UNIT VIII. DETERMINATIONOF SEX OF ORIGIN, NONGENETIC MARKERS AND BLOOD COMPONENT PROFILING Sex of Origin

SECTION 49. DETERMINATION OF SEX OF ORIGIN

49.1 Introduction from abnormalities of sex chromosome composition, the For some years, there has been interest in developing pro- correlation between phenotypic sex and sex chromatin body cedures which would enable the reliable determination of frequency in nuclei may not hold up. These cases are com- sex of origin of bloodstains and of cextain other human paratively rare in the population, but could lead to misinter- tissues such as saliva and hairs. In some cases, such a deter- pretation in medicolegal work. Discrepancies between mination could be informative and useful. Three general ap- phenotypic sex and chromatin body frequency may be ob- proaches have been used for this purpose, two of them cyto- served in (I) chromatin negative females; (2) chromatin logical, and the third having to do with the measurement of positive males; and (3) genetic mosaics. Phenotypic females sex hormone levels. The cytological techniques are con- who are chromatin negative are mainly of two types, those cerned with the determination of either sex chromatin (Barr with Turner's syndrome, and those with testih feminiza- bodies), or of so-called F-bodies (Y-bodies) in cells. tion syndrome. The former have various abnormalities, in- 49.2 Determination of Sex Chromatin cluding ovarian agenesis, and karyotype analysis reveals that they have only 45 chromosomes, the missing one being Sex chromatin bodies (Ban bodies) were discussed in sec- latter tion 1.2.4.4. These structures are characteristic of nuclei in an X. They are sometimes called "XO females". The certain cells from female mammals, including the nuclei of group are phenotypically female, although they exhiii various abno-ties of sexual development. Karyotype smooth muscle, adrenal cortex, and various epidermal cells. analysis shows that they are chromosomally male, i.e., that The structures may also be seen in polymorphonuclear they are XY and have 46 chromosomes. They are sometimes leucocytes from peripheral blood smears of females, where called "XY females". Males who are chromatin positive are they are sometimes called "drumsticks". These structures often those who have soalled Klinefelter's syndrome. They were fm observed in cat neurons (Ban and Bertrand, have 47 chromosomes and an XXY sex chromosome com- 1949), and later in other cells and other species (Barr, 1957 position, and they show a variety of abnormalities of sexual and 1960). The nature of sex chromatin in cells is discussed development. Genetic mosaics may be of either phenotypic by Moore (1%6a), and he has also described its discovery sex, and different tissues of the same individual may show and the earlier work in the field (Moore, I%&). Davidson differences in sex chromatin pattern. Cytologic sexing by and Smith (1954) first noted sex chromatin structures ("drumsticks") in the neutrophil leucocytes of peripheral means of chromatin bodies has been reviewed by Grob (1970). blood from females. These structures should be called "sex chromatin", or "X chromatin", though the latter term has not gained wide acceptance. Dr. Barr himself has discour- 49.2.1 Sex chromatin determinahion in blood and aged the use of the term "Barr bodies". bloodstnins Reports on the frequency of sex chromatin have varied, Postmortem blood does not appear to be very suitable for depending upon the tissue being studied. At least 100 cells sex chromatin determination, apparently because the leuco- must be examined according to most authorities, and some cytes undergo lysis and degradative changes quite soon after have recommended from 300 to 500. The variations may be death. Dixon and Torr (1956 and 1957) said that they had due to tissue differences, and to differences in various had no success with this technique in post mortem blood. technical factors. Other nuclear appendages may be mis- Schilling (1960) reported that post mortem blood was good taken for sex chromatin bodies by inexperienced observers for up to about 6 hours after death for this determination. as well. The somewhat subjective nature of sex chromatin Some reports have indicated limited suces with blood- determinations has been noted by Grob and Kupperman stains. De Bemardi (1959) studied bloodstains on hairs with (1%1), and they emphasized the importance of using some success. He said that at least 200 nuclei needed to be reproducible techniques and experienced scorers. examined. Davidson (1963) described a procedure for the In the earlier literature, a low frequency of ocmmnce of isolation of white cells from bloodstains, and examined a sex chromatin bodies was reported in cells from nond number of slides for chromatin bodies. Diagnosis of sex human males, but the general consensus of opinion some could not be made in every case, and he thought that the years ago seemed to be that nonnal male cells do not in fact results of the determination should be taken only as an in- exhibit the structures (Grob and Kupperman, 1%1). As was dication of possible sex. Ishizu et al. (1973) suggested that noted earlier, there is evidence (Ohno, 1966) that the sex chromatin body detemhations coupled with F-body deter- chromatin body structure represents an inactivated X chro- minations (see below) on the same material provided a more mosome (see in section 1.2.4.4). Thus, in people suffering defintive conclusion than either cytological method by Sourcebook in Forensic Serology, Irnrnunologv, and Biochemistry itself. caroff and Breton (1Wreviewed chromatin body 49.3 Y-Body (F-Body) Determinations sexing from a medicolegal point of view. In 1%. Caspersson et al. reported that metaphase chromosomes exhibited various fluorescence patte-rns after 49.2.2 Sex chromatin determination in epithelial cells and staining with fluorescent dyes, and it was suggested that the hair technique might provide a useful tool for chromosome map From a forensic point of view, the abity to determine sex ping. These experiments were done on nonhuman material. in epithelial cells is probably most applicable to post mortem Certain loci of the chromosomes were observed to bind tissues and to exfoliated epithelial cells in saliva stains and quinacrine diHCl preferentially. In 1969, Zech found that traces. Dixon and Ton (1956 and 1957) noted that chroma- the long arm of the human Y chromosome fluoresced tin bodies could survive for a few weeks in post mortem brightly after staining human chromosome preparations epithelial tissues, and thought that this determination might with quinacrine mustards. The ability to observe a small be of value in some cases, such as for example with a body fluorescent body in human cell preparations containing a Y part which had been recovered and submitted for identifica- chromosome, after staining with quinacrine diHCl, was tion. They buried fetal tissue from a female in soil, and quickly shown to be a property of a number of types of could still determine sex chromatin for about 4 weeks. cells. In buccal epithelium, leucocytes and cultured skin Holzer and Marberger (1957) obtained somewhat similar fibroblasts, a large proportion of the cells from males results in their tests on post mortem tissues. Edwards and (usually 25-50070) exhibited the fluorescent body (or Cameron (1%4) said that excellent preparations could be F-body) after suitable preparation and quinacrine di-HCl obtained from the mucosal epithelium of the urinary blad- staining (Pearson et al., 1970; George, 1970). der, but that bodies even a few days old could give mis- leading results. 49.3.1 F-BO~~determinations in dried blood and Sanderson and Stewart (1%1) described a rapid, easy bloodstains method for sex chromatin determination in buccal epithelial The current of opinion in the literature appears to be that cells using aceto-orcein staining. Radarn (1965) discussed the F-body deteminations can provide a highly reliable method preparation of buccal epithelial cells for chromatin body of dried blood sexing if the procedures are carried out skill- determination. Renard (1971) described techniques for the fully and if a sufficiently large number of leucocytes can preparation of epithelial cells from salivary and vaginal be obtained for counting. A number of different techniques materials and for chromatin body determinations. Gener- have been reported for these detednations. There is some ally, 40% of the nuclei of cells of female origin recovered variation in the estimate of the maximum age of a blood- from various stains were chromatin positive. Ando (1973) stain that is still suitable for sexing by F-body counting. examined buccal epithelial cells from salivary stains and In addition, storage temperature and conditions as well as traces for both Barr bodies and F-bodies (see below). The the nature of the substratum appear to affect the dts chromatin bodies were found to be less stable over the obtained. course of time than the F-bodies. Phillips and Gaten (1971) reported successful results with Hair root sheath cells have been found to be suitable bloodstains up to 10 days old on solid (nonabsorbent) sub- material for chromatin body sex determinations in many strata. About 86% of the leuuxytes from male blood cases. Dixon and Torr (1956) found that hair root sheath smears showed F-bodies, while the figure was only about cells provided excellent mat& in post mortem examina- 0.5Qo with female samples. In 1972, Phillips and Webster tions. Schmid (1967) described a technique for sexing single gave an improved technique in which 2 mM Mga2was em- head hairs from living persons. It was best if the hairs were ployed as an extraction medium instead of McRvaine's buf- examined at once, and not stored, in his experiments. Cul- fer. In 1974, Phillips and Gitsham gave a procedure for bertson et a[. (1%9) also obtained reliable detennhtions bloodstains on absorbent substrata, and reported the results from hair root sheath cells. In 30 people, 15 males and 15 of a series of blind trial studies. Extraction techniques were females, Montanari et al. (1%7) found that Barr bodies oc- not found very satisfactory for F-body deteminations. curred in 29k 5% of the nuclei of these cells from females, Scnqi11g the stain so as to cause a "dust" of bloodstain to while the values were very much lower in preparations from fall upon the slide was the preferred technique. Reliable sex- males (6i2Qo). Bassett (1978) described procedures for sex ing was not possible in all the stains tested, but could be chromatin determination in hair root sheath cells, and done in many cases. At least 100 nuclei had to be scored, reported generally reliable and satisfactory results. He noted they said. It was important to recognize, too, that readers the importance of having experienced examinem score the had to become experienced in examining these preparations, nuclei, and the possiiity of subjective error. He said fur- and that there was an element of subjectivity in the deter- ther that preparations with sex chromatin frequency in the mination. Miiller et ul. (1971) tested stains up to 32 days old range of 10 to 40% of scorable nuclei should be considered and always found a higher F-body score in male bloods than inconclusive. Males and females showed sex chromatin in in female bloods. 5% and 57%, respectively, of their scorable nuclei. Schwinger (1972) could detect F-bodies in stains up to 30 Moore (1966c) discussed the determination of sex chro- days old. Blaiek and Brh(19724 were able to determine matin in connection with medicolegal problems. the sex of bloodstains by F-body technique in stains 32 days Sex of Origin old. They said that the decay in the percentage of F-body stressed. Cotton was found to be a problemetical sub- positive nuclei from male stains was not a linear function of stratum for bloodstains for this procedure, when compared stain age. Aragones and Egozcue (1973) described a method with glass or nylon. It was difficult to obtain good results which was applicable to stains up to 90 days old on absor- from bloodstains on wood or leather as well. In contrast to bent or nonabsorbent surfaces. Brinkmann and Jobst (1973) other reports, there was no signifcant reduction in the Y in- found that 12.5% to 50% of cells from male bloodstains up dex of bloodstains as a function of age up to 138 days. In to 4 weeks old contained F-bodies. It was possible to diag- blind trial studies on casework materials, 65% of male nose stains stored for 28 months at room temperature as be- bloodstains could be correctly identified, while no stains of ing of male origin. Ishizu (1973) reported fmding from 49% female origin were misidentified as being male. Siblind to 88% F-bodycontaining cells in blood smears from males trial studies, with similar results, were reported by Wiore and only 0070 to 4% in those from females. Similar results et d.(1979b). were obtained with blood smears from newborns, and with Thomsen (1979) gave a brief review of bloodstain sexing cord bloods. Blood smears up to 5 months old from males by F-body technique, and said that it provided a useful tech- showed more than 30010 F-body positive cells. Schaidt and nique in bloodstain analysis. Kriiger (1973) found that blood smears could be sexed by Curran (1976) reviewed a trial court case in New York in F-body technique for up to 19 days when kept at room which opposing panels of experts disagreed over whether a temperature, but for up to 63 weeks if kept in a refrigerator. bloodstain on a jacket could be reliably sexed by cytological Bloodstains on textiles could be sexed for 8 weeks. Troger techniques. The stain was about 22 months old when the and Liebhart (1974) found that male bloodstains stored for tests were conducted. The trial court judge m this case held up to 15 months still showed detectable F-bodies. Thomsen that the tests to determine the sexual origin of cells in the (1975) gave an improved procedure for F-body determina- stain (sex chromatin and F-body) failed to meet the re- tions, the improvement consisting of a more effective fdter quirements for reliability and general scientific Wtance, combination for the fluorescence microscope. and the testimony was not admitted. Kringsholm et al. (1976) noted that they had found no overlapping between the sexes in the percentage of cells 49.3.2 F-Body determinations in other lissues showing F-bodies. The highest female counts were always There are a number of reports on sex determination by lower than the lowest decounts. They had not, therefore, F-body technique in hair root cells (Sakai, 1972; Ishizu, observed any "false positive" males. Occasionally, a male 1972; Brinkmann and Jobst, 1973; Triiger and Liebhart, blood could give a deceptively low F-body count, however, 1974; Kringsholm et al., 1977). Troger and Liebhart (1974) and this situation would result in a "false negative". A low found that hair root sheath cells were suitable only for up to F-body count had to be interpreted with caution, therefore, about 5 days with stored hairs, but Ishizu (1972) reported and could mean (1) a stain of female origin; (2) an older successful results with hairs up to 150 days old, as did stain that was of male origin; or (3) a stain of male origin Kringsholm et (11. (1977) with hairs up to 27 weeks old. from a male with an abnormally low percentage of F-body Nagamori (1978) reported successful F-body sexing deter- positive cells. Kringholm et al. (19' found that most minations in hairs without roots up to 4 weeks old, provided bloodstains stored at room temperature for up to 21 weeks they were kept dry. could be sexed by F-body technique. They did observe a few Cells from various tissues and organs, examined past false negative results. Thomsen (1978) extended these aging mortem, have been found suitable for F-body sexing deter- studies to include stains kept at 5" and at 55". False negative mination. Kovacs et d.(1972) obtained reliable resuits in results appeared earlier in stains kept at either of these two looking at 100 cells from brain, heart, liver, spleen and bone temperatures than in those kept at room temperature, but marrow in 16 male and 10 female bodies. Blaiek and Brb there were no false positives. It appeared, therefore, that (1972b) obtained simibr results with various tissues, and bloodstains retain their Y-body counts better at room tem- they showed further (1972~)that another fluorescent dye perature than at So, while the opposite appeared to be the called 1-[(6-chloro-2-methoxy-9-acridinyl)amino]-3-(di- case in the studies of blood smears (Schaidt and Kriiger, methylamino)-2-propanol diHCl worked as well as quina- 1973). crine diHCl. F-body determination has also been descni Schwinger and Troger (1977) carried out blind trial in the cells of cartilage, periosteum and marrow (Berghaus studies using two different, independent scorers. In 21 stains et a/., 1973) and in human tooth pulp (Sommermater, 1975). up to 4 months old, the results of the two readers were very Troger et al. (1976) found that Y-bodieswere detectable similar. Nanko (1980) has noted that F-body count in male in -cant percentages in buccal epithelial cells derived blood smears fvred in methanol declines over the course of from cigarette butts smoked by males. 10Vo of the male cells time, but the reason for the decline was not clear. stdl showed Y-bodies when the cigarette butts were 43 days Wigmore et al. (1979a) reported on their technique for old, while the female cells from the same source had never F-body determination in stains and on a series of blind trial shown counts higher than 6Vo. Y-bodies can be detected in sexing experiments. The subjective nature of the technique sperm cells (Sakai, 1972). Troger and Eienger (1977) was emphasized, and the importance of carrying out case did F-body analysis in vaginal epithelium to try and use their sample and control examinations on a "blind" basis was absence as a means of identifying the cells as being of female Sourcebook in Forensic Serologv, Zrnrnunology, and Biochemistry origin. They said that stains had to be less than 4 weeks old, 1,500 serum samples. The T/E ratio varied from 60 to 0.02. however, to justify such a conclusion. Analysis showed that values greater than 3.51 could be Ishizu et al. (1973) described a technique for the consecu- assigned as male with a high degree of probability. Predict- tive determination of F-bodies and sex chromatin in the ability of sex based on the T/E was not 100% accurate, same sample material, which included saliva, hair roots and however; several male sera showed abnormally low ratios blood. The results of the two determinations, they said, and several female sera showed abnormally high values. Un- were more definitive than those from either separate deter- fortunately, nothing was known about the history or state mination alone. of health of the donors. The ratio was next determined in about 250 bloodstains from 6 to 8 months old. Sex of origin was known, but the hormone level measurements were car- 49.4 Sex Hormone Level ried out on a blind trial basis. The predictability of sex was Determinations about the same as that found for the serum samples. It was The use of pregnancy-associated hormones as bloodstain noted that in the bloodstains, the T/E ratio separating males markers for blood derived from pregnant women was dis- from females at the same probability level was 1.0, rather cussed in section 8.2.1. The idea of using male and female than the 3.51 seen with the serum samples. 98.9% of blood- sex hormones (primarily testosterone and 17~estradiol)as stains from males had T/E ratios greater than 2.0, while indicators of sex in dried bloodstains has been around for 92.7% of stains of female origin had values below 1.0. some years, but there appear to be very few published Recently, Szendrdnyi and Foldes (1980) reported that they systematic studies on the subject. Among the most sensitive could successfully sex experimental bloodstains up to 6 (and, hence, desirable for this purpose) methods of the assay weeks old by determining the testosterone concentration by of lower MW compounds such as steroids in complex mix- RIA. Stains prepared from 32 males and 24 females were tures is radioimrnunoassay (RIA). This technique was dis- studied, and showed little change in testosterone concentra- cussed briefly in section 1.3.4.3. tion between 48 hours and 6 weeks of aging. The protein Many RIA procedures have been described for steroid concentration of the samples was also determined, and it hormones, like testosterone, in serum (see, for example, was recommended that the testosterone level be referred to Sheldon and Coppenger, 1977; Joshi et ul., 1979). In theory, the protein concentration when analyzing stains. Thus, these procedures should be adaptable to the analysis of testosterone levels expressed as pg4O mg protein in stains bloodstains, and one could try to relate the sex of origin of correctly predicted the sex of origin in all cases in the limited bloodstains to the levels of one or a combination of the ma- number of stains analyzed. jor sex hormones, especially testosterone, 17&estradiol and Other hormones have been assayed in dried blood by progesterone. RIA, but not for medicolegal purposes. Illig and Rodriguez The most extensive studies to date are those of Shaler de Vera Roda (1976) reported on an RIA technique for (1975). He adapted serum RIA procedures for testosterone determining thyroid stimulating hormone (TSH) in dried (T) and estradiol (E) to determinations in bloodstains, and blood spots. The method was apparently designed in order then measured the "T/E" ratios in a selection of samples. to enable samples from newborns to be forwarded to a dis- The ratio was first determined on a "blind" basis in about tant testing laboratory in a convenient way. Non-Genetic Markers

