84th SEMI-ANNUAL SEMINAR (Fall 1994)
Pasadena, California

D C Blakey, QPM MBA, Chief Constable, West Mercia Police Headquarters, Worchester, England, UK

The pressures on chief officers to detect the bulk crimes of theft and burglary; the pressures on police budgets; the detection techniques available; how forensic science compares in cost and effectiveness and the future uses of forensic science will be covered during the presentation of this paper.

Sergeant David R Asbbaugh, Forensic Identification Specialist, Royal Canadian Mounted Police

Friction ridge identification has already entered its second century of use as a method of personal identification but is only just approaching maturity as a science. Recent challenges to the current friction ridge identification philosophy, methodology and scientific basis have fostered review and modernizing of the science. The study of the uniqueness of friction ridges and their application to the identification process has been dubbed Ridgeology. Ridgeology advocates an evaluation in the aggregate of all available friction ridge details which were formed through differential growth or permanent damage. The Ridgeology presentation will give an overview of the premises of friction ridge identification and the need for an expert to understand the embryological and historical aspects of the friction skin. A few keystone research papers which lead up to modem friction ridge identification will be briefly reviewed. The identification process, the expert's role in court and the current position of only presenting positive identifications will be discussed.

Supervisory Special Agent JA Dizzino, FBI Forensic Science Research and Training Center, Quantico, VA

On April 19, 1993, approximately 80 individuals died in the Branch Davidian compound near Waco, Texas. As a result of an intense fire which made identification very difficult, the human identification process required a multidisciplinary scientific approach. The identification process began with crime scene processing and concluded approximately one and one-half years later with the conclusion of DNA testing procedures. Crime scene processing and the collection and preservation of human remains were critical to the identification process. The collection of pre-mortem medical, dental and fingerprint records was made difficult due to the diverse backgrounds of the deceased individuals.

After attempting all means of conventional human identification, DNA typing procedures were initiated on tissues gathered from the unidentified individuals. Reference DNA samples were gathered from around the world from blood relatives of deceased individuals for DNA comparison purposes. DNA typing procedures were employed to identify approximately one-half of the deceased individuals. The personal identification process utilized in the Waco incident requires the expertise of a wide variety of forensic scientists and provides a benchmark for personal identification in mass disaster situations.

Arie J, Zeelenberg, Head, National Fingerprint-Service, The Netherlands

Results of fingerprint investigations are presented as hard evidence. Conclusions are not expressed in grades of probability but as absolute certain, identifying one person at the same time claiming the exclusion of all possible other donors. It is known however that mistakes are made, false and overstretched identifications are, reluctantly, reported. For society the results of the process are the most important aspect of fingerprinting all others are secondary. Although numerous books and articles deal with fingerprints very little is written about this aspect and almost nothing is regulated. There is a number of good reasons to require guidelines, procedures and standards for the identification process in particular for latents on the edge of quality. In order to be able to guarantee the absolute outcome of any process in depth knowledge of all factors involved is vital if only to be able to control it. The definition of the process is also required for training of newcomers. The fact that the evidence has an empirical basis suggests that standards are maintained about how to do it. Scrutiny and verification of identifications requires known and com-mon standards. Since they don't exist a real challenge is virtually impossible (with the exception of apparent mistakes). As a result the dispute whether an identification is justified will likely end in a deadlock about taste or opinion. For the sake of fingerprinting it is doomed to be covered up with brotherly congeniality. The last but not the least reason to have well defined standards is the fact that we ask utmost confidence from society. This confidence can only be based upon well defined and public rules of the game applied by experts. With this in mind the presenter will look at indicators in order to establish how scientific we operate. It will be argued that the fingerprint community in general has failed to address this problem adequately. An outline of a complete alternative approach is presented.

John DeHann, California Criminalistic Institute, Sacramento

Many new techniques have arisen for the recovery of latent fingerprints that today allow almost every surface some chance of producing identifiable prints. Even in the presence of contaminants such as water, greases, food, soot, or sugars, prints can still be recovered. Leaves, bricks, fabrics, concrete, and adhesive films have all yielded prints. Thanks to the development, by scientists on both sides of the Atlantic and Pacific, of powders, stains, dyes, films, filters, and special light sources, the crime scene examiner stands of five times better chance of recovering prints than was the case just 15 years ago. Paralleling these technological leaps in finding prints, have been dramatic advances in comparing the prints recovered to data banks of suspect prints. Automated latent print and AFIS systems have made it possible to identify single prints from thousands of candidates in both state and regional records. This symposium will describe the advances in selective and sequential processing of evidence using both fluorescent (dyes and powders) non-fluorescent (dyes, stains, vacuum metal and powders) techniques, all of which are enhanced by innovative light sources. The functions and features of automated print systems of various types will be revealed along with discussion of what the future could hold for what remains the most individual form of personal identification.

K G Barnett, Birmingham Forensic Science Laboratory, England, UK

Techniques for the enhancement of footwear impressions will be reviewed with a particular emphasis on those made in blood. The methods used for both porous and non-porous surfaces by the Birmingham Forensic Science Service Laboratory will be discussed in detail. Casework studies will be used to show how these techniques have been used and developed in investigating serious crime. Some problems raised from their use in court will also be discussed along with a method of connecting the small amount of blood normally present in an impression with a particular individual.

