California Association of Criminalists

GENETIC MARKER TYPING OF NAF BLOODS
Rhodes, Carol N.; Schliebe, Stephan A.; California Laboratory of Forensic Science, 17842 Irvine Blvd., Suite 224, Tustin, CA 92680

The degradation effects of sodium fluoride on whole bloods are well known. However, frequently requests are received to compare a NaF-preserved evidence blood sample to a standard whole blood sample. A study examining the typability of the most common enzyme and serum protein systems utilized in crime laboratories will be presented.

SENATE BILL 809
Carleton, William, California Department of Justice, 6014 N. Cedar St., Fresno, California 93710

Senate Bill 809 requires that before a registered sex offender be released from prison that he provides a blood and saliva sample for testing of various genetic markers. Over four thousand samples have been analyzed for ABO, Lewis, PGM, and Pep A types. Statistical examination of this data is presented.

DOT IMMUNOBINDING TESTING FOR SPECIES IDENTIFICATION
Super-Mihalovich, J. and Sensabaugh, G.F., Forensic Science Group School of Public Health University of California Berkeley, California 94720

The practicability of using the dot immunobinding technique for species identification was investigated. Whole blood from various animals was tested using antibodies to immunoglobins IgG and IgM. The immunodot blot method can be used as a rapid and discriminating screening technique for he detection of human antigen present in whole blood.

IDENTIFICATION OF MN BLOOD GROUP ANTIGENS IN BLOODSTAINS USING WESTERN BLOTS
Calandro, L., and Sensabaugh, G.F.; Forensic Science Group, School of Public Health, University of California Berkeley, California 94720

We have previously described an enzyme immunoassay for M antigen in bloodstains (CAC Seminar, May 1986). We describe here a Western Blot procedure for the detection of glycophorins which exhibit the MN determinants. Bloodstains were extracted in a buffer containing SDS, glycerol, TRIS-HCI, and mercaptoethanol. The extract is electrophoresed by the SDS-PAGE technique. The transfer of the glycophorins to nitrocellulose is accomplished using the Western Blot technique. The MN blood type is determined by using staining procedure which involves the use of anti-M and anti-N monoclonal antibodies. A blind trial study of over 80 bloodstains has been completed. No incorrect results were obtained. The Western Blot technique has enabled us to correctly determine the MN blood type in blood stains stored frozen and at room temperature.

ABH PERCURSOR STRUCTURES IN SEMEN, SALIVA AND VAGINAL FLUID
McGinnis, M.D. and Scnsabaugh, G.F., Forensic Science Group, School of Public Health, University of California Berkeley, California 94720

A, B, and H blood group specificity is determined by the terminal sugar attached to a precursor carbohydrate structure on a protein or lipid carrier molecule. It has been shown that differences in sugar linkages on the precursor chain can be used to distinguish four types of structures. Further differentiation of the precursor is possible based on the extent of fucosylation; the chains are either mono- or difucosyl. Evidence suggests variation among body fluids with respect to precursor structure. Of forensic interest is the potential to distinguish mixtures of body fluids belonging to the same blood group. With an enzyme immunoassay we have used a panel of monoclonal antibodies, each possessing a distinctive affinity for a given precursor structure, to test semen, saliva, and vaginal fluid samples. Semen and saliva both show some variation in precursors detected.

THE SIMULTANEOUS PHENOTYPING OF ADA, AK, EAP, ESD AND PGM BY ISOELECTRIC FOCUSING
Kuo, Sze-Em, Los Angeles Police Department, 150 N. Los Angeles Street, Los Angeles, California 90012

A method is presented here that allows the simultaneous phenotyping of five enzymes in blood stains. Realistically in case work only four enzymes can be typed simultaneously with one sample application. The gels (180 x 130 x 0.25mm) are cast by the flap technique using a solution containing 7.7ml acrylamide stock (5.7%T, 3%C), 0.28gm MOPS, 1.7mg APS, 0.25ml pH4-6 Ampholine (R) and 0.25ml pH6-8 Ampholine. The gel is prefocused at 10ma limit to reach 1900V. Samples are placed midway between anode and cathode and focused for 80min. at 1900V. The isozymes are visualized by the methods used in the conventional electrophoresis methods.

