[Blood group expertises including related presumptive fathers (author's transl)]. / Blutgruppengutachten mit nahe verwandten Eventualvätern.

In four affiliation cases with two or three brothers involved as putative fathers, exclusions of non-fathers were obtained by the application of the so-called isoelectrofocusing systems (twice by PiM subtypes, once by PGM1, TfC and Gc1 subtypes) as well as by GPT and the extended Gm system. In three cases a discrimination between the involved brothers was possible, in a fourth case both brothers were excluded from paternity. Special problems of anthropological expertises and of biostatistics in cases with closely related putative fathers are discussed. Possibilities to refute false statements of the alleged men, claiming sexual intercourse of the mother with their own relatives in order to invalidate positive biostatistical evidence, are pointed out.

[Blood group expert evaluation: relation between the extent of testing and the reliability of paternity determination. Reflections on revision and guidelines]. / Das Blutgruppengutachten: Beziehungen zwischen Untersuchungsumfang und Zuverlässigkeit der Vaterschaftsfeststellung. Uberlegungen zur Neufassung der Richtlinien.

In affiliation cases a combined exclusion chance for non-fathers of 99.995% is obtained by the examination of well-established blood group systems. In complicated cases, i.e. if known putative fathers are unavailable, the biostatistical limits for the ascertainment of paternity are obviously very high. They have to be determined by court in each particular case. For routine cases the application of an extended basic blood group expertise, including 19 systems with an combined exclusion chance of 95.17% is considered to be sufficient. The analysis of 263 own filiation cases from 1979 to 1982 yielded an average realistic prior probability of paternity of 83.3%, in 52 many-man affairs even of 90.3%. A similar percentage (89.5%) was observed in 67 two-man affairs of contested legitimacy. Since the father is rarely found among men included at a later stage the rate of children without known father is estimated at 5-15%.

The efficacy of modern blood group genetics with regard to a case of probable superfecundation.

The reported case of probable superfecundation is apparently the most extensively tested of all published cases: More than 40 specificities of the HLA Loci A and B, 5 of the Locus C, Bf and GLO I, furthermore ABO, MNSs, DCcCwEe, K 1--6, Fya, Fyb, Jka, Jk5, Lu3, Hp, Gc, Gm 1, 2, 4, 5, 21, Km 1, C3, Tf including C-subtypes, Pi, Bg, ACP, PGM1 including the extended polymorphism by PAGIF, AK1, ADA, 6-PGD, EsD, GPT, GALT, CAII and Hb. A reciprocal exclusion for putative father 1 and putative father 2 to the twin 2 and the twin 1, respectively, has been reached in 7 blood group systems: HLA, MNSs, Lu, Hp, Tf, Bg and GPT. The random probability for the fatherhood of an unknown third man suitable for both twins is extremely low among whites (1 : 10(8)) or blacks (1 : 10(9)). The development of paternity investigations within the last two years, mainly basing on isoelectrofocusing techniques, is shortly reported. The following systems are preferably involved: PGM1 (4 common alleles), Gc (including 1F and 1S), Pi (3 common subspecificities of M), C6, Tf (3 common subspecificities of C), FUC, Apo E, C2.

A third common allele in the transferrin system, TfC3, detected by isoelectric focusing.

Further genetic heterogeneity of the transferrin system (Tf) was revealed by prolonged isoelectric focusing (PAGIF) of human sera on polyacrylamide gels (pH 4--6.5). One of the two common subtypes of TfC, designated previously as TfC1, is split into TfC1 and the new subtype, TfC3. The gene product of TfC3 has an isoelectric point between C1 and C2. In our sample (n = 252) seven phenotypes, TfC1, C2-1, C2, C3-1, C3-2, C3, and C1B2, were observed and the following frequencies calculated: TfC1 = 0.795, TfC2 = 0.155, TfC3 = 0.042, and TfB2 = 0.008. Family studies (n = 25) indicate an autosomal codominant mode of inheritance. The six TfC phenotypes are present after treatment of sera with neuraminidase and can be revealed in part by PAGIF and subsequent immunofixation.

Properdin factor b-polymorphism in the population of Hessen, Germany.

The polymorphism of properdin factor B (Bf, C3 proactivator) in a population sample from Hessen, Germany has been investigated by agarose gel electrophoresis and immunofixation. In 522 unrelated individuals seven different phenotypes were observed and the following allele frequencies calculated: BfS = 0.7998, BfF = 0.1772, BfS0.7 = 0.0163 and BfF1 = 0.0077. Investigations of 100 families with 198 children and 30 mother-child combinations support the assumed autosomal codominant way of inheritance.

Investigations on the PGMa1 polymorphism (phosphoglucomutase--EC 2.7.5.1) by isoelectric focusing.