SECTION 50. NONmGENETlC MARKERS AND BLOOD COMPONENT

50.1 Introduction certain specific non-genetic markers, such as HBsAg for ex- Over the years there has been occasional interest in ample, in a bloodstain would be valuable in an investiga- attempting to individualize blood using non-genetically tion, and could prove helpful if a comparison were possible determined components. Three general classes of compo- with the blood of the suspected depositer of the stain soon nents have been studied: (1) Components present in blood as afterwards. the result of environmental exposures or of the ingestion of specific agents. This class includes primarily antibodies and 50.2 Detection of Specific Components dm@. The antibodies may be directed against blood group of Blood antigens, against specific pathogenic agents to which the individual has been exposed, or to allergens. This class also includes the hepatitis antigens. (2) Components present in Among the first studies on the detection of spedic anti- blood as the result of normal metabolis processes. This class bodies in dried bloodstains are those of Kirk and his col- includes primarily metal ions and smaller MW metabolites. laborators in the early 1960's. In 1%3, Thornton and Kirk And (3) The serum proteins. Although se~umproteins are conducted studies on the detection of incomplete (Coombs genetically determined, and many are controlled by poly- reactive) anti-D in dried bloodstains up to 28 days old. They k morphic loci, the kinds of studies being discussed here found that the antibody was detectable, and that it persisted usually seek to fmd individuaI differences in the qualitative for at least several weeks in dried bloodstains was judged by or quantitative patterns of a large series of the proteins. its titer against Rh + type 0 cells. Leister and Kirk (1963) Two general approaches have been taken in these studies. reported on the detection of rheumatoid arthritis factor in In the first, a specific antigen, antibody, drug or component dried bloodstains using gamma-globulin treated latex par- is detected and/or quantitated. In the second, the pattern of ticles as a detection system. Dried, powdered blood kept in a series of components (usually serum proteins) in different vials was used for these experiments. They found that the individuals is examined. The individual components are not factor was detectable in dried blood up to 150 days old, and necessarily identified. Quantitative immunaelectrophoretic that blood with an initially high titer tended to retain its ac- techniques have been employed in a number of these kinds tivity in the dried state. The relationship of the factor being of studies. tested for in these studies to the Gm system (section 44) is A number of different factors are involved in determining not altogether clear. In 1%4, Leister et al. reported a series whether non-genetic markers and blood component profiles of experiments on the detection of syphilis antiiodies in are truly individualizing markers. In the case of spedlc dried blood. A microflocculation test (Kirk and Bemett, blood components, these include: (1) whether the presence 1939) was used in these studies. 520 dried blood samples up andlor quantity of the component is constant in an in- to 140 days old were tested, and the loss of activity was dividual over time, and over what length of time; (2) greatest in the first five days. Not every sample retained whether the presence and/or quantity of the component is activity equally well, but a higher percentage of initially detectable in dried bloodstains in a way that can be related strongly reacting samples gave positive results after 140 days to a whole blood sample; (3) the extent to which the pres- than of samples which had reacted more weakly at the ence or quantity of the component declines in dried blood- beginning. stains over time; and (4) what fraction of the population More recently, King, Whitehead and Werrett have done might be expected to exhibit the presence and/or quantity of studies on the detection of various antipamitic and the component detected. In the case of blood component allergen-associated antibodies in bloodstains. King (l974a) profiles, there are similar considerations: (1) whether inter- and King et al. (1975) descnied the detection of antibodies individual differences exceed intra-individual diffetences at to Mycobacterium tubemlosi~,Treponema pallidurn (non- a given time, and over the course of time; (2) whether pat- WIG "syphilis"), Virio cholera (cholera), Toxophma terns in bloodstains accurately reflect the pattern in whole gondii (toxopIasmosis), Trichomonas vagi'nafb (t&ho- blood from which the stain was derived; and (3) to what ex- moniasis), Candida albicmrs (candidiasis) and Toxocma tent the different patterns actually vary in a population. canis (toxocariasis). There were technical difficulties with Work on non-genetic markers is, for the most part, at an the Toxop~and Toxocma. Approximately 75 blood- early stage of development in this field. Additional studies stains were tested with the other five antigens using an in- will be needed to establish the individualizing value of many direct fluorescent antl'body technique. The majority of of the markers. There is not much doubt that the fmding of samples gave consistent positive and negative results in Sourcebook in Forensic Serology, Zmmunologv, and Biochemistry separate readings, but some gave ambiguous results. The be detected in a fragment of bloodstain equivalent to 10 pk' antibodies were stable in bloodstains for at least several of blood, and that this represented an amount approxi- months. In the sample of bloods studied, the discriminating mately 100 times larger than what was needed for detection. power for the five antigens was calculated to be 0.71, This work has since been extended to include detection of roughly comparable to the ABO blood group system. Anti- morphine (Mortimer et al., 1978a; Smith et al., 1980), bodies to Mycobacterium tuberculosk could be demon- digoxin (Mortimer et al., 1978b), and barbiturates (Smith, strated in semen if it was not more dilute than 1:lO. It was 1980) in bloodstains. noted that the antibody titer in a positive group of bloods King (1974b) examined the possibility of profiling a series behaves as a continuous variable, and that scoring samples of plasma constituents of clinical importance in the individ- as "positive" or "negative" was an oversimplification. It ualization of bloodstains. Data on the levels of Na+, K+ , was also noted, however, that the procedure lends itself to Cl-,Ca++,Mg++, protein, albumin, glucose, uric the profiling of a large number of different antigens to acid, urea nitrogen, cholesterol, SGOT (soluble glutarnic ox- which antibodies might be expected. In interpreting the aloacetic transarninase) and LDH (lactate dehydrogenase) results, caution must be exercised because there are some were sweyed in this study. Generally, the intra-individual cross reactions with related antibodies in a large series. differences were too close to the population variances to In 1976, Werrett et al. extended the antibody profiling make this "biochemical profile" of much individualizing studies to include allergen-associated antibodies. These anti- value. He noted, however, that better analytical methods bodies are of the IgE class (section 1.3.3.2), and require very might improve the prospects for the use of this kind of pro- sensitive detection techniques because the circulating levels filing in forensic serology. of IgE are very low compared to most other senun proteins 50.2.3 Hepatitis antigem (see in section 39). Commercially available RIA test kits for four allergens were used in the studies to detect the an- One of the best reviews of hepatitis serology may be tibodies in experimentally prepared bloodstains. It was found in the Nobel address by Blumberg (1977). The found that the antibodies could be detected in bloodstains discovery of a detectable antigen associated with hepatitis B on a variety of substrata, but that antibody activity declined grew out of studies on the Ag lipoprotein polymorphism more or less linearly to about 50% after 6 weeks. Using the (section 45.1). A precipitating antiserum was discovered in four allergen-associated antibodies, it was possible to the serum of a multiply transfused hemophiliac patient in discriminate six different blood stains on a blind trial basis, New York which reacted differently from the known anti- and the results correlated with the reported allergic condi- Ag serums. Tested against a panel of sera, the antibody tions of the donors. The results were considered to be reacted with the serum of an Australian aborigine, and the preliminary but promising, since the range of different au- antigen was given the name "Australia antigen", or "Au". tibodies which one assumes could be detected in bloodstains The antigen was found to be very stable in stored sera, and (and for which reagents are available) is quite large. was consistently positive or negative in the same person's King et al. (1976) reviewed the studies on antibody pro- serum collected in some cases over a number of successive filing of bloodstains. The promise and potential value of years. It was noticed that all Au(+) sera came from this approach were discussed. A large number of antirnicro- transfused individuals, and by the end of 1966 it had been bial and allergen-associated antibodies could potentially be realized that Au was associated with acute viral hepatitis. employed for this kind of profbg. An antimicrobial anti- Au could be t-ed by transfusion, and some of the body profde reflects an individual's environmental exposure people who received it in this way developed hepatitis. Some and vaccination history. The blood of full-grown adults can such people also developed anti-Au. This finding led to the be differentiated from that of younger children in this way mass screening of donor blood units for Au prior to any (King and Whitehead, 1975). In addition, menstrual blood transfusions, and rejection of the Au( +) bloods. The virus was found to have a much higher antibody level relative to responsible for hepatitis B is now known to contain the hemoglobin than the circulating blood of the same donor. Australia antigen on its surface, and it is now termed Allergen antibody profiles could give information about the hepatitis B surface antigen (HBsAg). The surface antigen geographical origins or history of an individual. Informa- can be removed to reveal a core antigen (HBcAg), and an- tion of potential value to an investigation might thus be pro- tibodies to both of these can be found in human sera. It vided from the examination of a bloodstain at a crime scene. turns out that HBsAg exhibits antigenic heterogeneity and a number of surface antigenic determinants (designated by 50.2.2 Drugs and plasma metabolites various lower case letters) are known to be associated with The same kind of RL4 techniques used to detect drugs in the virus. Dmochowski (1976) has given a thorough and serum can be adapted for measuring drug levels in blood- readable review of the entire subject of hepatitis, including stains. As noted in section 49.4, Shaler and his collaborators information about the surface and core antigens and their adapted RIA techniques for steroid hormone determination serology. HBsAg may be detected by a variety of immuno- to bloodstain analysis. They have extended these studies to logical techniques, immunodiffusion, crossed electropho- include the detection of a number of drugs as well. Shaler resis and complement fixation among them. Hemagglutina- (1975) said that dilantin, a drug used to treat epilepsy, could tion techniques, in which red cells or latex particles are Non-Genetic Markers coated with anti-HBsAg and used as the test system, have dividual differences could sometimes be seen. The technique also been described (see, for example, Archer, 1977). was found to be especially valuable for distinguishing Hp HBsAg can be detected in dried blood. Hoste et al. (1977) phenotype (see in section a),and the possibility of using the described a series of studies on a number of experimentally profiling technique to distinguish phenotypes in a number of prepared bloodstains using a "sandwich" RIA procedure different immunologically detectable polymorphic serum for detection. HBsAg could be detected in stains up to 6 protein systems simultaneously was discussed. Phillips months old, though more material was needed for testing (1974) looked at a somewhat different approach to serum with the older stains. The presence of HBsAg is a fairly protein profiling. The quantity of various serum proteins in stable characteristic, and can persist for years in people who the sera of different individuals is known to vary. It was have it. The incidence of the antigen is fairly low in Western thought that these differences might be exploited as individ- Europe and in this country, but much higher in some popu- ualizing markers using specific antisera and immunochem- lations. HBsAg is sometimes found in so-called caniers, ical quantitation. Antigen-antibody crossed electrophoresis who appear to be healthy. Hoste et al. (1977) suggested that was considered too variable for this purpose, and a nephelo- HBsAg could provide a useful nongenetic marker in blood- metric quantitation procedure using an autoanalyzer was ex- stains. Frazadegan et al. (1978) have described detection of plored. The pdure was found to be unsatisfactory for HBsAg in blood dried on fdter paper, although their interest bloodstain extracts and even to a great extent for older was not in bloodstain testing per se. If HBsAg subtyping hemolyzed blood samples because of interference by hemo- should become possible in bloodstains, an even greater globin. If suitable techniques could be devised for ridding -on . .. among individuals will probably be posslible. the samples of hemoglobin, Phillips said, the technique Richer et al. (1977) have shown, for example, that the fre- might be very useful, and he recommended further study. quency of the "e" antigen aswciated with HBsAg varies Grunbaum and Hjalmarsson (1976) bedout preliminary among different ethnic groups m the Montreal population. experiments on aqueous extracts of bloodstains using isotachophoresis (section 2.5.2). There were differences in 50.3 Serum Protein Profiling the patterns from four people, and these patterns did not A number of workers have looked at serum protein pro- change if the extracts were retested 24 hours later. files in different individuals using a variety of different tech- In 1976, Sweet and Elvins (1976a and 1976b) applied niques of separation and detection. The number of com- antigen-antibody crossed electrophoresis (which they call ponents resolved is dependent on the nature and resolving "crossed electroimmunodiffusion" or CEID-see in section capability of the technique selected. In addition, some of the 2.4.3.2) to the problem of distinguishing individual serum studies were canied out before the extent of serum protein protein pattern differences. By selecting and scoring those polymorphisms was fully recognized (Unit VII). In any case, peaks which showed completely different he@t ranges over if the proteins being profled are not identified, one can not several detembations among individuals being compared, be sure that one is not seeing an example of a known poly- it was possible to distinguish among ten persons in the morphic serum protein when a difference is detected. study. There were five males and fwe females in the test In 1953, Bernfeld et al. found differences in the serum group, and differences between males and females were protein profiles of a number of different individuals using noted in the patterns as well. There were a few serum pro- moving boundary electrophoresis in a Tilius cell (section teins which showed &nifkaut differences in amount be- 2.3.1). Brown and Kirk (1957) conducted similar experi- tween the sexes, and it was suggested that analysis of these ments on dried blood using paper electrophoresis. They might provide a probable sex of origin estimate in blood- could distinguish some but not all the stains in a series pre- stains. Whitehead (1973, in a letter prompted by the work pared from ten different people. Laudel et al. (1%3) could of Sweet and Elvins (1976a), pointed out some of the prob- detect differences m the immunoelectrophoretic patterns in lems awxiated with serum protein profiling as a means of the sera of a small number of people after carrying out elec- individualizing stains. He thought that considerably more trophoresis on cellulose acetate membranes and detecting work would be needed before profiling techniques would be the proteins with a horse immune anti-human serum. suitable for use m actual casework, and that genetic markers Whitehead (1969) and Whitehead et al. (1970) explored would continue to provide the principal means of individu- the possibility of using antigen-antibody crossed electropho- alization available for some time. Sweet (1977) responded to resis (burell technique-see in section 2.4.3.2) as a means Whitehead's comments in the same issue of Science, and did of finding individual differences in the serum protein pat- not, for the most part, disagree with them. terns. Studies with bloodstain ex&racts indicated that in-

References for Unit VIII

REFERENCES

Ando, K. 1973. Studies on the sex identification with human saliva and Dmochowski, L. 1976. Vitype A and type B hepatitis. Am. J. Clin. saliva stains by the analysis of exfoliated oral epithelial cells. Jpn. J. Leg. Pathol. 65 (suppl. to No. 5): 741-786 Med. 27: 356-367 0' Edwards, J.H. and A.H. Cameron. 1964. Postmortem nuclear sexing. hagones, J. and J. Egozcue. 1973. A method for sexiug My-old blood- Lancet 1: 328 J, stains in absorbent and nonabsorbent material Acta Genet. Med. Fanadegan, H., K.H. Noon and F. Ala. 1978. Detection of hepatitis-B Gemdol. U: 113-116 surface antigen in blood and blood products dried on fdter paper.

