Social and Behavioral Sciences Commons^™

A Novel Dual Configuration Dart/Uhplc-Ms/Ms System For The Rapid Analysis Of Psilocybin-Containing Mushrooms And Other Seized-Drug Submissions, Aron R. Jaffe

Master of Science in Forensic Science Directed Research Projects

The use and availability of controlled substances is highly dynamic. When a classical or novel psychoactive substance becomes popular, an influx of seized-drug evidence submissions closely follows, contributing to an ever-growing backlog of exhibits awaiting identification. The need for inclusive, rapid, and robust analytical methods is apparent, as demonstrated by recent growth in hallucinogen use. Psilocybin and psilocin, among other entheogens, have become increasingly popular and an increase of psilocybin mushroom seizures by law enforcement has followed. The structural similarity and thermal lability of many entheogens preclude their identification using many conventional analysis techniques. A DART and UHPLC were interchangeably …

Evaluation Of Direct-To-Amplification Cell Lysis Techniques For Forensically Relevant Non-Sperm Cells, Rhea Arya

Master of Science in Forensic Science Directed Research Projects

While efforts have been made to reduce the pervasive backlog of sexual assault evidence collection kits, the actual laboratory process remains very time-consuming due to the requirement of a differential lysis step before DNA purification, as well as intricate mixture analysis towards the end of the DNA workflow. Previous work in the Dawson Green laboratory at VCU has developed alternative solutions for differential extractions with sexual assault samples using both an in-tube and microdevice assay. Prior work led to the identification of an alternative sperm lysis method – alkaline lysis using 1M NaOH. However, the current lysis method used for …

Optimization Of Microfluidic Device Methodology For Optical Trapping Epithelial Cells In A Mixture, Brittney N. Hackworth

Master of Science in Forensic Science Directed Research Projects

Evidence containing biological mixtures can make DNA interpretation more difficult due to ambiguous allele calls and artifacts that may be present within electropherograms. Separating out these cell mixtures can be advantageous to the analysts’ analysis methods for identifying STR profiles with higher accuracy. A potential cell separation method that could be implemented in the beginning of the forensic DNA workflow is optical trapping.

This research utilized a 5W ytterbium linearly polarized laser split into two optical traps, paired with a microfluidic device to isolate spermatozoa and vaginal epithelial cells from a 1:1 mixture of vaginal fluid and 1:20 diluted seminal …

Analysis Of Forensic Relevant Cell Mixtures Using Cellular Autofluorescence And Fluorescence Activated Cell Sorting (Facs), Shayla Smith

Master of Science in Forensic Science Directed Research Projects

Biological mixtures containing DNA from more than one contributor can make DNA profile interpretation challenging, especially for ‘touch’ or trace biological evidence. One approach to improve this is by differentiating cell populations from separate individuals prior to DNA profiling by utilizing intrinsic features of cells such as size, shape, and biochemical profiles. The goal of this study was to develop a new method for detecting and separating contributor populations in biological mixture samples using flow cytometry, a rapid and non-destructive approach with an unexplored potential for forensic casework. To accomplish this, two different mixture systems were investigated (1) mixtures containing …

Investigation Into The Relationship Between Dna Content And Dna Yield Prediction And Presence Of A Multiple Contributor Cell Population In Trace Dna, Sarah Holton Roisch

Master of Science in Forensic Science Directed Research Projects

Due to advancements in DNA analysis technology and an increased ability to detect trace quantities of low template DNA, ‘touch’ or trace DNA has become a prevalent form of evidence for forensic investigations. Trace DNA is biological material left on a surface from skin cells after being touched or handled. The success rate of casework could be improved with new methods to non-destructively screen trace DNA to predict DNA yield and whether multiple cell populations are present in the evidence prior to STR analysis. Current methods of trace DNA analysis require quantifying the sample with qPCR followed by STR analysis, …

Horizontal And Vertical Transmission Of Microbes Associated With Blow Flies Of Forensic Importance, Gabrielle Alisa Burton

Master of Science in Forensic Science Directed Research Projects

Entomological and microbiological evidence can be used in the estimation of postmortem intervals (PMI). This study observes the horizontal and vertical transmission of the microbial community associated with blow flies of forensic importance. Variable region four (V4) of the 16S ribosomal DNA (16S rDNA) was amplified and sequenced from various life stages of blow flies (n=72) using dual-index high throughput sequencing strategy on the MiSeq FGx platform. No significant difference in bacterial community structure was observed between blowfly adults with carrion access and blowfly adults with no carrion access. Dysgonomonas, unclassified Proteobacteria, Escherichia, Ignatzschia, Providencia, …

