California Association of Criminalists

THE MULTISYSTEM ANALYSIS OF BLOOD STAINS
Brian G. D. Wraxall*, Metro Police, London & Beckman Instruments, Anaheim, CA Mark D. Stolorow, Michigan State Police

The combination of two or more polymorphic enzymes or proteins onto one electropherogram has been and still is a goal of many forensic scientists. Not only does it save time and require less equipment, but it enables the serologist to obtain far more information from a smaller blood stain. However, this goal has not been easily achieved because each genetic marker has typically been phenotyped under individual conditions of time, voltage, pH, and ionic strength.

Under a L.E.A.A. sponsorship, eight polymorphic enzymes and protein systems: PGM, EAP, EsD, GLO I, AK, ADA, Hp, aid Gc, have been examined with a view to phenotyping two or more of them in a single electrophoretic cell. Simultaneous separation of all these markers in only three cells has been achieved with no loss of activity and with an increase in detectability and reliability in many cases.

The results achieved under this contract will be shown; the problems of multisystem analysis and its application to the crime laboratory will be discussed.

MODIFICATION OF PGM ISOZYME PATTERNS IN SEMEN CONTAMINATED WITH SALIVA
G. F. Sensabaugh & E.T. Blake, U. C. Berkeley D. Northey, Contra Costa Co. Sheriff's Dept.

PGM typing of semen evidence in a rape case involving oral copulation yielded apparently discordant results; a PGM type 1 sample gave an electrophoretic pattern similar to the type 2-1 pattern. Further analysis showed that semen PGM type 1 patterns were altered in a similar way when semen was mixed with saliva; the "a" isozyme band was progressively degraded relative to the "c" isozyme band giving rise ultimately to a pattern where only the "C" band was visible. The rate of the pattern shift depended upon the proportions of the saliva-semen mixture. Additional experimentation using a mixture of red cell hemolysate (PGM type 1), saliva and semen (both PGM type 2) as an indicator system showed that both semen and saliva factors were required for "a" band degradation; neither semen nor saliva factors alone were active. Both semen and saliva factors were heat labile suggesting that both are enzymes; further biochemical characterization is required to clarify this issue. Several conclusions may be drawn at this point. First, PGM patterns in semen-saliva mixtures are subject to alteration; similar alterations may occur in other mixtures of physiological fluids. Second, the nature of the alteration is such that typing errors might be made; in particular, type 1 may be mistyped as type 2-1 and type2-l may be mistyped as type 2. The risk of error is minimized if the analyst recognized that the kind of pattern alteration is possible and makes interpretations accordingly,

PRE-AND POST COITAL QUANTITATIVE ACID PHOSPHATASE LEVELS
G. F. Sensabaugh, U. C. Berkeley

The human vaginal fluids normally contain low levels of acid phosphatase (AP) activity. Statistical analysis of 5 sets of quantitative data on endogenous vaginal AP levels reveals that the activity is log normally distributed; this allows the definition of significance thresholds for distinguishing normal and elevated vaginal AP levels. Parallel analysis of post coital AP levels shows that the decay of enzyme activity proceeds in at least two phases, an initial rapid phase in which 50-99% of the deposited activity is lost, and a second phase in which the activity loss is first order. The standard deviations of the post coital AP values are large; nevertheless, it has been possible to predict with considerable accuracy the proportion of post coital AP determinations in which the observed values fall in the normal vaginal range.

EXPRESSION OF GLYOXALASE IN HUMAN SEMEN
E. T. Blake & G. F. Sensabaugh, U. C. Berkeley

The Glyoxalase polymorphism has been shown to be a useful system for blood analysis. We have investigated the expression of this genetic marker in semen to assess its potential in the analysis of semen evidence. Glyoxalase activity is found in both sperm and seminal plasma. In sperm, the expression is identical to the expression in red cells. In seminal plasma, on the other hand, glyoxalase has an altered electrophoretic mobility and is degraded very rapidly. The mobility of red cell glyoxalase is altered by addition of a semen factor indicating that the altered mobility of the seminal glyoxalase is not an intrinsic feature of that enzyme. Based on its altered pattern in seminal plasma and on its level in whole semen, glyoxalase would appear to be of marginal value as a marker in the analysis of semen evidence.