Archer, A.C. 1977. An improved haemagglutiuation technique for the Lancet 1: 362-363 detection of hepatitis B, antigen. Med. Lab. Sci. 34: 345-350 George, K.P. 1970. Cytochernical differentiation along human chromo- Barr, M.L. 1957. Cytologic tests of chromosomal sex. Pmgr. Gynecol. 3: somes. Nature 226.8&8 1 131-141 Grob, H.S. 1970. Cytologic sexing, in: T.D. Steward (ed.), Personal Iden- Barr, M.L. 1960. Sexual dimorphism in interphase nuclei. Am. J. Hum. tif~ationin Mms DMets. Smithsonian Institution. Washington, D.C. Genet. U: 118-127 Grob. H.S. and H.S. Kuppexman. 1961. Experiences with technics of Barr, M.L. and E.G. Be-. 1949. A morphologic distinction between chromatin sex dacrmination. Am. Clin. Pathol. 36: 132-138 neurones of the male and female, and the behaviour of the nucleolar Grunbaum, B.W. and S.G. Hjalmarsson. 1976. Isotachophorcsis of satellite during accelerated nucleoprotein synthesis. Nature 163: 676-677 bloodstains. J. Forensic Sci. Soc. 16: 325-331

Bassat, W.A.G. 1978. Sex determination by sex chromatin identification Holzer, F.J. and E. Marberger. 1957. Zur mikroskopischen Gachle- in the hair root sheath. J. Gmad. Soc. Forensic Sci. 11: 221-228 chtsbestimmungan Leichenteilen. Dtsch. Z. Gesamte Gerichtl. Med. 46: Berghaus, G., U. Reifenberg and G. Dotzauer. 1973. Geschlechtsbcstim- 242-249 mung an Skeletteilcn. Nachweis der sogcnnanten Y-Fluorcs- Hoste, B., J. Brocteur. B. Donea and A. Andre. 1977. Identification of cenzkorperchen. Z. Rechtmted. 72 255-268 blood stains by non-genetic characteristics: the HBs antigen of viral Bernfeld, P., V.M. Donahue and F. Homburgcr. 1953. Characteristic in- hepatitis B. Forensic Sci. 10: 229-234 dividual electrophoretic patterns in humans. Proc. Soc. Q. Biol. Med. Illig, R. and C. Radriguez de Vera Roda. 1976. Radioiunologischer 83: 429-434 Nachweis von TSH in getrockneten Bluutropfen: mbgkhe Screening- Bkiek, J. and J. Bra. 19721. Detemination of cellular sex in old- blood Methode zur Entdcckung da Hyperthyreor bei Neugebornen. spots and in hair follicle cells with the a$ of anitline fluorochromes. Schweu. Med. Wochemchr. 106. 16761681 Soudni Lek. 17: 17-22 [incorporated in Cgk. Pml. 8 (1972)l Ishii, H. 1972. Sex identification of hairs by Yehromosome.J, Jpn. J. Leg. Bkiek, J. and J. Br;lza. 1972b. Dcmonstratjon of Y-chromosome in Med. 26: 40346 necroptic material by fluorescent methods. Cesk. Paol. 8: 45-49 Ishizu, H. 1973. Studies on sex identification of human blood and blood- Bkiek, J. and J. Brdza. 1972~.The method of sac determination in stains. Jpn. J. Leg. Med. 27: 168-181 necropsies with the aid of l-[(&chloro-2-metho~y-9-acridinyl) Ishizu, H.. K. Ando, M. Nobuhara and Y. Mikami. 1973. Sex identifica- amino]-3-(dimeth~0~2-propanoldihydrochloride. Cesk. Patol. 8: tion with a minimal sample by combining Y-chromatin identif~cation 108-1 10 and the X-chromatin detection method. Jpn. Leg. Med. n: Bhunberg, B.S. 1977. Australia antigen and the biology of hepatitis B. 287-294 1' Science 197: 17-25 Joshi, U.M., H.P. Shah and S.P. Sudharna. 1979. A sensitive and specific Brinkmann, B. and U. Jobst. 1973. Bestimmung des Kemgcschlcchts an auzymcimmunoassay for serum testosterone. Steroids 34: 354.6 biologischcn Supren. Forcnsischer Bcwciswcrt und Anwandungsbmich. King, L.A. 1974a. The identification of anti-parasitic antibodies in blood- 2.Rechtmd. 73: 14 stains using an indirect fluorescent antibody technique. J. Fore& Sci. Brown, C.L. and P.L. Kirk. 1957. Individuality of blood. Elec- SOC.14: 117-121 trochromatophoretic patterns of dry blood. J. Forensic Med. 4: 58-64 King, L.A. 1974b. The value of biochemical profling for the discrimina- Caroff, J. and J. Breton. 1966. Le sexc chromatinien-inttrft mtd- tion of bloodstains. I. Fo&c Sci. Soc. 14: 323-327 ico-ltgale revue gCnCrale. Ann. Med. Leg. 44: 98-105 King, L.A., D.J. Werrett and P.H. Whitehead. 1975. Antibody profhg Caspmson, T., S. Farber, G.E. Foley. J. Kudynowski, E.J. Modest, E. of bloodstains. Forensic Sci. 5: 144 Simonsson, U. Wagh and L. Zech. 1%8. Chemical differentiation along King, L.A., D.J. Wemtt and P.H. Whitehead. 1976. Antibody profwg metaphase chromosomes. Eap. Cell Res. 49: 219-222 of bloodstains. Forensic Sci. 8: 151-154 Culbertson, J.C., N.A. Brrslau, M.K. Moore and E. Eneel. 1969. Sex King, L.A. and P.H. Whitehead. 1975. The differentiation of an adult's chromatin determination from hair. J. Am. Med. Asoc. 207: -561 bloodstain from that of a child using an indirect fluorescent antibody Curran, W.J. 1976. Forensic medical science: the continued problems of technique. Forensic Sci. 6: 197-203 judges and juries. New Engl. J. Med. r)4. 1042-1043 Kirk. P.L. and C. Bennett. 1939. A rapid technique for syphilis testing Davidson, W.M. 1963. Leucocytes in blood stains, in: Fore& Im- with fmger blood. J. Lab. Clin. Med. 25: 86-88 munologv, Medicine, Pathology and Toaicologv, Rept. of 3rd Intl. Kov;lcs, M., L. H&yi and M. Sellyei. 1972. Y-body in cell nuclei of M&g, Int. Congr. Ser. No. 80, Excerpta Medica Foundation, parenchymatous organs. 2. Rechtsmed. 71: 104-107 Amsterdam & New York Kringsholm, B.. J.L. Thomsen and K. Henniagsen. 1977. Fluorescent Y- Davidson, W.M. and D.R. Smith. 1954. A morphological sex difference in chromosomes in hairs and blood stains. Forensic Sci. 9: 117-126 the polymorphonuckar neutrophil leucocytes. Br. Med. J. 2: 6-7 Kringsholm, B., J.L. Thomsen, K.T. Henningsen and E. Niebuhr. 1976. De Bernardi. A. 1959. Diawosi istomorfolomca di scsso su sannue- imbrat- Fluorescent Y-bodies in interphase nuclie: medicolegal aspects of false tante caklli. Minem Medicoleg. 7!k 7679 negative males. Forensic Sci. 7: 171-172 Dixon, A.D. and J.B.D. Ton. 1956. Sex chromatin as an acid to the iden- Laudel, A.F.. B.W. Grunbaum and P.L. Kirk. 1963. Individuality of tiflation of sex in forensic medicine. Nafure 178: 797 human whole dry blood by immunoelectrophorcsis on cellulose acetate. Dixon, A.D. and J.B.D. Tom. 1957. Sex determination of human tissues J. Forensic Med. 10: 57-64 from cell morphology. J. Forensic Med. 4: 11-17 Leiier, C.I. and P.L. Kirk. 1963. Rheumatoid arthritis factor. A sensitive Sourcebook in Forensic Serology, Immunology, and Biochemicstry

method for its detection for individualizing dry blood samples. J. Foren- Sheldon, P.E. and C.J. Coppenger. 1977. A rapid radioimrnunoassay for sic Med. 10: 157-161 serum testosterone. Steroids JO: 149-157 Lcister, C.I., J.I. Thomton and P.L. Kirk. 1964. Individualitaton of dry Smith, F.P. 1980. Detection of drugs in dried bloodstains. IV. Bar- blood samples. Demonstration of syphilis antibody. I. Forensic Med. biturates. Paper delivered at 32nd Ann. Meeting, Am. Acad. Forensic 11: 31-35 Sci., New Orleans, LA Montanari, G.D., B. Viterbo and G.R. Montanari. 1967. Sex detcrrnina- Smith, F.P., R.C. Shaler, C.E. Monimer and L.E. Emchetto. 1980. tion of human hair. Med. Sci. Lmu '1: 208-210 Detection of drugs in bloodstaim. 11. Morphine. J. Forensic Sci. 25: Moore. K.L. 1%6a. Introduction, in: K.L. Moore (ed.), The Ser 369-373 Chtvmotin, 14, W.B. Saundm, Philadelphia & London Sommermater, J. 1975. Contribution to nuclear sex determination by iden- Moore, K.L. The discovery of the sex chromatin, in: K.L. Moore tif~cationof the Y chromosome in human pulpal fibrocytes. Forensic (4.) TheSer Chromatin, 7-15. W.B. Saunden, Philadelphia &London Sci. 5: 167 Moore, K.L. 1966~.Sex chromatin and medicolegat problems, in: K.L. Sweet, G.H. 1977. Crossed electroimmunodiffusion and bloodstain in- Moore (ed.) The Ser Chtvmotin, 453-464. W.B. Saunden. Philadelphia vestigations. Science 198: 427 & London Sweet, G.H. and J.W. Elvins. 1976a. Human bloodstains: individualiza- Mortimer, C.E.. F.P. Smith. L. Errichetto and R.C. Shaler. 1978a. Detec- tion by crossed ekctroimmunodiffusion. Science 192: 1012-1014 tion of drugs in a bloodstain. 11. Morphine. Paper delivered at 30th Sweet, G.H. and J.W. Elvins. 1978. Studies by crossed electroimmuno- Ann. Meeting, Am. Acad. Forensic Sd.. St. Louis, MO diffusion on the individuality and sexual origin of bloodstains. J. Foren- Mortimer, C.E., F.P. Smith and R.C. Shaler. 1978b. Detection of drugs in sic Sci. 21: 498-509 a bloodstain. 111. Digoxin. Paper delivered at 30th Ann. Meeting, Am. Szendrhyi. J. and V. Folda. 1980. Mcssung des Tmosterongehaltes in Acad. Forensic Sci., St. Louis, MO Blutflccken nu kriminalistischen Geschlechtsbestirnmung. 2. Rechts- Miiller, H., E.M. Biihler, M.G. Voegelin and G.R. Stalder. 1971. Eie med. W. 263-267 neue Methode der Geschlechtsbcstimmung in Leukozyten aus Thornsen, J.L. 1975. Eioe verbcsscrte Methode zurn Nachweis des Y- ehgctrockneten Blutflccken. Schweb. Med. Wochenschr. 101 (32): Chromsoms in Blutspuren. 2. Rechtsmed. 76: 81-86 1171-1174 Thomsen, J.L. 1978. Influence of the temperature on the detection of Nagamori, H. 1978. Sex determination from plucked human hairs without fluorescent Y-bodies in blood stains. Forensic Sci. 11: 123-126 epithelial root sheath. Formsic Sci. Int. U: 167-173 Thomsen, J.L. 1979. Y-chromosome detection in bloodstains. Forensic Nanko, S. 1980. Decrease of Y chromatin frequency with time after fm- Sci. Int. 14: 104 tion of blood smear. Forensic Sci. Int. 15. 1-2 Thomton. J.I. and P.L. Kirk. 1963. Individualizing dry blood samples by Ohno, S. 1%. Single-X derivation of sex chromatin, in: K.L. Moore (4.) demonstration of the Rhesus antibody. J. Forensic Med. 10: 123-127 The Ser Chtvmotin, 113-128, W.B. Saunders, Philadelphia & London Tropr, H.D. and W. Eienger. 1977. Eie Methode zum Nachweis Purrson, P.L., M. Bobrow and C.G. Vosa. 1970. Technique for identify- von Scheidenepithclzekn. Beitr. Gerichtl. Med. 36: 109-111 ing Y chromosomes in human interphase nuclei. Noture 226: 78-80 Trager, H.D. and E. Liebhart. 1974. Zur zcitlichen Nachweisgrenze des Y- Phillips. A.P. 1974. The potential of nephelomctric immunoprecipitin Chromosoms an Blutspwen and Haaren. Beitr. Gerichtl. Med. 32: quantitation in forensic science. I. Formsic Sci. Soc. 14: 135-136 159-162 Phillips, A.P. and E. Gaten. 1971. Y chromosome fluorescence in blood- Triiger, H.D., E. Liebhart and W. Einmenger. 1976. Wer hat die stains. Loncet 2: 371-372 Zigarette geraucht? Bestimmung des m2ndchen Kerngeschlechts an Phillips, A.P. and C. Gitsham. 1974. The identification of male blood- Mundschleimhautzellen. Beitr. Gerichthd Med. 34. 207-209 stains by Y chromosome fluomccnce J. Forensic Sci. Soc. 14: 47-54 Wmett, D.J., L.A. King and P.H. Whitehead. 1976. The detection of Phillips, A.P. and D.F. Webster. 1972. Improved Y-chromosome fluore allergen-asochted antibodies in bloodstains. I. Forensic Sci. Soc. 16: ma in the presence of magnesium ions. J. Forensic Sci. Soc. 12: 121-126 361-362 Whitehead, P.H. 1969. Antigen-antibody crossed electrophoresis. A new Radam. G. 1965. Zum Nachweis von Speichelspuren. Beitr. Gerichtl. Med. means of characterizing blood? Proc. 5th Int. Meaing Forensic Sci., 23: 226-235 Toronto, in: Int. Microf~lrnJ. Leg. Med. q4): card 10 Renard, S. 1971. Determination of sex of exfoliated epithelial cells and its Whitehead. P.H. 1977. Crossed electroimmunodiffusion and bloodstain si@iuu1ce in forensic science. J. Forensic Sci. Soc. 11: 15-20 investigations. Science 198: 427 Richer, G.. D. Phaneuf. F. Boisvert, R. Gutvin and A. Vit. 1977. Dif- Wbiehead, P.H., S.S. Kind, P.A. Morris, M. Davia and R. Clecvley. ference in the distniution of e antigen among different ethnic groups in 1970. The examination of bloodstains by Laurell electrophoresis (an- a population of blood donors. J. CMud. Med. Asroc. 116: 757-759 tigcn/antiiy crossed electrophoresis). J. Forensic Sci. Soc. 10: 83-90 Sakai, K. 1972. Inv~onof fluorrsccnt body in the human cells. Jpn. Wire, R.. P. Dolton and P.H. Whitehead. 1979b. The sexing of I. Utvl. 63: 658-671 0' bloodstains using Y-chromosome fluorescence. Report of a "blind trial" Sanderson, A.R. aad J.S.S. Sterward. 1961. Nuclear sexing with accto- involving 195 casework bloodsrains. Tech. Note No. 113, Home Off. orcein. Br. Med. 1. 2: 1065-1067 Central Res. Establ., Aldemon. Reading, Bcrks., England Schaidt, G. and R. Kriiger. 1973. Die Gschlechtsbestimmung an Blut- Wire, R., D.J. Wemtt. L.A. Kiug, P.H. Whitehead and A. Ema. prokn und Blutspuren mibdicher Pmonen. Arch. Kriminol. 151: 1979a. The detection of Y chromosomes in bloodstains--a reevaluation. 4148 J. Forensic Sci. 24: 36375 Schiig. H. 1960. Zur Frage der Dauer der Nachwdsbarkeit da Ge- Zech, L. 1969. Investigation of mctaphase chromosomes with DNA-bind- schlechts an eingcsandtem Blut, insbesondere an Blut von Leichen, Dis- ing fluorochromes. Eap. Cell Res. 58: 463 serkation. Bedin (Humboldt), cited by 0.Prokop, Forensische Medizin, Berlin, 1% Schmid, W. 1967. Scx chromatin in hair roots. Cytogenetics6: 342-349 Schwinger, E. 1972. GdechtsbCSthOl~g Blutspum. 2. Rmhts- med. 10: 157-162 Schwiuger. E. and H.D. Troga. 1977. Wie sicher ist die Geschlechts- Bibliographic Notes to References for Unit VIU bcstimmung in Blutspuren? Beitr. Genchtl. Med. 35: 267-271 Shalcr, R.C. 1975. The individualization of forensically important Joponew Journol of LegoIMedicine (Jpn. J. Leg. Med.) has Japanese physiological fluids. FdReport, Grant #75-NI-99-0011.Nat. Inst. Law title Nippon Hoigoku %hi Enforcement Crim. Just., Law Enforcement Asshnce Administration, 5' Joponese Journalof Urology (Jpn. J. Utvl)has Japanese title Nippon U.S. Dcpt. of Justice, Washington. D.C. Hinyokika Gokkoi ZPrshi Index