Morphological And Autofluorescence Signatures For The Identification Of Vaginal Cells In Mixture Samples Containing Saliva And/Or Epidermal Cells, Hannah L. Burden

Master of Science in Forensic Science Directed Research Projects

In forensic casework the identification of body fluids at a crime scene can frequently play an important role in reconstructing the events that occurred. There are several bodily fluid identification methods to characterize unknown stains recovered as evidence. However, there has not been a reliable method for detecting the presence vaginal cells and/or differentiate it from other forensically relevant tissue types that may be recovered from a crime scene. The detection of vaginal fluid is important for forensic casework due to the abundance of sexual assault kits that are not only backlogged but continue to be submitted to the lab …

Assessment Of The Acidified Hydrogen Peroxide Method On Uncontrolled Fired Cartridge Case Samples, Kylie S. Richardson

Master of Science in Forensic Science Directed Research Projects

The use of fingerprints to identify individuals in forensic cases has been useful for many years due to the persistence and individuality of the prints. In addition, the presence of firearms in forensic casework is steadily increasing over the years. This study aims to expand upon prior research in the development and recovery of latent prints on fired, brass ammunition using an acidified hydrogen peroxide method. Although latent prints are left on the cartridge casing of ammunition during the handling and loading of ammunition into the firearm, it is hypothesized that the pressure within the barrel of the firearm during …

Comparison Of Cell Counts To Dna Yield In Mock Biological Evidence Samples, Jacob Sewell

Master of Science in Forensic Science Directed Research Projects

The implementation of rapid DNA analysis systems is of interest to forensic DNA casework, as this technology exhibits a sample-to-profile time of approximately two hours with limited handling of samples. This decreases the likelihood of contamination, and can potentially reduce DNA backlogs; however, current rapid DNA analysis systems do not execute one crucial step of the forensic DNA analysis workflow: human DNA quantitation. This step is necessary for DNA profiles to be compliant with the FBI’s Quality Assurance Standard 9.4, allowing DNA profiles to be uploaded and searched within the CODIS database. Coupling traditional qPCR methods with rapid DNA analysis …

Evaluation Of The Age Of Evidence In Mock Biological Touch Evidence Samples, Emma P. Hirst

Master of Science in Forensic Science Directed Research Projects

As more sensitive DNA profiling techniques have been developed, questions about how and when the DNA was deposited have become a driving issue in forensic casework. Image flow cytometry (IFC) offers a way to estimate the time since deposition (TSD) of trace DNA samples based on individual cells' morphological and autofluorescence properties. Previous work has shown that a TSD can be determined from a single contributor in touch samples. To determine if TSD could be correctly estimated for mixtures containing a fresh deposit (less than a week old) and an older deposit, this project characterized a series of samples that …

Determination Of The Trace Elemental Signature Of Drywall Dust And Its Implication As Forensic Evidence, Emilee Holmes

Master of Science in Forensic Science Directed Research Projects

Drywall is found in most homes built in the United States of America because it is cost effective, light weight, and easy to install. Its brittle nature makes it less resistant to impact which can cause dust to form if a force is exerted on it. Airborne dust can settle on anything or anyone around it and can be considered trace evidence for its ability to connect suspects, victims, and scenes; however, this type of dust is not emphasized as trace evidence and is not always collected due to a lack of understanding of their forensic signatures.The seven producers of …

Development And Validation Of Microrna Markers For Forensic Body Fluid Identification Using Rt-Qpcr, Jane Goble

Master of Science in Forensic Science Directed Research Projects

Body fluid identification is one of the first and most fundamental steps in the forensic DNA workflow. It allows analysts to determine where a sample might exist on an item of evidence, as well as the type of sample that they are dealing with. However, current serological methods have been documented to produce false or inaccurate results in some cases, and may consume large amounts of sample during processing. Molecular methods, such as the use of microRNA and microbial profiles have been introduced as a means to increase specificity of testing, and have even been combined into a singular assay …

Evaluation Of Cytokeratin-Specific Antibody And Autofluorescent Signals As Predictors Of Time Since Deposition For Saliva Samples, Dillon Ryder

