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Biomedical subjects

D J Werrett

Publications and source records attributed to D J Werrett.

At least 19 recordsLinked to original sources

Interpretation of DNA profiles using a computerised database.

An image analyser has been utilised to store data derived from autoradiography of hypervariable loci (both multi and single locus). By examination of the electrophoretic system, it is possible to assign standard deviations to bands and then to determine whether the bands 'match' each other. The problems associated with the practical applications of such a system are discussed. Improvements are suggested such as the adoption of internal markers.

DNA

The analysis of hypervariable DNA profiles: problems associated with the objective determination of the probability of a match.

A computerised system has been used to store DNA profiles from 3 hypervariable loci. This initial survey illustrates that band matching is only possible after analysis of the errors associated with electrophoretic systems. A number of databases have been constructed with the three probes investigated and two methods of frequency determination, 'binning' and 'sliding window' fitting, have been compared.

Autoradiography

An evaluation of the polymerase chain reaction method for forensic applications.

This paper describes the use of polymerase chain reaction (PCR) for amplifying small amounts of DNA obtained from samples of interest to the forensic scientist. A region of the HLA DQalpha (DQa) locus was amplified in DNA prepared from the following: hair roots, liquid blood, blood-stains, semen and vaginal swabs (semen free and semen contaminated). A population study was conducted using DNA from 78 unrelated individuals. The observed distribution of HLA DQa alleles varied from that reported for an American population but obeyed the Hardy-Weinberg equilibrium. Interpretation problems associated with the PCR technique are discussed.

Alleles

The evaluation of an ELISA for semen-specific 19-OH prostaglandin F1 alpha/F2 alpha using ex-casework swabs and stains.

The application of an enzyme-linked immunosorbent assay (ELISA) for 19-OH Prostaglandin F1 alpha/F2 alpha (19-OH PG F) to casework analysis of seminal contamination of swabs and stains is reported. The results are compared to those where the identification of semen was based on the presence of acid phosphatase and spermatozoa. Five hundred and one samples were analysed and there was good agreement between the results of ELISA and conventional methods. The determination of 19-OH PG F confirmed both the presence of semen where spermatozoa were absent and indicated semen was present when acid phosphatase and spermatozoa were both absent. The results indicate that 19-OH PG F represents a useful marker for the casework identification of semen and is particularly valuable where spermatozoa are absent.

Acid Phosphatase

Individual specific DNA fingerprints from a hypervariable region probe: alpha-globin 3'HVR.

A probe detecting a hypervariable region (HVR) 3' to the alpha globin locus on chromosome 16 has been used to produce DNA fingerprints. Segregation analysis has revealed multiple, randomly dispersed DNA fragments inherited in a Mendelian fashion with minimal allelism and linkage. The fingerprints are highly polymorphic (probability of chance association between random individuals much less than 10(-14]. The probe is, therefore, a powerful discriminating tool: it is envisaged that this probe will have forensic applications, including paternity cases, and will be informative in linkage analysis.

Alleles

Group-specific component: a review of the isoelectric focusing methods and auxiliary methods available for the separation of its phenotypes.

This review compares the major isoelectric focusing methods that have been published for the separation of group-specific component (Gc) phenotypes since 1978. The various parameters of gel composition, size, electrical and running conditions and sample application points are listed. More current auxiliary methods are also listed. These relate to the extraction of Gc from bloodstains and its identification after isoelectric focusing. Protocols are then recommended for the forensic analysis of Gc phenotypes.

Forensic Medicine

A comparison of the sensitivity of typing group-specific component (Gc) and the phosphoglucomutase (PGM1) locus in control and casework bloodstains by three isoelectric focusing methods.

The performance of typing group-specific component (Gc) in bloodstains by two isoelectric focusing methods followed by its detection with silver staining has been compared with an established forensic system of typing phosphoglucomutase (PGM1) locus phenotypes by isoelectric focusing (IEF) in 1 mm gels. For Gc typing ultra-thin isoelectric focusing (UTIEF) gels and immobilized pH gradient (IPG) gels were used. Both laboratory prepared stains and casework stains were examined. The Gc UTIEF method is approximately eight times more sensitive than the existing PGM1 1 mm IEF method for control and casework stains. However, on average, a larger amount of stain was taken from casework stains than control stains for each typing system. A total of 53 casework stains were examined. Comparable success rates of 62% and 64% were obtained for typing Gc on UTIEF gels and PGM1 by 1 mm IEF, respectively. A success rate of 55% was obtained for typing Gc on IPGs. Bloodstains that were over 200 days old were successfully grouped by all three methods.

Blood Stains

The use of isoelectric focusing (IEF) as a method for the combined phenotyping of erythrocyte acid phosphatase (EAP) and esterase D (EsD).