ABH Sumtor system (see Secretor system) bloodstain grouping by mixed agglutination: development of tech- ABO aptem: nique, 310-312 A subgroups: A, and A,, 266-268 smsitivty, 3 12 anti-A, in A, and A2B sera, 267 bloodstain grouping of A antigen by inhibition of hemolysis, 305 detamioation in bloodstains, 312-313 bloodstain grouping using formalin treated red cells, 312 quantitation of reactivity, 269-270 body fluid and tissue grouping: acquiredB in forensic serology, 316-317 subgroup A,. 268 development. 313 subgroupsweaker than A,, 268-269 of eanvax. 316 ABH antigeusas widely distributed cell surface structures, 279-280 by dution technique, 314-315 acquired B, 27S280 by fluorescent-labelted antiiytechnique, 315 animal blood ABH reapton, 280 of gdsViCmucosa, 316 anti-type XIV pncumococcal serum reactive antigen, 288 of hair, 320-321 antibodkc antCH and "anti-O", 272-273 heat stabilityof ABH antigensin tcah, 320 anti-A and anti-B, 270-272 by inhiiin technique. 313-314 anti-A from snails. 283 of inner ear fluid, 316 anti-H lectins, 273 by mixed agglutination technique. 314 in body fluids otha than serum, 273-274 of nails, 321-322 cross-reacting anti-A- and anti-Blike antibodies in 0 sera,276-278 problans and false reactions, 317-319 genetic control of. 271 by ~u hhib'ion technique,~ 315 n origin of "naturally occurring" antibodies, 270-271 of skin fragments and organ cells, 316 persistence in dried serum, 300 of sweat. 316 antigenicsfruaures, 289 of teeth. 322 application to disputed parentage testing. 296-297 of tooth tissue, 316 B subgroups or bts,270,271 of urine and urine stains, 316 bacterial ABH receptors, 279-280 (See&Secretor system) biochemicalgenetics, 290-293 Bombay phenotype, 275-276 blood group ma,287-288, (table) 291 carbohydrate chain-peptide linkage, 289 bloodsfain grouping: early studies, 297 charanerizationof ABH antigens, 287 Landsteiner-Richtertest, 297 cic-AB, 276 bloodstain grouping by elution: of ammoniacal extracts of bloodstains, destruction of blood group d~dtiesby enzymatic hydrolysis, 287-288 307 dismvay, 261 antiserum evaluation and selection, 3-310 dinniufion of phenotypes in U.S. populations, 324-327 detectabity of antigens in aging samples, 310 dried strum, 300 development of technique. 305-3M early serologkal studies. 261-264 enhancement of blood group antibody reactivity, 309-310 mzymatic dcgdation of ABH substances. 287-288 interfmnce by adventitious substances and false results, 308 factorsaffectingtyping success in stainsand post-mortem samples, 320 sensitivity, 307-308 group C, 277-278.279 technical development and modifications, 306-307 hair grouping, 32&321 thread technique (Howard-Martin), 307 inhaitauce: cis-AB, 276 bloodstain grouping by ferritin-labelled antibodies. 312 multipleaflclicmode, 264-265.(table) 267 bloodstain grouping by fluorescent-labelled antibodies, 312 nomenclature according to Wier, 277 bloodstain grouping by inhibition: "all or none" technique, 301 nomenclature standardization. 261-262 anti-H reagents and lectins, 304-305 numbm of antigenic sites on red cells, 270 detectsbiity of antigensin aging samples, 300-301 pamBombay phenotypes. 276 development of technique, ?OW01 physiwchenlical studies of antigen-antibody reaction, 274-275 gamma-globulin deviation, 305 plant ABH receptors, 279 Himfeld and Amzd-Kind technique, 302-303 plant, animaland baasABH-like antigens: structural studies, 289 Hobtechnique, 301,303 puriftcation of ABH antigens, 286-289 interpretation of results, 302 quautitatiw studies of a&utination. 269-270 nonspecific absorption and contamination, 302-304 quautitative studies of antigen-antlbody reaction, 274-275 sensitivity, 301 relationship of ABO, Secmor and Lmis genotypes to red cell and body use of 0(anti-A.B) sera, 300,301-302 fluid an?igcns, 295 bloodstain grouping by isoagglutinin detection (Lattes): development rrhtiooship to Ii antigens,369 of methods. 298-299 relationship to Secretor and Lewis, 2W-293 early work, 298 secrrtian of ABH in body fluids (seeSecretor system) irregularanti-A, and non-ABO antibodies, 299 solubleABH substances(see Secretor system) sensitivity and enhancement, 299 subgroups: quantitative designations, 275 aabiity and persistence of isoagglutinins, 299-300 (See aLFo A sub~oups,B subgroups) Sourcebook in Forensic Serology, Immunology, and Biochemistry

terminal carbohydrates involved in antigenic determinant, 287 separation of SAP and VAP, 166 type 1 and type 2 carbohydrate chain endings, 289 senun level elevated in prostatic carcinoma patients. 155 type 1 chain conversions to group substances, 293 substrates for, 162-163 type 2 chain conversions to group substances, 294 units of activity, 163, (table) 164 weak receptor types and unusual antibodies in serum of casework vaginal, 163-166 samples, 320 ACP (see Acid phosphatase) Wiener's conception of, 277-278.279 Acqllirrd B (see ABO system, acquirril B; ABO system, body fluid and Absorption-ehliin: tissue grouping) for ABO grouping of bloodstains (see ABO system, bloodstain group el Advator, 595 ing by elution) Activily alaio. 423 of "cold" agglutinins in bloodstains, 389 (See u&v detection, under specific ixmuyme name headings) methods of eluting blood moup antibodies from red cells, 310 AeylrSdhn acyl-hydrohr (seeCholinesterases) principles involved in evaliation of antisera for, 308-310 ADA (seeAdenosine deaminase) systematic studies on elution of blood group antibodies from blood- Addiwnt (see Complement) stains, 309-310 Adcuemmbohydrob (seeAdenosine deaminase) (See ahRh system, MNSs system, Kell system. Duffy system, Kidd AdcaorBcd- system) alleles and red cell locus, 457 Absorption-inhibin: application to disputed parentage, 461 for ABO grouping of bloodstains (see ABO system, bloodstain group biochemical properties of red cell ixmuymes, 460-461 ing by inhibition) biochemical properties of tissue enzymes, 458459 (See also Inhibition tests, Rh system, MNSs system, Kell system, Duffy daectabilityofisoenzymesinagingstains,461 system) detection of isomzyma in typing media and assay methods, 460, 461 Acetylcholine hydrolase (seeCholinesterascs) development of red all isomzymes in fetal blood, 461 Acetylcboliaeste- (seeCholinesterases) distributionsof phenotypes in U.S. populations, 462 N-scetylgrl.ctosamine: elarrophorctic patterns of phenotypes, 458 involvement in ABH antigen structure, 287 groupingin bloodstaias. 461 structure of, 287 grouping in body fluids and tissues. 462 Ache (seeCholinesterases) human tissue, 458-459 ACby (seeAntichymotrypsin) Linkage and chromosomal IocalizMion of ADA, 458 a,-Acid &coprotein: polymorphism of red cell enzyme, 457 in seminal plasma, 180,618 recognition of. 457 serum genetic variants, 617418 relationship of deficiency to combined immunodeficiency disease, Acid phospha~ 457-458,45940 ACP locus designations, 447-448 shtalleles. 457-458 assay techniques, 162-163 sulfhydryl effccts on red cell isoenymes, 460 biochemical properties of different isoenymcs, 44849 typing simultaneously with other isocmymes, 461 dye-coupling assay with naphthyl phosphates, 157 Admybtekhmc erythrocyte: application to disputed parentage, 450 additionalAK loci, 440 biochemical properties of isoenzymes. 449-450 appticafion to disputed parentage. 441 detectabity of isoenzyma in aging blood and bloodstah, 451452. biochemical properties, 440-441 453 dastabiyof isoenywsin aging blood and bloodstains, 442 distributions of phenotypes in U.S.populations, 454-455 detcaion reactions in typing media. 439 effect of buffers on electrophoretic patterns. 446 distributionsof phenotypes in U.S. populations, 443 electrophoretic patterns of phenotypes, 447 electrophoreticpatterns of phenotypes, 440 goodnessof-fit tests on population data, 446 grouping in bloodstains, 441 grouping in bloodstains, 450-452 grouping in body fluids and tissues, 442 inheritance and alleles, 445446 polymorphimo, 439-440 phenotyping methods, 452453 recognition of. 439 phosphotransferase activity of, 451 siknt alleks. 440 polymorphii, 445 species determination by isoenryme separation, 244 recognition of, 445 typing simultaneousiy with other kocnzyme systems,441442 silent alleles, 445446 Adla's td (see Catalytic tests, benzidine) species determination by isoenyme separation, 244 Ag sfstem (see Serumlipoproteins) typing simultaneously with other isornyme systems. 452-453 Am linkage relationships of ACP loci, 449 gcls for ekctrophoresis (see Electrophoresis) multiple loci determining, 44-448 studies on structure, 60 persistence of seminal ensyme in vagina (see Identification of semen, Ageof bbodsdm, determination of, 131-133 acid phosphatase) mthutkn: properties of enzyme in human tissues, 446-447 effect of red cell concentration, 299 prostatic (seeldentification of semen, acid phosphatase) enh-ent by high moleculai weight colloids. -51,352 red cell (seeAcid phosphatase, erythrocyte) enh~ncementby proteolyticenzymes, 51,353 salivarypolymorphism (seeSaliva, polymorphic isoenzymes) of humpn cells by animal sera, 236 seminal: apparent polymorphism, 167-168 w~haaisn.49-51 imrnunogenicity, 168 passive: covalmt linkage of soluble antigens to blood goup antibody, molecular heterogeneity, 167-168 231 purification and properties, 166-168 tanned red cells, 229-231 tartrate inhibition of. 165-166 of sensitized particla (see Smsitizcdparticles) (SeeakoIdentifmtion of semen, acid phosphatasc) tempaature dependence, 52 Index

Ambinin bmdimg (see Absorption-inhibition) Amykpectin (see Starch) Agglotinins: Amylase (see Starch) antibodies as, 49 A- immune response (see Antibodies, formation) avidity, 53 A, (see ABO system, para-Bombay phenotypes) as case of mediate hypersensitivity, 56 AK (see Adenylate kinase) description of procedure, 234-235 Ahhemhotnosfeme (seeGlutamic-pyruvic tansaminasc) identification of semen by (see Identification of semen, immunological) Albumin: sp#ies determination by, 234-235 characteristics, structure, genetic variants, 620-621 (see uhHypermsitivity) detectabiity in aging bloodstains (see Age of bloodstains, detennina- AO~CIIU.M~~serum: tion of) detection of incomplete antibodies with, 50,352-353 luaptonurir, 20 development of, 352 Aldoses (see Carbohydrates) inhibition of, for human species determination,227-229,230 Akxioe (see Complement) mixed antiglobulin technique for species determination, 231-232 A-Uke .n(igca (seeABO system,bacterial and pknt ABH receptors) and tanned red cells for species determination, 231 AUuliac phospbaIsiw an^^ hemoglobin (see Identification of blood, anti-human hemo- bone, liver and intestinal enzy'mes, 509 globim) description, 128-129 (See ohDetermination of species, immunological) as one of fw isoenzymes, 423 An(Cbod*s: origin of enzymes in plasma, 128-129 ABO (see A90 system) placental: in forensic diagnosis of pregnancy (see Identification of anti-pamitic and anti-allergen as nongenaic markers in blood. 669670 blood, diagnosis of pregnancy from bloodstains) biding by antigen-antibodycomplex. 278 genetic variation and polymorphism, 509-510 blocking (see Antibodies, incomplete) placental-like enzyme in cancer patient serum. 510 blood group (see under spec if^ blood group system names) salivary (see Identification of saliva) in body fluids other than serum, 273-274 of sen&, 509 cold, 52 in mrmof cancer patients, 129 complete, 49-50 Meks. formation,45 codominant, 30 Gm (see Gm system) defmed, 28 to human semen components (see Identification of semen, immuno- dominant, 27-28 logical) multiple, 32 "hypaimmune". 272 recessive, 28 "immune", 271 Mom, 601 immunoglobulinclass of blood group, 48 AGM (see a?-Macroglobulin) incomplete, SO, 352 Aloin tat (see Catalytic tests, aloin) "natural", 271 ALP (see Alkaline phosphatase, placental) as nongenetic markers in blood, 669-670 &ha,-Acid giycopmteia (see Add &coprotein) 0saum cross-rcaczing=ti-A- and anti-B-like, 276-278 &ha,-Antitrypsio (see Antitrypsin) profiling(see ~on~enctic markers) &hrM (see Macroglobulin) prof~gof in serum related to age of bloodstain donor, 130 &har-Mac@obulin. 595,617 Raga. 603 Alpha (a) Rtio (see Age of bloodstains, determination of) to saliva components (see Identification of saliva) Am markas, 607 SNagg. 603 Amboaptor (see Hemolysins) structure, 45-48 Amidopyrme (see Catalytic tests, amidopyrine) warm, 52 Amino rids: Anticbgmotrypsin, 5% amphoteric behavior of, 11-12, (illus.) 14 Antigen: as components of proteins, 11 Australia (see Australia antigen) generic structure, 11 autologous,44 structure of naturally occuning. 13 blood group: development in fetuses and children (see Genetic marker 3-AmiuophIbrlhydde (see Catalytic tests, luminol) systems, dwelopment in fetuses and children) AmpWes, 65 nomenclature conventions. 386 (=Amylases) (Seeahunder specific blood group systems) Amghss: conditions for antigenicity. 43-44 activity of AMY2in semen, 51 1 general nature, 43 assays with dyed starch substrates, 185-187 heterologous. 44 biochemical properties of the isoenzymes, 511-512 heterophile, 44 concentration in body fluids, 186 homologous, 44 distribution of phenotypes in U.S. populations, 51 1 nature of antigenic determinant, 44-45 dyed-starch test papers, 187 spenn cell (seeldentification of semen, composition of semen) electrophoreticpatterns of AMY, and AMY2phenotypes, 511 htigea-a~~tibodyaosP#l electrophoresis (see Immunoelectrophoresis, enzymatic properties, 187 crossed) genetic variation and polymorphism, 51&511 Anligen-~atibodyreactions: linkage relatiins of the loci and chromosomal localization, 511 ABO agglutinin-red cell reactions, quantitative methods, 269-270 pancreatic: presence in fecal material, 198 equilibrium treatment of. 50-51 recognition of amylase enyme, 184 temperature dependence of agglutinin biidiig to agglutinogen, 305 salivary and pancreatic iswnzymes, 510-51 1 astwo-stage event, 48-49 units of activity, 186-187 types. 48-49 Sourcebook in Forensic Serology, Immunology, and Biochemistry

