Forensic Science, 6 (1975) 5-7 0 Elsevier Sequoia S.A., Lausanne - Printed in The Netherlands
ABO(H) GROUP SUBSTANCES
FRANCISZEK
IN HUMAN INNER EAR FLUID
TRELA and BOZENA TUROWSKA
Institute of Forensic Medicine, Academy
of Medicine, Cracow (Poland)
SUMMARY Tests were carried out on 89 specimens of human inner ear fluid and the results compared with those of blood and salivary gland tests. It was found that ABO(H) group substances are present in the perilymph of secretors.
Blood group identification of human corpses or parts thereof may sometimes present serious difficulties. When non-hemolysed blood can be obtained from the cadaver, of course, the determination of the group to which it belongs poses no problem, but when hemolysis or processes of decomposition have set in recourse must be taken to indirect methods such as absorption, elution, etc. When the corpse has been dismembered, .an attempt may be made to determine the blood group from fragments of muscle, but as practice and experience have shown, experiments of this kind give only a small percentage of positive results [l] . While experimenting on the determination of alcohol in human perilymph, it occurred to us to find out whether the presence of blood group ABO(H) substances could be shown in inner ear fluid (perilymph and endolymph). If good results were obtained, then recourse might sometimes be made to this kind of experiment in cases of difficulty in determining to what group a cadaver belongs [ 2,3] . We have found nothing on this subject in the available literature. MATERIAL
AND METHODS
The study included 89 autopsy cases. Blood samples, the salivary glands and lymph from the inner ear were taken from the cadaver and tested serologically. Lymph from the inner ear was obtained by the method of Trela et al. [ 31. After penetrating the superficial osseous layers and part of the pyramids of the temporal bones at the site of the semicircular canals, and chiselling away further fragments of bone, the semicircular canals were opened. Perilymph and endolymph (250-350 mg) were taken by means of a glass pipette from each ear of the cadaver. The presence of ABO(H) group substances was determined by the absorp-
6
tion method, using anti-A and anti-B sera with a titer of 1 : 16 (serum obtained by diluting human serum to a titer of 1 : 128) and an extract of the seeds of Ulex europaeus at a titer of 1 : 4. In testing for the presence of A and B antigens, one drop each of anti-A and anti-B sera were placed in a test tube, and one drop of inner ear fluid added. In determining the presence of O(H) antigen, one drop of Ulex europaeus extract and two drops of perilymph were used. After leaving for 18 hours at a temperature of’4” C, tests were then made with standard A, B and 0 erythrocytes after 10 minutes. Complete inhibition of the agglutination reaction indicated the presence of the given antigen. Parallel control tests of blood samples and salivary glands were carried out, using the standard agglutination and absorption methods [4]. The salivary glands were examined in order to show whether the human cadavers came from secretors or non-secretors of ABO(H) group substances. The extract from salivary gland was prepared as follows: 2 g of the pieces a salivary gland were added to 1.5 ml saline and boiled for 15 minutes. The tissue was removed by centrifugation (3000 rpm for 10 minutes). One drop of extract was mixed with one drop of anti-A, anti-B serum at a titer of 1 : 16, 1 : 4 for anti-H phytohemagglutinin. The next step was analogous to the absorption test with perilymph fluid. RESULTS
AND DISCUSSION
The results of testing 89 specimens of blood, salivary glands and perilymph are shown in Table I. In 9 blood samples it was not possible to determine by direct methods the blood group to which the cadavers belonged, but using the method of Lattes and Holzer [4] it was shown that 2 cases belonged to Group 0, 4 to Group A and 3 to Group B. These results were confirmed by tests of the salivary glands and perilymph. TABLE I Results of ABO( H) group substance determinations human inner ear fluid.
