Biomedical subjects
Publications and source records attributed to D J Brock.
The value of maternal periconceptional vitamin supplementation in preventing the conception of fetuses with neural tube defects has yet to be assessed in a properly designed study. Avoidance of the birth of affected infants continues to rest on second-trimester maternal serum alphafetoprotein screening. The performance of screening programmes in achieving this objective is reviewed.
At eight weeks of pregnancy a couple were informed that the prospective father's mother had died of Huntington's disease (HD). There were no living affected members in the immediate family to confirm the diagnosis. By inspection of the local genetic register, it was established that it was indeed HD segregating in the extended family. Genotyping of the prospective mother and father, the father's unaffected father, and his unaffected maternal grandmother was carried out using a battery of polymorphic DNA markers, including a new probe which has a very low recombination rate with the HD locus. Analysis of DNA from a chorionic villus sample taken at 10 weeks of pregnancy showed that the fetus must have inherited a chromosome from its father's affected mother. Its risk of developing HD was 47%. If the genotype of the unaffected maternal grandmother was taken into account, the risk was reduced to 42%. Neither risk was considered acceptable by the prospective parents and the pregnancy was terminated at 12 weeks' gestation. Prospects for future pregnancies are good, with a 50% chance of having a child whose risk of inheriting the HD gene is less than 1.5%. In retrospect it was noted that although genotyping of the maternal grandmother had refined the fetal risk, it had also nearly contributed to an inadvertent and unwanted predictive test for HD on the father. This case makes the point that in prenatal exclusion testing, linkage information must be generated with considerable care.
Explore the source record for details and available documents.
Explore the source record for details and available documents.
Between 1977 and 1985 there was a 65% increase in births to women aged 35 or more in England and Wales, but only a 15% increase in all births. Two factors of roughly equal importance were responsible for this differential increase. Firstly, the proportion of older women (35-44) among all women of reproductive age (15-44) increased from 28% in 1977 to 31% in 1985; and, secondly, in the same period the fertility rate for women aged 35-39 increased from 18.2 to 24.1 per 1000 and for women aged 40-44 from 4.1 to 4.6 per 1000. The increased fertility rate among older women is not due to an extension of the reproductive period but to a delay in childbearing. This delay was seen in women married only once and also in those who had remarried. As prenatal diagnosis for the exclusion of chromosome abnormalities is customarily offered to older mothers the increased numbers of women aged 35 or more and their increased fertility rate have important implications for the provision of obstetric and laboratory services. There were 51,859 live births to women aged 35 and over in 1985; the projected figure for 2001 is 85,000. If the use of prenatal diagnosis continues to increase facilities for about 70,000 prenatal cytogenetic analyses will be needed in 2001.
First-trimester prenatal diagnosis of cystic fibrosis (CF) using linked DNA markers is usually only possible if there is an index affected child to establish the haplotype of the parental chromosomes. We describe a prenatal diagnosis where fibroblasts, cultured from the skin of a deceased affected child and then held in frozen storage for 3 years, were used as the starting point for tracking the CF gene. The fetus was diagnosed as a homozygous normal and the diagnosis confirmed by immunoreactive trypsin testing after birth. It was also possible to establish heterozygosity in the aunt of the affected child.
Prenatal diagnosis of cystic fibrosis by microvillar enzyme assay on amniotic fluid supernatant has been carried out on 258 sequential pregnancies with a 1 in 4 recurrence risk, all with known outcome. In general the three enzymes evaluated, gamma-glutamyltranspeptidase, aminopeptidase M and the intestinal isoenzyme of alkaline phosphatase, showed a high degree of concordance. However, there were two unusual patterns of microvillar enzyme activity; in seven cases a low gamma-glutamyltranspeptidase activity was associated with elevated values of intestinal alkaline phosphatase, and in ten cases there were isolated low values of intestinal alkaline phosphatase. The former pattern was found to be associated with cystic fibrosis in five cases, while the latter was associated with a normal outcome in all ten cases. A retrospective analysis of enzyme values suggested that the optimal system for minimizing false positives and false negatives was to define foetal cystic fibrosis as a sample where two of the three microvillar enzymes were below a cut-off of half the median value for the gestational week. If such scoring were applied to the cases where conventional microvillar enzyme patterns were observed, the false positive rate was 2.3% and the false negative rate 4.4% between 17 and 20 weeks of gestation.
