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PubMed1990

[Possibilities for individual identification of hair roots using DNA probes].

High-molecular weight DNA was extracted from hair roots. The obtained DNA was digested with Hae III and Msp I and transferred to nylon membranes by Southern blotting. The possibility of applying a probe for sex determination (pHY2.1) a "single-locus"-probe (YNH24) as well as a "multi-locus"-probe (M13) was examinated as a function of the amount of hair being supplied. In the laboratory experiment a single hair root was enough for a sex determination or individual identification. Because of methodical difficulties in practice this kind of profiling should only be done with several hair roots.

Adolescent↗
PubMed1998

Genetic variation in captive koalas (Phascolarctos cinereus): parentage determination and individual identification.

Highly repeatable randomly amplified polymorphic DNA (RAPD) markers were developed for parentage studies in the koala (Phascolarctos cinereus). Of the 25 RAPD primers screened, 5 (20.0%) produced 32 repeatable polymorphic RAPD bands (average/primer = 6.4 +/- 4.2). A high level of polymorphism was observed for each group of koalas (Featherdale, 71.9%; Lone Pine, 84.4%). All 25 koalas could be uniquely identified using either RAPD or microsatellite markers. Of the 32 RAPD markers generated in koalas, 25 were informative for parentage analyses. These RAPD markers successfully determined both parents to three offspring and a male parent to a fourth offspring. Paternity analysis (where the female parent is known) succeeded in assigning the correct male parent to seven offspring. Our RAPD-PCR method generates informative genetic markers that are useful for parentage determination and individual identification of captive koalas. This would provide genetic analysis to zoos and wildlife parks as a low-cost alternative to the more expensive microsatellite markers.

Animals↗
PubMed2004

Minisatellites and MVR-PCR for the individual identification of parasite isolates.

In recent years, a wide variety of biochemical and molecular typing systems have been employed in the study of parasite diversity aimed at investigating the level of genetic diversity and delineating the relationships among different species and subspecies. Parasite sequence-specific polymerase chain reaction (PCR)-based genotyping systems are among the most useful tools employed to date, because they can be applied to very small quantities of host-contaminated parasite material and, using repeated loci such as mini- and microsatellites, allow the identification and tracking of individual strains as well as the determination of allele and genotype frequencies in populations. Although minisatellites have been used very successfully to study parasite populations, in particular Trypanosoma brucei populations, there are some technical problems involved in the use of these markers. For example, minisatellite alleles tend to vary in a quasi-continuous fashion, making unambiguous allele identification difficult. The development of minisatellite variant repeat (MVR) mapping by the polymerase chain reaction (MVR-PCR) as a digital approach to DNA typing has overcome many of the drawbacks of minisatellite length analysis. The system assays the dispersion patterns of MVRs within minisatellite alleles, producing an easily interpretable code for each allele. This technique not only allows unequivocal allele identification but also reveals cladistic information that can be used to determine the possible genetic relationships among the different strains and subspecies. The MVR mapping technique has been applied successfully to minisatellites in the parasite Plasmodium falciparum to uniquely identify strains, and more extensively in Trypansoma brucei, where it was used to determine population structure and to examine the relationships among T. brucei subspecies, providing evidence for multiple origins of human infectively. In this chapter, the methods for genotyping of T. brucei parasites using both minisatellite allele length and MVR mapping are described in full and can be easily adapted to apply to mini-satellites in other parasites.

Animals↗
PubMed1990

Simplified method of odontograms for individual identification.

A computerized method of codifying dental lesions and treatment is presented to enable faster identification of victims of catastrophes. Each tooth is assigned a bidigital value. The first digit refers to the root and designates whether it is found to be perfect, damaged and/or treated, or absent. The second digit refers to the crown and is assigned according to the number of surfaces showing lesions or treatment. For the identification of corpses, missing individuals whose record reveals values less than or equal to those of the subject in every tooth (or of each quadrant in cases of doubt) are selected for consideration from the data base. Individuals who have even one tooth with a value greater than that of the subject are eliminated.

Dental Records↗
PubMed1995

Individual identification by means of conventional bitewing film and subtraction radiography.