BULLET-LEAD STUDIES � THE ASSASSINATION OF PRESIDENT KENNEDY
Dr. Vincent P. Guinn, U. C. Irvine

During the period of 1973-1975, the author made an instrumental neutron activation analysis (INAA) study of the composition of Western Cartridge Company Mannlicher-Carcano (MC) 6.5 mm bullet leads, from production lots 6000, 6001, 6002, and 6003 (the type of ammunition reportedly fired by Lee Harvey Oswald in Dallas on November 22, 1963). These studies revealed that this bullet lead was remarkably heterogeneous, particularly in antimony, from one bullet to another even from the same box of cartridges. For example, four specimens from lot 6002 ranged all the way from 24 ppm Sb to 949 ppm Sb (they were less heterogeneous in silver, ranging from 6.0 to 9.7 ppm Ag). On the other hand, individual bullets were found to be moderately homogeneous.

While this work was in progress, it became known that the FBI had used IMAA on the Dallas specimens, during May of 1964, but notified the Warren Commission that the results were felt to be inconclusive. Through a colleague, Dr. John Nichols, some 70 pages of the original FBI INAA data were obtained (in 1975), under the Freedom of Information Act. In a first study and recalculation of the FBI data, the author also felt that the results were inconclusive. However, more recently the author organized the data in a different fashion and treated the results statistically, with the surprising conclusion that the data definitely reveal (1) the presence of only two (MC) bullets, (2) the stretcher bullet and the fragments from Governor Connally's wrist matching one another, and (3) the fragments from President Kennedy's brain and the other frag-ments from the car matching one another. The numerical values, and the reason for the original opinion that the results were inconclusive, will be presented.

BLOODSTAIN INDIVIDUALIZATION: RESULTS OF COLLABORATION BETWEEN ALAMEDA COUNTY SHERIFF'S DEPARTMENT CRIME LABORATORY AND A UNIVERSITY OF CALIFORNIA RESEARCH LABORATORY
Patricia L. Zajac*, Alameda Co. Sheriff's Dept. Benjamin W. Grunbaum, U. C. Berkeley

The Alameda County Sheriff's Department Crime Laboratory and the University of California Environmental Physiology Laboratory at Berkeley undertook a cooperative program of research and development of bloodstain analysis methodology for practical forensic applications.

In 1973, prior to initiation of this cooperative effort, dried bloodstain analyses in this laboratory were limited to species determination and ABO typing. The laboratory now performs routine analyses for an additional eight protein and enzyme systems. The total number of blood and bloodstain analyses has increased from 87 in the year 1973-7U to 792 in the year 1976-77.

Methods that are currently used routinely in this crime laboratory for eight enzyme/protein systems utilizing cellulose acetate membranes will be outlined. The methods have been standardized for sequential and concurrent analysis under optimum conditions, recognizing the inherent problems of analysis of dried bloodstains.

In the crime laboratory, blood evidence is usually received in infinite stages of degradation and contamination. Unlike other physical evidence which remains relatively permanent in it composition, blood evidence continues to degrade. Nevertheless, the use of bloodstain evidence has a high potential value in criminal investigations. Problems associated with systematic analysis of dried bloodstains in casework will be presented.

ANALYTICAL AND PREPARATIVE ELECTROFUCUSING IN A FLAT BED SYSTEM
Verna Venson, LKB Western Instruments, Inc. Pleasant Hill, CA

The development of the technique of electrofocusing represents a major advance in the field of high resolution separations of proteins and other amphoteric macromolecules. Electrofocusing is an equilibrium technique in which amphoteric molecules are separated on the basis of their isoelectric points in a pH gradient. Methods are available for performing electrofocusing under analytical as well as preparative conditions. The resolving power of electrofocusing facilitates the separation of molecules which cannot be resolved using conventional electrophoretic techniques. The instrumentation used for electrofocusing in a flat bed system allows the evaluation of many samples within a relatively short experimental time period.