M A Green, Department of Forensic Pathology, University of Sheffield

In recent years there has been increasing international interest in forensic facial reconstruction for identification purposes. The growth of this interest has been accelerated by the development of readily available and relatively cheap computer technology. From late 1993 through the spring of 1994, in conjunction with staff of the Department of Archaeology in the University of Sheffield, I undertook a detailed review of the "state of the art" in the United Kingdom, Canada, and certain centers in the United States of America. A detailed literature search was carried out, and the history of facial reconstruction was reviewed. A postal survey of the opinions of archaeologists, mainly in the United Kingdom, was also undertaken, and their opinions incorporated into the report. This paper will review the methods currently employed by different practitioners in the field of facial reconstruction, and will offer suggestions upon standards of training, of quality assurance, and possible research avenues which could be explored in the future.

Pat Pitt, Superintendent, Hampshire Constabulary

Superintendent Pat Pitt's presentation will deal with the emergence of the AFR Consortium as an organization dedicated to the provision of a networked AFR service to 35 locations in England, Wales and the Channel Islands. The operational need that generated the project, ifs progress, technical and organizational challenges and deliverables will be discussed. The service will be described and statistics concerning usage and results will be presented. The future strategy for 'the development of the system, it's place in the wider world of AFR and it's migratory possibilities will be considered together with the organizational and technical lessons learned from the endeavor. The presentation will look briefly 'over the horizon' at the future of networked AFR systems and the business and operational benefits that are available both now and in the future.

Colin M. Livingstone, Police Consultant, Public Service Business, 1, New Square, Bedfont Lakes, 081 8184234

IBM is a committed supplier to the Police Market Place across the world. Along with its partner Sagem of France they supply AFR (AFIS) systems throughout the world. To date, the system they have installed in the United Kingdom, is one of the largest in the world. Together they are currently developing future enhancements to their AFR (AFIS) system. IBM are also developing other exciting solutions for the Police Market Place which include video for surveillance and enhancement.

Donald J. Johnson and Ronald R. Linhart, Los Angeles County Sheriffs Department Scientific Services Bureau, 2020 West Beverly Boulevard, Los Angeles, California 90057

The products of an abortion are often submitted as the only physical evidence of rape or incest. Recovering embryonic or fetal tissue permits the use of paternity testing to evaluate suspects. Here we report on the products of conception retrieved from induced (therapeutic) and spontaneous (miscarriage) abortions, using low magnification stereomicroscopy. By way of review, the embryonic period begins with the formation of the embryonic disc at approximately two weeks, and continues to the end of the eighth week; the fetal period extends from the ninth week until birth. Abortion methods vary with gestation age; however, the majority of case submissions are produced by instrumental evacuation of the uterus. This procedure results in the fragmentation of the embryo/fetus, embryonic/fetal membranes, placenta and umbilical cord, and decidua (uterine endometrium). Typically, abortion products are enclosed in a gauze sac immersed in blood; the wet weight of the sac and contents has ranged between 68.8 and 104.8 grams. The bulk of the specimens are admixtures of fine to sometimes coarse pieces of maternal and extraembryonic or extrafetal tissue-the percent weight of embryonic or fetal tissue proper has ranged from 1.3 to 9.4% (1.4 to 10.4 grams). With induced and spontaneous abortions, the types and amounts of tissues and organs recovered vary. However, the vertebral column has been consistently found in fetus cases. Extraembryonic/extrafetal tissues form supportive structures, which primarily are unincorporated into the embryo/fetus. Importantly, these tissues are derived from the zygote, and therefore genetically identical to the embryo fetus. The chorionic membrane with immature villi(extraembryonic structures) have been recovered in cases where the embryo proper was either not submitted or identifiable. Lastly, tissue samples have been obtained from intact fetuses aborted by the medical induction of uterine contractions during the second trimester.

S-B. Lee, M. Ma, K. Brown, B. McNamee, J. Bashinski and L. Gima, Department of Justice DNA Laboratory, Berkeley, CA 94710

Rapid, reliable extraction of DNA from blood for genetic typing is desirable in forensic laboratories constructing genetic databases that routinely examine several thousand samples per year. Furthermore a rapid, reliable extraction technique on hair samples is also desirable for forensic DNA analysts conducting casework. Amplified short tandem repeats (STRs) and mitochondrial DNA D-loop regions (mtDNA) provide useful genetic markers for identification. The objectives of this study were to 1) Evaluate a microwave mini-prep technique (.Goodwin and Lee. 1993 Biotechniques 15:438) for extraction of DNA from liquid blood, blood stains, single hairs and hair shavings and 2) amplify STRs and mtDNA from the microwave extracted DNA. Blood and hair samples in stain extraction buffer were incubated for 5-10 minutes at 85C, then microwaved for 10 seconds. Proteinase K is then added and the sample is incubated for 15 minutes. All samples were then subjected to two organic extractions and then the DNA was purified using a size exclusion column (Centricons). DNA visualized under UV was quantitated by comparison to standards. The DNA was transferred by capillary blotting to a nylon membrane and quantitated using chemilumi-nescence (ACES kit) as described (Zee, et al. 1995 AAFS Meeting). Thol, CSF1PO and TPOX were amplified from 5-10 ng of DNA, separated by acrylamide gel electrophoresis and detected by silver staining using Promega protocols. MTDNA was amplified using previously published protocols (Sullivan, et al. 1992. Int.J. Leg. Med. 105:83). Microwave extraction from blood yielded from 400-1200 ng per 50μl of blood. DNA extraction from 1-3 hairs yielded between 50-150ng. Amplification of STRs using 5-10 ng from every type of tested sample demonstrated the utility of microwave DNA extraction from blood and hair for PCR. mtDNA was successfully amplified from microwave extracted DNA and we are currently sequencing the PCR products to confirm these results. We are conducting further evaluation of the quality of the microwave extracted DNA for other PCR genetic marker systems (D1S80 and DQ alpha) and RFLP.