Parents at risk of bearing a child with cystic fibrosis, and who have no living affected child, often use prenatal diagnosis based on microvillar enzyme assay in second-trimester amniotic fluid samples. If enzyme levels are abnormal and the pregnancy is terminated, it is possible in principle to use the fetal tissues to establish the phase relationship of linked DNA markers for a subsequent first-trimester prenatal diagnosis. However, the probability of a fetus being affected after an abnormal microvillar enzyme test may be no greater than 80%. The strong linkage disequilibrium between haplotypes at the D7S23 locus and the cystic fibrosis gene may be used to increase this probability. If fetal tissues are homozygous for the 6.6-kb band defined by pKM.19 and PstI and also homozygous for the 2.1-kb band with pXV-2c and TaqI, the chance of being affected increases from 80% to between 95% and 97%. We regard this as being sufficiently certain for use in phase determination.
Forty-six British families, containing at least one child affected with cystic fibrosis, were typed for restriction fragment length polymorphisms (RFLPs) by the probes pmet H, pmet D, pJ3.11 and 7c22. Thirty-five (76%) were fully informative for prenatal diagnosis and carrier detection, while in the remainder prenatal exclusion of an affected fetus could be carried out in half the pregnancies. The frequencies of individual alleles did not differ between cystic fibrosis and normal chromosomes. However, the previously noted excess of one haplotype on chromosomes carrying the cystic fibrosis gene was confirmed.
Placental alkaline phosphatase (P-ALP) was measured by a specific monoclonal antibody-based immunoassay in plasma samples of 117 women who subsequently were delivered of an infant of birthweight less than 2.5 kg. P-ALP values greater than twice the normal median were found in 32% of maternal plasma samples from low birthweight cases in one series and in 35% in another series, while in normal outcome controls the corresponding value was 8%. The differences were highly significant. The proportion of low birthweight cases with elevated maternal P-ALP values appears to be very similar between 15 and 34 weeks gestation. At 16-18 weeks gestation there is a significant positive correlation (r = 0.40) between P-ALP and maternal plasma alpha-fetoprotein (AFP) values in low birthweight cases. The use of P-ALP assay in combination with AFP assay appears to improve the detection of pregnancies with subsequent low birthweight outcome.
Explore the source record for details and available documents.
Tissues from eight fetuses, diagnosed on the basis of amniotic fluid microvillar enzyme assay as having cystic fibrosis, were conserved in frozen storage for up to three years. Adequate samples of undegraded DNA could be extracted from small intestine, lung, and liver. DNA typing, with restriction fragment length polymorphisms tightly linked to the cystic fibrosis gene, showed all eight diagnoses to have been correct. Determining the DNA genotype of fetal material can also be used to establish the linkage relationship between markers and the cystic fibrosis gene, and will permit subsequent first trimester prenatal diagnosis for couples who have no living affected child.
An immunoassay based on a monoclonal antibody specific for acetylcholinesterase (AChE) was used prospectively over a two year period to screen second trimester amniotic fluids for fetal abnormalities. All 26 cases of spina bifida, three of which had normal alphafetoprotein (AFP) concentrations and ultrasound scans, were detected by the immunoassay. Four clear fluids, with abnormal AFP concentrations, had normal AChE titres and yielded normal outcomes. Some difficulties were encountered with both the AChE immunoassay and polyacrylamide gel testing when amniotic fluids were contaminated with old blood. However, the quantitative nature of the AChE immunoassay and its independence of operator experience make it an ideal adjunct to AFP assay for routine screening of amniotic fluids.