zone phenomena. 52 proof value of positive and negative results in medico-legal tests, (See& Agglutination, Precipitation, Complement Fiion) 86-87 Antisrmm: studies on properties of, 86 origin of the tm,222 studies on specificity and sensitivity of Takayama's procedure. &87 principles involved in waluation for bloodstain grouping by elution, Takayama's procedure, introduction in Europe, 86 408-310 Takayama's solutions for preparation of. 86 Antithrombin Jll, 595 variations in methods of reo oar at ion of. 85-86 ~I-Antitrgpsia: Takayama crystals (see BIA crystals, hemochromogen crystals) application to disputed parentage, 598 Teichrnann's crystals (see Blood crvstal tests. hematin cnstals). ~, biochemical properties, 597-598 Wagenaar's cry& (ABlood &tests, .acetoncchlorhemin nystals) in body fluids and tissues, 598 Blood group C (seeABO system) distn'bution of phenotypes in U.S. populations, 599 Blood gropp system (see under specific system names) geneticvariation and polymorphism, 5954% Blood group systems, genernl considerations. 391-392 recognition. 5% BMgroups, Winadesignation and nownrtptnre, 277 relationship of deficiency to pulmonary degenerative disease, 595, 597 Blood identification (seeIdentification of blood) subtypes of Pi M, 59&597 Body flnids and tkslles: typing in bloodstains. 598 ABO grouping (see ABO system, body fluid and tissue grouping) MMnt nomenclature and standardization, 597 acid phosphatases. 446447 AP (see Acid phosphaw) adenosine dearninasc isoenzymes, 458460,462 Arthas do,56 adenyhte kiiase grouping, 442 L-Aspartate:toxoglutnnte dnotnasferase (see Glutamic-pyruvic carboxylestenseisoenzymes, 472-473 transaminase) DIA, isoearymetypes in, 520 hpemia, 155 arrrasc D isoenzymes, 475 al-AT(see a,-Antitrypsin) glucose-6-phosphate dehydrogenase activity, 487 AT III. (see Antithrombin 111) glutamic-pyruvictransaminasc activity, 502 Atlrinron phenotypes (see Phosphoglucomutasc,PGM, alleles) glyoxalase I grouping, 497 ATP:AMP pbospbotransferree (see Admylate kinase) pcpsinogcn iwrmyma,517-518 Ans (seeAuberger antigen) pcptidasc A grouping, 508 Anberger antigen, 390 phosphoglucomutase grouping, 433-435 A~~ antigen, 611 iphosphogluconate dehydrogenase grouping. 493 (Seedso Hepatitisantigens) sex of origin of blood and tissues (see Sex of origin) Avidity (see Agglutinins) Bombay phenotypes (seeABO system) AzoospermiP, 155 Boosterimmune rrspow (see Antibodies, formation) Bordet, Jub Bur bodks (seeSex chromatin) Nobel laureate irnmunologirt, 221.233 Balk,45 photo, 5 Bebrhg, Emil A. von (see von Behring) BM,R.P., 73 Bedonesproteins, 604 Bus (see Scianna system) &nddine (see Catalytic tests, benddine) -me diitlyhdine (see Catalytic tests, benytidine dimethyl- CA (see Carbonic anhydraw) aniline) Cd (see Sid and Cad) Bernrtdn, Felix, 264 CAP (see Cynie aminopeptidase) Bf (see Complement components, properdin factor B) Cubohydnte% Bg (see~IClycoprotcinI) chemical defiition, 7 B, (see ABO system, para-Bombay phenotypes) structure and nomenclature. 7 Bkbricb d t (seeStains, histological) structure of maldoses, 8 Biiret metbod (see Proteins, assay methods) Cubonate hpdrdyrPe (see Carbonic anhydrase) B-like antigen (see ABO system, bacterial and plant ABH receptors) CIlboaic anhydme: Bloekiog antibody (see Antibody, incomplete) application of CAI, phenotypes to diiuted parentage, 482 Blood, content of gm& marker proteins in (see Genetic marker systems, biochemical propcrties of the isomrymes, 481482 content of proteins in blood, body fluids and tissues) CAIlocus variants. -1 Blood compolwnt profiling (see Nongenetic markers) CA,,locus variants and polymorphism. 481 Bkod aydd t*. distributions of CAI, phenotypes in U.S. Black populations, 482 acetone chlorhemin crystals,85 electrophoretic patterns of CAI, phenotypes, 482 as certain methods of blood identification, 79 electrophoretic patterns of red cell isoenymes, 480 crystaUographic measurements and, 79 genetic loci responsible for isoenzymcs, 480 early observationson blood crystals, 79 grouping CAI, phenotypes in bloodstains,482 hematin aystals: as basis for medico-legal test for blood, W-84 nomenclature of red cell isoenzyma, 480 effects of bloodstain age on. 84 phylogrnetic implications of red all isoenzyrnes. 482 effects of heating bloodstains on, 84 recognition of, 479 general description,80 red all isoenzpes,479 proof value of positive and negative raults in medico-legal teas, 84 substrate spccif~cityof red cell isoenrymes, 481 solution of Nippc (1912) and Kirk (1953), 84 typing of CAU phenotypes simultaneously with other isoempne variations in methods and preparation of solutions. 80, 84, (table) 83 syst-, 482 wiations in procedure. 80.84 Cubo~ensts: hemochromogcn crystals: early description of preparation of, 85 ckssification and properties, 469. (table)470 as medicolegal test for blood, 86 genetic interpretation of tissue and red cell isoenrymes, 472473, (table) pnparation with pyridine, 86 474 human tissue, 472473 structure and oxidation, 104 red cell: application to disputed parentage testing, 473 as test for blood, 103 biochemical properties of isoe~zymes,471472 pphenylenediamine, 110 distribution of estmD phenotypes in U.S. populations, 476-477 rhodamine B, 109-1 10 electrophoretic patterns of, 472 specificity and interpretation of results, 114116 electrophoreticpatterns of esterase D phenotypes, 473 - tetramethylbenddine:as basis of medicolegal blood test. 111 esteme D polymorphism, 471 structure, 112 esteraseD typing in body fluids and tissues, 475 o-tolidine: as basis of medicolegal blood test, 113 genetic variation of A esterases, 471 carcinogenicity of, 110 isoenzymes of, 469-47 1 suucture, 110 shnultancous typing of esterase D and other isoenzyme systems, 475 0-tohlidine. 110-1 11 salivary polymorphic (see Saliva. polymorphic isoenzymes) CcDobii, 9 ~xyphosphate(seeAcid phosphatase, substrates) Cmbroside (see Glywlipids) CYciaophantrl enzymesntigen (seeRegan isoenzyme) Cemlophsmin: &Cuolene. 12 deficiency and Win's disease,619 Cuotenoids (see Lipids) recognition, properties and polymorphism, 618-619 ca*, gendmNon, 101 Ccn*rl mucus (see Vaginal secretions) ~tcsts: Q. (see Complement components, C4) aloin: structure of barbaloin, 104 CbbsqaM(see Population genetics) as test for blood, 103 Chido (see Complement components, C4) amidopyrine, 110-1 11, (structure) 112 Cuorpromszine (see Catalytictests, chlorpromazine) bmddine: carcinogenicity, 107-108 Cboksterol. 11 certainty of positive test as test for blood, 105-107 Cholinc. 174 dangersdue to carcinogenicity. 108 (SeeaLFo Identifmtion of semen) introductionas mediwlegal blood test. 105 Cbolinesle~9cs: oxidation and structure of oxidation product, 106 of blood, 463 sensitivity, 107 classes of, 463 specificity. 105-107 as neurotransmitter, 463 structure, 106 recognition of, 463 substancesgiving false positive results, 105-107 red cell acetylcholinesterase, 463-464 benzylidine dimethylaniline, 11 1 serum pseudocholinesterase: alcohol inhibition and alcohol number, 465 chlorpromazine. 111-1 12 assay procedures and detection of E, variants, 468-469 2,7-diaminofluorene, 109 biochemical properties of isoenzymes, 467-468 odhisidine: as basis of medicolegal blood test, 110 chloride inhibition and chloride number, 465 carcinogenicity, 110 detection of E, phenotypes in bloodstains, 473474 structure. 111 dibucaine inhibition and dibucaine number, 464-465 diphaylamine, 112 distribution of El and Ea locus phenotypes in U.S. populations. eosin, 109 41-77 evidential (probative) value of positive results, 116 electrophoretic variants. 46546 fluorescin, 112 fluoride inhibition and fluoride number, 465 genddiscussion of basis of, 101 generaldescription, 463 guaiacum: wmpositimn of guaiacum blue, 102 genetic loci responsible for variants (E, and Ea). 466 composition of guaiacum resin, 101 genetic variation of, 464-467 early examples of use in medicolegal cases, 101-102 nomenclature and propaties of variants, 466 early studies on peroxidase activity of blood, 101 relationship of genetic variation to sucdnyldicholine sensitivity, 464 false positives, substances and materials giving, 102 screening procedure for E, variants, 468-469 of historical interat as mcdicolcgal blood test, 103 silent alleles at El,46647 interpretation of results with bloodstains, 102 standard assay conditions, 464 medicolegal blood identification and, 101-102 typing of Ea locus electrophoretic variants in bloodstains, 475 sensitivity. 102-103 ChromPtognphk methods: leucocrystal violet: as basis of mediwlcgal blood test, 108 for blood identification (see Identification of blood) structure, 108 for wncentration of diluted blood samples, 119-120 leucomatachitegreen: as basis of medicolegal blood test, 108 for identification of semen (seeIdentification of semen) specificity and sensitivity, 1W109 for identification of urine (see Identification of urine) StNctUrC, 108 for separation of blood from debris, 119-120 luminol: as basis of medicolegal blood test, 112-1 14 ~roantogpphy: chemiluminescence reaction mechanisms, 115, 116 gel filtration, 14-15 chcmiluminacence reactions, 114 as method for estimation of MW of proteins, 16 structure, 113-114 ion exchange, 14 peroxidase activity of blood, 101 Chromosomes: peroxidase activity of hemoglobin. 101 in cell division. 25-26 pcroxidase reaction, generakd, 101 diploid number, 26 peroxide. 109 genetic map of human, 36-37 phenolphthalin: evaluation of, as medicolegal test for blood, 104-105 haploid number, 26 oxidation of, as test for "oxida&', 103 homologous, 26 preparation of reagent for blood test, 104 sex determination and, 26-27 sensitivity, 103-104 Gronic g~nulomntousdi(CGD), relationship to Kell blood group specificity, 104 system,370-371 Sourcebook in F0rem.c Serology, Immunology, and Biochemktry

Cbronic obstrorlke pnlmonvy dii, relationship to a,-antitrypsin development of precipitin test, 221-224 deficiency, 595,597 double immundifusion asforensic species test, 225 CI (see Stains, histological) double immundifusion technique development, 224-W Cibrbmn blau F3C-A starch, 185 external influences effects on precipitin test, 225-226 CID (see Combiied immunodeficiency disease) fluorerent antibody technique, 237-238 Co (see Colton system) heating of bloodstains, effect on precipitin test, 226 "Cold" aggln(lnins(see Ii system) hemolysins, 235-236 and bloodstains (seeAbsorption~lution) immunoeledrophoresis, 238 Colton system. 390 mixed antiglobulin technique, 231-232 Combined immnnodefieieaey dipense, relationship to ADA deficiency, opthiation of precipitin reaction conditions, 227 457458,459460 origins of prccipitin test, 221-222 Complement: phylogenetic relationship and antibody cross-reactivity, 241 cascade, 55, (ius.) 56 phytoprecipitins, 237 components, 54-55 precipitation-inhibition, 226 dacription of, 54-55 pracipitin test applied to game killing cases. 223 pathways, 55 pdpitin test, early studies, 221-224 unit of, 55 prccipitin tat for differentiating meats, 223 Complcmcnt components: prepamtion of precipitating antisera, 223-224 C2.614-615 protein concentration estimation in bloodstain extracts, 227 C3.615 quantitative precipitation techniques, 238-239 C4,61%16 ring test, 224 C6. C7 and C8,616 sensitized colloidon particles, 231-233 polymorphism of, 614 sensitized latex particles, 233 properdin factor B, 617 serum-hemoglobin precipitation, 236-237 Coppkment fiXltiOn: single immunodiffusiontechnique development. 224 description of, 55,233 specificantiseraagainst human and animal blood, 222-224 immunologicalspecies of origin test, 234 SUbStratUm effects on pepitin test, 225-226 Chccotntioo of dilute blood samples (see Chromatographic methods) tanned red cell hemagglutination technique, 231 cl AetMor, 5% tanned red cell technique for discrimination of closely related spe- clh(see cl Activator) cia,239 Coombsamm(see Anti-human globulin serum) turbidiwtry, 238-239 Coombs tat (see Anti-human globulin serum, detection of incomplete isoenryw separation for, 244 antlwes with) miaometry, 215-217 Copper binding proteio (see Ccruloplasmin) cases of historical intmst, 216-217 Cp GFee MOP-) (See aLPo Red blood cell, measurement) CPK (see Crcatinephosphokinase) odor (see Determination of species, chemical methods of historical Crrrthe pbosphoLinrrc(see Identification of semen) interest) ChaAmmooizatioa(see Determination of species, immunological cross- tissues*244 dons) Di (wDigo system) Chawedons of species aotinm (see Determination of species, immu- 2.7-Dhminotlooreoe (see Catalytic tests, 2,7-diaminofluorene) nological) o-Dhlw&c Crmrd ekctmiioodiffuaion (see Irnrnunoelectrophoresis, crossed) as catytic test reagent (see Catalyric tats) CIJP~tloepp,339 as reagent for detection of Hp-Hb complexes (see Haptoglobii, de- C$ C3,C4, C6.0,CII (see Complement components) tcction in typing media) C~~BCuoioopeptidnec (see Identification of blood, diagnosis of preg- h.aa#iac. . bhe (see Naphthanil Diazo Blue B) nancy from bloodstains) Dbpborrs: ~ocbmwb, rednctsse(see Diaphoases) biochemical properties, 519-520 description and propenies, 5 18 Ihfs test (see Catalytic tests, guaiacum) DL43 phenotype distribution in populations, 520 Db(seeSativa, polymorphic protein systems) multiple gmdcloci and polymorphism, 518-519 Ddect.biWy of geode hersin rging sampks (see Genetic marker sperm diaphorase, 519 systems, detcctabiity in aging samples) (see Amyhc) lMamht&nof specks: -Dib.aine, 464 chemical methods of histowinterest, 215 (&eahCholincstemes) fibrin pkte method. 243 Dkgo system, 389-390 gamma globulin deviation. 237 Dimethyi-pdmpbcnyl phosphate, 469 hemoglobin denaturation with alkali, 243 (see ahCarboxylest~) hanoglobin sqmahns for, 244 Dipko- (see Catalytic tests) historid, 215-217 Didmhthindex (see Population genetics) immunological: adjuvants in antiserum preparation, 224 MrpoW PMntrsc-g: aggluthh in animal sera as basis of, 236 ABO system, -297 agglutination of human cells by animal sera, 236 adenosine deaminase system, 461 auaphylaxis, 234-235 adenylatekinasc system. 441 anti-human globulin serum inhibition, 227-229.230 A8 system. 613 auti-human hemoglobin sera. 227 carbonic anhydrasc I1 system, 482 bloodstain washing, effects on precipitin test, 226 Duffy system. 373 complement fixation. 233-234 early developments in ABO applications,296 cross over immunodectrophoresis, 225 erythrocyteacid phosphatasc system, 450 cross-reactions of antisera to related species, 239-241 aterase D system, 473 680 Index

galactose-1-phosphate uridytyl transferase variants, 515 EkehophorKk genetic marker systems and. 257-258 agar gel: development of technique, 60 glutamic-pyruvic transamiwe system, 502 ekaroendosmosis, 60 glyoxaiase I system, 4-97 blood identification using (see Identification of blood, electrophoretic Gm and Km systems, 608 methods) group specific component, 584-585 cellulose a&te membrane, development of technique, 6041 haptoglobim system. 577 crossover (see Immunoelectro~horesis,cross over) hemoglobin variants, 555 description-, 58 HLA system, 629-630 immunofition (see lmmunofition electrophoresis) KeU system, 371-372 mechanisms. 58-59 Kidd system. 375 paper, 59 Lutheran system. 387 polyamyhmide gel: development of technique, 61-62 MNSs system, 342-343 disc electrophoresis, 61-62 P system, 385 flat elelectrophoresis, 62 peptidase A system, 507 of serum proteins, (ius.) 567 phosphoglucomutase system, 430 starch gel, development of technique, 59-60 dphosphogluconate dehydrogenase system. 492 Ekdnwmtrts&(see lmmunoelectrophoresis,cross over) polymorphic proteins of saliva, 635 El&. S.D.. 224 primary or fiorder exclusions, 257 Elation of bloodgroup antibodiesfrom red cells, 310 probabiity of exclusion, 258 Emwine Cornmipaion (see Enzymes, nomenclature) Rh system. 361-362 EazywJ: secondary or second order exclusions, 257 cofacton and coenzymes, 19 semorsystem. 297,313 inhibiton, 18-19 transferrin system, 593 Kinetics of enzyme-catalyzed reactions, 16-19 DNA: multiple molecular forms of (see Isoenzymes) as the genetic material, 21 nomenclature. 16 replication, 23.28 origin of the term. 184 structure. 21-23,&27 (See&under specific enzyme names) Do (see Dombrock system) Eolin: DiMtdcin's bodnus (see Vaginal secretions) as mc blood test reagent (seeCatalytic tests, eosin) Dombrock sogtem, 390 as hiiolo~calstain (see Stains. histolostical) Dmp, as nongenetic marken in bloodstains, 670 ~ry(hrocyle-ridpbo~h~~rse(k~cid phosihatase, erythrocyte) Dnfk -em. 372-373 ESD (seeCabxylesterases, red cell) di.&iution of phenotypes in U.S. populations, 378-379 E600 (see ~ieth$-pnitropehnyl phosphate) Dyebiding metbod (see Proteins, assay methods) Estcrrr D (see Carboxylesterases, red cell) Estemes: EAP (seeAcid phosphatase, erythrocyte) categories of. 463 Enmar, ABO gonphg of (see ABO system, body fluid and tissue as one of fiisoenymes, 423 grouping) of semen and sperm (see ldentification of semen) LC. [Eazymc CommWaml (see Enzymes, nomenclature) species determination by isoenzyme separation, 244 LC. 1.1.1.27 (see Lactate dehydrogenase) (See& Carboxylesterases, Cholinesterases, Carbonic Anhydrase) E.C. 1.1.1.44 (see dPhosphogluwnate dehydrogenase) w,12,668 LC. 1.1.1.49 (see Glucose-&phosphate dehydrogenase) Exdoaions(see Disputed parentage testing) LC. 1.6.2.2 (see Diaphorases) LC. 1.6.43 (seeGlutathione reductase) Fatty acids (see Lipids) LC. 1.11.1.9(see Glutathione peroxidase) Fast Blne B (see Naphthanil DioBlue B) E.C. 1.15.1.1 (see Superoxidedismutase) Fast Gund B, 157,158 E.C. 1.163.1 (see Ceruloplasmin) Frrt Red AL (see Naphthanii Diazo Red AL) E.C. 2.6.1.2 (see Glutamic-pyruvic transaminase) Frrt Red RC (seeNaphthanil Diazo Red RC) E.C. 2.7.1.1 (see Hexokiiase) F-bodies: E.C. 2.7.33 (see Crcatine phosphokinase) description, 666 LC. 2.7.43 (see Adenylate Ki) as sex of origin indicator in bloodstains. -7 E.C. 2.7.5.1 (see Phosphoglucomutase) as sex of origin indicator in tissues, 667668 E.C. 2.7.7.12 (see Galactose-1-phosphate uridylyl transferase) Fedmaterial (see Identifition of fecal material) E.C. 3.1.1.7 (see Chobesterases) Fm#in.IPbelkdamwies for ABO grouping, 312 E.C. 3.1.1.8 (see Chohesterases) a,-Fetoprotdn(m ldentification of blood, fetal and infant blood diag- E.C. 3.13.1 (see Alkaline ph0sphat.W nosis) E.C. 3.13.2 (see Acid phosphatase) Fdgcn st.in (see Stains, histological) E.C. 3.13.18 (see Phosphoglycolate phosphatase) Fiiuia plate method: E.C. 3.2.1.1 (see Amylases) for menstrual blood identifiion. 123-124 LC. 3.4.1.1 (see Leucine aminopeptidase) for species determination, 243 LC. 3.5.4.4 (see Adenosine deaminase) Fibrinolyticsyst~m,122-123,125 E.C. 4.2.1.1 (see Carbonicanhydrase) (See uku ldentification of blood, menstrual blood) E.C. 4.4.1.5 (see Glyoxalase I) EluoKsce~: E.C. 5.3.1.9 (see Glucose phosphate isomerase) saliva stains(see ldentification of saliva) Ebrlieb, Paul: seminal stains(see ldentification of semen) Nobel laureate immunologist, 221 urine stains (see ldentification of urine) photo, 5 ~o-nt-hbe~cd antibodies, for ABO and MN grouping, 312 Ek&oiinoodWlrsion (see Immunoelectrophoresis) Fluorescin (see Catalytic tests, fluorescin) Sourcebook in Fomnsic Serology, Immunologv, and Biochem&try