O.R. Gibb, P.C. Hau, J. Dunlop and Y.F.Mclaen, Police Forensic Science Laboratory, Dundee, Scotland

DNA profiling is typically carried out using Single Locus Probes (SLP's) where polymorphism is due to highly variable repeat unit number in minisatellite DNA. MVR typing utilizes differences discovered within the repeat units themselves. Using a specialized form of PCR (Amplification Refractive Mutation System or ARMS) it is possible to generate a diploid MVR code from extracted DNA. The effect of varying the amount of template DNA and the, number of PCR cycles was studied to find optimum amplification conditions. DNA was extracted from blood, hairs, saliva and semen stains and MVR typing was performed. No single set of amplification conditions was found to be optimum. MVR maps were produced from each of the sample types studied, including a clear legible code from a single hair root. The advantages and disadvantages of MVR typing in comparison with the DNA profiling techniques in current use are discussed.

M. Piucci, D. Butler, M. Buoncristiani, K. Konzak and S-B. Lee; Department of Justice DNA Laboratory, Berkeley, CA 94710

The expected result for heterozygotes in restriction fragment length polymorphism analysis (RFLP) generated with the restriction enzyme Hae III using variable number of tandem repeat probes (VNTR) is the detection of a two band pattern representing two different sized alleles. We have infrequently observed more than the usual two bands for heterozygotes in Hae III RFLP analysis (particularly at locus D10S28). These 'extra bands' usually come in pairs, are reproducible and consistent for a given sample and are always smaller than the primary band(s). These results suggest that this phenomenon is not due to incomplete enzyme digestion (partials), mixtures of samples, or 'extra bands' due to unstripped probes from previous hybridizations. This phenomenon has been observed for control DNA samples (K562 cell line) and DNA extracted from blood or semen stains. The 'extra bands' produced are not equimolar with the primary bands (usually less hybridization signal is detected for the 'extra bands'). We are evaluating the possibility that these are the result of a relaxation of the Hae 111 endonuclease specificity for the sequence GGCC to include similar but specific sequences. Purposeful overdigestion (with orders of magnitude greater enzyme concentration than our current protocol) can produce similar 'extra band' patterns, but because this was not a systematic result for every set of samples, we believe there may be other factors involved. We have also observed that DNA extracted from reference blood samples stored as liquid in preservatives (as opposed to stored as a stain) produce 'shadow bands' that are predictably detected just below the primary diagnostic band. This band often appears around 150 base pairs below the diagnostic band, again primarily with D10S28. This band is often unresolvable from the diagnostic band, especially in longer autorad exposures. It is worth noting that the size of this difference is approximately the same as the amount of DNA wound around nucleosomes, which are known to be hypersensitive sites.

Yiwen Wang(1), Jingyue Ju(1), Brooke Carpenter(1), Jeanette Atherton(2), Richard Mathies(1), and George Sensibaugh(2)

(1) Department of Chemistry, (2) Forensic Science Group, School of Public Health, University of California, Berkeley, 94720

The implementation of DNA profile databanks will require a DNA typing technology and information management system that can accommodate the typing of many hundreds of samples per day. This requirement defines to some extent the markers chosen to establish the database. With this consideration in mind, we have investigated the use of capillary array electrophoresis (CAE) for the high throughout typing of polymorphic short tandem repeat (STR) loci. We demonstrate here typing at the TH01 locus using a two-color labeling system in which the allelic ladder is labeled with one fluorescent dye label and the test sample is labeled with a second. Labeled PCR products are detected using a laser excited confo-cal fluorescence scanning system. Near baseline resolution is achieved for the 4-bp repeat allelic products in 20 minute runs using capillaries filled with 0.8% (w/v hydroxyethyl-cellulose,(BEC), 0.5X TBE pH 9.3, and 1μM 9-aminoacridine. The 9.3 and 10 alleles, which differ by a single base, can be distinguished in isolation but could not be resolved in a 9.3,10 heterozygote. With this exception, genetic typing at the TH01 locus is unambiguous; typing results can be translated by software directly into database files. A very large number of polymorphic STR loci are known and use of five or more, singly or in multiplex, can provide high discriminatory power.

