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PubMed2003

Analysis of vaginal swabs for paternity testing and marker-assisted selection in cattle.

Blood is the standard source for DNA analysis, but requires venipuncture of cows by veterinarian and tedious and costly DNA extraction. A procedure was developed for sampling of vaginal cells from cows, establishing a cell lysis protocol using robotics, and applying fluorescent analysis of genetic markers. Two insemination technicians collected vaginal cells from 254 elite Israeli Holstein cows located in 152 herds using commercial Catch-All sample collection brushes. Cells were lysed in a 400-microliters solution, and 5 microliters was used as template for polymerase chain reaction (PCR). Sensitivity of the PCR was enhanced using only 1 microliter of lysed cells. Eight markers of the International Society of Animal Genetics paternity panel were amplified in four separate PCR. ILSTS039, a marker for a quantitative trait loci on BTA14, was amplified in a separate reaction. Genotypes from one to nine genetic markers were obtained for 253 out of 254 samples, and 244 cows had genotypes for at least three markers (96%). Cows that did not inherit either paternal allele for at least two loci were considered not to be daughters of the sire listed. Fifteen cows met this criterion, for a paternity misidentification rate of 6.25%. The frequency of allele 225 of ILSTS039, which was associated with increased milk fat content, was 11.1% in the bull-dam population, similar to the 12% found in the cow population in Israel. The use of vaginal cells for genetic analysis is accurate, as demonstrated by replicated analysis and by comparison to individual and population analysis based on DNA derived from blood.

Alleles↗
PubMed1995

A validation study of four single locus probes (MS1, MS31, MS43 and g3) in a Korean population--further evaluation for paternity testing.

Hypervariable tandem repetitive regions in human DNA are proving to be increasingly useful for genetic analysis in humans. We chose four single locus probes (SLP; MS1, MS43, MS8 and g3) for a validation test among Koreans. The specimens were from 216 unrelated individuals and 33 paternity inclusion families. Extracted DNA from EDTA blood was restricted by Hinfl and electrophoresed in 0.7% agarose gel, transferred and hybridized with chemiluminescent probes. Heterozygosity was over 90% by all of the probes. Total numbers of unassignable mutant bands from 33 paternity inclusion cases were 5, and the highest mutation rate was determined in probe MS1(0.045). The probability of having the same DNA band between two unrelated individuals was 5.7 x 10(-10) when four SLPs were used at the same time. The data presented here on allele frequencies and mutation rates provide preliminary data supporting the validity of these probes in paternity analysis and forensic investigators in the Korean population.

Alleles↗
PubMed1998

Paternal exposure to pesticides and congenital malformations.

OBJECTIVES: A case-referent study with 261 matched pairs was carried out in 8 hospitals of Comunidad Valenciana, Spain, to assess the relation between occupational exposure to pesticides and selected congenital malformations. In this paper, the results concerning paternal exposure are presented. METHODS: The parents of the case patients and the referents were interviewed to collect information about exposure to pesticides and potential confounding variables. Detailed information on direct involvement in the handling of pesticides was collected for the interviewees involved in agricultural activities during a previously defined period in relation to conception and pregnancy. Exposure data were reviewed by 2 experts who assigned ordinal scores for the probability and intensity of exposure to pesticide classes and active ingredients. RESULTS: The dichotomous analysis of exposure (absent, present) yielded some increased risks, although not statistically significant, for aliphatic hydrocarbons [adjusted odds ratio (adjusted OR) 2.05, 95% confidence interval (95% CI) 0.62-6.80], inorganic compounds (adjusted OR 2.02, 95% CI 0.53-7.72), and glufosinate (adjusted OR 2.45, 95% CI 0.78-7.70), and a significant association for pyridil derivatives (adjusted OR 2.77, 95% CI 1.19-6.44). The analysis based on the experts' scores (2 levels of exposure) showed some consistent associations for these compounds. CONCLUSIONS: This research indicates a possible risk of congenital malformations for paternal exposure to some pesticides, notably, pyridils, aliphatic hydrocarbons, inorganic compounds, and glufosinate. It did not find an increased risk for paternal exposure to pesticides in the classes of organophosphates, carbamates, organochlorines, chloroalkylthio fungicides and organosulfurs. These findings warrant further investigation.

Agriculture↗
PubMedDate not supplied

Autonomy and paternalism in medicine.

The role of ethics in medical practice is now receiving close scrutiny, so it is timely that ethical concepts, such as autonomy and paternalism, be re-examined in their applied contexts. As neither autonomy nor paternalism has a current universally accepted meaning, their significance varies in both ethical and clinical discussion. Of the two, autonomy has moved further from its original moral context, to the extent that it often now signifies no more than a person's expressed intention. Paternalism, characterised as the antithesis of autonomy, is widely thought not to have any role in medicine. The transforming effects of illness, which may radically alter a person's decision making capacity, are commonly ignored.