The determination of phosphoglucomutase (PGM1) phenotypes was performed by isoelectric focusing on samples from 1678 unrelated individuals from Hessen, Germany. Ten common phenotypes are considered as gene products of four alleles at the PGM1 locus with the following frequencies: PGMa1(1) = 0.6305, PGMa2(1) = 0.1844, PGMa3(1) = 0.1320, and PGMa4(1) = 0.0530. Twenty-two different mating types were observed in 113 families with 202 children. The segregation of the phenotypes in the offspring supports the assumed way of autosomal codominant inheritance. The example of a silent allele (PGM0(1)) as well as a rare variant (PGM7(1)) is reported.

Transferrin: evidence for two common subtypes of the TfC allele.

Evidence is presented for an extended polymorphism of human transferrin (Tf). Three common phenotypes were observed among TfC individuals after isoelectric focusing of sera on polyacrylamide gels. They are explained in terms of two subtypes of the TfC allele, tentatively designated TfC1 and TfC2. The distribution of the phenotypes Tf C1, C2-1, and C2 provides a good fit to the Hardy-Weinberg equation. In our population sample (n = 942) the following frequencies were calculated: TfC1 = 0.8195, TfC2 = 0.1720, TfB2 = 0.0064, TfB1-2 = 0.0016, and TfD1 = 0.0005. Family studies (n = 112) indicate an autosomal codominant way of inheritance. The observed subheterogeneity is detectable in purified transferrin after isofocusing and subsequent immunofixation. The subtypes are still present after treatment of sera with neuraminidase.

PiT: a new allele in the alpha 1-antitrypsin system.

In a genetic investigation of the population in Hessen, Germany, we found a family with a new, rare allele in the Pi system (alpha 1-antitrypsin). According to electrophoretic analysis and isoelectric focusing patterns, it is designated PiT. A pedigree study suggests autosomal codominant inheritance. The serum concentration of six heterozygous carriers of this allele (phenotype M1T or M2T) revealed normal alpha 1-antitrypsin levels.

An improved method for the identification of Gc1 subtypes (group-specific component) by isoelectric focusing.

The occurrence of common subtypes of the Gc1 gene in a German population (n = 261) was investigated by isoelectric focusing on thin layers of polyacrylamide in the pH range 4-6. Six common phenotypes designated Gc 1S, 1F, 1F-1S, 2-1S, 2-1F, and 2 are considered as gene products of three common alleles with the following frequencies: Gc1S=O.603, Gc1F=0.125, and Gc2=0.272. The sum of the allele frequencies of Gc1S and Gc1F corresponds with that of the 'old' Gc allele. Family investigation are in agreement with an autosomal codominant way of inheritance. the method employed provides an identification of the subtypes within 4 h without using anti-Gc-serum for immunoprecipitation.

PEP A9, a new, unstable variant in the peptidase A system.

A rare peptidase A variant, tentatively designated PEP A9, was observed in six members of a German family, indicating autosomal codominant inheritance. The electrophoretic mobility is similar to that of PEP A 3-1, but it has very low in vivo stability. There is no apparent association with a disease state. A simple and sensitive staining reagent for PEP A was found in o-phthalaldehyde.

Isoelectric focusing of rare transferrin (Tf) variants and common TfC subtypes.

TransferrinC (TfC) subtypes were determined by isoelectric focusing (PAGIF) on samples from 90 carriers of the TFB and TfD alleles. In all cases of CB and CD heterozygotes only one of the two common subtypes of the TfC allele, TfC1 or TfC2, was observed. This is considered strong support for the hypothesis of two common alleles at the Tf locus. The different isofocusing patterns of rare B and D variants are compared with the results obtained after agarose gel electrophoresis (AGE).

Investigations on the polymorphism of glyoxalase I (EC 4.4.1.5) in the population of Hessen, Germany.

The phenotypes of glyoxalase I (GLO) were determined in a random population from Hessen (Germany) by high-voltage agarose gel electrophoresis. The gene frequencies in 1150 unrelated individuals were 0.4391 for GLO1 and 0.5609 fro GLO2. Rare phenotypes were not observed. The segregation of phenotypes in 50 families and 32 mother-child combinations supports the assumed autosomal codominant inheritance. The possibility of a simultaneous typing for GLO, esterase D (EsD), and carbonic anhydrase2 (CA2) on one gel is discussed.

Evidence for two additional common alleles at the PGM1 locus (phosphoglucomutase--E.C.: 2.7.5.1). A comparison by three different techniques.

