Regulation of expression of carbohydrate blood group antigens.

The carbohydrate antigens associated with the human ABO and Lewis blood group systems are excellent models for the study of the genetic regulation of glycoconjugate biosynthesis because their expression on erythrocytes and in saliva has been thoroughly investigated in terms of classical genetics and the chemical structures and pathways for the formation of the antigens are now well understood. The primary protein products of the blood group genes are believed to be the glycosyltransferase enzymes that complete the biosynthesis of the determinants. The important controlling factors still to be elucidated are the genetic and environmental influences leading to the tissue specific expression of these antigens. The 3 types of regulation mechanisms discussed in this review are those arising: 1) from the specificity requirements of the glycosyltransferases encoded by the blood group genes; 2) from the competition or co-operation of glycosyltransferases encoded by genes at the same or independent loci; and 3) from the existence and tissue distribution of glycosyltransferases with related, but not identical, substrate specificities.

Identification of a subset of group B donors reactive with monoclonal anti-A reagent.

Two red blood cell (RBC) units labeled group B were returned to the source blood center after they had been retyped as AB by a transfusion service. The discrepancy could be reproduced, but only with the use of the transfusion service's reagent, Ortho Diagnostic's anti-A Bioclone (a licensed, blended, murine monoclonal anti-A reagent). RBCs from 35 of 3,458 random group B donors (1%) reacted with the monoclonal anti-A after immediate centrifugation. Reactivity was associated with high serum levels of B-gene-specified transferase and was caused by the MH04 component, a potent anti-A capable of detecting some examples of Ax RBCs. It is probable that the potency of MH04 permitted detection of low levels of A determinants synthesized by the donors' unusually strong B-gene-specified transferase. Transfer of N-acetylgalactosamine by B-gene-specified transferases, reported in vitro, has not been detected previously in vivo. Use of highly sensitive monoclonal reagents may result in clinically ambiguous blood grouping results.

Hepatitis C-related chronic liver disease among asymptomatic blood donors in the north west of England.

In the first 19 months of screening, the North Western Regional Transfusion Centre (RTC) tested 224,000 consecutive blood donors for antibody to hepatitis C virus (anti-HCV) by second generation enzyme immunoassay (EIA). Of these, 366 repeatedly reactive samples were referred for confirmatory testing at Manchester Public Health Laboratory (PHL). There, the initial EIA was repeated, together with two further EIAs. All the referred samples were subjected to a confirmatory line immunoblot (RIBA-II). Reverse transcription followed by the polymerase chain reaction (RT-PCR), in order to detect viral RNA, was performed on selected samples. Among the donors, 61 accepted offers for medical review and were assessed for risk factors, clinical findings and results of standard liver function tests. Of these donors, 53 proceeded to liver biopsy. The overall prevalence of confirmed positive donors was 0.04%. Main risk factors identified included intravenous drug abuse in 31 (51%) donors and prior blood transfusion in 12 (20%) but a risk factor was not apparent in 11 (18%). Viraemia, detected by RT-PCR, could be predicted with a high degree of accuracy by means of the readily available and simpler screening and confirmatory tests (EIA and RIBA-II). Established chronic hepatitis was demonstrated in 90% of the liver biopsies. A trend towards worsening histological findings accompanied increasing concentrations of serum transaminase. Even so, many donors with normal transaminase values had abnormal biopsies including those showing chronic active hepatitis (CAH). These findings indicate that a substantial proportion of previously unrecognised asymptomatic persons with established chronic liver disease exists among North Western blood donors.(ABSTRACT TRUNCATED AT 250 WORDS)

Enzymes involved in the biosynthesis of glycoconjugates. A UDP-2-acetamido-2-deoxy-D-glucose: beta-D-galactopyranosyl-(1 leads to 4)-saccharide (1 leads to 3)-2-acetamido-2-deoxy-beta-D- glucopyranosyltransferase in human serum.