R. Rubin, T. Worley, D. Hanzel, E. Mansfield, Molecular Dynamics

The closing this summer of the Barnwell, South Carolina radioactive waste site to California and other states has accelerated interest in non-isotopic DNA detection methods. The new era of fluorescence imaging offers a versatile, high throughout, quantitative complement to chemiluminescence for non-radioactive methods. For human identification analysis, we have recently used the Molecular Dynamics Fluorlmager Scanning System for D1S80 typing and STR multiplex analysis in polyacrylamide gels and single-copy Southern blot detection of RPLP's, Additionally, we have used the Fluorlmager System's 96-well microplates scanning capability for high throughout quantitation of total DNA in human samples. For the PCR-based assays, D1S80 and STR, the Fluorlmager detects either fluorescein labeled or unlabeled PCR products. The unlabeled assays require just a 10 minute post-electrophoresis stain in SYBR Green I (Molecular Probes) with no destaining step. In either case, wet gels are scanned eliminating any drying, fixing or blotting time. In RFLP analysis we have detected target DNA in as little as 100 ng (0.05 attomole) genomic DNA using a single locus probe to D2S44 (YNH24) (Promega Corp.). We adapted a chemiluminescent protocol using alkaline phosphatase coupled to the probe. The fluorescent substrate, AttoPhos (JBL Scientific) is substituted for a chemiluminescent substrate eliminating the use of film. Typical scans on the Fluorlmager System require under 5 minutes. Scanning occurs off-line of electrophoresis allowing the scanner to be shared by several tabs which processes many gels or blots in parallel. Fluorescence offers a greater dynamic range and more linear response than film used in autoradiography or chemiluminescence. Images are digitized and become available immediately for quantitative image analysis using Molecular Dynamics ImageQuaNT and Fragment Analysis software programs. Both image analysis packages completely preserve the integrity of the raw data files while allowing optimal on screen contrast of dark and weak spots over the entire four order dynamic range of the instrument.

N.M. Duda, *J. W. Sehumm; *C. Sprecher, *A. Lins, and S.B.Lee.
Department of Justice DNA Laboratory, Berkeley, CA 94710; Promega Corporation, 2800 Woods Hollow Road, Madison WI 53711

Polymorphic short tandem repeats (STRs) are valuable genetic markers because they exhibit easily detectable variation in the number and content of repeats among individuals. They are particularly useful for forensic applications because DNA extracted from evidentiary materials is often degraded. Since STRs are relatively short (139 to 327 bp) and smaller target sequences are more readily amplified than larger ones, analysis of degraded DNA samples is possible.The objective of this study was to optimize a multiplex PCR co-amplification approach for the determination of STRs and gender identification using the amelogenin gene (Nakahori, et al. 199 1, Akane, et al. 19 1, Garcia, et al. 1994. CAC Meeting presentation). The triplex containing the STR loci THO 1, TPOX and CSF1PO was co-amplified with different pairs of amelogenin primers, separated by polyacrylamide gel electrophoresis and detected by silver staining using previously described protocols (ProMega) except for the following change. For the co-amplification with the first primer pair described by Garcia et al. (1994), we evaluated different temperatures for annealing. Different amelogenin oligonucleotide primers yielded varying results. The primers previously described by Garcia et al. (1994) produced 540 and 356 bp fragments for the X and Y sequences respectively when amplified alone. Although the amelogenin products could be co-amplified with TH01 or CSF1PO, they were not successfully co-amplified with the triplex. Differences in the optimal temperature of annealing between the amelogenin primers (57°C and 71°C) and the suggested optimal temperature for the STR primers (64°C) may explain the lack of co-amplification. A second amelogenin primer pair (provided by ProMega) produced 212 and 218 bp fragments for the X and Y sequences respectively. While these were successfully co-amplified with the STR triplex, the close proximity of the X band (212 bp) and the largest TH01 allele (203) suggests a potential for typing error if shadow bands and/or extra band artifacts are produced. We have therefore designed and synthesized a new set of amelogenin primers which produces 552 and 547 bp fragments that have the same optimal temperature of annealing as the STR triplex (64°C) and that do not overlap any of the STR alleles in size. Current evaluation of the efficiency of co-amplification using these new primers is underway. Future goals include evaluation of other STR loci and the detection of amplified alleles by fluorescent methods.

Thomas T. Noguchi, M.D., Professor of Forensic Pathology, Los Angeles County , University of Southern California Medical Center, 1200 North State Street, Los Angeles, CA 9033-1084

Due to the malpractice crisis in the 1970s, the medical community has realized the need for quality assurance (QA) and peer review of medical practitioners. The accreditation program, conducted by the Joint Commission on Accreditation by the Health Organization, is the most comprehensive inspection and accreditation program of the healthcare system encompassing space and equipment allocation, personnel and training program, safety and infection control and meeting with all required laws and regulations.

The QA program should be conducted in a context of total quality management of the institution with a plan for improvement based on an actual outcome study, thus the autopsy has once again become an important documentation and QA instrument. Currently there has been an emphasis on quality assurance programs with documentation of the peer review results. People are the most important resources in an organization. Each person should be accounted on the basis of their proper professional conduct. For the individual professional staff, the report or work product is regularly reviewed by the peer review members. Their assessment as to adequateness of the description, interpretation, and timeliness of the report is also recorded. In the healthcare system, the professional staff association has mandatory membership which is required for physicians to practice in any hospital, they must be credentialed into the department/specially field. The certifying body is the Credential Committee. The Author is currently Chairman of the Credential Committee in the Department of Pathology, at LAC+USC Medical Center. The attending staff is required to renew the membership by submitting an application for re-appointment every two years. For the Active Staff, in addition to the required Category I credit units (25 hours per year) and updating the specially board certificate, the member should not have any disciplinary action or any malpractice action against them. If such action resulted in payment or settlement, it will be carefully reviewed. The patient records and the report prepared by the attending staff is peer reviewed by the peer members and documentation as to the competency must be on file prior to giving the re-appointment. The National Practitioner Databank has been established to keep track of credentials, medical licenses, delineation of staff privileges and disciplinary actions throughout the United States, and a consorted effort of risk management along with quality assessment and value improvement program coupled with stronger actions taken against to physicians who have failed to meet the standards of medical practice set by the state medical licensing boards has virtually eliminated major malpractice program. The members of the California Association of Criminalists and forensic community may consider this valuable lesson on the evolution of the QA and peer review process in the healthcare and adopt the similar program to the further advancement of forensic sciences.