Lysates of erythrocytes, leukocytes, lymphocytes, and extracts of sperms were investigated for the PGM1 isozymes by three techniques: starch gel electrophoresis, high voltage thin-layer agarose gel electrophresis, and thin-layer isoelectric focusing on polyacrylamide gel. On starch, only the well known common phenotypes 1, 2-1, and 2 were demonstrable. On agarose, different distances of the two main cathodal bands (a,b) among the phenotypes 2-1 were noted. Furthermore, on agarose, some types considered as homozygous on starch gel had a single, sharp banded pattern while others were broad and blurred. Optimal separation was achieved by isoelectric focusing on polyacrylamide gel. In 291 leukolysates, 10 different phenotypes were identified. These are considered as gene products of 4 different common allesles at the PGM1 locus as suggested by preliminary family investigations. In a random population from Hessen these four alleles had the following frequencies: PGM1al 0.6186, PGMa21 0.1718, PGMa31 0.1426, and PGMa41 0.067, The preliminary designation a1, a2, a3 and a4 was chosen as the assumed polymorphism was demonstrated on acrylamide and agarose. The sum of the frequencies PGMa11 and PGMa31 (the gene products of which have apparently the same electrophoretic mobility on starch) is similar to the frequency of the old PGM11 allele (0.757) in Caucasoids, PGMa21 and PGMa41 have a frequency of 0.2388 corresponding with the frequency of the old allele PGM21.

A new procedure for the determination of transferrin C (Tf C) subtypes by isoelectric focusing. Existence of two additional alleles, Tf C4 and Tf C5.

Isoelectric focusing (IEF) is widely used for the determination of serum protein polymorphisms in forensic investigations. Because of non-genetic heterogeneity of some proteins and of protein-ligand dissociation observed after IEF standardization of procedures is necessary. In this article the authors point out the main difficulties encountered in studying the transferrin subtype polymorphism and describe a new and sensitive method. Two new Tf C subtypes were observed: Tf C4 in North American, Bolivian and Brazilian Indians, Tf C5 in Black Americans, both with polymorphic allele frequencies. In a Pyrenean population the allele frequencies for Tf C1, Tf C2 and Tf C3 were quite similar to those observed in a German population sample.

Investigations on the polymorphism of sperm diaphorase in man. Evidence for a third common allele, SD.

The polymorphism of sperm diaphorase (SD) was investigated in 141 unrelated persons from Hessen, Germany, by high voltage thin-layer agarose gel electrophoresis (Age) and thin-layer isoelectric focusing on polyacrylamide gel (Pagif). In addition to the three known common phenotypes SD 1, 2-1, and 2, two further phenotypes with the preliminary designation SD 3-1 and SD 3-2 were discovered. This polymorphism can thus be explained in terms of three alleles, SD1, SD2, and SD3 segregating at an autosomal locus. The allele frequencies calculated from the five different phenotypes SD 1, 2, 2-1, 3-1, and 3-2 are: SD1 = 0.7553, SD2 = 0.2234, and SD3 = 0.0213. As we also found SD activity in female reproductive tract tissues (ovaries, oviducts, uterus), the term 'gonadal diaphorase' (GD) appears to be applicable.

Occurrence of Su in a German family.

Investigating a paternity case we observed a pretended opposite homozygosity of the S and s alleles between a mother and her child. The presence of the rare allele Su in this family could be demonstrated with the aid of dosage studies and blood group testing of the mother's parents. 22 additional genetic marker systems, including the HLA-system were examined, a biostatistical calculation was performed as well. The results obtained from these testings were in accordance with the hereditary rules and the posssibility of ilegitimacy or of children's substitution could be excluded. Only Caucasoid ancestry was known to the family. In addition, family members showed another rare blood group marker, the allele GPT0.

Immunogenetic markers in patients with Graves' disease.

110 carefully characterized Caucasoid patients with Graves' disease were tested for HLA class I and class II antigens. Compared with Caucasian controls (n = 193), the frequencies of HLA B8, Cw7 and DR3 were significantly increased (pc less than 0.05). In the subgroups with and without exophthalmos, HLA A3 exhibited a negative but insignificant association with the eye involvement, while A19 and Cw2 showed positive, however even weaker correlations with eye disease. HLA DR5 was associated with relapsing thyrotoxicosis, whereas HLA DR7 and B12 were negatively correlated with relapse. These results confirm the positive correlation of HLA B8 and DR3 with Graves' disease and reveal a not yet observed association with Cw7. Reported correlations of antigen frequencies with eye disease and relapsing thyrotoxicosis could not be confirmed. Other previously unknown, however subtle differences in disease subgroups were observed.

Report of a working group for the European Pharmacopoeia. Collaborative assay on human albumin of different origin.

A collaborative assay was conducted by 9 laboratories on 31 samples of human albumin which were in clinical use. It was the object of the study to establish test systems which would differentiate between albumins of venous or placental origin. The properties examined for this purpose were: appearance, total protein, haem, polymers, alkaline phosphatase and blood group substances. Additional tests such as for beta-thromboglobulin and citrate were included; pyrogenicity, however, was excluded because this was under study for all plasma proteins at that time. Results obtained were in satisfactory agreement both between laboratories and between samples. They, therefore, enabled the verification of a number of correlations in the test systems. The evaluation did not allow, however, the differentiation of the samples in relation to their origin. The results were, therefore, regarded as a tool to define the upper limits of acceptance for human albumins corresponding to the quality prescribed by the European Pharmacopoeia.