An analysis of simulated post- and antemortem (p.m. and a.m.) victim radiographs was performed. Existing pairs of bitewing radiographs randomly sampled from a large population of adolescents were used. Two categories of individuals were included based on dental therapy conditions (none or few simple fillings). Subtraction images were performed of pairs of bitewings originating from the same individual (identical images) and different individuals (non-identical images) within each group. Four observers were asked to match the bitewings and to assess the subtraction images with respect to identity based on the score system: 1 = 'eliminated', 2 = 'possible', 3 = 'likely', or 4 = 'certain'. Three observers each mismatched the film radiographs among two out of 12 individuals without fillings. True positive identification (identified by scores 2 + 3 + 4) of subtraction images were made in 10-12 individuals out of a possible 12. The range of false positive (FP) and false negative (FN) scorings was 3-15 and 0-2, respectively. All observers were capable of correctly matching bitewings originating in the 12 individuals with amalgam fillings. By use of subtraction radiography, 12-14 identical images out of a possible 15 were assessed correctly. The range of FP and FN scorings of subtraction images in this group was 1-4 and 1, respectively. Defining only score 4 as positive identification, the sensitivity of the subtraction technique decreased from approximately 0.90 to 0.70 in both categories of individuals, while specificity increased to almost 1.00 from 0.82 vs. 0.96 in the categories with and without dental restorations. Bitewing radiographs from single individuals within a group of individuals were sufficiently identical to allow for valid identification by a strict criterion. The subtraction technique may add to the subjective matching of radiographs as a screening test in victim identification.

Adolescent↗
PubMedDate not supplied

Highly polymorphic minisatellite DNA probes. Further evaluation for individual identification and paternity testing.

Reliable and reproducible protocols have been developed for the routine DNA fingerprinting of individuals using the highly polymorphic minisatellite DNA probes 33.15 and 33.6. Comparison of DNA fingerprinting from 50 individuals has generated further data on the level of band sharing in the DNA fingerprints of unrelated individuals, as well as the number of bands scorable in individuals. These results are consistent with previous studies. The occurrence of mutant bands in offspring has been examined in over 100 families. Further support is presented for the Mendelian inheritance of minisatellite loci and for lack of significant allelism and linkage between different variable DNA fragments detected in a human DNA fingerprint.

Alleles↗
PubMed1996

Cellular reservoirs of HIV-1 in the central nervous system of infected individuals: identification by the combination of in situ polymerase chain reaction and immunohistochemistry.

OBJECTIVES: The majority of HIV-1-infected individuals manifest a plethora of central nervous system (CNS) diseases unrelated to opportunistic infections, including AIDS dementia complex, encephalitis, and various other disorders of the CNS. The present study sought to evaluate the cellular reservoirs and expression patterns of HIV-1 in brain tissue to gain further understanding of HIV-1 neuropathogenesis. DESIGN: CNS tissue, obtained post-mortem from 22 patients with AIDS and four HIV-1-seronegative controls, was analyzed. METHODS: CNS samples were evaluated using a combination of in situ DNA polymerase chain reaction (PCR), reverse transcriptase (RT)-initiated in situ PCR, and immunohistochemistry. By utilizing this triple-staining methodology, HIV-1 proviral DNA and HIV-1-specific mRNA can be identified at the single cell level. RESULTS: HIV-1 was detected in all 22 AIDS brain specimens and in none of the four brains from HIV-1-seronegative individuals. The most commonly infected cells in AIDS brains were microglia cells and macrophages, but variable levels of HIV-1 infections were demonstrated in many of the major histological cell types within the CNS, including neurons, microvascular endothelial cells (MVEC) and astrocytes. The presence of HIV-1-infected cells was not uniform with infected cells unevenly distributed throughout the brain parenchyma. The degree of HIV-1 mRNA expression varied from 39-65% of the cells in the CNS harboring HIV-1 provirus. Choroid plexus and MVEC exhibited relatively high levels of productive infection. CONCLUSION: These findings demonstrate that several cell types in the CNS, in addition to microglia or macrophages, may become infected with HIV-1 in vivo with variable levels of HIV-1 mRNA expression. The diverse cellular reservoirs for HIV-1 in the CNS may be critically linked to the molecular mechanisms involved in HIV-1 neuropathogenesis. In addition, in vivo infection of MVEC, and possibly cells in the choroid plexus, may directly contribute to penetration of the blood-brain barrier by HIV-1.

Adult↗
PubMed1977

National individual identification of horses.

Methods of equine identification including signalment, blood typing tattooing and freeze marking are discussed. A new system of individually identifying horses with an unalterable freeze mark is proposed. Unalterable numerical and alphabetical symbols have been developed to apply a registration number to the animal.

Animal Identification Systems↗
PubMed1992

Non-acid glycosphingolipid expression in plasma of an A1 Le(a-b+) secretor human individual: identification of an ALeb heptaglycosylceramide as major blood group component.

