Biomedical subjects

P Wiegand

Publications and source records attributed to P Wiegand.

At least 19 recordsLinked to original sources

PubMed2005

Short amplicon STR multiplex for stain typing.

We developed a short tandem repeat (STR) typing kit based on DNA database systems that are included in, for example, the Interpol Standard Set of Loci recommendations (i.e., TH01, VWA, D3S1358, FGA) and the gender typing system Amelogenin. Two different multiplex sets were tested using the fluorescent dyes FAM, JOE, and VIC. The PCR results were compared to the commercially available AmpFISTR Blue kit, which contains the STRs D3S1358, VWA, and FGA. The advantage of our multiplex compared with the Blue kit was the generation of shorter amplicons (<200 bp) and the higher combined power of discrimination.

Amelogenin↗

PubMed2003

A very long ACTBP2 (SE33) allele.

Analysing a buccal swab we found a long allele in the STR system ACTBP2. For confirmation we sequenced the isolated PCR product and found a sequence structure common in alleles of type III. Based on the repeat array the new allele is assigned as allele "49".

Alleles↗

PubMed2003

Population genetic comparisons of three X-chromosomal STRs.

The X-chromosomal short tandem repeats (STRs) DXS6800, DXS101 and DXS8377 were analysed in male and female population samples from Germany and Austria using a PCR multiplex approach. We investigated 135 family trios from Innsbruck (Austria) and surrounding areas and 50 families and further male and female samples from Ulm (Germany) and surrounding areas. The comparisons of the allele frequencies gave similar distributions for Innsbruck and Ulm although minor variations were found for some alleles. Additionally, some differences were found when comparing the allele frequencies of the male and female samples independently. The forensic efficiency values demonstrate that especially DXS101 and DXS8377 are highly informative markers for kinship analysis and deficiency cases. Based on the investigated meiotic events no new mutations were detected.

Adolescent↗

PubMed2002

Somatic mutations at STR loci--a reason for three-allele pattern and mosaicism.

Two families are analysed in which one of the parents exhibited a three-allele pattern at the ACTBP2 locus. Since the alleles were obviously segregated independently to the children, a generalised mosaicism must be assumed involving at least two tissues in one of them and at least four tissues in the other one. The intensity of the PCR amplified alleles in both three-allele individuals indicate an occurrence in a very early embryonic stage. Occurrence was most probably due to a single step mutation in both cases. Forensic implications would include paternity testing as well as stain analysis.

Alleles↗

PubMed2002

Unusual presentation of central nervous system relapse with oculomotor nerve palsy in a case of CD56-positive acute myeloid leukemia following allogeneic stem cell transplantation.

Allogeneic stem cell transplantation (allo-SCT) plays an important role in the treatment of infants and children with acute myelogenous leukemia (AML). Leukemic relapse after allo-SCT is responsible for a high rate of treatment failure. Extra-medullary relapse (EMR), without involvement of bone marrow, is rare compared to medullary relapse. CD56, the neural cell adhesion molecule, may contribute to the higher frequency of CNS relapse in CD56-positive AML. We observed an isolated EMR on the oculomotor nerve of a 17-month-old girl 12 weeks after cord blood transplantation (CBT), who was transplanted because of CD56-positive AML. Diagnosis of relapse was suspected clinically and confirmed by magnetic resonance imaging (MRI), and fluorescence-activated cell sorter (FACS) and chimerism analysis of cerebrospinal fluid (CSF). Therapy consisted of intra-thecal chemotherapy, CNS irradiation, and systemic immunomodulation by cyclosporin A (CsA) and basiliximab withdrawal. Twenty-one months after relapse, the patient shows full remission of symptoms and previously described oculomotor nerve infiltration.

CD56 Antigen↗

PubMed2001

Assessment of shooting distance on the basis of bloodstain analysis and histological examinations.

A 28-year-old man was shot using a pump-gun. The main question to be resolved was whether the biological stain pattern on the suspect's trousers, and in particular the bloodstains, can provide evidence to assess the shooting distance between the suspect and the position of the victim's body. The biological stain pattern (i.e. bloodstains and brain tissue) showed backspatters from the shot entrance wound on the back of the head, while the victim was lying face down and the suspect was standing close behind his head.

Adult↗

PubMed2001

Less is more--length reduction of STR amplicons using redesigned primers.

PCR primers closely flanking the repeat region were redesigned to reduce the amplicon length of the selected STRs down to approximately 100 bp for the shorter alleles (loci HumTH01, D10S2325, DYS19 and DYS391). Highly degraded DNA (e.g. formalin-fixed tissue) and very low amounts of DNA could be more successfully typed using the new redesigned primers compared to the established sequences generating longer amplicons.