INTER AND INTRA-INDIVIDUAL VARIATION IN THE LEVELS OF ABO BLOOD GROUP SUBSTANCES IN HUMAN SEMEN
Blake, Edward T.; Cook, Charles E.Jr., Forensic Science Associates, 1400 53rd Street Emeryville, CA 98608; and Bashinski, Jan S., Oakland Police Department

The ABO genetic marker system is one of the most useful genetic markers in the analysis of sexual assault evidence due to the stability of the antigens in the dried state and the high levels of antigen material found in the semen of secretors. The central problem in interpreting ABO antigen data in mixed stain samples is to determine the significance to be placed on the absence of particular antigens and/ or their relative levels. The answers to such questions require knowledge of the amount of semen in a specimen together with the amount of antigen activity normally found in the semen of secretors. The results of this work support our previous findings that the ABO antigen levels in semen are log normally distributed as are the ratios of A and B antigen to H antigen. The distribution statistics provide a basis for setting guidelines to interpret the absence of antigen activity and the significance of antigen ratios. These studies also demonstrate that the intra-individual variation and the variation within an individual over time is small compared to the population variation. Thus, reference semen specimens are useful in cases where an unusual donor is suspected,

INTER AND INTRA INDIVIDUAL VARIATION IN THE LEVELS OF ACID PHOSPHATASE AND P30 IN HUMAN SEMEN
Blake, Edward T.; Cook, Charles E., Jr., Forensic Science Associates, P.O. Box 8313, Emeryville, CA 94608; and Bashinski, Jan, Oakland Police Department. California Association of Criminalists Semi-Annual Seminar, Palm Springs, California, October 9-11 1986

Central to the interpretation of data from sexual assault evidence material is an understanding of the relative amounts of semen in sexual assault specimens. Knowledge of the amount of semen on vaginal swabs can provide clues as to post coital interval and knowledge of the relative semen dilution in stain extracts can aid the interpretation of the absence of ABO antigens. Acid phosphatase (ACP3) and p30 are potentially useful for estimating semen levels. We have surveyed semen specimens from 439 individuals. The results of this analysis reveal that ACP3 and p30 levels in semen are log-normally distributed. The distribution statistics provide a basis for selecting a normalization value for estimating semen levels in evidence specimens. These studies demonstrate that the intra-individual variance is significantly less than the population variation and is relatively invariant over time. Thus in any particular case a reference semen specimen can provide additional information if an unusual donor is suspected.

INTOXILYZER 5000
Carleton, William, California Department of Justice, 6014 N. Cedar, Fresno, California 93710

Since September, 1985, the Fresno Regional Laboratory has been using the Intoxilyzer 5000 with computer interface. The data base has been examined in order to compare alcohol levels in breath samples with the alcohol level in blood samples which were taken at nearly the same time. A comparison is given of that data. Also a brief description of the data management system with respect to the Intoxilyzer 5000 will be given.

PYROLYSIS PRODUCTS OF STRUCTURE FIRES
DeHaan, J.D., BATF, San Francisco Lab Center, Bldg. 233, Treasure Island, CA 94130, and Bonarius, Kristin, Institute of Forensic Science, Oakland, CA 94607

Common floor-coverings (nylon carpet, indoor-outdoor carpet, Astro-Turf, vinyl flooring, jute pad, foam rubber pad and polyurethane pad) were exposed to a real-life fire environment consisting of an ordinary combustible fuel load in a normal size room. Samples of these floor coverings were recovered immediately after extinguishment and were subjected to lab testing via heated head space and charcoal trap/CS2 extraction. Astro-Turf and polyurethane padding were found to present a significant volatile hydrocarbon signature after such exposure. In a second test, several floor coverings were burned in a room fire involving the use of three common petroleum products. None of the floor coverings presented significant interference to identification of these products in the post-fire debris. The difficulties and value of reproducing full-scale fire environments and their effects on combustibles will be discussed.