A 2-acetamido-2-deoxy-beta-D-glucopyranosyltransferase (N-acetylglucosaminyltransferase) that catalyses the transfer of 2-acetamido-2-deoxy-D-glucose from UDP-2-acetamido-2-deoxy-D-glucose to terminal nonreducing beta-D-galactosyl residues in disaccharides, oligosaccharides, glycoproteins, and glycolipids has been detected in human serum. The preferred acceptors are those with beta-D-galactosyl residues linked (1 leads to 4) to the subterminal sugar residue. Activity is greatest when the second sugar residue is 2-acetamido-2-deoxy-D-glucose but beta-D-Galp-(1 leads to 4)-D-Glc and beta-D-Galp-(1 leads to 4)-D-Man are also good acceptors. Compounds with beta-D-galactosyl groups linked (1 leads to 3) to 2-acetamido-2-deoxy-D-glucose are relatively poor acceptors and beta-D-Galp-(1 leads to 6)-D-GlcNAc is inactive. Oligosaccharides substituted with an L-fucose unit on the beta-D-galactosyl unit or on the adjacent sugar residue failed to accept 2-acetamido-2-deoxy-D-glucose. Similarly, glycoproteins with L-fucose or sialic acid substituents are less effective acceptors before removal of these sugars to expose more beta-D-galactosyl end-groups. The transferred 2-acetamido-2-deoxy-beta-D-glucopyranosyl residue is cleaved from the enzymic reaction products by the N-acetyl-beta-D-glucosaminidase from Jack beans. Methylation analysis of the products of 2-acetamido-2-deoxy-D-glucosyl transfer to N-acetyllactosamine [beta-D-Galp-(1 leads to 4)-D-GlcNAc] and lactose [beta-D-Galp-(1 leads to 4)-D-Glc] revealed that the terminal, nonreducing D-galactosyl group in both these acceptors had been 3-O-substituted with 2-acetamido-2-deoxy-D-glucose. Hence, the enzyme in human serum catalyses, in the presence of Mn2+, the reaction UDP-GlcNAc + beta-D-Galp-(1 leads to 4)-R leads to beta-D-GlcpNAc(1 leads to 3)-beta-D-Galp-(1 leads to 4)-R + UDP and is a UDP-2-acetamido-2-deoxy-D-glucose: beta-D-galactopyranosyl-(1 leads to to 3)-2-acetamido-2-deoxy-beta-D-glucopyranosyltransferase.

UDP-N-acetyl-D-galactosamine as a donor substrate for the glycosyltransferase encoded by the B gene at the human blood group ABO locus.

The properties of the enzyme in the serum of blood group B individuals that catalyses the transfer of small amounts of N-acetyl-D-galactosamine to H-active precursor structures were compared with those of the blood group B gene-associated alpha-(1----3)-D-galactosyltransferase and with the blood group A gene-associated alpha-(1----3)-N-acetyl-D-galactosaminyltransferases in the serum of blood group A1 and A2 individuals. The biosynthetic products formed by the enzyme in B serum were identical with the A-active structures synthesised by the A1 and A2 gene-associated alpha-(1----3)-N-acetyl-D-galactosaminyltransferases but the enzyme differed from the A1 and A2 transferases in its apparent Km for UDP-N-acetyl-D-galactosamine, its heat susceptibility, its failure to bind to Sepharose 4B, and its adsorption to H-active sites on group O red cell ghosts under conditions which bind the B transferase but fail to adsorb the A1 and A2 transferases. The correlation between the levels of alpha-(1----3)-D-galactosyltransferase and alpha-(1----3)-N-acetyl-D-galactosaminyltransferase activities in all the group B serum samples tested, the maintenance of the same ratio of activities after successive cycles of binding to group O red cell ghosts, the retention of the ability to convert blood group O to A-active cells after treatment of the serum with Sepharose 4B, and the failure to detect any comparable activity in group O serum samples tested under the same conditions indicated that the enzyme in group B serum that utilises UDP-N-acetyl-D-galactosamine to make blood group A-active structures is the B gene-associated alpha-(1----3)-D-galactosyltransferase.

Characterisation of a blood-group A-active tetrasaccharide synthesised by a blood-group B gene-specified glycosyltransferase.

The B gene-specified alpha-D-(1----3)-galactosyltransferase, isolated from the serum of a blood-group B individual, was used to catalyse the transfer of N-acetyl-D-galactosamine from UDP-N-acetyl-D-galactosamine to the blood-group H-active trisaccharide 2'-fucosyllactose. The biosynthetic product had blood-group A activity and its structure was confirmed as alpha-D-GalpNAc-(1----3)-[alpha-L-Fucp-(1----2)]-beta-D-Galp-(1--- -4)-D-Glc by methylation analysis and high-resolution 1H-n.m.r. spectroscopy. This tetrasaccharide was structurally and serologically identical with that made from the same donor and acceptor substrates when the blood-group A gene-specified alpha-D-(1----3)-N-acetylgalactosaminyltransferase was used as the enzyme source. The enzyme encoded by the B gene at the blood group ABO locus thus has overlapping donor substrate specificity with the enzyme encoded by the allelic A gene, and this property confers upon the B gene-specified alpha-D-1----3)-galactosyltransferase the potential to synthesise blood-group A-active structures.

The genetic and enzymic regulation of the synthesis of the A and B determinants in the ABO blood group system.

Possible genetic models for the inheritance of the ABO blood groups are discussed in terms of the glycosyltransferase enzymes which complete the synthesis of the A and B determinants. Recent immunologic evidence in support of the allelic status of the ABO genes is reviewed. Results are presented of experiments which demonstrate that the B gene associated alpha-3-D-galactosyltransferase can be used to synthesis blood group A determinants.

A blood group Sda-active pentasaccharide isolated from Tamm-Horsfall urinary glycoprotein.