Beneficence↗
PubMed1999

Hemopoietic damage and induction of leukemia in offspring due to preconception paternal irradiation from incorporated plutonium-239.

Preconception paternal irradiation has been implicated in a localized excess of childhood leukemia and non-Hodgkin's lymphoma close to a nuclear reprocessing plant. Other epidemiological studies, however, threw doubt on the validity of this hypothesis. Experimental evidence implicating preconception paternal X rays in the development of lung and skin cancers has also been questioned. In this study, (239)Pu (0, 128 or 256 kBq kg(-1)) was injected into male CBA-H and DBA-2 mice 12 weeks before they were mated with CBA-H and C57BL/6 females. Their offspring were assessed for hematological status at 6 to 18 weeks of age or were treated with either 3.3 Gy whole-body gamma rays or methylnitrosourea (MNU, 50 mg kg(-1)) and monitored for onset of malignancies of the lymphoid and hemopoietic system. As a group, offspring were normal hematologically, but up to 35% of individual mice had femoral cellularities and numbers of spleen and fibroblastoid colony-forming cells outside the normal range. Exposure of the offspring to radiation or MNU significantly increased the rate of incidence of lymphoid and myeloid leukemias. Simulation of the experiments with preconception gamma irradiation indicated that damage to the spermatogenic stem cell was an important factor. It is concluded that preconception paternal irradiation can influence susceptibility of offspring to a subsequent exposure to a carcinogen.

Animals↗
PubMed2000

Population studies and validation of paternity determinations by six microsatellite loci.

A single locus system of 6 microsatellite markers was evaluated for paternity testing. A nonradioactive method based on peroxidase labeling of a DNA probe was used to estimate the allele frequency of markers D1S216, D3S1217, D7S480, D9S157, D13S153, and D16S422 by genotyping 1134-1698 chromosomes. The number of detected alleles were 22, 15, 23, 10, 16, and 19, respectively, and the allele frequency varied from 0.001 to 0.317. The genotype of 87 families, consisting of mother, father, and child was determined. The probability that a random individual will give a positive paternity was evaluated. We conclude that the markers can be reliably typed and give sufficient and reliable information for paternity testing.

Adult↗
PubMed1998

[Application of VNTR D17S30 locus polymorphism in the paternity test].

A sensitive and rapid PCR-based technique was adopted to genotype the VNTR D17S30 locus. It was confirmed through the genetic analysis of 20 normal families that the inheritance of D17S30 locus coincides with Mendelian law as simple co-dominant. Retrospective analysis of 100 paternity cases demonstrated that D17S30 locus could be used in forensic paternity test in our country. The exclusion probability estimated from allele frequencies of D17S30 locus (74.04%) does not differ significantly from the observed rate of exclusion (80.00%) in these cases. In all excluded paternity cases there are two in which the exclusion evidence is solely provided by the D17S30 locus.

Chromosomes, Human, Pair 17↗
PubMed2002

Medical paternalism serves the patient best.

It seems obvious that in a post-modern, constructivist world where meaning and value systems are often subjective and relative, any absolutist view is likely to be questionable. This is more so if it relates to ethics, the foundations, interpretation and application of which have been and continue to be much debated. So, in addressing the proposition, my efforts were directed at identifying a position that would mediate polarity. I examined the contention that the doctor, because he is better informed, may claim greater acuity and powers of judgment, and its defences against the charge of interfering with individual liberty and autonomy through various arguments such as the harm principle, the welfare, the principle of legal moralism and the appeal to uncertainty. While there is some validity to the arguments proposed, absolute paternalism would seem incompatible with respect for individual rights. How satisfactory, then, is the paradigm shift from paternalism to the independent choice model where the doctor presents neutral statistics as little biased as possible by his own views and judgments and leaves the decision making entirely to the patient or his/her relatives. This clearly had its limitations too. As with much of human experience, the answer would seem to rest in mediating the happy mean. Recognising a distinction between autonomy (self-determination) and independence (total freedom of choice without any interference) allows for a model of qualified independence or "enhanced autonomy" (Quill & Brody, 1996). This is predicated on doctor-patient dialogue, exchange of ideas/views, negotiation of differences, and sharing power and influence for the common purpose of serving the patient's best interest. This model would seem to be a responsible and effective approach to management of clinical dilemmas, as well as one that in its pluralistic approach is consistent with fundamental moral and philosophic propositions. It is by no means flawless, but in an imperfect world, there can be no perfect solution; constant negotiation with the realities--however uncomfortable--is an inescapable fact of life. Actions are right in proportion as they tend to promote happiness: wrong as they tend to produce the reverse of happiness. (J S Mil, Utilitarianism) On that supposition, I submit that guided paternalism is arguably what serves the patient best.