Total non-acid glycosphingolipids were isolated from plasma of an A1 Le(a-b+) secretor individual with Refsum's disease (phytanic acid storage disease). The glycolipids were separated into 11 fractions by open column chromatography and by HPLC. The fractions were analyzed by thin-layer chromatography and tested for different blood group A activities as well as blood group Le(a )and Leb activity. The fractions were structurally characterized by proton NMR spectroscopy and FAB mass spectrometry and in selected cases by EI mass spectrometry of the permethylated and permethylated-reduced derivatives. Degradation analysis was performed on partially permethylated or permethylated-reduced alditol acetates. The dominating blood group compound was found to be a blood group A active type 1 chain difucosylheptaglycosylceramide. Other blood group compounds were identified as a blood group A active type 1 chain monofucosylhexaglycosylceramide, a blood group Leb hexaglycosylceramide, a blood group H active type 1 chain pentaglycosylceramide, and a globotetraosylceramide (the P-antigen). The presence of a Le(a) glycosphingolipid and blood group A type 3/4 chain structures were also found by immunostaining. Glucosyl-, lactosyl-, and globotriaosylceramides were the dominating short chain compounds. The amount of phytanic acid incorporated into the monoglycosylceramide fraction was found to be less than 5% of the fatty acids.

Animals↗
PubMed2005

Subtyping mtDNA haplogroup H by SNaPshot minisequencing and its application in forensic individual identification.

Sequence variation of the hypervariable segments (HVS) I/II of mitochondrial DNA (mtDNA) and the haplogroup affiliation were determined in a sample of 271 Italian subjects. This analysis showed that 42% of the individuals could be ascribed to H, the most frequent haplogroup in European Caucasian populations. This fraction was then screened for specific single nucleotide polymorphisms located in the coding region to identify H subclades H1-H15. We set up two multiplex polymerase chain reactions and specific SNaPshot assays to investigate the frequency distribution of these subgroups in our population sample and to examine their usefulness in discriminating among commonly shared HVS I/II sequences. This allowed the assignment of a large portion of the mtDNAs ( approximately 70%) to specific subhaplogroups, with H1 and H5 being the most represented. About two-thirds of the individuals sharing common HVS I/II sequences were subdivided and ascribed to specific H subhaplogroups with a significant reduction of the frequencies of the most common mtDNA haplotypes. Haplogroup H subtyping could thus be extremely useful in forensic identification when many samples have to be analysed and compared, avoiding excessive time-consuming and labor-intensive sequencing analysis.

Complementarity Determining Regions↗
PubMed1990

Hypervariable polymorphic VNTR loci for parentage testing and individual identification.

Three kinds of variable number of tandem repeat DNA probes (VNTR: pYNZ22, pYNH24, and pYNZ2) showing hypervariable polymorphisms were studied. Allelic frequencies and their confidence intervals among Japanese individuals were obtained. Co-dominant segregation of the polymorphism was confirmed in family studies. Two a priori probabilities were calculated for each VNTR locus: exclusion probabilities for an alleged father/mother/child trio and for an alleged parent/child duo, and probabilities of matching of genotyped two unrelated individuals or two siblings. Availability as well as highly discriminating polymorphic pattern of VNTR loci makes it potentially very useful for forensic and human genetic purposes.

Alleles↗
PubMed1994

The reliability of skull/photograph superimposition in individual identification.

The accuracy of video superimposition methods for identifying unknown human skulls was examined. Three identified human skulls were each compared to 97 lateral view and 98 frontal view "mug shot" photographs using two television cameras, an electronic signal mixer, and a video monitor. The skulls were not from individuals represented by the photographs. All comparisons were done without using anterior dentition. The results found that 9.6% of the lateral view and 8.5% of the frontal view superimpositions were classified as a consistent fit based on the criteria that were identified. The incidence of false matches was reduced to 0.6% of the sample when a frontal view and lateral view photograph of the same individual were both compared to one skull. It was concluded that without anterior dentition, skull/photograph superimposition is reliable when two or more photographs, clearly depicting the facial features from different angles, are used in the comparison.

Cephalometry↗
PubMed1998

The CCR5 receptor acts as an alloantigen in CCR5Delta32 homozygous individuals: identification of chemokineand HIV-1-blocking human antibodies.

The chemokine receptor CCR5 is the major coreceptor for infection by macrophage-tropic R5 HIV-1. A 32-bp deletion in the gene coding for CCR5 (CCR5Delta32) occurs with a frequency of 10% in the Caucasian population and results in a receptor protein that is truncated and not expressed at the cell surface. CCR5Delta32 homozygous individuals are apparently normal but resistant to infection with R5 HIV-1. In two individuals homozygous for CCR5Delta32, who had been repeatedly exposed to CCR5-expressing blood cells through sexual activity, we have identified antibodies to CCR5 that bound specifically to the surface of CCR5-expressing cell lines. Serum from these individuals, in contrast to serum from CCR5(+/+) individuals, competed with radiolabeled RANTES for binding to the CCR5 receptor and inhibited infection of peripheral blood mononuclear cells with R5, but not X4, primary isolates of HIV-1. The identified human antibodies to CCR5 define an alloantigen that may cause allograft rejection in a mismatch situation even in individuals with no history of blood transfusions or i.v. drug abuse.

