Prenatal exclusion testing for Huntington''s disease: a problem of too much information.

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.     

Improvement of visual function with glare testing after photorefractive keratectomy and radial keratotomy

PURPOSE: To evaluate the effect of a glare source on visual function in patients after photorefractive keratectomy and radial keratotomy. METHODS: Thirteen patients (22 eyes) who underwent photorefractive keratectomy and 20 patients (40 eyes) who underwent radial keratotomy were evaluated in this cross-sectional study. LogMAR visual acuity and contrast sensitivity were measured. Pupils were measured with the Rosenbaum card. A halogen/tungsten glare source approximated the luminance of headlights of an oncoming car at 100 feet. RESULTS: In the photorefractive keratectomy and radial keratotomy groups, pupils were significantly smaller (P<.01) and the pupillary clearance of the ablation zone in photorefractive keratectomy and the clear zone in radial keratotomy were significantly larger under the glare condition (P<.01). In the photorefractive keratectomy group, visual acuity and contrast sensitivity under the glare condition were significantly higher than in the no-glare condition (P = .02). In the radial keratotomy group, contrast sensitivity under the glare condition was significantly higher than under the no-glare condition (P = .001 to .003). CONCLUSIONS: After photorefractive keratectomy or radial keratotomy, the traditional glare source constricted the pupil and partially masked the optical aberrations, which resulted in an improvement in visual function. A "pupil-sparing" aberration test is needed for evaluation of visual function after refractive surgery.     

Rapid review (partial rescreening) of cervical cytology. Four years experience and quality assurance implications.

AIMS: To determine the sensitivity of the partial rescreening method of rapid review for internal quality control of cervical cytology; to determine which staff members are most suited to undertake it; and to investigate the cell patterns of false negative smears previously detected by the method. METHODS: As a prospective study 9517 cervical smears were partially screened by four cytotechnologists using the 'step' method prior to conventional screening and the results compared with the final report. As a retrospective study 62 false negative smears that had been identified by the method over four years were reviewed. RESULTS: A detection rate for dyskaryosis of 86% (range 82-91%) was achieved. Sixteen abnormal smears were missed on conventional screening that had been detected by prescreening. Review of the 62 false negatives revealed three patterns: (1) scanty abnormal cells; (2) abundant dyskaryotic cells presenting as "microbiopsies"; and (3) abundant, readily recognisable abnormal cells. CONCLUSIONS: Partial rescreening enables the detection of errors due to both fatigue and misinterpretation. In this laboratory the method has, together with targeted full rescreening, reduced the false negative report rate from 5.0% to 0.4%. For laboratories using a rapid review method to reduce false negative reports, a prescreening trial is recommended in order (1) to select the most effective review method and the staff most suited to undertake it; and (2) to determine the laboratory's sensitivity with the method, as this is required for accurate estimation of the false negative rate.     

A molecular genetic service for diagnosing individuals with familial hypercholesterolaemia (FH) in the United Kingdom

A genetic diagnostic service for familial hypercholesterolaemia (FH) has been established over the last 4 years in the Clinical Molecular Genetics Laboratory at Great Ormond Street Hospital for Children NHS Trust (GOSH), London. In total there have been 368 referrals; 227 probands and 141 family members, which have come from a number of lipid clinics and from general practitioners. FH is caused by mutations in the low-density lipoprotein receptor gene (LDLR) and these are analysed by SSCP, DNA sequencing and direct assays. The clinically indistinguishable disorder, familial defective apolipoprotein B100 (FDB) is caused by one of three mutations in the apolipoprotein B100 gene (APOB) which are analysed by direct assays. Mutations predicted to be pathogenic were found in 76 probands, 67 in LDLR (23 previously undescribed) and nine in APOB. The mutation detection rate was 53% in paediatric probands, 32% in adults with a 'definite' FH diagnosis (tendon xanthoma positive) and 14% in adults with a 'possible' FH diagnosis (tendon xanthoma negative). The predicted loss of sensitivity that would result from reducing the number of exons tested has been assessed, and a molecular screening strategy suitable for UK patients is proposed. A similar strategy may be useful for other countries where genetic heterogeneity results in a wide mutation spectrum for FH.     

Spatial accuracy of 3D reconstructed radioluminographs of serial tissue sections and resultant absorbed dose estimates

Many agents using tumour-associated characteristics are deposited heterogeneously within tumour tissue. Consequently, tumour heterogeneity should be addressed when obtaining information on tumour biology or relating absorbed radiation dose to biological effect. We present a technique that enables radioluminographs of serial tumour sections to be reconstructed using automated computerized techniques, resulting in a three-dimensional map of the dose-rate distribution of a radiolabelled antibody. The purpose of this study is to assess the reconstruction accuracy. Furthermore, we estimate the potential error resulting from registration misalignment, for a range of beta-emitting radionuclides. We compare the actual dose-rate distribution with that obtained from the same activity distribution but with manually defined translational and rotational shifts. As expected, the error produced with the short-range 14C is much larger than that for the longer range 90Y; similarly values for the medium range 131I are between the two. Thus, the impact of registration inaccuracies is greater for short-range sources.     

DNA based prenatal testing for the skin blistering disorder epidermolysis bullosa simplex

Epidermolysis bullosa simplex (EBS) is a skin fragility disorder in which mild physical trauma leads to blistering. The phenotype of the disorder is variable, from relatively mild affecting only the hands and/or feet, to very severe with widespread blistering. For the severest forms of EBS there is a demand for prenatal diagnosis which until now has involved a fetal skin biopsy in the second trimester. The identification of mutations in the genes encoding keratins K5 and K14 as the cause of EBS opens up the possibility of much earlier diagnosis of the disease. We report here four cases in which prenatal testing was performed. In three of the cases the genetic lesions were unknown at the start of the pregnancy, requiring the identification of the causative mutation prior to testing fetal DNA. In two of the four cases novel mutations were identified in K14 and in the two remaining families, a previously identified type of mutation was found. Fetal DNA, obtained by chorionic villus sampling or amniocentesis, was analysed for the identified mutations. Three of the DNA samples were found to be normal; a mutant K14 allele was identified in the fourth case and the pregnancy was terminated. These results demonstrate the feasibility of DNA-based prenatal testing for EBS in families where causative mutations can be found.     

Clinical patterns of × linked agammaglobulinemia in Malaysian children

X linked agammaglobulinemia (XLA) is rarely reported from developing countries especially from South East Asia. It appears that × linked agammaglobulinemia is less common in certain ethnic groups. It is very uncommon in black people in USA and South Africa. In multiracial Malaysia we have documented five XLA in Malays and Indians but not in the Chinese that constitute about 31% of the population. First degree relatives afflicted with XLA or other primary immunodeficiencies occurred more often in our study. All showed lung involvement although the etiologic organisms involved were atypical, being Gram negative.