Polymorphism of the human phosphoglycolate phosphatase in Northrhine-Westphalia (F.R.G.) and its application to paternity testing

Blood samples from 563 unrelated German and from 110 Turkish individuals living in the Düsseldorf area were studied for phosphoglycolate phosphatase polymorphism. The distribution of the observed phenotypes in the population genetic study did not diverge from the expected values according to Hardy-Weinberg law. In our series, the gene frequencies were calculated as follows: (a) Germans: PGP1 = 0.851, PGP2 = 0.118, PGP3 = 0.031, (b) Turks: PGP1 = 0.973, PGP2 = 0.018, PGP3 = 0.009. The assumed autosomal codominant mode of inheritance was confirmed by the examination of 109 mother-child pairs and by analysis of 70 cases of disputed paternity. The plausibility to exclude German non-fathers from paternity is 12.78%.     

The problem of single parent/child paternity analysis--practical results involving 336 children and 348 unrelated men

In a certain amount of paternity investigations, only DNA from child and alleged father is analyzed, thus increasing the possibility of false paternity inclusions. The aim of this study was to determine how many wrong paternity inclusions could be detected in a rather small geographical area comparing empirical results from 336 children and 348 men (13-15 STRs were investigated per person). This comparison between each child and all unrelated men (i.e. all putative fathers from the other cases) with an especially designed computer program resulted in 116,004 man/child pairs. Less than three excluding STRs were found in 1666 child/unrelated man pairs (1.44% of the comparisons). At least one unrelated man with only two or less STR mismatches could be determined for 322 children (95.8% of all investigated children). In 26 comparisons no STR mismatches between a child and an unrelated man were detected, thus at least one and up to three "second father(s)" under 350 men could be found for 23 children, if the mother is excluded. Paternity probabilities between 95.475% and 99.996% were calculated. Our results underline the difficulties in motherless paternity cases using only STR analysis and advise great precaution in assigning verbal predicates such as "paternity proven" in those investigations.     

PCR typing of DNA fragments of the two short tandem repeat (STR) systems upstream of the human myelin basic protein (MBP) gene in Danes and Greenland Eskimos

DNA from the double short tandem repeat (STR) system MBP (locus 18q23-pter) was amplified by the polymerase chain reaction (PCR) and the two polymorphic repeat systems were separated by cutting with the restriction enzyme NlaIII. The lengths of the DNA fragments of the two MBP STR systems MBP-A and MBP-B were analyzed by vertical electrophoresis in polyacrylamide gels followed by silver staining. DNA samples from 112 unrelated Danes, 140 unrelated Greenland Eskimos, and 88 Danish mother/child pairs were analyzed. The distributions of MBP phenotypes were in Hardy-Weinberg equilibrium in both the Eskimo and Danish populations. Significant differences were observed between the distribution of fragments (‘alleles’) in Greenland Eskimos and in Danes. The allele MBP-A7 was considerably more frequent in Eskimos (0.2214) than in Danes (0.0775) and also the allele MBP-B9 was considerably more frequent in Eskimos (0.225) than in Danes (0.06). Strong gametic associations were found between fragments from the MBP-A and MBP-B series in both Danes and Eskimos. Some of the associations were different in Danes and Eskimos. In the 88 Danish mother/child pairs, the segregation of the MBP genotypes were in accordance with a genetic model of co-dominant inheritance and no mutation was found. Two MBP STR regions with irregular structures were sequenced. One fragment had a single base G to A transition at position 124 in the primer binding region between the MBP-A and MBP-B regions. In the other fragment, a deletion starting at position 117 and including the primer binding region between the MBP-A and MBP-B regions was found.     

Calculation of likelihood ratio y/x in diagnosis of paternity using computer methods.

A new computational method using a Monte Carlo technique is described for the calculation of plausibility of paternity in blood group systems. In this study gene frequencies of a blood group system are simulated by the range of the seven digit random numbers. By using a Monte Carlo method, four random numbers are generated and converted into paternal and maternal genotypes. Then the genotype of the child is determined according to the law of inheritance, and finally genotypes of the father, mother and child are converted into phenotypes. Repeating this process more than one hundred thousand times, the phenotypic frequencies of child-mother-father combinations (trio) and the likelihood ratio of paternity in any blood group system are calculated for all phenotypic combinations of the trios. This method is much easier than methods reported previously, and is sufficiently accurate.     

Polymorphism of isoenzymes in preserved muscle tissues

In muscles preserved in formalin enzymes were not found to be active. In muscles treated by ethanol the ESD, GLO, GPT and PGP enzymes were active. The best results were obtained in the case of acetone treatment. The phenotypes ESD, GLO, GPT and PGP in tissues corresponded with the ones in the comparative blood samples.     