A NEW DIMENSION WITH "MIKROSIL" CASTING MATERIAL
Barber, Dave and Cassidy, Frank, California DOJ, 820 Francis Botello Rd., Goleta, CA 93117

Mikrosil, the silicone rubber casting material designed for forensic applications, is widely accepted as one of the best casting materials for the replication of toolmarks, impressions, gun barrels, etc. One drawback of the material for forensic applications is its viscosity. The possibility of altering the physical properties of Mikrosil in order to decrease the viscosity while still retaining its desirable replicating characteristics was investigated. It was found that it was possible to dilute the Mikrosil with one of the Dow Coming oils and still obtain excellent reproducibility of the surface in question. The testing program and the recommendations will be presented.

BAREFOOT IMPRESSIONS - A PRELIMINARY STUDY OF IDENTIFICATION CHARACTERISTICS
Laskowski, G.E.; and Kyle, V.L., Regional Criminalistics Laboratory, 1431 L Street, Bakersfield, California 93301

Footprint impressions of one hundred and seven male adults ranging in age from nineteen to sixty-seven years were recorded and examined. Included in this study were foot impressions from a pair of monozygotic twins. The impressions were recorded and photographed. A series of measurements were then recorded and converted into a set of indices which essentially are width vs. length ratios of prominent features of the human foot. These indices were then correlated to yield probability values for use in this study and for comparison to data published by previous investigators Qamra, Cassidy, and Robbins. Friction ridge minutiae were not considered in this study; however, crease marks, well impressions, and toe step measurements were considered in the study but not incorporated in the probability values due to the unique aspect of these features and the inability to convert these features to mathematical indices. These features introduce a subjective nature to the analysis scheme. This study utilizes the combined index probabilities of foot impressions so that the data can be used to assign a given probability to a particular foot impression without clear "individual" features, so that it can be linked to the person who left the impression.

POSTMORTEM RECOVERY OF DUSTY FINGERPRINTS USING THE ELECTROSTATIC DUST PRINT LIFTER
Ojena, Steven M., Contra Costa County Sheriffs Department, 1122 Escobar Street, Martinez, CA 94553

In a test situation dusty fingerprints were placed on human skin. These invisible fingerprints were successfully lifted using an electrostatic dust print lifter. The conditions for lifting a dust print from skin will be discussed as well as photography of the resultant lift.

BURIED BODIES
Chisum, Jerry, California DOJ, 2213 Blue Gum Avenue, Modesto, CA 95351 and Sheldon, Karen

The location of a grave site and the excavation of the remains can cause the criminalist considerable problems. This is partly because a number of "myths" about how to find a grave and partly because of a lack of knowledge about how to proceed. We will discuss various methods available for locating a grave, methods for documenting the grave contents in three dimensions, and how to best excavate the body. We will also discuss crime scene management and management of resources pertinent to a major site. We will illustrate with examples from a recent case.

PRIMER COMPOSITION AND GUNSHOT RESIDUE
Matty, William, California Department of Justice, 1500 Castellano Road, Riverside, California 92509

The testing of samples for the presence of gunshot residues has increased significantly in the past few years. This testing usually involves the identification of metallic elements known to be found in ammunition primers. These elements and their purpose in the primer will be discussed.