Igbal S. Sekhon, Department of Justice, Fresno

Pharmacia LKB was our only source for the carrier ampholyte, pH 5-6.5, we have been using to make ultrathin polyacrylamide gels for PGM-1 typing. The lots of this product supplied lately caused problems in the interpretation of results. An alternate carrier ampholyte, pH 5-7, manufactured by Serva was tried. We faced polymerization problems with this Serva product. Efforts to overcome these problems will be discussed.

Joseph Varlaro, B.S. and Rebecca Reynolds, Ph D., Human Identity Group, Roche Molecular Systems, Inc., 1145 Atlantic Avenue, Alameda, CA 94501

Determining the gender of an evidentiary sample can be an important part of casework analyses. Gender information can serve to separate biological evidence from two people who share the same DNA type(s) but differ by sex. In the case of DNA typing of sexual assault evidence, gender information can also serve as confirmation that the resulting DNA type obtained from the sperm fraction is of male origin. The reverse dot blot gender assay we have developed relies on amplification of a small, polymorphic region of a homologous zinc finger protein locus designated ZFX and ZFY on the X and Y chromosomes, respectively. Subsequent gender determination is based on the detection of a sequence polymorphism between ZFX and ZFY PCR products using immobilized sequence specific oligonucleotide probes. The gender of the sample donor is assigned based on the pattern of X and/or Y probes that is visualized following colorimetric detection. An advantage of this assay is the ability to couple the amplification and typing of the gender product to the AmpliType HLA DQ and AmpliType PM systems, as well as to other PCR-based marker systems. Additional data to be presented will include sensitivity and mixture studies, as well as the analysis of actual casework samples using the described gender assay.

Donald T. Jones, San Bernardino Sheriffs Scientific Investigation Divisions, 200 South Lena Road, San Bernardino, California, 92415-0056

The allele frequencies of five polymorphic proteins used in forensic casework were determined for 100 Mexicans. Esterase D, phosphogluco-mutase I, erythrocyte acid phosphate, adenosine deaminase, and adenylate kinase were studied using isoelectric focusing and zonal electro-phoretic methods. Alleles frequencies were: EsD 1 0.845, EsD 2 0.150, EsD 5 0.005; PGM 1+ 0.415, PGM 1- 0.365, PGM 2+ 0.1 10, PGM 2- 0.1 10, AcP A 0.260, AcP B 0.720, AcP C 0.015, AcP R 0.005, ADA 1 0.960, ADA 2 0.040, AK 1 0.985, and AK 2 0.015. Chi-square analysis showed no significant deviation from Hardy Weinberg equilibrium at the 0.05 level.

Robert S. Blackett, Arizona Department of Public Safety, Northern Regional Crime Lab, Flagstaff, Arizona

A sexual assault case submitted to the Arizona DPS Crime Lab involved the utilization of a zucchini squash. The alleged weapon was recovered from the scene and included. No semen was found, but blood was visible on one end. Typing included identification as "human" and PGM subtyping (IEF) with the victim's blood standard (1+1-). Unstained zucchini were obtained as controls and PGM subtyped. The study was expanded to include other potential vegetable sex assault "weapons." PGM was subtyped for yellow crookneck squash, cucumber, onion, celery, carrot, broccoli, and asparagus. All, like zucchini, showed strong anodal activity, and only asparagus gave any banding in the human 2+2- 1+1- region. The study demonstrated that human PGM subtypes are readily discernable from vegetable PGM subtypes.

Lyn M. Feredy, Forensic Science Service, Newbury, Berkshire

The Forensic Science Service introduced the DNA technique-Short Tandem Repeat (STRs) into casework as of August 15th 1994. This first system comprises four loci which have been tested internationally and will be subsequently enhanced with further loci. The talk will address the application of STRs to casework in these early days and also look forward to the use of a more discriminating second system.

Dr T.M. Clayton, Forensic Laboratories, Wetherby, England

On 19th April 1993, a fire broke out which engulfed a heavily fortified compound in Waco, Texas. This was the headquarters of a religious sect calling themselves the 'Branch Davidians'. The blaze resulted in the deaths of over 70 of the occupants of the compound which was razed to the ground. Afterwards, the authorities were faced with the difficult task of trying to positively identify over 70 bodies, many of which had been charred and extensively fragmented. In an attempt to identify, genetically, the victims, bone and tissue samples were submitted for DNA profiling along with blood samples from relatives of the deceased individuals. The genetic identification program was hampered by the poor state of the tissue samples and by the fact that many of those killed belonged to family groups and as such, a number of the bodies were genetically related. The relatively new PCR-based technique of STR profiling was used in the identification program. The STR analysis relied upon a 'multiplex' test which simultaneously analyses four tetrametric STR loci and a PCR-based sex test (gender test) both of which were developed by the FSS. The tests were shown to be fast, accurate, reliable and robust. The multiplex test also proved highly successful in terms of both its ability to cope with highly degraded samples and discrimination power allowing a high number of positive identifications to be made. This exercise also provided valuable information concerned with the processing and analysis of forensic samples in an advanced state of decay.