Alleles↗
PubMed2006

School health nurses and substance use among adolescents - towards individual identification and early intervention.

Adolescents' health is today threatened by the use of alcohol and other psychoactive substances. It is therefore important to develop interventions related to substance use in school health care. The aim of this study was to examine the empowering or risk background factors related to substance use among adolescents, and the ability of school nurses (PHN) to identify these factors and to provide needed individual early intervention. The data were collected by semistructured questionnaires completed by 14- to 18-year-old adolescents (n = 326, response rate 79) and PHNs (n = 10) in 2004. The adolescent questionnaire consisted of items related to the respondents' background and Adolescents' Substance Use Measurement (ADSUME). Following individual consent, adolescents' ADSUME responses were sent to the PHNs for intervention. The PHNs assessed the adolescents' empowering background factors and intervention using the questionnaire, and 70% (n = 228) of their answers matched the adolescents' answers. The data were analysed with the SPSS software using the chi-squared test, Fisher's exact test, kappa coefficient and agreement percentages. Substance use among adolescents was associated with parental support, mother's education and smoking, the adolescents' knowledge about substances, peer support and hobbies. The PHNs' assessments regarding supportive background were not in agreement with the assessments of adolescents who were using hazardous substances. One-fifth of the adolescents received the brief intervention, although many of them might have needed extra support and follow-up on the basis of their ADSUME results. The research findings can be generalized only for alcohol use, because only 3% of the study informants used substances other than alcohol. Further research is warranted concerning PHNs' ability to identify hazardous substance use and to ensure preventive early intervention and requisite support among substance-using adolescents in order to improve evidence-based health promotion.

Adolescent↗
PubMed2004

Rapid individual identification by minisatellite variant repeat (MVR)-PCR at D1S8 locus using "exponential law".

We describe an efficient and simple minisatellite variant repeat mapping by PCR (MVR-PCR) method based on the assignment of the tandem array of 29 bp repeating units into a-type, t-type and 0-type (a rarely appearing unamplified unit), from the first repeat unit position (code position 0) of 293 bp in D1S8 locus. After microchip electrophoresis of PCR product amplified from the target DNA of a human hair root, each rung of the ladder at the position of 293 + 29 n bp (n: code position) was detected and the type of repeating unit was determined, i.e., aa, a0, tt, t0, at and 00. The peak area of the rungs from PCR product decreased with increase in code position. We found for the first time that the logarithmic plots of the peak area against the code position showed a linear relationship, which implied that peak areas decrease exponentially. The present method was successfully applied to identify 37 individuals using only a hair root as a biological specimen. This "exponential law" is expected to be an effective tool in forensic science.

Base Sequence↗
PubMed1995

Individual identification of flood victims by DNA polymorphisms and autopsy findings.

On September 23rd 1993 Genova was flooded by heavy rainstorms and 4 people disappeared, including an elderly couple. Four days later a partially skeletonized body was found floating near the coast. No visual identification was possible. Autopsy findings were consistent with the medical history of a possible victim. DNA was extracted from a muscle sample and compared to paraffin embedded prostatic gland fragment taken by surgery. A positive identification could be made. On October 11th the body of a decomposed and partially skeletonized female was found. The visual identification was also uncertain and no clinical records were available. A blood sample from the son was obtained for maternal identification by the polymerase chain reaction.

Aged↗
PubMed2002

Individual identification of disguised faces by morphometrical matching.

The reliability of a morphometrical matching method for identifying disguised faces was examined experimentally using a computer-assisted facial image identification system. The 2D right oblique facial images of three target persons disguised with sunglasses, cap and gauze mask were each compared with each of the 3D facial images of 100 subjects, yielding 900 face-to-face superimpositions. The average perpendicular distance between the facial outlines and the average point-to-point distance of the corresponding landmarks in the 2D image of the disguised face and the 3D facial image, were calculated. As a matching criterion, the sum of the values of the average perpendicular difference of the facial outlines and the average point-to-point difference between the corresponding landmarks was used (abbreviation: average difference). The range of the average difference was 2.3-2.8mm for the same person (a match) and 4.0-14.6mm for different (non-matching) people, respectively. The ranges for matching and non-matching faces did not overlap. Even the 3D facial images of the non-matching person showing the closest value of average difference to the average difference for the matching person could be excluded easily. It was concluded that the morphometrical matching method can reliably identify disguised faces and the results produced by this method could be easily understandable by a court of law.

Adult↗