Decision Making↗
PubMed2001

[Study on application of CODIS loci to excluding paternity].

OBJECTIVE: The study was carried out on application of CODIS loci (FGA, vWA, CSF1PO, TH01, TPOX, D3S1358, D5S818, D7S820, D8S1179, D13S317, D16S539, D18S51 and D21S11) in 100 cases of excluding paternity. METHODS: The PCR amplified products of Profiler Plus and Cofiler amplification kit were injected into a capillary on the ABI PRISM 310 Genetic Analyzer. GeneScan software analyzed the collected data, which can then be imported into Genotyper software for genotyping of alleles. RESULTS: In the group of Mother-Child-Alleged Father, more than 3 STR loci incompatibilities between alleged father and child were found in all observed cases, the mean incompatibility was 6.63; In the group of Alleged Father-Child, 94.0% of all cases observed was found more than 3 STR loci incompatibilities, the mean incompatibility was 5.01. CONCLUSION: The results showed that CODIS loci had good application to excluding paternity, and the choice of hyperpolymorphic markers in investigation, which were valued by DP, H and PE, had direct relation with the augmentation of incompatibilities in excluding paternity.

Adult↗
PubMed1999

[HLA-DRB1 typing by PCR-sequence specific primers for paternity determination].

HLA-DRB1 typing by PCR-SSP technique was first used for paternity testing. The analysis of 42 paternity cases shows that this identification method is simple, fast and reliable and has high exclusion probability of paternity (66.3%). This method can also be applied to the study of transplantation, HLA associated disease and anthropogenesis.

Forensic Medicine↗
PubMed1975

[Blood group opinion and judicial judgement 1. 250 paternity cases 1965-1970 (author's transl)].

In 250 paternity actions (one-man one-child affairs 1965-1970) without exclusion by blood group opinion, which include 48 cases involving persons from abroad, 41 men acknowledged the paternity or withdrew the suit during the process. 32 cases were judged by default. 174 men were judged of child support according to the former 1717 BGB, 13 of these after an anthropological expertise. In 2 cases the anthropological opinion gave an indication of nonpaternity; accordingly, the court dismissed the action. In 157 cases the plausibility of paternity according to Essen-Möller's formula was calculated by the expert and dealt with in the blood group opinion. It appears, however, that the W values had no remarkable influence on the decision of the court.

Anthropology↗
PubMed2003

Novel paternity testing by distinguishing parental alleles at a VNTR locus in the differentially methylated region upstream of the human H19 gene.

Conventional PCR-based genotyping is useful for forensic testing but cannot be used to determine parental origins of alleles in DNA specimens. Here we describe a novel method of combined conventional genotyping and PIA typing (parentally imprinted allele typing) at a minisatellite region upstream from the H19 locus. The PIA typing uses two sets of primers and DNA digested with methylation-sensitive Hha I enzyme. The first amplification produces only the methylated fragment of paternal H19 allele, and the second detects polymorphism in the minisatellite. Hence, this distinguishes paternal and maternal alleles by difference in the DNA methylation. Furthermore, the polymorphism in this polymorphic locus was examined using 199 unrelated Japanese and 171 unrelated Germans, their polymorphism information content being 0.671 and 0.705, respectively. Feasibility of this typing is demonstrated for six families, and the usefulness is shown by application to paternity testing.

Alleles↗
PubMedDate not supplied

[Mono- and dizygotic twins in forensic paternity testing in practice at the Department of Forensic Medicine (Silesian Academy of Medicine, Katowice) in the years 1996-2003].

Giving an opinion on disputable paternity, concerning monozygotic twins in practice at the Department of Forensic Medicine (Silesian Academy of Medicine, Katowice) demonstrated their ideal agreement according to examined genetic markers possible. Even the mutation, which was revealed using the RFLP-VNTR method was the same for both twin sisters. In the case of dizygotic twins a firm differentiation of paternity index and probability of paternity was proved. This was the consequence of independent features segregation in first, reductive meiotic division. While the rare, out-ladder allele 16 at the CSF1PO locus was transmitted to both twins: a daughter and a son by the putative father.

DNA Fingerprinting↗
PubMedDate not supplied

[Paternity analysis in deficiency cases with related putative fathers: simulation of a deficiency analysis in 27 families].