DETERMINATION OF PGM AND EAP TYPES BY ISOELECTRIC FOCUSING
Joan Provost, Acadiana Crime Lab, LA

A method for determination of EAP and PGM types simultaneously is presented. Equipment used is manufactured by LKB Instruments and consists of a constant power source, electrophoresis tank and focusing lid. LKB Ampholines pH 5-7 and 7-9 were used to prepare a pH 5-8.5 gradient polyacrylamide gel which is a 2mm thick slab.

Samples are applied to the gel as extracts of dried stains. Electrofocusing is performed at 25 watts with maximum settings of 1600 volts and 50 m. amps for 2-1/2 hours. The gel is then developed on one end for EAP activity and on the other for PGM activity.

Isoelectric focusing has several advantages, some of them being reduced aging effects in EAP stains, concentration of dilute stains, reduction of time of analysis and the possibility of increased discrimination offered by the PGM system when typed by electrofocusing.

DYNAMICS, KINEMATICS AND MECHANISM OF INJURY IN AUTO CRASHES
A. W. Siegel, Trauma Research Group, Encino, CA

The mechanism of injury in auto crashes is grossly affected by the nonuse of restraint. As an introduction, an ultra-slow motion series of 30 mph sled crashes with anthropometric dummies simulating occupant kinematics (body motion) and differences in injury patterns will shown. An overview of the statistical relationship between all crashes and those that produce minor to fatal injuries will be discussed. Special films will be utilized to show the types, methods and deployment of experimental and actual passive restraints (airbags). Advantages and limitations of the air bags will be discussed.

Collision research studies have shown that an impact rarely occurs or results from a single cause. An interrelated chain of events leads to the impact, although, one "key event" maybe considered as the "point of no return" for each crash. Given driver alertness, reaction time, vehicle response time, roadway friction and other factors, a collision will occur before the "key event" is considered the "Pre-Impact Phase". Those after are the "Impact Phase", and events after the vehicle has come to rest are the "Post Impact Phase". With proper scientific analysis, one can understand and influence collisions and injuries. One simply cannot change the inevitability of a specific collision after the "key event".

This paper is concerned with a detailed analysis of the Dynamics of Vehicle Crashes, Occupant Kinematics and the Mechanism of Injuries. As in many activities, collision patterns are repeatable and predictable; therefore scientific methodology can be applied to study, analyze and influence future impact sequences.

All too often the Forensic Scientist is confronted with the question of "who was driving" in a fatal or sole survivor crash. The paper will use high speed films and slides to illustrate and present the fundamentals of collision and collision injury analysis, procedures and methodology.

SOAP SCUM AS EVIDENCE
N. L. Fort, Ventura Co. Sheriff's Dept.

Gas chromatographic, infrared spectrophotometic, microscopic, and spectrographic tests were conducted on stains present on the clothing of suspects in a homicide which occurred in a shower stall at the county jail. The results were compared to the results of similar tests conducted on samples of soap residue collected from several areas of the shower stall as well as samples of soap used in the shower. The results of all tests were consistent with the suspect stains being soap scum.

SYNTHESIS AND CLEANUP PROCEDURES FOR 4-METHYLUMBELLIFERONE PHOSPHATE
E. Blake & G. F. Sensabaugh, U. C. Berkeley

Methylumbelliferone phosphate (MUP) is the most popular substrate used for the visualization of red cell acid phosphatase activity on electrophoresis plates. Commercial preparations of MUP vary considerably in purity. Some preparations are contaminated with appreciable amounts of methylumbelliferone, the fluorescent product of MUP hydrolysis by the enzyme. These contaminated preparations have an undesirably high background fluorescence that significantly reduces the sensitivity of visualization.

Contaminated preparations of MUP can be cleaned up by simple solvent precipitation steps. Alternatively, MUP is readily synthesized de novo from methylumbelliferone and phosphorus oxychloride according to the procedure of Fernley and Walker (Biochem S. 97. 1965, 95-103). Handouts with a full description of cleanup and purification procedures will be available.

MICROCRYSTAL TESTS ON DEVELOPED THIN LAYER SPOTS
Duane L. Mauzey, Calif. DOJ-Riverside

Thin layer chromatography (TLC) has long been used in controlled substance analysis for effecting separations and as an aid in the identification of unknowns. As an identification tool, its usefulness has been limited by its poor ability to separate homologs and some analogs under customary laboratory practice. To increase the usefulness of TLC, methods have been devised for the direct application of selected microcrystal tests to commonly encountered controlled substances subsequent to their separation and detection on TLC plates. Drugs to which this technique has so far been found applicable include: amphetamine, methamphetamine, heroin, cocaine, phencyclidine and some barbiturates.