in blood samples, salivary glands and .~
Group determined from cadaver blood samples
No. of cases tested
Salivary glands
Perilymphatics -
Group substance present not present
Group substance not present present
A B 0 AB Complete hemolysis
39 15 20 6 9
34 13 16 3 9
5 2 4 3 -
32 12 16 3 9
7 3 4 3 -
Total
89
75
14
72
17
On the basis of the results of the salivary gland tests it was possible to show that out of 89 cases 14 were non-secretors of group substances, while in 75 cases the group could be established without doubt. The results are similar to the frequency of secretors ABO(H) substances in the Polish population (Se = 83.94%, se = 16.06%) according to Turowska et al. [ 51. Out of 89 specimens of inner ear fluid, A, B or H group substances could be demonstrated in 72 cases, and the results of these determinations agreed with those of the salivary gland tests. In 3 cases secretors did not exhibit group substances; the reasons for this were not established. The other 17 specimens of perilymph tested came from non-secretors, and the negative results of the tests again agreed with the salivary gland results. These results indicate that antigens of the ABO(H) groups appear in the perilymph, and may be demonstrated in secretors by means of the absorption method using anti-A and anti-B sera with the low titer of 1 : 16 and anti-H serum at a titer of 1 : 4. It appears that these observations may sometimes be of use in the identification of dismembered corpses or in difficult cases by determining the blood group.
REFERENCES 1 B. Popielski and J. Kobiela (eds), Medycyna Sadowa. PZWL, Warszawa, 1972. 2 S. Iwankiewicz, J. Kornobis and W. Mikulewicz, Alkohol etylowy w przychlonce ucha wewnetrznego. Arch. Med. Sad. Krym., 1(1973) 129-132. 3 F.M. Trela, Z. Marek, A. Halama and Z. Grochowska, Poziom alkoholu we krwi i przychlonce pobranych ze zwlok. Arch. Med. Sad. Krym., 24 (1974) 65-67. 4 F.J. Holzer, Untersuchungen iiber die gerichtlichmedizinische Verwertbarkeit der Ausscheidung von Blutgruppensubstanzen. Z. Gerichtl. Med., 28 (1937) 1, 3, 234-248. 5 B. Turowska, Z. Marek and K. Jaegermann, CzestoSi: wystepowania poszczegolnych tech grupowych w populacji polskiej. Acta Med. Pol., (1975) in press.
ABO(H) GROUP SUBSTANCES
FRANCISZEK
IN HUMAN INNER EAR FLUID
TRELA and BOZENA TUROWSKA
Institute of Forensic Medicine, Academy
of Medicine, Cracow (Poland)
SUMMARY Tests were carried out on 89 specimens of human inner ear fluid and the results compared with those of blood and salivary gland tests. It was found that ABO(H) group substances are present in the perilymph of secretors.
Blood group identification of human corpses or parts thereof may sometimes present serious difficulties. When non-hemolysed blood can be obtained from the cadaver, of course, the determination of the group to which it belongs poses no problem, but when hemolysis or processes of decomposition have set in recourse must be taken to indirect methods such as absorption, elution, etc. When the corpse has been dismembered, .an attempt may be made to determine the blood group from fragments of muscle, but as practice and experience have shown, experiments of this kind give only a small percentage of positive results [l] . While experimenting on the determination of alcohol in human perilymph, it occurred to us to find out whether the presence of blood group ABO(H) substances could be shown in inner ear fluid (perilymph and endolymph). If good results were obtained, then recourse might sometimes be made to this kind of experiment in cases of difficulty in determining to what group a cadaver belongs [ 2,3] . We have found nothing on this subject in the available literature. MATERIAL
AND METHODS
The study included 89 autopsy cases. Blood samples, the salivary glands and lymph from the inner ear were taken from the cadaver and tested serologically. Lymph from the inner ear was obtained by the method of Trela et al. [ 31. After penetrating the superficial osseous layers and part of the pyramids of the temporal bones at the site of the semicircular canals, and chiselling away further fragments of bone, the semicircular canals were opened. Perilymph and endolymph (250-350 mg) were taken by means of a glass pipette from each ear of the cadaver. The presence of ABO(H) group substances was determined by the absorp-