Antony van Leeuwenhoek, Paul de Kruif, Brian J. Ford, Linda A. Wraxall, California Criminalistic Institute, DOJ, 4949 Broadway, Room A104, Sacramento, CA 95820

Antony van Leeuwenhoek has come in person to talk about his lifelong interest in the invisible world that surrounds us all, and the studies that convinced him that the widespread belief in "spontaneous generation" (the formation of life from inert materials) was groundless. Others had tried to use microscopes before, but Leeuwenhoek was the pioneer of high power microscopy. He was the true founder of microbiology, and the first to use a microscope for genuine forensic purposes. He will explain his observations of the tiny animals he saw-protozoa in rainwater, bacteria in ground pepper suspensions, etc.--and his work on blood. And he will naturally include mention of his extensive correspon-dence with the Royal Society of London, his first scientific audience who so much admired his research effort extending over fifty years.

Christopher Rogers, M.D., Lakshaman Satbyavagiswaran, M.D., Department of Coroner, 1104 No. Mission Rd., Los Angeles, CA 90033

On January 17, 1994, the San Fernando Valley sustained an earthquake of magnitude 6.7. The earthquake occurred at 4:31 a.m., when most people were in bed. Thus, in spite of extensive damage to buildings and roads, the death toll was relatively low. This presentation will cover the mortality estimates for the earthquake. The initial estimate of 57 deaths is an underestimate for several reasons. Also, the presentation will cover the damage sustained to, the Coroner's facility during the earthquake and methods for reducing the damage in future earthquakes.

Steve Dowell and Judy Myers Suchey, Los Angeles County Coroner

Southern California, with its densely populated metropolitan areas lying next to remote desert and mountainous terrain, gives rise to numerous skeletonized forensic cases. Many of these cases relate to serial homicides; the efficient analysis of such cases in terms of identification and establishment of cause of death is vital. A team approach is needed whereby the criminalist, forensic anthropologists, and forensic dentist collaborate with the forensic pathologist. Recovery techniques are as important as analytical techniques in the forensic science center. Communication between workers in adjoining regions may be necessary to solve the cases. This paper focuses on selected forensic cases which illustrate this creative process.

Ronald L. Moore, Orange County Sheriff-Coroner

The identification of subjects or the association of items of evidence with subjects or other items is often a critical part of a criminal investigation. There have been tremendous advances recently in two of the major means of human identification: fingerprint development and comparison, and DNA profiling. However, it is possible for the canny criminal to avoid leaving either fingerprints or biological material suitable for DNA profiling at the scene of the crime. There is another type of evidence available which is even harder to keep from leaving behind: scent. Human scent is the volatile products of bacterial decomposition of skin cells. Due to the unique composition of skin and bacterial complement from person to person, it is believed that human scent can be as individual as either fingerprints or DNA. Also like these samples, scent is invisible. While we have produced devices to detect the presence of fingerprints or DNA, we have yet to develop a human scent detector. Fortunately nature has saved us the trouble having already developed a powerful and sensitive human scent detector in the canine. Canines have evolved a very sensitive sense of smell for finding food, choosing mates, and avoiding enemies. Using this ability they can be trained to find and follow human scent. The well trained canine can then learn to find the source of a particular scent, which has several specific applications to forensic practice, such as selecting a subject from a line up using a scent article from a crime scene. Scent may be collected and preserved and used for evidence at a later date. Scent collection and preservation techniques reflect the composition of scent and scent sources, i.e. volatile, skin cells, and bacteria. Scent evidence can be a powerful and successful tool to add to the forensic arsenal. The forensic professional should keep the possibility of using scent evidence in mind when investigating the scene of a crime.

Detective Chief Inspector Olive Wolfendale & Detective Inspector Geoff Keeling, Greater Manchester Police

Greater Manchester Police needed to increase forensic awareness amongst police officers who attend crime scenes. The speakers will describe how this problem was approached as a partnership venture between G.M.P. and the U.K. Forensic Science Service. The outcome was an integrated training package including a video, trainer's guide and supporting information sheets.

Rick Morehead, Chris Linton, and Alan Spilkin, Restek Corp., 110 Benner Circle, Bellefonte, PA 16823

The analysis of specimens for the presence of ethanol and other volatiles is one of the highest volume tests in forensic laboratories. High throughput and specificity for ethanol are two critical issues in developing an assay for ethanol. Two new stationary phases have been developed which resolve ethanol from all of the other commonly encountered volatile organics. Each column resolves the desired components under isothermal conditions, with an analysis time of under 3 minutes. The presence of ethanol or other intoxicating volatile substances in forensic specimens carries significant legal implications. The identification and quantitation of ethanol should be performed using methods that are as specific and sensitive as possible. GC has been used as a testing procedure for ethanol in forensic specimens for over two decades. The applicability of WCTO capillary columns for blood alcohol has been limited by the inability of any single stationary phase or column to completely resolve ethanol and all of the other commonly occurring volatile compounds. The resolution of volatile compounds such as alcohols, ketones and aldehydes can be optimized through the process of selectivity tuning. In selectivity tuning, the chemical composition of the stationary phase is varied in a systematic manner to produce the greatest differences in relative retention times of the components of interest. This paper will describe the process of column development for analysis of blood alcohols and chromatographic testing to ensure column performance. The analysis of blood alcohols by dual capillary column GC will be demonstrated, using static headspace sampling.