DETECTION OF DRUGS IN HAIR USING RADIOIMMUNOASSAY
Annette M. Baumgartner & Peter F. Jones, The Aerospace Corporation

Detection of drugs in hair and possibly the determination of characteristic drug use histories for individuals would provide a valuable aid in hair individualization. We have shown that it is possible to detect opiate metabolites in single human hair using the commercially available morphine radioimmunoassay kit. Extracts prepared from the hair of morphine-treated mice and from heroin users at a drug clinic contained nanogram levels of morphine per milligram of hair (a single human hair). It was also found that the mouse hair retained measurable drug levels for at least two months after the last injection, while urine and serum analysis of humans is useful only 1 to 72 hours after intake. Our preliminary data indicate a direct relationship between drug metabolite concentration in the hair and total drug use. Sectional analysis of hair suggests that the distribution of drug or metabolite along the length of hair can indeed reveal information regarding the time of drug use, thus providing a characteristic drug profile.

NOTES ON CURRENT PRACTICE IN DRUG SCREENING
Duane L. Mauzey, Calif. DOJ-Riverside

Analytical data from the literature has been abstracted for all controlled substances. The data has been organized by Controlled Substance Schedule, by color test and by UV spectral group. This compendium facilitates the rapid screening of solid dosage material for the presence of controlled substances. Preliminary identification of the most likely controlled substance present is quickly achieved. A discussion of the mechanism of the common color tests will also be included in the presentation.

The compendium is expected to be ready for publication sometime this summer.

THE CRIME LABORATORY APPROACH TO THE CLANDESTINE LABORATORY
J. Raymond Wells, Los Angeles Sheriff�s Dept.

The synthesis of illicit drugs has become a major problem to law enforcement. The clandestine laboratory is the main source of these illicit drugs. During the past two years the Los Angeles County Sheriff's Crime Laboratory has handled in excess of 70 clandestine laboratories. Based upon these experiences the laboratory developed methods to help in the investigation of the clandestine laboratories. Since the majority of clandestine laboratories investigated have involved the drug Phencyclidine (PCP), the discussion will be limited to the various facets of its production. However, much of this information will also be beneficial for the investigation of the clandestine production of other drugs such as amphetamine, MDA, LSD, methaqualone and others.

Further objectives are to familiarize criminalists with the hazards of chemicals, symptoms of persons under the influence, and how police, fire and the criminalistics laboratory must work together to combat the increasing danger of illicit drugs and their manufacturing.

NEUTRON ACTIVATION ANALYSIS OF SHOTGUN PELLETS
Norman R. Wallis*, LA Medical Examiner/Coroner Vincent P. Guinn, U. C. Irvine; W. Jack Cadman, Cal State Univ. Los Angeles

Neutron activation analysis has proved to be useful in the comparison of bullet-lead fragments with recovered fired or unfired bullets to establish whether various specimens do or do not have a common manufacturing origin.

In this study, the NAA method has been applied to shotgun pellets of many different known manufacturing brands specifically for Sb, Ag, and Cu composition. Samples of about 30 mg are scraped from individual shotgun pellets and irradiated for 40 seconds in the UC Irvine Triga Mark II reactor at a thermal neutron flux of 2.5 X 1012 n cm-2 s-1, allowed to decay for 40 seconds and then counted for 40 seconds. The gamma ray spectrometry is carried out with a 90 cm3 Ge (Li) detector coupled to a 4096-channel pulse-height analyzer. The spectrum is placed on magnetic tape for analysis. Later, the peaks of interest (498 keV of 124ml sb, 658 keV of 110Ag,, and 1039 keV of 66Cu) are read out.

The results obtained with the shotgun pellets are similar to those obtained with bullet lead: the three elements normally detectable are Sb, Ag, and Cu, and the concentration of each varies greatly from one brand to another. In the shotgun pellets studied, Sb concentration varied from 0(+/- 49) ppm to 6.43%. The Ag concentration varied from 7.6 (+/- 1.1) ppm to 100.3 ppm. Cu concentration varied from 0 (+/- 16 to 502 ppm.