FobCkakmmgmt, for protein assay, 15 Kidd antigens, 375 Foramcla mtigca (seeAntigens, hcterophik) MNSs antigens, 344.345 FOUR (seeHLA system) phosphoglucomutase isoenzymes, 432-433 Frnctose, as marker for semen identification, 175 Rh anti-, 362-363 crL-Fbcose: development in fetuses and children: adenosine deaminase, 461 involvement in ABH antigm structure, 287 admylatekinase, 441 suucturc, 287 Duffy antigens, 373 hnao55 (seeCarbohydrats) #uramic-pyruvic transamitme, 50t FP (seeDuffYsystem) Kidd antigens, 374 Lewis antigens. 329 Gahclohhac. 514-515 Luthaaa antigens, 387 geneticvariants and deficiencies. 5 15 P antigens. 383 relationship of deflcimcics to cataracts,515 peptidasehenqmes, 507 N-&e N-&e (seeAcetyf-galactosamine) phosphoglucomutase. 430 Gabctosel-pbosphmtenridytyl Uuufewe: bphosphogluconate dehydrogenase, 492 application to disputed parentagetesting, 515 disputed parentage testing and, 257 biochemical properties. 515 table of, 259 distribution of phenotypes in U.S. populations, 514-515 tabk of dimhimtion indexes. 259 electrophoretic patterns of phenotypes. 514 table of probabilities of exclusion of paternity, 259 genetic MMtion and galaaos~mia.513-514 w recognition and metabolic role. 512-513 developmat as a science,20 C.bctostmh (see Galactoscl-phosphate uridylyl tansferase) origin of the term, 20 CALK (seeGalactokii) (See&Inheritance) CII-I-PUT (see Galactose-1-phosphate uridylyl transferase) Genotype, 27 GALT (seeGalactose-1-phosphate uridyIy1 transferase) Gkma ruin (see Stains. histological) C8qbddcs(see Glycolipids) G1 (seeSaliva, polymorphic protein systems) C;mod,Arcblbr# E., 20 GWD(see Glucoscb.phosphate dehydrogenase) Clrbic mamsm, ABO tgping (see ABO system, body fluid and tissue CU) (seeGlyoxiilasc I) grouping) 1,4cr-D.CIncamghemobydrdrPc(seeAmylases) GBC (seeComplement components, properdin factor B) -ppnos*l, 9 Gc (see Group specific component) (~PCkcose-l.ddiphoop~~-~~~~-l~osphate pbosphotmnsfer- Gd (see GIuwsed.phosphatedehydrogcm~) me (seePhosphoglucomutasc) Gd filcntkn (see Chromatography) Gloam+phospbnte debydrogcarpc: Gcnfrequency, 39 biochemical properties of isoenzymes, 486-487 CaeQif code. 24-25.30 detection in typing media, 481 Cactiemap of humehroooroms, 36-37 distribution of phenotypes in U .S. populations, 489 Gem& nrpping. 33 ekarophomtic patterns of common phenotypes, 488 GeDetkmwkersatenrr geneticvariation in red cell enzyme, 483-484 applications in forensic serology, 257-258 grouping in bloodstains, 487 blood and body fluid individualization using. 258 inactivatingeffect of red cell stroma, 487 claws of, 257 metabolic rolein cell. 484-485 content of proteins in blood, body fluids and tissues: adenosine de- nomenclature of red cell genetic variants. 484-486 aminasc, 462 recognition of, 483 admylate kinase, 442 red cell geneticmiants of, 484-486 a,-antiuyPsin. 598 rehionship of genetic variation of red cell enzyme to induced hemo- concentrationrangesof serum proteins in mm,563-564 lytic anemia, 483-484 estaasc D,475 relationship of X-linkageof Gdand Lyon hypothesis, 486 glucoseb.phosphatedehydrogmax, 487 sPlivary polymorphic (see Saliva, polymorphic isoe~yma) glutamic-py~victransaminase, 501-502 in sm~n and spermatozoa, 487 6,-glycoprotein I, 621 X-lintage of Gd locus. 486 glyoxalascI, 497 Ghcoaephoapbr(e bomennc, 515-516 Gm and Km,609-610 DGhcor69borpbrle.NADP+-l-oaidoml~ (see Glucose-6-phos- group spcdfic component, 5&(, 585 P- deb-) haptoglobin. 569.579 Ghmaic~ (seeGhrtamic-pyruvic transaminase) hemoglobin. 545 cM..llcg+- kmuyme assays in diagnosis of deficiency types. 490,502 aaivityinbodyfluidsandtirmes.502 a,-macroglobulin, 617 apparent quantitative genetic variation, 502 pcptidasc A. 508 appktion to disputed parentage testing, 502 biochemicalpropaties of kcnzymes, 501-502 detection reactionsin typing media, 501 transf~.593 distribution of phenotypes in U.S. populations, 502 deteanbi in aging samples: ABO antigens, 3aJ-301,310 electrophoretic pattansof phenotypes, WY) ABO iFoaggtutinins, 299-W gactic variation and polymorphism. 499 admosine dePminase boenzymes, 461 groupingin bloodstains, M2 admylatekinasc ism,4-42 phmotyping procedures. MO-501 Duffy antigens. 373 recognitionof. 499 erythrocyte acid phosphatascisoensymes,451452,453 silent alleles. 499.502 aterase D isoenzyms,475 ChUhmpaow.516 Ken antigens, 372 Ghathione ductme, 516 Glycerophosplutes (see Acid phosphatase, substrates) content in serum,569 Glycin&cb~-sfycoprotdn(see Complement components, properdin detection in typing media: with dpicblood test reagents for Hp-Hb factor B) complex, 578 Glycoupids odbkidiue as reagent for. 110 cerebrosides and gangliosides, 7 use of guaiacol for, 103 as ckssof lipids, 7 dism'bution of phenotypes in U.S. populations, 579-580 3salClycopmtd. (see al-Antitrypsin) eicctrophomic patterns of common types and subtypes, 571 BrClycoprotdn I, 621 electrophoreticpatterns of wiants. 573 ~2-clycoprotdn Ll (see Complement components, properdin factor B) genetic variation and polymorphism. 569-573 GlyoxnbPI: occumaa in body fluids, 579 application to disputed parentage testing, 496-497 physiobgid role, 569 assay procedurrs. 495 cecoguition of, 569 biochemical properties of isoenrymes, 4% stnrauralgene duplication. 575-577 detection reactions in typing media, 495496,497 structure: of Hp-Hb complex, 574-575 distributions of phenotypes in U.S. populations, 497498 polypeptide chains. 575 ekctrophoretk patterns of phenotypes, 4% subunit, 573-574 genetic variation and polymorphirm, 4% subtypes of Hp 1,570 grouping in bloodstains, 497 typingin bloodstains,577-579 groupingin body fluids and tissues, 497 HYdy-WebbergeqmWwbm (see Population genetics) recognition of, 495 H)(seeHemoglobin) typing simultaneously with other isoenzymesystems, 497 HBsAs (see Hepatitis antipens) Glyouhsen,495 HCC (see Human chorionic gonadotropin) Gm -em: He (see hlNSssystem. asociated antigens) application to disputed parentage testing. 608 HesUog, effects on bloodstain component solubility, 84,226 assianment of factors to IgG subclasses, 604 HemrlCglotinrtbainhibition (see inhibition tests) distribution of phenotyp& in U.S. populations, 610 Iiemtoporphyrin (see Identification of blood, spearal tests) factors and nomendature, 6014M. (table)602 ~~(seeStains, histological) genes and expression in cells, 604 &me aud kmrtineomponnds: genetics and inheritance, 605406 description. 77-79 nonmarkm. Kt4405 internlationships,82 rrcognition of, as inherited factors, 601 structureand nomenchture, 77-79,81 typing in bloodstains, 608609 structure of heme. 77.80 typing in body fluids and times, 609-610 H-&l!mw, 34 typing methods, 607608 -: GP1 (seeGtucose phosphate isomerax) application of genetic variants to disputed parentage testing. 555 GPT (see Glutamic-pyruvic transaminase) biochemical genetics, 554 GPX (see Glutathione peroxidase) content in blood. 545 Group SpCdfir component: diUri'bution of phenotypes in U.S. populations. 560-561 application to disputed parentage testing, 584-585 electrophoretic and isoelcctric focusing methods of characterization, biochemical properties. 584 554-555. 5%-557 distribution of phenotypes in U.S. populations, 586587 fctal:alkalidenaturationcharact-, 129 electrophoretic pattam of phenotypes, 582 as marker for fetal and infant blood (see Identification of blood, genetic variation and polymorphi, 581-583 fetal and infant blood diagnosis) methods of phenotyping, 583-584 structure,547-549 nomenclature of vahts, 583 as geneticmarker system, 545 occurrence in body fluids, 585 gcnaic valiants. 549-553 physiological role, 584 nomenclature of ets,553 recognition of, 581 origia of the name,93 serum concentration of proteins. 584 serum precipitation of (see Detamhation of species, immunological) subtyping of Gc 1,583 sicklecell. 550 typing in bloodstains. 585 sicklecell and balancedpolymorphism, 558 variants, 581-583 speciesdevrmination by alkali denaturation, 243 variation in patterns in newborn and infant blood, 130 specks detemhation by separation of, 244 as vitamin D biding protein, 584 spearal properties. 89-91 Group double combiwtion wthod, 31 1 spectral propertis as means of blood identification (see Identification GSR (see Glutathione reductasc) of blood, spectral tests) Garhrnm (see Catalytic tests, guabxm) structuralgene duplication, 554 structure embryonic and fetal, 547-99 EI.ir: of polypeptide chains, 545-549 ABO souping of (seABO system) temmcric, 545, (f-) 549 isoenzymes in hair roots, summary,637 thrte-dimensional,547 polymorphic structural proteins of, 637 typing of variants in bloodaains, 555,558 HSPlotgpe, 601 variants diUri'bution in world populations, 558-559 ~toglobim: Hemglobin F (see ~emo~lobiif&) as antibody against streptococcalsurface antigen, 569 Hemlysh% application to dsputed parentage testing, 577 antibodiesas, 49 biochemical genetics, 575-577 in complement fmation reactions, 55 chloroform extradon procedure for bloodstains, 578-579 speciesdeemhation and. 25-236 classification of variants, 570 rmeprtitk .nt&m. 670471 Sourcebook in Forensic Serology, Immunologv, and Biochemistry