Dean M. Gialamas, California Laboratory of Forensic Science, 17842 Irvine Boulevard, Suite 224, Tustin, California 92680

Using a simple, yet entertaining, geometric puzzle as a focus of attention, the not-so-simple task of case approach will be addressed. Solving this puzzle and analyzing and interpreting items of evidence require a similar approach. If one understands that looks can be deceiving, then that individual is another step closer to keeping an open mind. An open mind will avoid "pigeonholing" an interpretation and allow consideration to all possibilities, even the unexpected. Specific casework examples from crime scene investigations, bloodspatter interpretation, sexual assault evidence analysis, DNA, arson and GSR will be addressed to sway the unwary.

Brian Burritt, CA Department of Justice DNA Lab, Berkeley; Louis A. Maucieri, California Criminalistic Institute, Sacramento

This study reviews several TMDT spray reagents, including 8-hydroxyquinoline (8-HQ) which was suspended from use because of uncertainty about possible chemical hazards. The work assesses the suitability of these reagents for the test procedure, and recommends the resumption of TMDT with 8-HQ as the reagent of choice. This allows law enforcement the flexibility to apply a test in the field in cases where metal objects have been handled. Types of cases would include theft of objects (tools, materials); assault (holding / concealing knife, pipe, etc.); or shootings (handgun traces on victim or subject. Recent literature on chemical hazards excludes 8-HQ from the carcinogen and toxin status it was suspected to possess. Moreover, several OTC pharmaceutical products containing 8-HQ at or above the TMDT concentration, and intended for use on injured skin, were discovered. Therefore, the most recent hazard studies and the continued use of 8-HQ in non-prescription antiseptic preparations shows it is suitable for use in TMDT investigations.

Gerald Vale, DDS, JD, Forensic Odontologist, Los Angeles County

Even in an era of increasing use of DNA evidence, the teeth remain an important means of identification when decomposition, burning or mutilation defeat other techniques. Because of their extreme durability relative to other body tissues, teeth are often among the last body parts to remain, making them available for study using either DNA analysis or other methods. This presentation illustrates various means of establishing identity by dental evidence.

Peter Lamb, Forensic Science Service, Huntingdon, England

The identification of animal hairs and the comparison of human hairs have posed problems for the Forensic Scientist for a considerable time, Even when there is no doubt that hairs match each other, the scientist must question the value of that evidence and advise lay people of its potential impact. The presentation will outline the research carried out by the speaker and highlight the results which enable weighting of the evidence. The methods involve light microscopy, color matching, sectioning, electron microscopy and electrophoresis.

James G. Bailey, Los Angeles Sheriff Criminalistic Lab, 2020 W. Beverly Boulevard, Los Angeles, CA 90057

The US Federal Trade Commission has denned rayon as: "a manufactured fiber composed of regenerated cellulose, as well as manufactured fibers composed of regenerated cellulose in which substituents have replaced not more than 15% of the hydrogens of the hydroxyl groups." However, various international groups such as ISO and BISFA have stated that: "this term [rayon] is no longer acceptable for labeling goods containing these fibers, which must now be designated cupro, viscose, or modal." Unlike other synthetic fibers, analysis by infrared spectrophotometry does not distinguish between these different types of regenerated cellulose fibers. This paper discusses three microscopical tests which will allow discrimination of these sub-classifications: fiber cross section, birefringence, and isotropic refractive index. The 37 fibers tested were mainly from the NBS/CTS Reference Collection of Synthetic Fibers, with the addition of a sample of "cupro" obtained from the FBI Forensic Science Research and Training Center in Quantico, Virginia.

J. D. DeHann, California Criminalistic Institute, Sacramento, CA 95820

On the evening of September 21, 1988, a fire destroyed the home of Patti and Scott Berns and their 18 month old triplets, in a suburb of Ft. Worth, Texas. First responders found Scott on the front lawn and rescued the infants through a bedroom window. Patti was found unconscious in a rear bedroom and was taken to the hospital where she died 2 days later. A thorough joint investigation by local and BATF investigators revealed two separate points of origin in the structure-one in the living room sofa where Scott said he was sleeping and one in the attic above the kitchen. Neither the original investigators nor the experts retained during trial could find any connection between the two fires or a possible accidental cause for the attic fire. Patti has suffered burns and yet was found in a room which had not sustained any fire damage. Her lack of response to medical treatment was not normal for a smoke inhalation victim and the possibility arise that she had been poisoned prior to me fire. In 1991, Scott Berns was tried for the murder of his wife. There had been a history of marital problems and witnesses reported an attempted shooting, a physical assault, and an emotional confrontation between Scott and Patti in the hours preceding the fire. There were numerous indications of impending separation. A detailed reconstruction of the fire was only possible due to the extensive documentation by photographs and diagrams. The intentional origin of the separate fires occurring at the same time (and not as a sequence of one to another) was conclusively established for the jury. The actual mechanism of Patti's death and its link to the fire was unfortunately not established with certainty. This paper will review the case and demonstrate the reasoning behind the origin-and-cause determination using the extensive document record available. The mystery of who (or what) killed Patti Berns wail be explored.