Possible pitfalls in motherless paternity analysis with related putative fathers

Nowadays, more and more paternity cases are carried out investigating only child and putative father, mostly for economical or private reasons. Usually, reliable results can be obtained and the putative father can be included or ruled out with a high certainty. Considerable problems might arise when a relative of the biological father is investigated as being the putative father. In this study, we investigated 164 persons from 27 families creating artificial deficiency cases using the AmpFlSTRIdentifiler kit, which amplifies 15 STRs simultaneously. We analyzed 93 child/biological father pairs and the corresponding uncles, respectively the brothers of the biological fathers. The average paternity probability for the biological father was 99.9699% (paternity index (PI): 3321.26); only in three cases the results were under 99.9%. In five out of 125 child/uncle pairs no STR mismatches were found and paternity probabilities between 99.9726% (PI 3652) and 99.9970% (PI 33,545) were calculated. The average number of excluding loci was 3.4, but in 31.2% of the cases only zero, one or two mismatches were found. When both putative fathers were genetically typed, the biological father usually had a statistically higher paternity probability. Nevertheless, the differences between probabilities for father and uncle were only small. These results show that a reliable investigation of deficiency cases (i.e. child and putative father) seems to be more difficult than generally assumed. Especially in cases with an unknown familiar background and/or when investigating foreigners for immigration purposes, the laboratory expert should include the mother, increase the number of investigated loci or include a second method such as RFLP-analysis, some serological systems or typing of X-chromosome specific STRs to further ascertain the results.     

Molecular analysis of the human esterase D gene ESD(*)Q0(yonago) responsible for incompatibility in a Japanese paternity case

In a Japanese paternity test, an alleged father was excluded only by reverse homozygosity of esterase D (ESD) phenotypes (mother, ESD 1; child, ESD 1; alleged father, ESD 2) out of 43 classical and DNA markers investigated. To solve the aberrant inheritance of the ESD phenotypes observed between them, fragments for all eight coding exons amplified by polymerase chain reaction (PCR) were subjected to DNA analysis. The child and alleged father shared a null allele, originating from ESD(*)1. It was characterized by having TGA for the stop codon instead of TCA for serine at codon 63. Thus, the sharing of a rare null gene, ESD(*)Q0(yonago), increased the probability of paternity.     

Differences in tissue distribution of HV2 length heteroplasmy in mitochondrial DNA between mothers and children

Sequence analysis of HV2 in mitochondrial DNA has been performed as a tool for forensic identification, in addition to that of HV1. HV2 contains length heteroplasmy, which shows high variability within an individual or in maternal relatives. In this study, we used cloning analysis and PCR direct sequencing to compare, between mothers and their children, HV2 length heteroplasmic profiles in different tissues. For two mother-child pairs, different types of variant distribution were observed by cloning analysis. In pair 1, length heteroplasmic patterns in most tissues were similar (predominantly 9 and 10Cs variants), but different length heteroplasmic levels, with shifts in predominant genotype, were observed for some hairs in both mother and child. In pair 2, genotype distribution was similar for all tissues, with a predominant 8Cs genotype, but varying in the proportion of minor component. The proportion of one minor length variant (9Cs) in blood from the child was significantly higher than that from the mother, but the proportions of minor components (7 and/or 9Cs) in other tissue samples decreased from mother to child. Moreover, we could confirm that sequence types of PCR products were reflected by the distribution of length variants, which were observed especially in high proportion, in cloning analysis. Our results reveal variable changes in length heteroplasmic level in various tissues between generations. Variability between tissues, especially among hairs, within an individual would result in complicated differences in genotype distribution between maternal generations, and correlate with longer length of Cs for predominant variants.     

Single-strand conformation polymorphism of hyper-variable regions HV1 and HV2 of human mitochondrial DNA: detection by silver staining

Human mitochondrial DNA from 50 trios consisting of mother (M), child (C) and father (F) was PCR amplified with primers flanking the hyper-variable regions, HVR1 and HVR2. The amplified products were then fractionated under non-denaturing conditions, silver-stained and compared by single-stranded conformational polymorphism (SSCP). In all but one case, mother and child displayed identical patterns, which could be promptly distinguished from that of the father. For the remaining cases, either set of primers was sufficient to resolve the familial ties. In no instance, M displayed alleles different from those of C within each trio, demonstrating that no false exclusions occurred. The SSCP approach proved to be a robust technique suitable as a preliminary screening in cases requiring identification of multiple samples.     

Corporal Punishment and Kids: How Do Parent Support and Gender Influence Child Adjustment?