Master of Science in Forensic Science Directed Research Projects

In a forensic investigation, STR profiles generated from biological evidence can frequently be informative to identify an individual or provide an investigative lead. However, STR typing does not provide information to answer activity-level questions which can be highly important to a criminal case, such as the Time Since Deposition (TSD) of biological evidence. Determining the relative time in which a biological sample was left at a crime scene would provide additional information regarding the timeline of a crime and a person of interest’s relationship to the scene. One proposed method of estimating TSD is to use flow cytometry to analyze …

The Analysis Of Commercially Available Kratom Products In Richmond, Virginia, James H. Fleming Jr

Master of Science in Forensic Science Directed Research Projects

Kratom is a novel psychoactive substance that has gained popularity within the past ten years. Originating from Southeast Asia, the leaves of the Mitragyna speciosa tree contain two principal alkaloids, mitragynine and 7-hydroxymitragynine, that play a key role in opioid-like effects. Twenty-nine kratom products were obtained from tobacco shops in the Richmond, Virginia area, including powders, teas, capsules, extracts, and a carbonated beverage. Samples were analyzed using Direct Analysis in Real Time-Mass Spectrometry (DART-MS) for kratom alkaloids, labeled ingredients, and other possible organic compounds. Inductively Coupled Plasma-Optical Emission Spectroscopy (ICP-OES) was used to quantitate aluminum, arsenic, copper, iron, magnesium, nickel, …

Optimization Of A Microfluidic Device For Cell Capture Using Optical Trapping, Samantha C. Pagel

Master of Science in Forensic Science Directed Research Projects

Biological mixtures which result from evidence produced in sexual assault cases are one of the most common types of mixtures observed in forensic biology casework and have contributed to evidence backlogs across the country. In the forensic community, this has highlighted the need for a more efficient cell separation method at the beginning of the DNA analysis workflow than the currently accepted method. One promising novel front-end cell separation technique is optical trapping. In heterogeneous mixtures, optical tweezers have been demonstrated to manipulate and isolate cells of interest from other cell types.

For this research, a range of 5 to …

Assessing The Detectability Of Cannabinoid Analogs (Delta-8 Thc, Delta-10 Thc And Cbd) And Their Major Metabolites In Six Commercial Cannabinoid Urine Screening Kits, Ashley A. Pokhai

Master of Science in Forensic Science Directed Research Projects

Background:

There has been an exponential surge in the presence and use of cannabinoids since the federal legalization of hemp (Agricultural Improvement Act of 2018), which involves the legalization of medical use, adult recreational use, and decriminalization of cannabis in several states. This growth is not only attributed to ∆9-tetrahydrocannabinol (∆9-THC) and cannabidiol (CBD), the most abundant phytocannabinoid components of cannabis and hemp, respectively, but with many other emerging THC analogs. Structurally, these analogs are similar to ∆9-THC, yet very little information is available about their potency and even less information is available regarding their detectability using commercially available cannabinoid …

Optimization And Comparison Of Methods For Separation Of Spermatozoa From Superabsorbent Polymer-Containing Forensic Evidence, Hannah Wells

Master of Science in Forensic Science Directed Research Projects

Sexual assault is currently one of the most prevalent crimes that affects victims of all ages. Forensic DNA analysts often confirm proof of contact in sexual assault cases through the identification of spermatozoa and subsequent STR profiling. Diapers and other feminine hygiene products, such as maxi pads and ultrathin pads, are types of superabsorbent polymer-containing evidence that complicate the process of DNA analysis due to the trapping of the spermatozoa in the gel-like matrix. In this study, a comparison of methods was performed to determine which extraction technique produces the highest sperm cell and DNA yield and generates a usable …

Using A Non-Destructive Method To Estimate The Number Of Contributors And Predict Dna Yield In Touch Dna Mixtures, Christin Lee

Master of Science in Forensic Science Directed Research Projects

Touch DNA samples are low templates and often are mixtures, making them typically one of the most difficult types of evidence to interpret. Short tandem repeat analysis is a standard method to estimate the number of contributors and DNA yield for each contributor; however, there are some limitations. Applying a non-destructive pre-DNA analysis technique that provides information about the touch DNA may improve the casework process. Antibody probes to hormones such as testosterone and dihydrotestosterone (DHT), and proteins including cytokeratins (CK) inside epidermal cells, have been demonstrated to produce a personalized signature for human touch cell populations. Testosterone, DHT, and …