Further work entailed analysis of different scrappings from one pellet, different pellets in a cartridge, and different cartridges in a box to determine homogeneity of the elements of interest. Results show that shotgun pellets are very homogeneous, and it appears to be readily feasible to ascertain whether a shotgun pellet, or pellet fragment recovered from a victim, is or is not of the same brand as any one of the suspected ammunitions.

IDENTIFICATION OF GASOLINE SOURCE BY LEAD ALKYL COMPARISON USING GAS CHROMATOGRAPHY WITH ELECTRON CAPTURE DETECTOR
Hubert A. Frank, Calif. DOJ-Salinas

Lead alkyl profiles of gasoline can provide three to seven parameters which can be of use in gasoline comparison. The method has been used in identifying leaking gasoline tanks by comparing lead alkyl profiles of samples collected from sewers, soil and asphalt. The method is applicable in arson cases where samples are collected from mini-pools, soils or rugs where heat and extreme evaporation has not substantially altered the lead profile. Vendor tanks generally have individualized lead profiles due to batch mixing. Some major brand gasolines have distinct lead alkyl profile class characteristics. The absence of lead can also be a significant factor. Experimental conditions: Column: 6 ft.-20% 1,2,3-Tris 2-Cyanoethoxy Propane (TCEP) on Chromosorb W 8100 . Column Temp: 70O; Injector: 200O; Detector: 200O; Flow Rate: 25cc/min.

THE USE OF THE STRONG-CAMPBELL TEST IN THE SELECTION OF EVIDENCE TECHNICIANS
Trevor White & John Thornton, U.C. Berkeley

This paper addresses the failure of many law enforcement agencies to adequately screen the personnel they assign to collect evidence at crime scenes. Police officers are assigned who lack the aptitude, the training, or the interest required for the conscientious performance of evidence-collecting duties; evidence is overlooked or mishandled. Although acknowledging the importance of aptitude and training, the research focuses upon interest as a determinant of performance. Using the Strong-Campbell Interest Inventory, a sample population of competent crime scene technicians was profiled. Also profiled were sample populations of non-technician patrol officers and crime scene technicians selected without regard to competence. The results are encouraging. They provide insight into the personalities of crime scene technicians and suggest that the Inventory would be useful as a selection tool.

THE INDIVIDUALITY OF NEW FIRING PINS AS SEEN IN THEIR IMPRESSIONS
Rory Shanahan & Donald F. Nelson*, D.S.I.R., Auckland, NZ

As this concept is still being explored in court by defense counsel in New Zealand the individuality of new and unused firing pins was reexamined by comparing the firing pin impressions produced by military 9mm pistols (Browning manufactured in Canada by Inglis) and 7.52 mm L1AI rifles (Small Arms Factory, Lithgow, N.S.W., Australia). Some of the pins studied were consecutively produced. Each firing pin produced impressions which were distinguishable from the impressions produced by the other firing pins.

SHOTGUN PATTERNS
Donald F. Nelson* & Rory Shanahan; D.S.I.R., Auckland, NZ

This is a "Pre-computer" approach to measuring test-fired shotgun patterns by counting the shot striking within concentric circles. By plotting the results obtained, the scene pattern can be assessed in order to determine the range at which a particular shot was fired.

EXPLODING BULLETS - AN ANALYSIS
F. A. J. Tulleners, Calif. DOJ-Riverside

A description of the Velet exploding bullet, its composite parts and method of test firing.

THE COMPARISON OF TESTFIRINGS FROM CONSECUTIVELY BUTTON RIFLED .22 CALIBER RIFLE BARRELS
John E. Murdock, Contra Costa Co. Sheriff's Dept

The Ithaca, Marlin, Mossberg, and Remington Firearm Companies provided the gun barrels used for this research. Five button rifled 22 caliber rifle barrels numbered 1, 2, 3, 7 and 10 selected from a consecutively manufactured series were received from Marlin, Mossberg, and Remington. One barrel of this same type was provided by Ithaca.

