simple, closed, semi-automated blood processing system, with excellent recovery of progenitor cells. Units are cryopreserved and stored in the vapour phase of liquid nitrogen. Blood samples from mothers and cord blood donations are tested for the UK mandatory red cell and microbiology markers for blood donors. Donations are typed for HLA-A, B and DR at medium resolution (antigen split) level using sequence-specific oligonucleotide probing and sequence-specific priming techniques. The selection of collection hospitals on the basis of ethnic mix has proven effective, with 41.5% of donations derived from non-European caucasoid donors. Bacterial contamination of collections has been dramatically reduced by implementation of improved umbilical cord decontamination protocols.

Blood Banks

Review of counselling in a transfusion service: the London (UK) experience.

Donor (and recipient) counselling within the Transfusion Service in the UK has grown in volume and complexity over the last 10 years. The addition of new tests for donated blood and the growth of bone marrow transplantation have increased the demands on counselling staff. New initiatives, such as the HCV look-back programme, have required an extension of the skills and knowledge of staff involved in counselling.

Blood Donors

The London Cord Blood Bank.

Cord blood (CB) contains a sufficient number of haemopoietic progenitor cells and can be used as an alternative to bone marrow transplantation. To date, a number of transplants using cord blood-derived stem cells from HLA-identical siblings and unrelated donors have been successfully performed. However, only approximately 30% of patients have an HLA-identical sibling while the rest are reliant on finding an HLA-compatible unrelated donor. This is particularly difficult for patients from ethnic minorities since the majority of unrelated bone marrow donor volunteers are from Caucasoid ancestry. The London Cord Blood Bank was established to provide an additional source of stem cells for all patients requiring bone marrow transplantation.

Blood Banks

Remineralization of root surfaces demineralized in solutions of differing fluoride levels.

The beneficial effects of fluoride on enamel have been well documented. However, limited data are available concerning the amount of fluoride required for beneficial effects on tooth root. Although studies have shown that fluoride inhibits root demineralization, the aim of this study was to investigate the location, extent and amount of remineralization on root dentin substrates after demineralization has occurred. The root surfaces of extracted human teeth were demineralized in a pure chemical buffer containing varying concentrations of sodium fluoride. After this lesion initiation, the same root sections were then placed into a remineralizing solution. The root sections were characterized after demineralization, and again after remineralization, by polarized light microscopy (PLM) and microradiography (MRG). Lesion depths after the demineralization phase were found to be inversely proportional to the fluoride concentration. When fluoride was present, bands or lines within the body of the lesion were observed with PLM and MRG. Using quantitative MRG, variations in mineral content and distribution were recorded. Examination of the root sections after the remineralization phase showed remineralization to have occurred on the remaining mineral and not on organic matrix devoid of mineral. The amount and location of mineral deposition may be of great significance in the arrestment and treatment of in vivo root surface caries.

Buffers

Prenatal diagnosis of Kell blood group genotypes: KEL1 and KEL2.

OBJECTIVE: Our purpose was to devise diagnostic test(s) that determine fetal KEL1 and KEL2 genotypes. STUDY DESIGN: KEL1 and KEL2 polymorphisms are due to a single C to T base substitution at nucleotide 698 of exon 6 of KEL. This allowed us to develop two polymerase chain reaction tests that distinguish KEL1/1 and KEL2/2 homozygotes and KEL1/2 heterozygotes. The first test uses a Bsm I restriction fragment length polymorphism in a genomic deoxyribonucleic acid polymerase chain reaction product containing the single base polymorphism, and the second test uses allele-specific primers to distinguish KEL1 and KEL2 genotypes. These tests were applied in a blind study to 15 amniotic fluid deoxyribonucleic acid samples. The corresponding KEL1 and KEL2 fetal red blood cell phenotypes were determined serologically. The tests were also applied to two families in which the mothers had antibodies to KEL1. RESULTS: In all cases results of analysis of Kell genotypes from the amniotic fluid deoxyribonucleic acid samples agreed with the fetal red blood cell Kell phenotypes. The tests were also successfully used to determine fetal Kell genotype by use of peripheral blood deoxyribonucleic acid. CONCLUSION: Two polymerase chain reaction-based tests can be used for prenatal diagnosis of KEL1 and KEL2 genotypes; these procedures should prove useful in the proper management of Kell-sensitized pregnancies.

Amniotic Fluid

Platelet transfusion.

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Antigens, Human Platelet

Erythropoietic suppression in fetal anemia because of Kell alloimmunization.

OBJECTIVE: Our purpose was to test the hypothesis that maternal anti-Kell alloimmunization produces fetal anemia by erythroid suppression. STUDY DESIGN: Erythropoiesis in 11 anemic fetuses from maternal anti-Kell alloimmunization was compared with that in 11 fetuses where the mother was alloimmunized to RhD; each was matched for hematocrit, gestational age, hydrops, and perinatal outcome. Comparisons of the difference were performed by either paired t or Wilcoxon tests. RESULTS: The anti-Kell group had reduced reticulocytosis (p = 0.007) and erythroblastosis (p = 0.045) and lower amniotic fluid bilirubin concentrations (p = 0.02) in comparison with the anti-D group. No correlation was found between hematocrit and reticulocytosis in the anti-Kell group, whereas the anti-D group had a significant linear relationship (r = 0.63, p < 0.05), indicating a progressive reticulocytosis in response to the degree of anemia. CONCLUSION: These findings suggest that erythroid suppression, rather than hemolysis, is the predominant mechanism in producing fetal anemia related to maternal Kell alloimmunization. Fetal blood sampling is the investigation of choice in the evaluation of anemia related to maternal Kell alloimmunization, because reduced hemolysis means amniotic fluid bilirubin concentrations correlate poorly with anemia.

Erythroblastosis, Fetal

Prenatal determination of human platelet antigen type using DNA amplification following amniocentesis.

OBJECTIVES: To demonstrate that fetal human platelet antigen (HPA1) type can be determined, without the need for fetal blood sampling, by amplification of fetal DNA from amniotic fluid cells using polymerase chain reaction and allele specific oligonucleotide hybridisation. DESIGN: Oligonucleotide DNA primers were designed to amplify a portion of the platelet glycoprotein GpIIIa gene which spans the site of the single base change which differentiates HPA1a from HPA1b. Specific oligonucleotides were designed to hybridise either to the amplified HPA1a allele or to the HPA1b allele. Amniotic cells were used as the DNA template both directly and following formal isolation of DNA. Fetal HPA1 type, determined by this method in fifteen pregnancies not at risk of perinatal alloimmune thrombocytopaenia, was compared to typing of fetal blood obtained following cordocentesis. The methodology was then used to HPA type the fetus in two pregnancies at risk of the disease. SETTING: Department of Molecular Biology and Centre for Fetal Care, Queen Charlotte's Hospital. SUBJECTS: Fifteen women undergoing amniocentesis and fetal blood sampling for other indications and two women at risk of perinatal allo-immune thrombocytopaenia whose partners were heterozygotes. RESULTS: In the 15 control cases and the two clinical cases, determination of fetal HPA1 type from amniotic fluid cells agreed with typing of fetal blood. There was no difference in the efficiency of amplification from amniotic fluid cells directly or from isolated DNA. CONCLUSIONS: Fetal HPA type may be reliably determined by amplification of DNA from amniotic fluid cells, eliminating the need for fetal blood sampling or immunoglobulin administration when the fetus is HPA1a negative.

Amniocentesis