Human Tamm-Horsfall urinary glycoprotein from an individual of the blood group Sd(a+) phenotype was tritium-labelled by treatment with galactose oxidase and sodium boro[3H]hydride and was then digested with endo-beta-galactosidase. A series of dialysable, labelled fragments was released from which a pentasaccharide was isolated that strongly inhibited the agglutination of Sd(a+) red cells by human anti-Sda serum and hence contained the Sda determinant structure. Reduction, methylation analysis and sequential exo-glycosidase digestion established the structure of the pentasaccharide as: GalNAc beta(1 leads to 4)[NeuAc(2 leads to 3)]Gal beta(1 leads to 4)GlcNAc beta(1 leads to 3)Gal

A human dispermic chimaera first suspected from analyses of the blood group gene-specified glycosyltransferases.

The red cells of a normal male blood donor, K.S., were first grouped as B but he was found to lack anti-A in his serum. Closer investigation revealed that his red cells had very weak A activity, demonstrable only by absorption and elution of anti-A. He is a non-secretor of ABH and a secretor of Lea. Blood group A-, B and H-gene specified glycosyltransferases were detected in his serum. In contrast to the finding of a B antigen of normal strength on his red cells, the B transferase in his serum was only about 30% of the normal level and, despite the very weak A activity of K.S's red cells, the A transferase level was about 50% of that found in the serum of group A individuals with normal strength of A antigen. Moreover, the A transferase on the basis of its pH optimum, Km values for donor and acceptor substrates, activation by divalent cations, isoelectric focusing profile and capacity to convert O to A-active cells, was characterized as the product of an A1 gene. A family study showed that K.S's wife is group A2 and that they have two sons, one group A2 and the other group B. The group B son is assumed to have inherited a B gene from the propositus but the level of B transferase in the son's serum is three times as high as that in his father's serum. The wife of the propositus and his group A2 son have normal A2 transferases in keeping with their A2 red cell status. The A2 son therefore appears to have inherited an A2 gene from his mother but neither the A1 nor the B gene shown to be carried by his father. The distribution of transferase activities in K.S's red cells differs from that in his serum. A level of B transferase within the normal range was found in his red cell membranes but a very low level of A transferase was detected. The discrepancies between the serum transferases and ABO red cell group, together with the pattern of inheritance within the family, led to a suspicion of chimaerism. This was confirmed by the finding of fibroblasts with the female 46XX karyotype in cultures of the propositus' skin. These results suggest that K.S. is a dispermic chimaera with two different cell lines of the genotypes BO and A1O or A1A1. The group A2 son is assumed to have inherited an O gene from his father. It seems probable that K.S.'s bone marrow and reproductive organs are comprised predominantly of the XY cell line which carried the blood group BO genotype whereas his skin and other tissues which contribute the A1 transferase to his plasma, are partly made up of the XX cell line which carries the blood group A1O or A1A1 genotype.

Evidence that the gene for tuberous sclerosis is on chromosome 9.

Linkage analysis was undertaken in nineteen families with tuberous sclerosis by use of 26 polymorphic markers. All affected members fulfilled strict diagnostic criteria and unaffected members were rigorously investigated to confirm their status. Maximum lod scores were 1.20 for adenylate kinase 1 (AK1) at zero recombination and 3.85 for the ABO blood group at zero recombination (confidence limits 0-0.10). These findings support the assignment of the gene for tuberous sclerosis to the distal long arm of chromosome 9.

Early discharge and risk for postnatal depression.

OBJECTIVE: To determine whether early discharge (< 72 hours) after childbirth increased the risk for women developing postnatal depression. DESIGN: Prospective cohort design consisting of an initial interview, and six-weekly assessments for 24 weeks using a self-report questionnaire and the Edinburgh Postnatal Depression Scale (EPDS). Women discharged within 72 hours were compared with the remaining women. SETTING: Tertiary referral hospital in western Sydney, New South Wales, 1993. PARTICIPANTS: All 749 women delivering over a three-month period were recruited. Of the 522 participants, 425 women completed the study. MAIN OUTCOME MEASURES: Women scoring > 13 on the EPDS on two or more occasions were considered potential "cases" of postnatal depression. The diagnosis was confirmed using the Structured Clinical Interview for DSM-III-R disorders (SCID). RESULTS: Of the 153 women (36%) discharged early, 22 women (14.4%) developed postnatal depression over the study period compared with 20 of the 272 women (7.4%) who had standard length of stay. Women who were discharged within 72 hours had a significantly increased risk for developing postnatal depression (odds ratio [OR], 2.12; 95% confidence interval [CI], 1.07-4.21). This risk persisted when other sociodemographic, obstetric and psychosocial risk factors were controlled for in a logistic regression analysis (OR, 3.06; 95% CI, 1.22-7.69). CONCLUSION: Women planning early discharge after childbirth should be carefully assessed before discharge and follow-up should be rigorous. The potential to develop postnatal depression should be considered in all women choosing early discharge from hospital.