Body Fluids↗

PubMed2000

Microsatellite structures in the context of human evolution.

Six microsatellite - or short tandem repeat (STR) - systems with uniform repetitive sequences (HumTH01, HumCD4, HumFES/FPS, HumF13B, HumTPO, HumLPL) and three compound repeat systems (HumVWA, HumFIBRA, D21S11) were used, including data from the literature, to determine genetic distances among eight populations worldwide. The TH01- and VWA homologous loci in nonhuman primates (chimpanzees, gorillas, orangutans, rhesus monkeys, ring-tailed lemurs) were compared and found to be shorter than in humans. Microsatellites of lower complexity were most efficient for the separation of major ethnic groups. The loci of higher complexity showed a leveling of the diversity differences among populations, which could be attributed to higher mutation rates.

Animals↗

PubMed1999

D18S535, D1S1656 and D10S2325: three efficient short tandem repeats for forensic genetics.

Three short tandem repeat (STR) polymorphisms characterized by PCR product length < 175 bp were investigated. D18S535 and D1S1656 contained a 4 bp unit as basic repeat motif, D10S2325 a 5 bp unit. The heterozygosity rates were 0.76 (D18S535), 0.88 (D10S2325) and 0. 90 (D1S1656), leading to a combined discrimination power of 0.9999. In contrast to D10S2325 and D18S535, which showed a homogeneous repeat array without any variation in the repeat motifs, repeat length and sequence variation was found for D1S1656. Robust typing results could be observed for all three STRs using highly degraded DNA.

Alleles↗

PubMed1999

German shepherd dog is suspected of sexually abusing a child.

A rare case of provoked anal penetration of an 11-year-old boy by a male German shepherd dog was confirmed by the results of morphological, serological and molecular genetic investigations. These results were of great importance to refute the suspicion on two adults. Some serious doubts remained in the version of the course of the event as presented by the boy. Some weeks later when confronted by a psychologist, the boy admitted having deliberately stimulated the dog manually and caused the animal to penetrate him.

Anal Canal↗

PubMed1998

Population genetic diversity in relation to microsatellite heterogeneity.

Nine populations (Germans, Turks, Moroccans, Ovambos, Ugandans, Chinese, Japanese, Papuans, and Australian Aborigines) were investigated using six microsatellite systems (HumCD4, Hum F13B, HumFES/FPS, HumTH01, HumVWA, and D21S11), so-called STRs (short tandem repeats). Allele frequency data and sequencing results were used to compare the population genetic diversity among these populations. The genetic differences varied depending on the STR applied. According to the systems investigated, we defined three categories of STR microvariation: LOMs (low microvariation systems), INMs (intermediate microvariation systems), and HIMs (high microvariation systems). LOMs (STRs: CD4, FES, F13B, TH01) are characterised by a number of repeats between 5-15 and a stable repeat sequence. INMs and HIMs each showed an increasing number of repeats and additional sequence variation in the repeat motifs. The rate of new mutations was associated with the extent of microvariation. The reconstruction of phylogenetic trees led to a clustering in an early split of the African populations followed by further branching of the Asian/Melanesian and the Caucasian groups.

Africa↗

PubMed1998

Tetranucleotide STR system D8S1132: sequencing data and population genetic comparisons.

In the present investigation of the D8S1132 locus 31 selected alleles were sequenced. In total there were 9 distinguishable alleles found to increase in size by regular 4 bp increments from 134 to 170 bp with a repeat array following the pattern (TCTA)n TCA (TCTA)n. One-third of the sequenced alleles exhibited an altered repeat sequence TCTG TCTA at the 3' flanking region of the repeat array. A nomenclature for the designation of D8S1132 alleles is proposed on the basis of this sequence data and in accordance with the ISFH recommendations. The allele distribution of the D8S1132 locus has been investigated in three German populations (Halle-, Münster-, and Wiesbaden area) with frequencies ranging from 0.004 to 0.24. No deviation from Hardy-Weinberg equilibrium could be observed. The heterozygosity was 0.83 and the discrimination power 0.96 for the Halle population.

Alleles↗

PubMed1997

Genetic variation at five STR loci in subpopulations living in Turkey.

Five short tandem repeat (STR) systems HumVWA, HumTH01, HumCD4, HumF13B and HumFES were investigated in 2 subpopulations living in Turkey (Laz Turks and Kurds). The population genetic data were compared to a Turkish population sample from the Adana area. A closer genetic relationship was found to the Laz Turks than to the Kurdish sample which was also confirmed by phylogenetic tree reconstruction with seven populations from three major ethnic groups (Caucasian, Asian and African). In contrast to the Laz and Adana populations the Kurdish sample showed relatively low heterozygosity values and deviations from Hardy-Weinberg equilibrium in four of the five systems.