Lynne D. Herold and William F. Leo, Los Angeles County Sheriffs Department, Scientific Services Bureau, 2020 W. Beverly Blvd., Los Angeles, CA 90057

In 1987 a construction crew initially unearthed human skeletal remains 6 inches below a sidewalk adjacent to a 40+ year old house while reinforcing the foundation. An investigative team continued the gravesite excavation over the next 2-1/2 days revealing a complete, supine, clad female skeleton partially encased in cement. The cement encasement was different than the overlying sidewalk cement. The apparent bottom boundary of the grave, about 20 inches below the surface, was lined by outstretched clothing items. A cigarette pack, clearly found within the grave above the lower boundary but below the body, was identified as a test market product issued for a limited time in 1974 allowing approximate dating of the grave to some 14 years earlier. Toolmarks located on the recovered bones showed the deceased to be the victim of multiple stab wounds. Toolmarks on some of the finger bones suggested defense wounds to the hand. The cement encasement had apparently been poured over the victim soon after death. The cement contained a negative impression of much of the victim's upper body including the face and right hand. Silicone based polymers were used to make positive casts of the hand and face. A composite drawing of the face was publicized leading to a possible victim name. The positive cast of the right hand revealed identifiable ridge detail in the right thumb and interdigital area of the palm. A positive identification of the murder victim was made by a comparison of photographed ridge detail from the handcast right thumb to a fingerprint card. In addition, the face and hand molds showed corroborative evidence of the stabbing and defense wounds by preserving the soft tissue trauma. Following successful identification of the victim, determination of the cause of death and dating of the incident, a suspect was arrested. The integrated and collaborative efforts of multiple experts were fundamental to the successful resolution of this 1974 murder.

Frederic A. Tulleners, Criminalist Supervisor, California Criminalistic Institute

In June 1994 the Office of National Drug Policy Control (ONDCP), a White House Cabinet Level Office was requested by the Office of Management and Budget (0MB) to review and evaluate two systems in the market that are utilized to automate the identification of bullets and cartridge cases from shootings and homicides. The two systems are DRUGFIRE, sponsored by the Federal Bureau of Investigation (FBI) and BULLETPROOF made by Forensic Technologies and sponsored by the Alcohol, Tobacco and Firearms (ATF). An independent committee was formed consisting of experts in Firearms Identification, Imaging Algorithms, Optics, Computers and Life Cycle Costs. This committee met in August 1994 and spent a week at the ATF facility evaluating BULLETPROOF. After this, the report was compiled and presented to the participating parties in September 1994. This paper will describe the committee dynamics, their methodology and conclusions. Note: Presentation of this paper will be subject to the approval of 0MB and ONDCP.

R. Cheeseman, 4844 Camino Roberta, Bonita, California, 91902; Lisa DiMeo, San Diego County Sheriffs Crime Laboratory, 3520 Kurtz Street, San Diego, California, 92110

Fluorescin was considered by Maucieri and Monk (1992) to be potentially useful as a luminol replacement for blood stain detection. Re-enacting many aspects of the Maucieri and Monk work on fluorescin we were able to confirm their results. In direct comparisons of similarly blood stained surfaces the Huorescin reagent was at least as sensitive as the luminol system. Our task was to develop and improve the fluorescin reagent into a practical field system for the detection of latent blood stains. One of the advantages of fluorescin is that it does not have some of the OSHA hazards associated with luminol and other blood detecting reagents. However, one of the fluorescin disadvantages as a field system is that, unlike luminol's single reagent application, it requires an application of fluorescin followed by an application of H2O2 for optimal results. The double reagent application greatly increases the probability of the blood stain running. This was particularly troublesome on vertical non-porous surfaces. By using a commercial thickener and an adequate delivery device, we were able to overcome the reagent running problem. Xanthan gum, an exocellular heteropolysaccharide produced via a fermentation process, helps prevent reagent running. This affords the crime scene photographers greater opportunity to document the blood stain patterns as evidence as well as being investigative.

Iqbal S. Sekbon, Nancy D. McCombs, and Rodney. H. Andrus

Pharmacia-LKB was our only source for the carrier ampholyte, pH 5-6.5, we had been using to make ultrathin polyacrylamide gels for phos-phoglucomutase (PGM) typing. The lots of this product supplied recently caused problems in the interpretation of the results. An alternate carrier ampholyte called Servalyt pH 5-7 manufactured by Serva was tried. The gels containing Servalyt accomplished the separation of the four alleles resulting in the successful genetic typing of all ten common phenotypes. However, we found gels poured using Servalyt were sensitive to polymerization. Based on our experimentation and observations, use of fresh ingredients particularly ammonium persulphate, which we use as an initiator agent for chemical polymerization, help in polymerization. Mixing the gel solution slowly, degassing prior to the addition of a chemical polymerization initiator agent, and slightly increasing the amount of initiator agent are recommended. Two other changes were made in our original method which were helpful. The cooling platen temperature was increased from 4degC to 6.5degC and a chemical separator was used in the gels. The increase in the platen temperature reduced the run time to one hour and forty minutes. The use of a separation agent EPPS (N-[2-HydroxyethyUpiperazine -N-B-propanesulfonic acid];HEPPS) helped to increase the distances between the diagnostic bands resulting in better resolution.