Development Of A Loop-Mediated Isothermal Amplification (Lamp) Assay For Detection Of Mirnas, Alisha B. Whelan

Master of Science in Forensic Science Directed Research Projects

Body fluid identification serves a vital role in contextualizing biological evidence received in forensic laboratories. While several fluids can be presumptively detected using immunological or catalytic tests, these tests are susceptible to false results and have limited sensitivity. True confirmatory testing is still somewhat limited in forensic laboratories. With the advancement of DNA typing technologies, laboratories can develop DNA profiles from trace amounts of sample, increasing the demand for sensitive and accurate body fluid detection. This project proposes using loop-mediated isothermal amplification (LAMP) as a method to rapidly detect microRNAs (miRNAs) that are differentially expressed in various body fluids with …

Stability And Variation Within Donors Of Mirna Markers For Body Fluid Identification, Kelsey R. Price

Master of Science in Forensic Science Directed Research Projects

Traditional enzymatic tests for body fluid identification are prone to false negatives, false positives, and several body fluids do not have confirmatory or reliable presumptive tests. Therefore, molecular-based tests may be more reliable, such as messenger RNA (mRNA), microbial DNA, or microRNA (miRNA) assays.

miRNAs are small, noncoding RNAs whose main function in-vivo is the regulation of protein expression by selectively suppressing the translation of their corresponding mRNAs. They also lack a poly-A tail, and because of this, their small size, and their association with other molecules such as the RISC protein complex, they are more robust than other RNAs, …

A Quantifiler Trio-Based Hrm Mixture Screening Assay For The Quantstudio 6 Flex Qpcr Platform, Dayanara A. Torres

Master of Science in Forensic Science Directed Research Projects

At present, the forensic DNA workflow is not capable of providing information about the contributor status (single source vs. mixtures) of evidentiary samples prior to end-point analysis. This shortcoming can exacerbate the challenges inherent to mixtures and low-template DNA samples. Provided additional sample information earlier in the workflow, protocols could be implemented to mitigate these challenges. High-resolution melt (HRM) curve analysis is a technique used to detect genetic variation in DNA fragments and in the last decade has been evaluated for use in differentiation of samples by genotype and/or contributor status. To this end, a proof-of-concept HRM assay using the …

A Comparison Of Cyanoacrylate Fuming Followed By Mbd Dye Staining And Gun Bluing For Development Of Latent Fingerprints On Cartridge Cases, Jamie Zaleta

Master of Science in Forensic Science Directed Research Projects

The development of latent fingerprints on cartridge cases is a desirable ability for the forensic laboratory, offering the potential to provide probative evidence as to the handler of whole ammunition or fired cartridge cases discovered at the crime scene. Previous studies have focused primarily on intentionally-laid fingerprints on cartridge case surfaces, which do not represent samples likely to come across the desk of a forensic analyst. This study focuses on naturally-handled ammunition and the success rates associated with latent fingerprints produced in a more realistic set of circumstances. A comparison was performed of two fingerprint development methods—gun bluing and MBD …

Classification And Differentiation Of Forensically Relevant Cell Populations Using Imaging Flow Cytometry, Arianna D. Decorte

Master of Science in Forensic Science Directed Research Projects

Forensic casework relies heavily on DNA profiles which may be time-consuming to generate and difficult to interpret when biological mixtures are present. Additionally, there are significant challenges in using sub-source data alone to answer the activity-level questions often most pertinent in criminal cases. Source level information can add critical probative value, however, methods that can provide information as to the source tissue of evidentiary cell populations are limited. There remains a need for new methods that can differentiate between and classify various cell populations, particularly for vaginal and epidermal tissue, since there are no conventional serological techniques specific to these …

Determination Of Cotinine Concentration In Urine By Liquid Chromatography Tandem Mass Spectrometry, Allison Beckett

Master of Science in Forensic Science Directed Research Projects

Tobacco use can cause lasting health conditions which can lead to death. Tobacco exposure comes from obvious sources like smoking or chewing tobacco, however these are not the only administration methods. Secondhand smoke exposure can affect even those who choose not to expose themselves to tobacco, which also opens them to the health issues that come along with tobacco use. Tobacco contains many different compounds, one of which is nicotine (Kataoka, 2009). Although nicotine is the most abundant compound in tobacco, it does not stay in the body for long periods of time. Nicotine metabolizes into cotinine which is more …