Heterozygosity,29 identifhtion of blood cells, 96-97 Hexolchsc, 516517 isolation and identification of white cells, 96-97 meld,Ludwig, 254 "reconstitution" of red cells from bloodstains, % Histocompatibility(see HLA system) transfer methods for bloodstains, 96-97 fM (see Hexokinase) origins, 74-75 HLA sgstem: papachromatographicmethods, 119 A and B locus antigens, M25 spectral tests: effects of environmental influences on bloodstains, W-W antigens in semen, 630 general considerations,89 application to disputed parentage testing, 629430 general features of visible spectra of hemoglobin and derivatives. biochemical propertics of antigens, 628-629 90-91 C locus antigens, 626-627 hcmatoporphyrin, different methods of preparation, 94 D and DR locus antigens. 627428 hematoporphyrin fluorescence as medicolegal test for blood, W definitiin of 624 hcmatoporphyrin fluorescence properties, W genetics, chromosomal localization and linkage relations, 625426 historical development, 89-93 lymphocyte defmed antigens, 627 transformation of hemoglobin into derivatives and their spectral mixed lymphocyte culture (MLC) test, 627 properties, 90-93 recognition and general description, 623424 thin-layer chromatographicmethods. 119 serologically defmed antigens, 624-625 Mcatiecrtion of fetal mrtcrhl: testing and typing procedures, 628 biibin, 197 typing in bloodstains, 630 characterization of bacteria, 197 H-tlke antigen (see ABO system, plant and bacterial ABH receptors) coprostanol. 198 Hob,Fnoz Josef (see Unit IX) microscopical examinations, 197 Homozygos&, 29 parasites, 197 Honor autotoaleus. 43 t~~biilin~gen~and ~robilins,197-198 Hp (see Haptoglobin) IdcnMatioa of snlipP: HPL (see Human placental lactogen) alkaline phosphatase, 183-184 H-type pdpii.ntiscR. 53-54 amylase: as basis of medid@ test for saliva, 184-185 HII(see MNSs system,associated antigens) dysd-starch wthods, 185-187 Humlla cborionic gomdotropin, detection by sensitized particle tech- dyed-Wh test p-, 187 nique. 233 anti-human saliva serum, 187-189 (See &o Identification of blood, diagnosis of pregnancy from blood- fluorescence of saliva stains, 189 stains) microscopical methods, 189 H~nrurpbcmt.l hctogcn (see Identifition of blood, diagnosis of preg- nitrite, 183 nancy from bloodstains) salts and low MW compounds composition, 195 Hv markers, 607 thiocyanate, 183 HypmeIltivity: IdenMiatiomof menun: delayed,56 add phosphatase: apparent polymorphism of seminal enzymes, 167-168 bundate, 55-56 assay techniques, 162-163 description, 155 Id (see Intcr-u-trypsin inhibitor) dismvcry, 155 IdeotiRation of blood: immunogddty of seminal enzymes, 168 age of bloodstain donor correlated with senun antibody profde, 1130 independence of antigenic and catalytic sites, 168 anti-human hemoglobin: anti-Hb F for fetal blood identification, 118 as marker for seminal fluid, 155-157 an- to Hb Aand Hb F. 117-118 mokcuhr heterogeneity, 167-168 medicolegal tests for identifying dried blood, 117-1 18 persistence in seminal stains, 162 prepamtion of antiserum, 117 wrsktence in vagina after semen deposition, 161-162 catalytictests for (see Catalytic tests) purifkation and properties of seminal enzyme, 166-168 crystaltests (see Blood crystal tests) quantitative assays in stains, 159-160 diagnosis of prcgnaacy from bloodstains: cystine aminopeptidase, 128 rocket electrophoresiswith specific antibody. 171 kucine aatinopeptidase, 128 seminal and vaginalenzyme relationship, 166 placental alkaline phosphatase, 128-129 separation of SAP and VAP, 166 pregnancy-associated proteins. 127-128 of test, 157-159.163-166 pngnancy hormones, 126-127 subs&ratcsfor the enzyme. 162-163 dectrophoretic methods, 120 tanrate inhibition of, 165-166 faal and infant blood diagnosis: fetal hemoglobin, 129 units of activity, 162-163, (table) 164 a,-fetopmtein. 129-130 aminoacids, 178 variations in Gc pattern, 130 choline: dctectcd chromatographically. 174-175 heating tat, 120 dCtcaed by crystaltest, 172-173 history, ancient and mediaeval, 73 chromatogmphicmethods, citric acid. 175 menstrual blood: fibrinolytic properties. 122 spennine and choline, 174-175 glycogen-containing epithelial cells in, 121-122 composition of sanen: enzymes, 180 LDH hemyme profde as diagnostic of, 125-126 (See crLw,under specificenzyme names) methods based on fibriuolytic activity, 122-124 seminal plasma proteins, 179-180 microscopical and histological methods for, 121-122 sperm antigens, 179 preparation of antisera against, 124 table. 181 toxicity bioassays for, 124-125 marine phosphokinase, 175-176 microscopical methods: histological staining of blood cells from blood- crystal tats: Barberio test, 173-174 staiaextracts,*97 Flor~ll~e test, 172-173 history and general discusion, 94-96 Nkderhnd and Peltzer tests. 174 Puranen's test, 174 interpretation of precipitin pattans. 57-58, (ius.) 59 diamiocoxidase, I77 earliest observations, 224 early chemical tests of historical interest, 149-150 single. 224 . tluorescenceof seminal stains, 178 description, 57 y-seminoprotein, 177 linear, 57 immunologicak anaphylaxis methods, 171-172 radial, 57 anti-human semen sera, 169-171 Immunodcebophorrsis: anti-p30, 180 cross over, 62-63 anti-seminal acid phosphatax, 171 in precipitation test for species determination, 225 complement fucation rest. 171 crossed. 63, (ifflus.) 64 y-scminoprotein, 177 quantitative, 63-65 kaic dehydropenase-X, 176 rocket dcctrophoresis: description, 63, (ius.) 64 p30 protein. 180 in identifmtion of semen with anti-seminal acid phosphatare, 171 potato phytagglutinio inhibition. 178 and serum group proteins, 564,566 seminal fluid esteraSes, 176-177 of sawnproteins, (iius.) 567 sperm cell cstcrase~.176-177 simple, 62 spermatozoa: dcsuuction of substratum, 151 and spedes determination, 238 uramination of stains directly for, 151 lmmmnofiution cketrophonsis. histological stainingof. 151-152 description. 63 historical material, 150 and serum group protein typing, 566 morphology and its forensic implications, 154155 ImmunogbbPiins: persistence in vagina (see Identification of semen, spermatozoa, clasm of, 46,48 swivalinvagioa) genetic markm of (see Grn system, Km system, Am markers. Hv separation of cells from substrata, lSQl51 markers) swival in vagina, 152-154 nomenclature, 46-48 transfer techniques, 151 properties of, 46.48 spermine: detected chromatographically, 174-175 structure (seeAntibodies, structure) detected by crystal test. 173-174 el-lna (seecl-Activator) detected mymatically. 178 Inbornmomof w(.bolism, 20 detected by FuchsTokuoka reaction, 178 Incomplete penelnnec, 30 Idmdi6ationof ark ladopknol oxidure (seeSuperoxide dismutase) chloride, 191 Inkitmce: chromatographic methods, 194 crossing over, 32-33,35 composition of urine: chloride, 191 independent assortment, 29-30,33 matinine, 193 linkage. 32 indican, 193 patterns in populations (seePopulation genetics) phosphate, 191 pedigrees and pedigree symbols, 39. (illus.) 40-41 table, 1% segregation, 27-28,32 urea, 193 sex-linked, 33-38 meatinhe: Jaffe test, 193 dosage compensation. 34 Sakaguchi test, 193 X-chromosome inacthtion, 34.38 Salkowskitest, 193 XX-dated, 33-38 fluorcscmce of urine stains, 191 simple mendelian patterns, 27-30 indican, 193 Inhibition of prrdpin (see Determination of species, immunological) microscopicalmethods, 191 Inhibitiontests, blood group substances in body fluids: odor, 191 A, and A, ceb in anti-A tests, 286 phosphate, 191 inhibitive titer, 285 urea: crystaltcsts, 192 techniques. 285-286 pdimethykminobedehyde reaction, 192 titration of body fluids, 285-286 dimthylaminocinnamaldehyde(DMAC)reaction, 192-193 lnhi'bition-ti(m(ion (see ABO system, bloodstain grouping by inhibi- enzymatic test with urease. 192 tion, Holzer technique) xanthydrolreaction. 191-192 Inter-drypsh inbibitor, 595 IdiolJpe, 601 kliwslmh reaclion, 184 w,Sm~m, 46 loouchagccbmnulogmphg (seeChromatography) igD, smam, 46 Iros-bmdiig protein (seeTransferrin) IgE: isodcctric focusing, development of technique, 65 asmediator of anaphylaxis. 56 Isoclaymcs: structure, 46 content in blood, body fluids and tissues (see Genetic marker systems, I%;,~CtlI~, 4647 content of proteins in blood body fluids and tissues) IgM, -. 46 development in fetus and children (see Genetic marker systems, de- Ii sg.stem,389 velopment in fetuses and children) Inl (see later-a-trypsin inhibitor) discovery of. 423 lmronc response (seeAntibodies, formation) in hair roots, 637 Immunity: molecular bases for, 423 cellular, 45 polymorphic forms in saliva (seeSaliva, polymorphic isoenzymes) humoral, 45 simuhancous typing of two or more systems, 432, 441-442, 452-453, lmmnnodiffwbn: 461,475,482,493,497,508 double: description, 57-57 Isoprecipii. 61 1 development of technique, 224-225 Isoprene, 12 Sourcebook in Forensic Serologv, Immunologv, and Biochemistry

Isotschophorrsis. 65 Lotus, 273,282 btvpe, 601 Bophocatpus, 282 Lsozymes (see Isoenymes) Uk,273.282-283.304 anti-M. Iberis MIoTa, 338 Jay sntigen (see P system, Tja) anti-N: Bauhinia, 338 Jk (see Kidd system) Viciagraminwe. 338 Dolichos: reacton with animal blood cells, 237 Kdl w: structural studies, 289 antibodies, 371 Lotus antigen in saliva. 283 application to disputed parentage testing, 371-372 purification and biochemical properties. 283 bloodstain grouping. 372 St~auralstudies, 289 complexities, 369-370 La (see Lewis system, in body fluids) daectabiity of antigens in aging samples, 372 Lcllriae .miaopeptidPJe (see Identification of blood, diagnosis of preg- distribution of phenotypes in U.S. populations, 376-377 nancy from bloodstains) genetics. 370-371 Lean, base. 108 Js (Sutter) antigens, 369 Lcwcompooad (see Leuco base) K and k antigens. 369 Lncoapstnl *kt (see Catalytic tests, leucocrystal violet) Kp antigens. 369 Leacocyte: KpCantigen identity with Levay. 370 recovery from bloodstains, 630 numerical nomenclature, 370-371 (See aLw White blood cdl, HLA system) relationship with chronic granulomatousdisease (CGD), 370-371 Leucomnbchite green (see Catalytic tests, leucomalachitegreen) Ketoses (see Carbohydrates) IMvisSyspE Kidd sgstcm, 373-375 antigenic structures, 290,291,293 distribution of phenotypes in U.S. populations, 378-379 antiscra. 331-333 Km -.: assignment of gene locus symbols, 386 appbonto disputed parentage testing, 608 biochrmical genetics. -293,333 factors and nomenclature. 607 body fluid grouping, 3 19 markers, (table) 602 in body fluids. 330 typing in bloodstains, 608-609 complexities,33CL331 typing in body fluids and tissues, 609-610 discovery and development, 329 typing methods, 607408 distribution of phenotypes in U.S. populations, 334 KO&, RoM, 221 gcndcs, 333 genetics and relation to Se/se and ABH. 329-330 Lantigem (see Leain,Lorus antigen) LeCand ~e~,33 1 LA (gpe HLA system) red cell antigens, 330 haatedehydrogemse: relationship to ABO and secretor systems, 290-293 genetic variation, 5 17 relationships of body fluid and red cell types, (table) 332 isoenzyme patterns. 125 relationship to Ii antigens, 389 isoenzyme pattems for identification of mensrual blood (see Identifi- relationship of red cell Lewis type and secretor status, 329 cation of blood, menstrual blood) relationship of secretor and Lewis genotypes to red cell and body fluid as one of fm discovered human isoenzymes,423 antigens, 295 reaction catalyzed by, 125 stain typing of. 333 species damnhation by isoenzyme separation. 244 type 1 chain conversionsto group substances, 293 subunit structure, 125 type 2 chain conversionsto group substances, 294 subunit structure of isocmymes, 423 Lipi X-bcmymc for semen identification (see Identification of semen) chemical defmition. 7 Lrto-N-focoknose 1,288 structure and nomenclature, 7-1 1 -0-Nhcopeataose n, 288 Lipoprotdn (see Serum lipoproteins) Lrtoyl-gbtathione lyrsr (see Glyoxalase I) LlSS (see Low ionic strength solutions) SLacto~bthkoemetby+gIyod Ig.s (imaidng) (see Glyoxabse I) Lon ionic strength soations, enhancement of agglutination by, 51, 363 Lmbteiner,Kul: Lawry metbod (see Proteins, assay methods) discovery of ABO blood groups, 261 Liin's pap8h. 353 Nobel laureate immunologist. 262 Lp Salem (see Serumlipoproteins) photo, 254 Lominot (see Catalytictests, luminol) L.nda&m-Richter test (see ABO system, bloodstain grouping) Mherm sgstcm. 386387 LAP (see Leucine aminopeptidase) Lyon hppothk (sInheritance, sex-linked, X-chromosome inactivation) Lmes, Leone, 255 (See&Unit IX) a,M (see a,-Macroglobulin) Lath proadme (see ABO system, bloodstain grouping by isoagglutinin arMrroglobalin, 595 detection) AL-M marker. 617 La system(see Serum lipoproteins) polymorphism, 617 LDH(see Lactate dehydrogame) Xm marker, 617 Lcctin: Major hictoeomp.tibilitycompkx (see HLA system) anti-A:Dolichos, inhibition by secretor saliva, 282 W e debydmgmwe,423 Phaseohu inhibition by secretor saliva, 283 Maltose, 9 anti-B: Evonyrnus. 283,305 M.gGfinw8Id stah (see Stains, histological) anti-H: Qs?iacs, 273,282-283 Meiosis. 26,31 Elythrina, 282 Mdibi, 289 Laburnum, 273.282-283.305 Memory immune response (?Antibodies, formation) Index

Mendd, Gwr: nL (seeLnvissystem, in body fluids) convibutions to classical genetics, 20 NoagcllPetie markem: photo, 5 antibodies in blood. 669-670 Mensbd blood (see Identification of blwd, menstrualblood) classes of and description, 669 Metcbnikoff, Elic, 221 drugs in bloodsrains, 670 QMethplumbdtifergt phosphate (seeAcid phosphatase, substrates) hepatitis antigens, 670471 hlr (see MNSs system, variants of M and N) plasma ions and metabolites, 670 MIiC(see HLA system) smunprotein profiling, 671 MichwlieMenten kinetics, 16-19 Nmddc dds(see DNA, RNA) Micrometry(see Determination of species, micrometry) Nnckoddes. 21.23 Mkmcopial methods for blood identification (see ldentification of Nlldcotidar. 21.24 blood, microscopical methods) NomP, G.H.F., 22,241 Miltenbwgersntigcas(see MNSs system, associatedantigens) Mito&, 26,31 Oph-Matuhui pbenomcnou, 278 Mhrcd@utiuation: 0,(seeABO system, para-Bombay phenotypes) for ABO grouping of bloodstains (see ABO system, bloodstain group- Or (see a,-Acid glycoprotein) ing by mixed agglutination) Wb,M J.B.: different wanins of the term. 52 biography, 74 Mixed dl aggiuti&n reaction (MCAR), 311-312 early investigations on blood identification, 74-75 *(see MNSs system, vhtsof M and N) vhoto (see Unit IXl MNSs mm: &mid (see a,:~cidglycoprotein)

antisera and kctins. 338 -e (see Catalytic tests. o-Widinel' application to disputed parentage testing, 342-343 Or(Lo4olW.e (see~atal~ tests, o-tolidine) associated antigens. 338 (Mho-tohidk (see Catalytic tests, o-toluidine) biochemical genetics, 341-342 Mophorpbork-monoarter pbospbohydrohoc (alkaline optimum) (see bloodstain MN grouping by fluorescent-labelled antibodies, 312 Alkaline phosphatasc) daeaabity of antigens in aging samples. 344,345 011ebt&ny, 0.. 224-225 distribution of phenotypes in U.S. populations, 346-347 genetics. 335-336 P mm: heterozygous advantage and MN. 338 application to disputed parentage testing, 385 linkage of MN and Ss, recombmation and mutation, 335-337 biochemistry and bibchemical genetics, 383-385,386 MN antigens: amino add sequence structures, 342-343 bloodstain groupind, 385 anti-N reaction with M cells. 340 discovery and development. 381 anti-N reactions with Mdsand stains, 344-345 Q antigen, 383 bloodstain grouping by elution and mixed agglutination, 344 serology, 381-383 bloodstain grouping by inhibition, 343-344 Tjaantigen. 381-382 chemical studies, 339-340 Pa (seeSaliva, polymorphic protein systems) discovery, 335 a,-PAC (see Identification of blood, diagnosis of pregnancy from blwd- Ssantigens: bloodstain grouping of, 345 -1 discovery,335 PAPP (see Identification of blood, diagnosis of pregnancy from blood- U antigen, 337 -) &ts of M and N. 337-338 Pur-pkoykncdinminc (seeCatalytic tests, pphenylencdiamine) Vw typing in bloodstains, 346 Pb @e Saliva, polymorphic protein systems) Mdcerrhr &vhg (see Chromatography, gel fdtration) P.C.E. (see Chotinestcmcs, serum pseudocholinesteme) Morgan, Tho- Hunt: Pedigm (see Inheritance) Nobel lamein genetics, 20 Peancy mtigca (seeKell system) photo. 5 PEP (seePeptidascs) Mom,ss genetic map diceunit, 33 P-, 517-518 Mucins. 339 Peptuns Mucoids, 339 application of PEPA polymorphism to disputed parentage testing. Mncopolpslcrbarida, 339 507 "Mwem" hnzyme typing (see Isoenzyma, simultaneous typing biochemical properties of kcmyma, 507 of two or more systems) detection reactions in typing media and assay, 504,506 Mutation, 38 distribution of PEPA phenotypes in U.S. populations, 508 W (seeMNSs system. variants of M and N) electrophoreticpatterns of PEPA phenotypes, 508 Mydoma proteins, 604 geneticvariation of PEP A, B, C, D. S507 MyoLiarrP (sef Adenykte kinase) pwping PEPA in bloodstains. 507-508 pouping PEPA in semen. 508 N-AatyEnepd rid,structure, 10 linkage relations and chromosomal localization of PEPloci, 507 NADH-metbcmoglobii mdue(rsc (see Diaphorases) multiple genetic loci determining isoenzymes. 503-504 NaphthanilDhzo Blue B, 157,159 red cell and tisue isoenzymes, 503-504 Naphth.nn Diazo Red AL, 157.158 dative electrophoretic mobities of different isoenzymes. 504 Naphthanil DioRed RC, 157,159 substratespecifidtics of different isoenymes, 505 Naphthol AS phosphates (see Acid phosphatase, substrates) typing PEPA simultaneously with other isoenzyme systems. 508 Naphthol ydlow S. 174 Pcrh~~pc~t.n~ph~~thr~~,10 Naphthylphosphates(see Acid phosphata~e,subnrates) (See ahSteroids) N-cwtyl-Neunarinie acid, structure, 10 Pcriodierid,destruction of blood group receptors, 308 Nitrite (see Identification of saliva) Pcroxidrr: pNhpbenyl phosphate (seeAcid phosphatase, substrates) activity of hemoglobin (see Catalytic tests, peroxidase activity) Sourcebook in Forensic Serology, Immunologv, and Biochemistry