Alleles↗

PubMed1997

DNA typing of epithelial cells after strangulation.

DNA typing was carried out on epithelial cells which were transferred from the hands of the suspect onto the neck of the victim. In an experimental study 16 suspect-victim combinations were investigated for estimating the typing success. Alternatively to an attack against the neck, the upper arm was used for "strangulation". PCR typing was carried out using the short tandem repeat systems (STRs) HumCD4, HumVWF31A (VWA) and Hum-FIBRA (FGA) and the success rate was > 70% for all 3 systems. In most of the cases mixed patterns containing the phenotype of the suspect and the victim were obtained. In a case where strangulation was the cause of death, epithelial cells could be removed from the neck of the victim. The DNA pattern of the suspect could be successfully amplified using four STRs, demonstrating the applicability of this approach for practical casework.

Adult↗

PubMed1996

Detection and significance of adenoviruses in cases of sudden infant death.

Respiratory tract infections have been thought to act as a trigger mechanism in sudden infant death. In 118 autopsy cases of infant death, paraffin-embedded or frozen lung tissues were investigated by means of a nested polymerase chain reaction (PCR) to detect adenovirus (AV) DNA. The primers used are general primers and allow the detection of most pathogenic adenoviruses with high specificity and sensitivity and independently of devitalization of viruses or degradation of viral DNA. For the investigation three groups were established: there were 13 cases of unnatural death, 78 cases of natural death without histological signs of interstitial pneumonia, and 27 cases with interstitial pneumonia. The first group was AV negative. In the group without interstitial pneumonia AV was detected in 10.2% of the cases. In the group with interstitial pneumonia the frequency of AV detection was almost 26%. The results obtained demonstrate an association between interstitial pneumonia and detection of AV DNA, indicating that AV may play an important part in pulmonary infection in infants. Histological evidence of interstitial pneumonia was not observed in all AV-positive cases, perhaps because nonspecific virus-related changes occurred only in early stages of viral infection. Comparison of the AV frequency in SIDS (25%) and non-SIDS cases (4%) indicates an association between pulmonary AV infections and sudden death. These results support the working hypothesis of respiratory infections acting as a trigger mechanism in sudden infant death.

Adenoviridae↗

PubMed1996

HumFES/FPS and HumF13B: population genetic data from north Italy.

DNA extracted from 119 unrelated individuals was analysed by the polymerase chain reaction at the polymorphic microsatellite loci HumFES/FPS (n = 115 individuals) and HumF13B (n = 119 individuals). The samples were collected from Caucasians living in the area of Milano (northern Italy). After horizontal polyacrylamide electrophoresis, 8 alleles were observed for HumFES/FPS, and 5 for HumF13B. Testing for Hardy-Weinberg equilibrium showed no significant deviation. The allele frequency data were compared with a German and a Turkish population sample.

Alleles↗

PubMed1996

[DNA degradation in formalin fixed tissues].

The intensity of DNA degradation in fixed tissues is dependent on the fixation solution and the fixation time. The aim of this study was the investigation of DNA degradation over fixation times of up to 70 days in different tissues (muscle, brain, liver, bone) and with different formalin concentration (2%, 4%, 8%; unbuffered). An additional test was performed to see whether the fixed tissues could be individualized using PCR analysis. The smallest amounts of DNA were extracted from liver and brain and the largest from muscle and bone. The amount of DNA that could be extracted decreased with increasing formalin concentration, while at the same time DNA degradation increased. With the PCR-VNTR system HUMTH01, all fixed samples could be typed regardless of the fixation time and the formalin concentration.

Brain↗

PubMed1996

Population genetic comparisons among eight populations using allele frequency and sequence data from three microsatellite loci.

Eight different population samples (Moroccans, Ovambos, Papuans, Australian aborigines, Germans, Turks, Japanese and Chinese) were studied using the tetranucleotide short tandem repeat systems HumTHO1 (THO1), Hum VWFA31 (VWA) and HumACTBP2 (ACTBP2). Ten alleles were differentiated in THO1, 11 alleles in VWA and 28 alleles in ACTBP2. THO1 showed 1 bp deletions in the repeat region, VWA sequence and structure variations of the 4-bp repeat motif and ACTBP2 sequence, structure and length variations in the repeat array and deletions/insertions (1-6 bp) in the flanking regions. A phylogenetic tree was constructed (UPGMA method) leading to branches which grouped Germans and Turks, Japanese and Chinese, and Papuans and Australian aborigines.

Asian People↗