6
tion method, using anti-A and anti-B sera with a titer of 1 : 16 (serum obtained by diluting human serum to a titer of 1 : 128) and an extract of the seeds of Ulex europaeus at a titer of 1 : 4. In testing for the presence of A and B antigens, one drop each of anti-A and anti-B sera were placed in a test tube, and one drop of inner ear fluid added. In determining the presence of O(H) antigen, one drop of Ulex europaeus extract and two drops of perilymph were used. After leaving for 18 hours at a temperature of’4” C, tests were then made with standard A, B and 0 erythrocytes after 10 minutes. Complete inhibition of the agglutination reaction indicated the presence of the given antigen. Parallel control tests of blood samples and salivary glands were carried out, using the standard agglutination and absorption methods [4]. The salivary glands were examined in order to show whether the human cadavers came from secretors or non-secretors of ABO(H) group substances. The extract from salivary gland was prepared as follows: 2 g of the pieces a salivary gland were added to 1.5 ml saline and boiled for 15 minutes. The tissue was removed by centrifugation (3000 rpm for 10 minutes). One drop of extract was mixed with one drop of anti-A, anti-B serum at a titer of 1 : 16, 1 : 4 for anti-H phytohemagglutinin. The next step was analogous to the absorption test with perilymph fluid. RESULTS
AND DISCUSSION
The results of testing 89 specimens of blood, salivary glands and perilymph are shown in Table I. In 9 blood samples it was not possible to determine by direct methods the blood group to which the cadavers belonged, but using the method of Lattes and Holzer [4] it was shown that 2 cases belonged to Group 0, 4 to Group A and 3 to Group B. These results were confirmed by tests of the salivary glands and perilymph. TABLE I Results of ABO( H) group substance determinations human inner ear fluid.
in blood samples, salivary glands and .~
Group determined from cadaver blood samples
No. of cases tested
Salivary glands
Perilymphatics -
Group substance present not present
Group substance not present present
A B 0 AB Complete hemolysis
39 15 20 6 9
34 13 16 3 9
5 2 4 3 -
32 12 16 3 9
7 3 4 3 -
Total
89
75
14
72
17
On the basis of the results of the salivary gland tests it was possible to show that out of 89 cases 14 were non-secretors of group substances, while in 75 cases the group could be established without doubt. The results are similar to the frequency of secretors ABO(H) substances in the Polish population (Se = 83.94%, se = 16.06%) according to Turowska et al. [ 51. Out of 89 specimens of inner ear fluid, A, B or H group substances could be demonstrated in 72 cases, and the results of these determinations agreed with those of the salivary gland tests. In 3 cases secretors did not exhibit group substances; the reasons for this were not established. The other 17 specimens of perilymph tested came from non-secretors, and the negative results of the tests again agreed with the salivary gland results. These results indicate that antigens of the ABO(H) groups appear in the perilymph, and may be demonstrated in secretors by means of the absorption method using anti-A and anti-B sera with the low titer of 1 : 16 and anti-H serum at a titer of 1 : 4. It appears that these observations may sometimes be of use in the identification of dismembered corpses or in difficult cases by determining the blood group.
REFERENCES 1 B. Popielski and J. Kobiela (eds), Medycyna Sadowa. PZWL, Warszawa, 1972. 2 S. Iwankiewicz, J. Kornobis and W. Mikulewicz, Alkohol etylowy w przychlonce ucha wewnetrznego. Arch. Med. Sad. Krym., 1(1973) 129-132. 3 F.M. Trela, Z. Marek, A. Halama and Z. Grochowska, Poziom alkoholu we krwi i przychlonce pobranych ze zwlok. Arch. Med. Sad. Krym., 24 (1974) 65-67. 4 F.J. Holzer, Untersuchungen iiber die gerichtlichmedizinische Verwertbarkeit der Ausscheidung von Blutgruppensubstanzen. Z. Gerichtl. Med., 28 (1937) 1, 3, 234-248. 5 B. Turowska, Z. Marek and K. Jaegermann, CzestoSi: wystepowania poszczegolnych tech grupowych w populacji polskiej. Acta Med. Pol., (1975) in press.