During the last few years, the number of privately ordered paternity investigations has increased considerably. Probably due to financial reasons in more and more cases only the putative father and the child are investigated. Additionally, very often only one method, such as STR analysis, is employed. This raises the question whether such a reduced analysis leads to reliable and clear results when investigating cases with related putative fathers. We investigated 165 individuals from 27 families using the AmpFlSTRIdentifiler multiplex PCR and calculated the paternity probabilities of the children to their biological fathers, uncles, grand fathers and other relatives. In more than 30% less than three exclusions between child and relative were detected. In five cases no exclusions were found between child and uncle, always leading to paternity probabilities >99.9%. These results show that the calculation of high probabilities (>99.9%) does not necessarily lead to the accurate conclusion of fatherhood. In many of our cases misleadingly the brother of the real father or another close relative would have been declared to be the biological father.

Chromosomes, Human, Y↗
PubMed2005

SNPs and MALDI-TOF MS: tools for DNA typing in forensic paternity testing and anthropology.

DNA markers used for individual identification in forensic sciences are based on repeat sequences in nuclear DNA and the mitochondrial DNA hypervariable regions 1 and 2. An alternative to these markers is the use of single nucleotide polymorphisms (SNPs). These have a particular advantage in the analysis of degraded or poor samples, which are often all that is available in forensics or anthropology. In order to study the potential of SNP analysis in these fields, 41 SNPs were selected on the basis of following criteria: conservation, lack of phenotypic expression, and frequency of occurrence in populations. Thirty-six autosomal SNPs were used for genotyping 21 inclusionary and 3 exclusionary paternity cases. The behavior of 5 X-chromosome SNPs was analyzed in a French representative population. Our approach to SNP typing is a multiplex PCR based amplification followed by simultaneous detection by primer extension (PEX) analyzed by Matrix Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS). The selected autosomal SNPs showed independent inheritance and gave clear results in paternity investigation. All X-SNPs were useful as both paternity and identification markers. PEX and MALDI-TOF MS, with their high sensitivity, precision and speed, gave a powerful method for forensic and anthropological exploitation of biallelic markers.

Chromosomes, Human, X↗
PubMedDate not supplied

[STR and RFLP polymorphism in paternity determination based on family investigation].

This paper shows a paternity examination of a deceased putative father carried out on the basis of his family investigation. Statistical evaluation proved the defendant's paternity using STR and RFLP analyses with the probability of more than 99,999% (PI > 100 000). The restriction analysis has proven considerably more informative than the amplification analysis, which means seven RFLP markers correspond with the similar paternity indices of the fifteen STR markers.

DNA Fingerprinting↗
PubMed2005

[Diversity of paternity index with different allele frequencies].

OBJECTIVE: To evaluate the diversity of combined paternity index (CPI) of multiple STR loci when different population allele frequencies was used to calculate the paternity index. METHODS: CPI of 13 CODIS (combined DNA index system) loci for 108 trio cases and 108 duo cases selected randomly were calculated by using five Chinese Han population allele frequencies, respectively. RESULTS: The CPI range for trio cases and duo cases were 2077.63-50897711626.46 and 25.12-2998685141, respectively. When different population allele frequencies were applied to the same case, the ratio of maximum CPI and minimum CPI, which was more than 100, for trio cases and duo cases were 20 cases (19.52%) and 13 cases (12.04%), respectively. CONCLUSION: The variation of CPI value of the CODIS loci was obvious with different allele frequencies. To prevent the error causing by uncertain allele frequencies, a conservative CPI value should be calculated in paternity testing.

Alleles↗
PubMed1991

[Paternity test with single locus DNA probes].

Seven kinds of DNA probes recognizing hypervariable DNA loci were applied to 28 cases of paternity test, involving two cases in which the putative fathers had died. The combinations of probe and restriction enzyme are as follows; MR24/1-HinfI, 3'Globin-PvuII, Ha-ras-PvuII, Mucin-PvuII, D2S44 (pYNH24)-MspI, D17S30 (pYNZ22)-MspI, D1S57 (pYNZ2)-RsaI. The reported number of the alleles are 37, 39, 5, 10, 33, 15, and 5, respectively. Those probes lie on different chromosomes except D1S57 on 1p and Mucin on 1q21. Exclusion probability (EP) and paternity index (PI) were calculated from the allele frequencies in Japanese population reported by Yokoi et al. Cumulative EP from 7 DNA probes was 0.999932, and cumulative PI ranged from 7.3 X 10(6) to 947. Also, cumulative EP from 17 kinds of conventional blood group markers (CBGM) was 0.9776, and cumulative PI ranged from 1290 to 0.11. Total EP from 7 DNA probes and CBGM was 0.999998478. Cumulative PI from 7 DNA probes were 5 to 2,000,000 times higher than that from CBGM. The single locus hypervariable DNA polymorphisms are considered to be informative for paternity test.

DNA Probes↗