general reaction (see Catalytic tests, peroxidax reaction) Po-e gd deetrophorrsk (seeElectrophoresis) as one of fm isocnzymes, 423 Polymorphicproteinsin hair (seeHair) salivarypolymorphic(see Saliva, pol~morphicisoawmes) Potymorphic proteins h stivP (see Saliva, polymorphic protein systems) Pmx#c (see Cataytic tests, peroxide) P0)ymorphism: Pg (see Pepsinogms) asbasis for discrimination in populations. 39 PGD(see CPhosphogluconate dehydrogenase) defmition, 38-39 PGM (see Phosphogluwmutase) as population genetic phenomenon, 39 PCP (see Phospho~b~~hte~hos~hw) Pop~ltntion,frquenfies of genetic markm in a (See United States pop ph (Asaliva,&iiorphic ;rot& systems) ulations, frequencies of distniution of genetic markm) Pk.dpbth.kia phosph.tc (see Acid phosphatase, substrates) Population geneticp: Pk.olpbth.lia tcsl (seeCatalytic tests, phenolphthalin) Chi-square goodn~f-fitM, -1 pk.otypc, 27 discrimination index, 42 pPk~).lcnwlbmine(see Catalytictests, pphen~lenediamine) Hardy-Weinberg equilibrium, 394 Phemyhtoaoria, 20-21 probabiity of identity, 42 Pbmyipbosph.te(see Acid phosphatase, substrates) significe (usefulness) of genetic marker systems, 41-43 Pborphrte. colorimetric determination of, 162 PorphJlins: Pborplmtidk rid,structure. 9 heme and hematin compounds, 77-79 Phospb.tayldcrinhiw, structurr, 10 structure,types,nomenclature, 77-78 6-PboupLo-~+~0~NADP+-&xidorednetrv (decarboxylatiig) (see R (see Saliva, polymorphic protein systems) CPhospho@uconatedehydrogenase) R protcims, 595 Pborpboglaeomatrr: Precipiia: application to disputed parentage testing, 430 antGbactaiaiantibodies. 221 biochemical properties, 427.429 antibodiesagainstblood and body fluids, 221 biochemical studies on the isoenzymes, 429-430 antibodiesagainst milk proteins. 221 detectability of isoauymes in aging blood and bloodstains, 432-433 antibodiesagainst ovalbumin, 221 daeaion reactions in typing media, 431 as antigen-antibody reaction distribution of phenotypes in U.S. populations, 436-437 antitoxio antibodies, 221 effect of PMS conantration in detection donmixture, 432 mechanism, 53 electrophoreticpatternsof phenotypes, 428 optimization of reactant concentrations, 227 grouping in bloodstains. 431432 prepadon of precipitating antisera. 223-224 grouping in semen, vaginal secretions and other tissues, 433-435 quantitativeprocedures, 53-54 . . linkagerehtionshipsof PGM loci. 427 PdpWkm-inhibStion (see Ikmmmhon of species, immunolo@cal) PGM alleles. 425-426 Prrdpitin (at: PGM*alleks. 426427 for detrrmination of species of blood (see Determination of species, PGMI alleks, 427 immunological) FGMIalkks by isoe1ccttic focusing, 426 for identifkation of saliva (see Identification of saliva, anti-human PGMl silent alleles. 425-426 sativasaum) polymorphism. 425426 for identification of semen (see ldentifcation of semen, immunological) problems with bloodstain grouping, 433 Plseipitins,antibodiesas. 49 reaaion wchanisn, 430 Pmgimcy, forensic diagnosis of (see Identification of blood, diagnosis mwgnition. 425 of pregnancy from bloodstains) specisdeterminationby isoenzyme separation. 244 Pmgnmey-~~&~~Ia&coprotdns (see Identifibtion of blood, diag- subtypii of PGM, isocntyws, 426,432 nosis of pregnancy from bloodstains) typingsimultanmusly with other hoemyme systems. 432 Pmgemqrodotcd phsmr. proteins (see Ideatifion of blood, diag- &Pborph*Colllce dehydmgemsc: nosis of pregnancy from bloodstains) application to disputed parentage. 492 hgnmcyeproteh (see Identification of blood, diagnosis of biochemical properties of isoenzymes. 492 pregnancy from bloodstains) distribution of phenotypes in U.S. populations,494 Pmpaeey hormoacs (see Identification of blood, diagnosis of preg- geaaic variation and polymorphism: classes of genetic variants, 488 nancy from bloodstains) deticicnt activitydtsand silmt alldes, 490 Pmgemq zone protein (see identification of blood, diagnosis of preg- nomenclature. 4-92 nancy from bloodstains) normal activity variants, 488-490 PlimvJrimrmw response (see Antiies, formation) properties of miants, (table) 491 ProbahMty of cxddoa (see Disputed parentage testing) grouping in bloodstains. 492-493 ProbahPitgof ideaditg(seePopulation genetics) grouping in body fluids aud tissues. 493 Prodon B-t Red MZBS starch, 185 inaaivotion of enzyme by NADP and red cell stroma, 492 Pro- of Moodcomponen(s(see Nongenetic markers) linkagerelations of PGD. 492 Pmkop, Otto. 255 mgnition, 488 RosEltic dd pbospb.tpc (see Identification of semen, acid phasphatasc) typingsimultaneoustywithother~esystems,493 Pm&d&s. anti-A from snails. 283 (see Lipids) Ro(dn~~, 24-25.29 Pkspbglgcohle plospb.t.lw, 520 Pmlcb phpbobex~~ imacrer (seeGlucose phosphate isowrase) assay methods, 15 -bb, species determination using, 237 estimation of concentration in bloodstain extracts (see Determination (SeeakoLectins) of species, immunological) PbytoprrdpitiaP(see Determination of species, immunological) forcesinvohred in structure, 12-14 Pi (see crI-Antim) primary structure, 12 PL (see AIkaline phosphatase, placental) purification. 14-15 Phraionsud metaboptes(see Nongmetic markm) qUaternSry StNUUre, 14 Pm (see!?ah, polymorphic protein systems) secondary structure. 12 Index

specific activity. I5 supp~as,deletiom and modifiers, 356 tertiary structure, 12 variationsin C (rh') antigen. 353-354 Protopotphgrin IX,80 vPriatiom in E(rh")and e (br") antigens. 354 Pseadoeboliawtensc(see Cholincsterasa) Rho* B (see Caulytic tests, rhodamine B) PtJntim (see Amylaw) RIA (see Radioiimmunoassay) Poriat bms, 21-22 Rickt,

semen, 284-285 Sgd (see Saliva, polymorphic isoemymes) sperm cells, 284 Shlirrid (seeN-ucetyl-Neuraminic acid) Momin H level with different anti-H. 282-283 Sid aud Cd,390-391 application to disputed parentage testing. 297,313 Sidaophilin (see Transferrin) biochemical genetics. m293 Si,Viorio (see Unit IX) distribution of phenotypes in U.S. populations, 328 6P~hoglacolutcdehydrogensse (see at Phosphogluconate dehy- inheritance,281-282 dros-1 relationship of secraor and Lmis genotypes to red cell and body fluid SI.actoyi-@~~~mctbyI&oxnl lymc (iimerizing) (we Glyoxalase I) antbas,295 Sm (see Scianna system) relationship to ABO and Lewis. 290-293 SOD (see Superoxide dhutase) secretor gene symbols, 282 Sohddiue. 465 sew: Sdsnine. 465 ABH substances in (see Secretor system, ABH substances in body fluids) Spcdcsof ow(see Determination of species) ABO antibodiesin, 273 composition of, salts and low MW compounds, 1% Spermdiaphowc (see Diaphorases) (See &o Identification of scmea,composition of semen) Spernntozoa: content of genetic marker system proteins in (see Genetic marker sys- detection as means of seminal stain identification (see Identification of tem, content of proteins in blood, body fluids and tissues) Sane?, spermatozoa) enzymes (see Identification of semen, composition of semen) DIA, isoenzymes in, 519-520 genetic markers in (see Body fluids and tissues) discovery, 1%) genaic marker typing in (see Body fluids and tissues) morphology (seeIdentification of semen. spermatozoa) identification (see Identification of smren) structural (cytological) profiling (see Identifdon of semen, spemra- phosphogiucomutaw grouping of, 433-435 tom) rotei ins of (see Identification of semen, composition of semen) Spcrmiw, 174 uystd tests (see Id~n~cation of semen,crystal tests) (Seeahldentification of semen) Scminrl wid pImpb.1.s (see Identification of semen, acid phosphatase) SphinIcolipids (see Lipids) SendbW(see Hemobins) Sphgomydin stmd~~re,10 scudtkdprutida: Sphingo&w stmctufe, 10 colloidon particles, 23 1-233 st.inshistologial: hex particles. 233 Bieb~ichscarlet. %, 100 ~rceuancytest based on. 233 for blood ails in bloodstains (see Identification of blood, microscopical !&&on ifblood from deb* (see ~hromato~ra~hicmethods) methods) Semm group wm,concentration ranges of proteins in serum, 563-564 CI [colour index], 97 (Seeako Srmmproteins) eosin, 97,100 Sam Iboprotcias: Feuken, 97 A~s$A: application to disputed parentage testing, 613 gad,97 recognition and polymorphism, 611413 -97 rdationship of studies on to A#ralia antigen, 61 1,670 hematoxylin. 97.100 clasification and properties, 61 1 MayGriinwald. 97 Ld system,613 prepadon of Graham. % Lp system, 613-614 for spermatozoa in seminal stains (see Identification of semen, sperma- Sam orotck tozoa) antigbcity of heat denatured, 226 table of. 98-99 basisof metic variation in. 564 St.rch: electrophoretic and irnmunoelatrophoreticbehavior of, (illus.) 567 amylopectin. 187.188 as genetic markm, 563-564 amylose, 187,188 methods of separation and chaacterhtion, 564,566 dye-lioked assubstrate for amylase. 185-187 nature and ciassification of, 563-564 SWgel elpbord(see Electrophoresis) nomenclature and propaties summary,565 StPcLiodiae raction, 184 profiles, related to bloodstain age (see Age of bloodstains, dctnmina- St& tion of) cholesterol, 11 profiting of. 671 as classof lipids, 9 (See&Nongenaic markers) esuadiol, 12 (See ako Saum FOUP systnns) steroid nudeus structure. 10 SemrndckDts. 56 testosterone, 11 !3mm ions aud mct8boMes(see Nongenetic markers) Sm(seeMNSs system, U antigen) Sct (see Saliva, polymorphic hnymes) Subpups of A (AW ABO system) Scxcbromrtim: SucBgldlchdhe, 464 in cells andtissues as sex of origin indicator. 666 (SeeoLFo Cholinesterases) description and occumnce. 665 (see Carbohydrates) as related to Xchromosome inactivation, 34.38.665 Superoxidedlsmutae, 512 (See& Inheritance, sex-linked, X-chromosome inactivation) Suthcrlsnd, Willhm mnbar,74 as sex of origin indicatorin bloodstains, 66546 Sutte m@m (see KeU system) Sex Lanoome knlmanrrrmcnts, 668 Slmwllonhm (seeSuccinyldicholine) Sar of ow: smrt: of blood by srmm protein profJing, 671 ABO grouping(see ABO system,body fluidand tissue grouping) cytological methods: F-bodies. 646-668 composition. 1% sa chromatin,665-666 identification. 199 by determination of sex hormones, 668 Syutcay. 427 Index

T antigen: ABO system. 324-327 in the Hiibcnu-Thomsm-Friedenreichphenomenon, 339 adenosine deaminav system. 462 as original designation of Lea,329 adenylate kinase system. 443 TUBS, 45 a,-antitrypsin, 599 Takayamst, -0.86 carbonic anhydrase 11 system. 482 (Seealso Unit IX) cholinesterasevariants, 47W7 Tanned red cdh Duffysystem. 378-379 absorption of proteins by, 52,229-331 erythrocyte acid phosphatase system, 454-455 closelv related soccies differentiation and. 239 esteraseD system, 476-477 ~uono;nic&(seeDetermination of immunological) galactose-I-phosphate uridylyl transferase variants, 514-5 15 TBC (see Thyroxinebiding a-globulin) glucm-6.phosphate dehydrogenase system, 489 TBPA (seeThyroxine bindkg ;realbun& glutamic-pyruvic transaminase system, 502 Tc (seeTranscobalamin) glyoxak I system, 497-498 Tars: Gm and Km,610 ABO antibodies in, 274 mupspecific component, 586-587 identification of, 199 haptoglobin,579-580 specific protein in, 199 Hb variants, 560-561 Tccth: Kell system, 376-377 ABO grouping of tissue from (see ABO system, body fluid and tissue Kidd system, 378-379 grouping) Lewis system. 334 phosphoglucomutasegrouping in tissues of, 435 MNSs system, 346-347 (SeeoLFo Body fluids and tissues) Peptidase A system, 508 Teiehmlran, L., 80 phosphoglucomutase system. 436437 (SeeafsoUnit 1X) dpho~phogluconatedehydrogenase system. 494 Testosterone. 11,668 Rh system. 366-367 TetRwtbyl benddine (seeCatalytic tests, tetramethyl bmzidine) secretorsystem, 328 Tetnzdinm oxklase (seeSuperoxide dismutase) traasfmin, 594 Tf (seeTransfenin) UIPilS: Thin siDlch gels for dcetrophoretic iPocozyme typing. 431 of acid phosphatase activity (see Acid phosphatase, units of activity) Tbiocylaste (see Identification of saliva) of amylase activity (seeAmylases, units of activity) Thomrn phenomenon. 339 Urn (see Identificationof urine) Thomen-Fritdenreicb antigen, 339 Umse (seeIdentification of urine) ThymolphthrkiD phosphate (seeAcid phosphatase, substrates) UlidiDe monophosphste kiu=: Thyroxine biidii a-globalin, 619-620 polymorphism, 518 Thyroxine biidii pdbamin, 619 species determination by isoenzyme separation, 244 Tir. U h of agglutinatingantiserum, 53 ABO grouping (seeABO system, body fluid and tissue grouping) of precipitating antiserum. 53 wmposition (see Identification of urine, composition of urine) Titration-inhibin (see ABO system, bloodstain grouping by inhibition, identification (seeIdentifition of urine) Hirszfeld and Amzel-Kind technique) U.S. poplrhdions(seeUnited States populations) Tj' antigen (seeP system) o-Tolidime (seeCatalytic tests, o-tolidine) V8ginal dd pbospbltrie (seeAcid phosphatase, vaginal) o-Tolnii (seeCatalytic tests, o-toluidine) V~scarliom: TnnscobJ.min, 619 ABO antibodies in, 274 Tnnsferrio: cmicalmucus, 169 application to disputed parentage testing, 593 wmposition of, 168-169 in body fluids and tissues. 593 enzyws and proteins in, 169 distribution of phmotypa in U.S. populations, 594 epithelialcells: glycogen content of, 121-122, 168 electrophoretic patterns of variant gene products, 590 staining with iodine, 121 genetic variation and polymorphism, 589-590 identification of, 168-169 nomenclature of variants. 590 intravaghal pH, 168-169 589 recognition of, phosphoglucomutase grouping of, 433-435 structure and biochemical properties, 592 relationship of pH, vaginal flora and glycogen in epithelial cells, subtypes of Tf C, 590 168-169 typing in bloodstains, 593 vaginal flora. 168-169 variants arranged according to electrophoreticmobility, 591 vaginal infections. 169 Triglycede structure. 9 van Dan's tat (seeCatalytic tests, guaiacum) (See also Lipids) VAP (see Acid phosphatase, vaginal) Triple bonded agglu(hrtion technique. 31 1 Vuhbkexpmsivity, 30 VW.min B12Mlldiig pmteio (seeTranscobalamin) Uhknhuth,Paul, 221-222 V i D biding protein (seeGroup specific component) (See IX) ulw Unit von Behring, Emit A.. 221 Ultracentrifugation: as method for estimatingprotein purity and MW,16 Wbi blood cell, identification in stains (see Identification of blood, as protein purification technique, 15 microscopical methods) Ultmviokt [280/26@Jprotein ~osy(seeProteins, assay methods) White patch enymc (seeSuperoxide dismutase) UMPK (seeUridine monophosphate kinase) Wi,AhderS., 254 United States popnktions, frequeades of dinbution of genetic marker Wibn's dbaoc,619 phenotypes: Wwtmm. 391 1 Sourcebook in Forensic Serologv, Immunology, and Biochemistp I a,-X (see Antichymotrypsin) Yt sgrrtem. 390 Xb (see Identification of blood, diagnosis of pregnancy from blood- -1 Zeta potcntisl (seeAgglutination, mechanism) Xm (see a2-Macroglobdh) ZaDc phenomena (see Antigen-antibody reactions) Zwiscbenferwnt (see Glucosed.phosphate dehydrogmase) Y-bod& (seeF-bodies) Zsuwgm~,423

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