Previous research on corporal punishment has failed to consider the interaction of parent support and parent gender in predicting child outcomes. The current study examined whether parental support moderated the effects of corporal punishment on child outcomes (i.e., depression and aggression), and more specifically, whether the gender of the supportive parent moderated the effects of punishment from the opposite-sex parent. Results differed depending on the gender of the punishing and supportive parents, suggesting that parental support can be a protective factor in child outcomes but only under certain conditions. Mother support moderated the effects of father punishment on child depression but not child aggression. High corporal punishment by father was related to more child depression at both high and low levels of mother support. High levels of mother support only seemed important (i.e., children were less depressed) at low levels of father corporal punishment. In contrast, father support moderated the relationship between mother corporal punishment and child aggression but not depression. Children with high father support showed less aggression across all levels of mother corporal punishment. At low levels of father support, child aggression increased as mother corporal punishment increased. For depression, mother corporal punishment was positively related to child depression regardless of level of father support. These findings suggest differential effects for mother and father support and have implications for the treatment and prevention of negative outcomes in children who are physically punished by their parents.

"New" phenotypes in the human red cell isozyme system ADA

Two rare ADA phenotypes were observed in a German mother and her child. These phenotypes may be due to the allele ADA *9 previously found in Bulgaria.     

Polymorphism of plasminogen in Tuscany (Italy)

The distribution of PLG phenotypes in the population of Tuscany (Central Italy) has been investigated by means of isoelectric focusing followed by immunofixation of desialyzed sera. In a random sample of 383 unrelated healthy blood donors registered at the Hospital of Pisa, three common phenotypes, PLG A, A-B, and B, and two rare variants were found. The allele frequencies calculated in our study were: PLG*A = 0.6749, PLG*B = 0.3225, and PLG*rare = 0.0026. The theoretical exclusion rate in cases of disputed paternity is 17.42%.     

Immunohistochemical determination of maternal and child blood groups (ABO) in mature placental tissue

Using the indirect immunoperoxidase technique (PAP method), ABO characteristics of mother and child were correctly identified in tissue specimens from 10 mature human placentas. In one case, a weak infantile A reaction was overlooked in the agglutination test but correctly identified by immunohistochemistry. In accordance with the weak expression of ABO characteristics in cord blood, immunohistochemical labeling of infantile erythrocytes with monoclonal and human antibodies, as well as Ulex europeaeus agglutinin I (UEA I), was less pronounced than that of mature erythrocytes. Labeling of the chorionic vessel endothelium, in contrast to that of adult endothelial tissue, was negative with anti-A or anti-B but, regardless of the infantile blood group, pronounced with UEA I. Regular identification of the blood groups was possible in decomposed placental tissue stored at room temperature for 1 week, but not in tissue stored for 2 or more weeks.     

Significance of additional RFLP single locus probes analysis after STR based determination of sibship

Autosomal STR typing alone seems to be no sufficient tool for resolving deficiency cases (e.g. cases of questioned paternity or half-sibships). Therefore, we investigated whether the additional analysis of RFLP single locus probes can improve the solution of such complicated kinship cases. We analyzed 207 children and men from 101 families using the AmpFlSTRIdentifiler multiplex PCR kit and three RFLP single locus probes. A comparison between each child and all unrelated men resulted in 11,023 man / child pairs. Less than three excluding STRs were found in 125 child / unrelated man pairs (1.13%). Additional analysis of RFLP results reduced the number of ambiguous cases to 35. Half-sibling pairs were simulated using STR results from 20 cases with high paternity probabilities (group 1) and relatively low paternity probabilities (group 2). Using a commercially available computer program we calculated probabilities for 778 half-sibling pairs. In 35 pairs (4.49%) half-sibling probabilities over 90.0% could be calculated. Additional investigation of RFLP single locus probes did not lead to a more reliable evaluation of these results. The combined investigation of autosomal STRs and RFLP single locus probes can satisfactorily solve deficient paternities but does not contribute to the solution of questioned half-sibships.     

The role of family conflict and marital conflict in adolescent functioning

Recent research has suggested that general family conflict is more disruptive to child functioning than is marital conflict. We hypothesized, in contrast to earlier work, that marital conflict will contribute unique variance to our understanding of child functioning but only when it occurs in front of children. One hundred and forty-six mother/adolescent pairs served as participants. Mothers completed measures of marital and general family conflict. Both mothers and teachers completed measures on child functioning at two points in time separated by 1 year. Consistent with the earlier findings, general family conflict was more predictive of child adjustment problems than marital satisfaction. However, marital conflict occurring in front of the child was equally predictive of child problem behaviors as was general family conflict.     