The effect on individuality of the crowning operation was evaluated by using the Ithaca barrel and the three barrels numbered 10 in the series of five received from the other manufacturers. An inter-comparison of test firings following successive recrowns revealed the minimal effect of this mechanical operation upon individuality. The firings from each barrel could still be identified upon individuality. The firings from each barrel could still be identified with firings from the same barrel.

The consecutively rifled barrels numbered 1, 2, and 3 from Marlin, Mossberg, and Remington were used for the following research. Ten plain lead standard velocity Remington .22 long rifle caliber bullets were fired through each barrel. In general, the comparison between the first, second, and third firing from any one barrel failed to result in an identification. The third, fourth, and fifth test firings from any one barrel could, however, be identified as having been fired through the same barrel. The intercomparison of test firings from barrels 1, 2, and 3 of each set revealed that there was absolutely no carry-over of family-type striations, thus demonstrating the individuality of button rifled .22 caliber barrels made by Marlin, Mossberg, and Remington.

AN EVALUATION OF THE FBI LABORATORY'S GUNSHOT RESIDUE ANALYSIS PROGRAM
John W. Kilty, FBI Laboratory, Washington, D.C.

The FBI uses neutron activation analysis (NAA) to do bulk elemental analysis of lifting mechanisms for antimony and barium, components of many primer mixtures. Number of samples analyzed, number of cases where significant results are obtained, the form of the expert opinion, results of test firings and problems encountered when a Laboratory does not have control over the samples submitted for analysis will be discussed. Some logistical problems encountered with high case load and speedy trial rules will be mentioned.

PHYSICAL EVIDENCE IN THE BOMBING DEATH OF PHOENIX NEWSMAN DON BOLLES
R. Gieszl, L. Haag & A. Raphael, Phoenix (Arizona) P.D.

In June of 1976 investigative reporter Don Bolles was mortally injured when an explosive device detonated under his automobile. Bolles, who was investigating land fraud in Arizona, succumbed to his injuries 11 days later.

This presentation summarizes the laboratory efforts and findings in determining the identity of the explosive used, the method of attachment to the vehicle and the means of initiation employed.

From the identity of the various components utilized in the device, valuable investigative leads were provided which subsequently linked a suspect with the crime.

A 3 FILAMENT HEADLIGHT
F. A. J. Tulleners, Calif. DOJ-Riverside

A description of a safety headlight with 3 filaments that may be encountered on 1970 vintage automobiles.

THE APPROACH OF THE NEW ZEALAND FORENSIC SCIENTIST
Rory Shanahan, D.S.I.R., Auckland, NZ presented by Donald F. Nelson, D.S.I.R. Auckland, NZ

The forensic scientist in New Zealand is not self-starting in any particular investigation, but waits to be consulted by the Police.

Laboratory findings can be used by the police to guide their investigations, to clear the innocent or to indicate guilt in court proceedings.

Although called by the prosecution the forensic scientist is in court to tell the whole truth and present impartial evidence to assist the court. His evidence should be clear to laymen. The challenge of problems arising from daily work can extend the best scientists and there are research opportunities in methods, interpretation and specific problems.

THE FORENSIC SCIENTIST'S OBSERVATIONS
Donald F. Nelson, D.S.I.R., Auckland, NZ

Examination of samples brought to the laboratory may (a) reconstruct what happened (b) indicate a crime(c) trace suspect or victim (d) confirm or disprove statements (e) test a theory (f) associate a suspect with a scene or a victim.

Often the task is to decide whether two samples could have a common origin and, if so, to determine the likelihood that an observed agreement is random. Simple and complex techniques are available.

Assessing the relevance of laboratory observations requires knowledge of principles of identification, experience, background knowledge and common sense. Experiments must have adequate standards and controls.

QUALITY ACHIEVEMENT IN CRIME LABORATORY WORK
Lowell W. Bradford, Consultant, San Jose

Discussion of current day professional problems, causes, outline of standards of practice and the prognosis.

CORRELATION OF INTOXILYZER MODEL 4011 WITH WHOLE BLOOD ANALYSIS
M. Breen, D. Clardy, E. Rhodes & K. Inman Orange County Sheriff's Department

(no abstract available)