ANALYSIS OF GUNSHOT PRIMER RESIDUE COLLECTION AND SWABS USING FLAMELESS ATOMIC ABSORPTION SPECTROPHOTOMETRY: A REEXAMINATION OF EXTRACTION AND INSTRUMENT PROCEDURES
Havekost, D.G., Koons, R.D. and Peters, C.A., FBI Laboratory Washington D.C. 20535

Methods of extraction of gunshot residue (GSR) swabs for determination of Sb, Ba, and Pb by flameless atomic absorption Spectrophotometry (AAS) have been studied. Optimum extraction parameters were determined using recovery studies of Sb, Ba, and Pb added to swabs at amounts representative of actual GSR and handblank levels. Two difficulties with most extraction-AAS procedures are incomplete extraction of Sb and incorrect Ba results arising from improper matching of sample and standard matrices prior to AAS determinations. Utilization of standards made by spiking analyte elements onto swabs and extraction along with samples by an efficient procedure minimizes errors in Sb, Ba, and Pb determination.

GUNSHOT RESIDUE IDENTIFICATION BY PARTICLE ANALYSIS, AN OVERVIEW OF THE ESSENTIAL ELEMENTS OF AN EFFECTIVE S.E.M.-E.D.X. ANALYSIS PROGRAM
Sprague, Anthony E.; Hinkley, Robert L.; and Cooper, Robert, Alameda County Sheriffs Criminalistics Laboratory, 15001 Foothill Blvd., San Leandro, California 94578

Electron Microscope-Energy Dispersive X-ray particle analysis is a well established method in forensic science. The Alameda County Sheriff's Crime Laboratory has utilized this methodology exclusively for the determination of gunshot residue on evidence handlift samples since 1978. The validity of this method and its importance to the investigative and judicial processes has been proven by an effective gunshot residue analysis program in Alameda County. Some of the key factors of this successful program are reviewed and illustrated.

ADVANCED SEM INSTRUMENTATION FOR MICROSCOPIC COMPARISONS AND AUTOMATED GUNSHOT RESIDUE (GSR) ANALYSIS
Owens, Tony, and White, Robert S.

SEM-based systems have been developed to aid the forensic scientist in two common tasks - the identification of (GSR) and the microscopic comparison of two samples. The GSR package is a fully automated, microprocessor-controlled system utilizing a back scattered electron detector and a conventional EDS system. The automated search covers a 1cm² sample area divided into 65536 independently accessible coordinates approx. 40 x 40μm in size. The location coordinates of each positively identified GSR particle may thus be stored in memory and automatically relocated at a later time. Automated GSR searches of multiple samples are currently being implemented. The SEM comparison system utilizes an additional deflection coil and a dual stage module, allowing real-time comparison of two samples using various image types. The dual stage module is quickly interchangeable with the conventional stage and allows independent rotation and x-y translation of the two samples. The additional deflection coil allows user selected areas in both samples to be rastered alternately at TV rates in order to generate effectively simultaneous images of the two samples. Examples of the uses of each system will be presented.

THE CONCENTRATION AND ISOLATION OF GUNSHOT RESIDUES FOR PARTICLE ANALYSIS
Sugarman, Loren A., Orange County Sheriffs-Coroner, Criminalistics Laboratory, 550 N. Flower St., Santa Ana, California 92702

An efficient, cost effective method of concentrating GSR particles for SEM/EDX analysis has been developed, using commercially available, disposable materials. A 25mm diameter adhesive sample is concentrated to a 5mm area on a polyester membrane filter having a 0.4 micron pore size. Particles are released from an adhesive coated disc of polypropylene by dissolving the adhesive with trichloroethylene in a micro-centrifuge filter tube. After centrifugation, the light-particle fraction is aspirated from the top of the solvent. The density separation is repeated using bromoform. High density particles are deposited onto the filter by centrifuging the unaspirated solvent through the membrane. Sample preparation requires 15-20 minutes. Recovery efficiency, evaluated by FAAS, was determined relative to the total Pb and Sb from the aspirate and filtrate fractions. 90% of the total GSR was recovered on the membrane, the remainder was found in the aspirate.