Apparently contradictory homozygosity of gc- and esd-markers in three generations (author's transl)]

In a paternity case we found contradictory blood group antigens in the serum- and enzyme-polymorphisms Gc and EsD of the child, her mother and grandparents. A possible accidental exchange of the child could be excluded by biostatistical calculations of the probabilities of motherhood, fatherhood and parenthood, and the descent from the parents was proven in both generations. The serological findings may be explained by the concurrent existence of silent genes or of genes that cannot be determined as yet. The genetic information Gc degrees descended from the grandfather and that for EsD degrees from the grandmother. Both were transmitted from the mother to the child. Thus, the case could be cleared by statistical considerations only.     

Evaluation of the Reliability of DNA Typing in the Process of Identification of War Victims in Croatia

Abstract:  Aiming to estimate the frequency of various types of errors that can occur in the large-scale process of identification, we identified and compared genotypes of 911 parent–child pairs in the database of 3498 relatives of people that disappeared during the 1991/1992 war in Croatia. Genotypes of 891 pairs (97.8%) were matching, while 20 pairs did not match in one or more loci. Reanalysis of these samples revealed that out of 1822 analyzed genotypes, one genotype was completely wrong, and two genotypes had one wrong allele because of human errors. Five genotypes had a single wrong allele due to either polymerase chain reaction or electrophoresis errors. In five genotypes mutations were the cause of mismatch. Genetic inconsistencies with parentage were found in four "fathers" (4.2%) and three "mothers" (0.36%). As the majority of observed single-locus errors were caused by nonhuman errors, all databases produced with similar technology would probably have comparable level of errors.     

Discrepancy between Mother and Child Perceptions of Their Relationship: II. Consequences for Children Considered within the Context of Maternal Physical Illness

Discrepancies between parent and child perceptions of parenting have been well established. The current study addressed whether discrepancies in one dimension of parenting, the parent-child relationship, are associated with child adjustment difficulties concurrently and longitudinally. In particular, it was hypothesized that greater discrepancies in mother and child perceptions of parenting would be associated with child adjustment difficulties. Furthermore, it was hypothesized that families experiencing a stressor—in this case, maternal infection with the human immunodeficiency virus (HIV)—would experience larger discrepancies than families in which the mother is noninfected and that maternal HIV infection would moderate the association between discrepant views and child adjustment. Participants were 183 African-American women (61 HIV infected; 122 noninfected) and one of their noninfected children. All participants were from the inner-city area of New Orleans. Results indicated that discrepancies in mother and child perceptions of their relationship was associated with mother and child reports of externalizing behavior problems concurrently and longitudinally. In addition, discrepancies were significantly higher in families experiencing maternal HIV infection than in the noninfected group. However, with one exception, maternal HIV status failed to moderate the relationship between discrepancies in perception and child adjustment, indicating that discrepancies play a similar role in both types of families. Clinical implications of the present results are discussed.     

Qualitative Exploration of the Influence of Domestic Violence on Motherhood in the Perinatal Period

The perinatal period may include domestic violence (DV). In this study, the perinatal period extends from the start of pregnancy to the end of the child’s second year of life. DV places the safety of both mother and child at risk and undermines the mother–child attachment bond. In this article, we look at the challenges for motherhood in a context of domestic violence during the perinatal period (DVPP). In semi-directed interviews, 17 mothers shared their stories of DVPP. Interview questions focused on the trajectory of DVPP, perceptions and occupation of motherhood within two years of childbirth and the associated challenges. Data were analyzed using thematic analysis and discussed in a feminist conceptual framework and a parental theoretical model. DVPP negatively influences the experience and practice of motherhood, in addition to increasing parental responsibility to protect the child. Three main themes were noted: 1) A parental experience impoverished by the context of DVPP, caused by an increase in tasks, a weakened bond with the child and a loss of self-confidence; 2) Parenting responsibility in a context of DVPP is expressed as greater vigilance and the need to protect the child from violence; and 3) Parenting practices: the children as targets and victims of violence, resulting in additional child needs that mothers must meet. The results underscore the need for early detection of DVPP and a trauma-informed approach in health services. Specialized support services should be provided to help mothers cope with violence.

     

Use of deoxyribonucleic acid (DNA) fingerprints for identity determination: comparison with traditional paternity testing methods--Part II

Six red blood cell (RBC) antigen systems, coupled with human lymphocyte antigen (HLA) phenotyping, were used to establish paternity on 28 mother/child/alleged-father trios. Samples were subsequently examined using the deoxyribonucleic acid (DNA) fingerprinting test with the multilocus Jeffreys DNA probes 33.6 and 33.15. In 27 of 28 paternity cases, the DNA fingerprinting test results supported and enhanced the results of RBC and HLA typing by resolving disputed paternity cases conclusively. One discrepancy between conventional serological methods and DNA analysis is discussed.