Impact Of 1,4-Dithiothreitol (Dtt) On Microbial Community Structures Associated With Human Semen Samples, Christian R. Blaise

Master of Science in Forensic Science Directed Research Projects

Body fluid identification (BFID) is one of the most important steps in forensic science as it provides important context to how DNA analysis should be performed on biological samples found at the scene of a crime. Many scientific methods on BFID have been developed over the years with respective benefits and flaws. One emerging method in recent years is microbial signature based BFID. This method uses the same DNA extract that is utilized for human DNA profile generation for microbial analysis. Although DNA is extracted from all body fluid samples without any additional steps, extraction of human DNA from semen …

Identification Of The New Synthetic Opioid Bucinnazine And Its Metabolites Using Liquid Chromatography Tandem Mass Spectrometry (Lc-Ms/Ms), Karissa N. Resnik

Master of Science in Forensic Science Directed Research Projects

In the past 20 years, the rate of opioid use increased drastically in the United States. The country has experienced an enormous number of deaths caused by opioid overdose, opioid use disorder (OUD), as well as other harms as a consequence of the high numbers of opioid prescriptions. Fentanyl, a well-known synthetic opioid, has been a contributor to the opioid crisis since 2013 due to its popularity in clinical and illicit use. It has a high potency, which makes it an effective analgesic but a dangerous illicit substance. A new synthetic opioid, bucinnazine, has recently become a new black-market opioid …

Differentiation Between Peripheral Blood And Menstrual Blood Using Mirna Markers, Anaya Valentine

Master of Science in Forensic Science Directed Research Projects

Abstract

In forensic investigations, body fluids can provide crucial information and is helpful for corroborating the circumstance of the case. For cases of sexual assaults or homicides, being able to differentiate if the blood is peripheral blood or menstrual blood is important. Peripheral blood can be indicative of a traumatic event, while menstrual blood is of a natural cause. Currently, serology based methods are used for body fluid identification, however, their lack of sensitivity and specificity remains an issue. MicroRNAs (miRNAs) are a class of non-protein coding nucleic acids that are able to be co-extracted with DNA, and their small …

Development Of An Analytical Method And Sample Preparation Technique For The Analysis Of Sulfur-Containing Fentanyl Analogs By Uplcmsms, And The Application To Forensic Science, Amber D. Budmark

Master of Science in Forensic Science Directed Research Projects

In February 2018, the US DEA Agency released a statement of the emergency scheduling (Schedule 1) of all fentanyl analogs not already regulated by the CSA due to an alarming increase in overdose deaths linked to synthetic opioids. Fentanyl analogs are pharmacologically similar to fentanyl, but often more potent. This increased potency can create problems with proper dosing of fentanyl analogs leading to untoward effects including an increase in overdoses and deaths. Since 2018, there has been a 38.4% increase in illicitly manufactured fentanyl overdose deaths leading the CDC to make available the FAS Kit and Emergent Panels containing previously …

The Efficacy Of Recovering Latent Fingerprints From Fired Cartridge Cases By Way Of Cyanoacrylate Fuming Combined With Basic Yellow Dye Staining, Lily Mantz

Master of Science in Forensic Science Directed Research Projects

This experiment attempted to tackle the long-standing struggle to consistently recover latent fingerprints from fired cartridges. With an emphasis on realistic handling of high volume of samples combined with Cyanoacrylate fuming followed by Basic Yellow dye staining, a total of 740 cartridges were examined. These included fired and unfired brass and nickel case 9mm Luger and 40 S&W cartridges. The cartridges were divided into five different groups; each were treated differently prior to cyanoacrylate fuming and dye staining, which was conducted in the same manner for the groups. The groups included the positive and negative controls, loaded, cycled, and fired. …

The Impact Of Latent Print Processing On Firing Pin And Breechface Impressions During Firearm Examination, Brittany B. Harrington

Master of Science in Forensic Science Directed Research Projects

Latent prints can be a valuable source of forensic evidence when solving a crime. They can verify if a person was at a specific scene, identify unknown individuals to connect them to a scene, and help to corroborate eyewitness accounts. Latent prints, however, are not always visible until they have undergone enhancement or visualization techniques. When fired cartridge cases are suspected of containing latent prints, they are brought in for latent print processing before any firearm analysis is performed. As a result, these cartridges are often coated in various residues or dyes when they arrive for firearm examination. In response, …