The simultaneous isoelectric focusing (IEF) in polyacrylamide gels (PAG) of erythrocyte acid phosphatase (EAP) and esterase D (EsD) allows the poor discriminating power (DP) of EsD to be usefully combined with a highly discriminating system EAP, such that a joint DP of 0.766 was achieved compared with PGM IEF DP 0.756. Focusing was carried out in a centrally flattened gradient containing ampholines (pH 4-6 and 6-8) and the chemical spacer 3-(N-morpholino) propanesulphinic acid (MOPS). It enabled the identification of six EsD phenotypes including the recently discovered EsD5 isozymes. The application of this method to casework bloodstains is discussed.

Acid Phosphatase

New information from bloodstains.

The traditional philosophy behind the grouping of bloodstains in the Forensic Science Laboratory is described. It is emphasised that while these blood-grouping methods are highly discriminating in the sense that they may distinguish one person's blood from hundreds, or thousands of others, such information may be of limited value in many crime investigations. The reason for this is that such information does not normally relate to physical features of the individual with which we are all familiar such as sex or age. Recent research work is described in which efforts are being made to deduce from small bloodstains information which indicates the donor's sex, age, ethnic origin and perhaps clinical history.

Adult

The detection of Y chromosomes in bloodstains--a reevaluation.

A method has been described for detecting Y chromosomes in the leukocytes of human bloodstains prepared on a variety of substrates. The factors that influence the proportion of chromosomes exhibiting a Y spot (the Y cell index) in a bloodstain are considered, including the subjective nature of assessment of the Y chromosome fluorescence, the substrate, and the age of bloodstain. In contrast to previous workers no decay in Y cell index with the age of the stain was observed. The results of a blind trial involving stains derived from case work, where from other evidence there was no doubt as to the sex of the donor, are presented. Sixty-five percent of the male bloodstains were correctly identified and no females were wrongly reported as male.

Blood Stains

Allergy profiles from bloodstains.

The radioallergosorbent test (RAST) has been adapted to measure specific IgE antibodies in bloodstains. By incubating several allergen discs simultaneously with one stain extract, an allergy profile can be readily obtained from less than 50 mul of dried blood.

Allergens

Antibodies and sperm survival in the female tract of the mouse and rabbit.

Rabbit and mouse spermatozoa from male and female tracts have been examined for their species-antigenic surface character, and for adherent antibodies, by double immunofluorescence techniques. Mouse spermatozoa from the ductus deferens showed an area over the acrosome which was positive to anti-mouse serum that had been absorbed with some male mouse somatic tissues including blood, but those from the uterus and oviduct were not stained. Spermatozoa from the uterus were shown to have an antibody coat on the acrosome, with anti-mouse IgG, but those from the ductus deferens and oviduct did not. Rabbit spermatozoa were more variable but their activity was similar: ejaculated spermatozoa sometimes already had antibody of male origin; the majority of the spermatozoa arriving early in the uterus were coated, but in general those that attained the oviducts were not coated. The results are interpreted as evidence for selection by the female tract of a small antigenically different population; the majority of spermatozoa are rejected and/or destroyed.

Animals

Antibody profiling of bloodstains.

The detection and measurement of antibodies in bloodstains represents a departure from the classical blood grouping used routinely in forensic serology. The antibodies in question are those associated with atopic allergy and infections by micro-organisms and other parasites. The production of these antibodies is not primarily under genetic control but reflects an individual's reaction to his environment. This paper describes the explicit information which may be obtained from an antibody profile, the high discriminating power and other advantages of the method.

Age Factors

Exclusion of a man charged with murder by DNA fingerprinting.

DNA fingerprinting was used to demonstrate that two murder-rapes committed in 1983 and 1986, respectively, were connected. The probability of chance association of the fingerprint was calculated as 5.8 x 10(-8). The man who had been charged with the murder was excluded because his DNA fingerprint did not match sperm DNA fingerprints obtained from swabs and clothing attributed to the two victims.

Autoradiography

A comparison of the isoelectric focusing patterns of red cell phosphoglucomutase separated on an ampholine, ampholine/separator and immobilised pH gradient.

The isoelectric focusing patterns of red cell phosphoglucomutase (PGM1) separated on Ampholine, Ampholine/separator and rehydratable immobilised pH gradients have been compared. The sharpest and most intense zymograms have been observed on immobilised pH gradients provided that Ampholine was added during the rehydration of the gel. The addition of ampholine in the rehydration of Immobiline plates has been shown to improve the sharpness and intensity of the zymogram obtained and the potential of immobilised pH gradients for PGM typing has been demonstrated.

Ampholyte Mixtures