This paper reports further study of the identity and function of a protein shown to be elevated in serum from cystic fibrosis (CF) patients and clinically normal heterozygotes. Monoclonal antibodies, specifically recognizing the tentatively named cystic fibrosis antigen (CFAg), were produced. Immunoaffinity purification of CFAg from several sources revealed two components: 11 x 10(3) and 14 x 10(3) Mr protein. cDNA clones corresponding to each protein have been isolated. Data-base comparisons of the deduced amino acid sequences suggest that both genes encode related but distinct calcium-binding proteins. We propose the name calgranulin A and B, for the 11 x 10(3) and 14 x 10(3) Mr components, respectively. It is clear from the assignment of the calgranulin genes to chromosome 1 that neither is the product of the mutant CF gene, which maps to chromosome 7. We have used the monoclonal antibodies to study the tissue distribution of the two proteins in a wide-ranging immunohistological survey. Where possible the pattern of expression was confirmed by RNA blot analysis. Strong calgranulin expression in granulocytes was confirmed. In addition to myeloid cells, a restricted subset of normal stratified squamous epithelia were found to be calgranulin-positive. These included tongue, oesophagus and buccal cells, the last of which has been shown to have altered calmodulin activity in CF patients. Using indirect alkaline phosphatase staining, tissue sections of lung, pancreas and skin (normally considered sites where the CF defect is expressed) were not calgranulin-positive. However, by indirect immunofluorescence, nasal polyp sections showed weak patchy calgranulin expression in some epithelial cells, and stronger, higher frequency expression when such cells were briefly cultured.(ABSTRACT TRUNCATED AT 250 WORDS)
IgG, IgM and albumin levels in sera from neonates with open neural tube defects (NTD), other congenital malformations, and matched controls were measured, as were levels in sera from their mothers. Neonates with NTD had significantly lower serum IgG levels than 22 control neonatal sera. On the other hand, the mothers of these neonates with NTD had elevated serum IgG levels resulting in a striking imbalance in the mother/neonate IgG ratio. In contrast to IgG, IgM levels in maternal and neonatal sera from the NTD group were within normal values. The maternal albumin levels were however significantly increased reflecting hemoconcentration. In sera from 3 neonates born with other congenital malformations (hydrocephaly, caudal appendage and sacral tumor) the only significant changes were higher IgM levels and lower mother/neonate IgM ratios. These results are discussed in the context of materno-fetal immune responses, placental transfer of plasma proteins and the possible involvement of immunoglobulins in fetal malformation.
Serum samples from cystic fibrosis homozygotes and heterozygotes contain elevated levels of a protein, known as cystic fibrosis antigen, which is synthesized primarily in granulocytes. We have produced monoclonal antibodies against cystic fibrosis antigen and developed a sensitive two-site immunoassay. Antigen levels were evaluated in serum samples from 50 cystic fibrosis homozygotes, 34 heterozygotes, 60 healthy controls and 25 disease controls. Simultaneous measurement was made of another granulocyte-derived serum protein, lactoferrin, and also of the acute-phase reactant, C-Reactive Protein. Attempts to use the concentrations of cystic fibrosis antigen in serum to distinguish cystic fibrosis patients from heterozygotes were unsuccessful, even when these concentrations were expressed as a ratio with lactoferrin or with C-Reactive Protein. However, examination of the ratio of cystic fibrosis antigen to lactoferrin in serum samples from cystic fibrosis heterozygotes suggests that there is some specific association between this antigen and the cystic fibrosis gene.
Two different methods of prenatal diagnosis of CF are currently available. Both are usually only suitable for couples with a 1 in 4 risk of bearing an affected child. The method of choice is first trimester diagnosis on a chorionic villus sample using linked DNA probes. The advantages are: 1. The very high degree of accuracy of diagnosis, 2. The opportunity of first trimester termination of pregnancy when the fetus is affected. The disadvantages are: 1. The need for a blood sample from an index affected child to establish phase, 2. The requirement for early workup of the nuclear family to ascertain informativeness, 3. The chance that some families will be uninformative or only partially informative for available DNA probes. An alternative form of diagnosis is that using microvillar enzyme testing on second trimester amniocentesis samples. The advantages are: 1. It is suitable for all pregnancies with a 1 in 4 risk of CF, 2. No prior workup of the family is required, 3. Assays can be completed within one hour of receipt of the sample. The disadvantages are: 1. The accuracy of diagnosis leaves some scope for error, 2. If termination of pregnancy is indicated this can only be carried out at about 18 or 19 weeks of gestation.
We have observed seven pregnancies at risk for fetal cystic fibrosis where second-trimester amniotic fluid microvillar enzyme activities presented an unusual pattern. Low gamma-glutamyltranspeptidase and borderline alpha-glucosidase values were associated with normal aminopeptidase M and intestinal alkaline phosphatase values. All seven pregnancies went to term; five of the seven infants were affected with cystic fibrosis.