Single source DNA profile recovery from single cells isolated from skin and fabric from touch DNA mixtures in mock physical assaults

The ability to obtain DNA profiles from trace biological evidence is routinely demonstrated with so-called ‘touch DNA evidence’, which is generally perceived to be the result of DNA obtained from shed skin cells transferred from a donor's hands to an object or person during direct physical contact. Current methods for the recovery of trace DNA employ swabs or adhesive tape to sample an area of interest. While of practical utility, such ‘blind-swabbing’ approaches will necessarily co-sample cellular material from the different individuals whose cells are present on the item, even though the individuals' cells are principally located in topographically dispersed, but distinct, locations on the item. Thus the act of swabbing itself artifactually creates some of the DNA mixtures encountered in touch DNA samples. In some instances involving transient contact between an assailant and victim, the victim's DNA may be found in such significant excess as to preclude the detection and typing of the perpetrator's DNA. In order to circumvent the challenges with standard recovery and analysis methods for touch DNA evidence, we reported previously the development of a ‘smart analysis’ single cell recovery and DNA analysis method that results in enhanced genetic analysis of touch DNA evidence. Here we use the smart single cell analysis method to recover probative single source profiles from individual and agglomerated cells from various touched objects and clothing items belonging to known donors. We then use the same approach for the detection of single source male donor DNA in simulated physical contact/assault mixture samples (i.e. male ‘assailant’ grabbing the wrist, neck or clothing from the female ‘victim’, or being in transient contact with bedding from the ‘victim’). DNA profiles attributable to the male or female known donors were obtained from 31% and 35% of the single and agglomerated bio-particles (putative cells) tested. The known male donor ‘assailant’ DNA profile was identified in the cell sampling from every mixture type tested. The results of this work demonstrate the efficacy of an alternative strategy to recover single source perpetrator DNA profiles in physical contact/assault cases involving trace perpetrator/victim cellular admixtures.     

Law And Sovereignty

How is it possible that the idea of sovereignty still features in legal and political philosophy? Most contemporary political philosophers have little use for the idea of ‘unlimited’ or ‘absolute’ power, which is how sovereignty is normally defined. A closer look at sovereignty identifies two possible accounts: sovereignty as the fact of power or sovereignty as a title to govern. The first option, which was pursued by John Austin’s command theory of law, leads to an unfamiliar view of law and the state, which was justly criticised by H. L. A. Hart. The second option, leads to a paradox, because under this view sovereignty is both limited and unlimited. Hence, this argument shows that law and sovereignty are actually incompatible. Where there is law there is no sovereignty, and where there is sovereignty there is no law.     

Optimal storage conditions for highly dilute DNA samples: a role for trehalose as a preserving agent

DNA extraction from trace samples or noninvasively collected samples often results in the recovery of low concentration solutions of DNA that are prone to DNA degradation or other loss. Because of the difficulty in obtaining such samples, and their potentially high value in wildlife and forensic studies, it is critical that optimal methods are employed for their long-term storage. We assessed the amplification yield of samples kept under different storage conditions with the addition of potential preserving agents. We stored dilutions of known concentration human placental DNA, and gorilla fecal DNA, under four conditions (+4 degrees C, -20 degrees C, -80 degrees C, dry at room temperature), and with three additives (Tris EDTA (TE) buffer, Hind III digested Lambda DNA, trehalose). The effectiveness of the treatment methods was tested at regular intervals using qPCR to assess the quantity of amplifiable DNA, and a PCR assay of a larger 757 bp fragment to evaluate the quality of that remaining DNA. The highest quantity of DNA remained in samples stored at -80 degrees C, regardless of storage additives, and those dried at room temperature in the presence of trehalose. Surprisingly, DNA quality was best preserved in the presence of trehalose, either dried or at -80 degrees C; significant quality loss occurred with -20 degrees C and +4 degrees C storage.     

The role of DNA in criminal indictments in Israel

In their investigations of criminal cases, law enforcement agencies rely heavily on forensic evidence. Numerous studies have examined the scientific and technological advancements of DNA testing, but little evidence exists on how the availability of DNA evidence influences prosecutors' decisions to move cases forward in the criminal justice system. We created a new database by juxtaposing data from the Forensics Division of the Israel Police, which recorded the presence (or not) of DNA profiles in criminal cases (n = 9862), and data on the indictment decision for each case (2008–2019). Rates of indictments are computed for each case, and trend lines are used to present variations in the rates of indictment decisions with and without DNA profiles. Approximately 15% of all criminal cases without DNA presented to the prosecutor's office are subsequently prosecuted, compared with nearly 55% of cases with DNA profiles. The presence of DNA evidence influences the prosecutor's decision to move a case forward in the criminal justice system. Utilizing a scientific approach to prosecute offenders is a welcome development; however, DNA evidence is not infallible, and caution must be exercised in regard to DNA's overuse in the legal system.     

Trace DNA success rates relating to volume crime offences

In this study, 252 trace DNA samples (from handled surfaces) from 201 burglary, robbery and drugs cases were compiled to assess success rates and to interpret the value of trace DNA evidence in volume crime investigations. The average amount of DNA recovered from the trace DNA samples collected was 1.7 ng. Full or major (12 or more alleles) profiles were recovered from 14% of samples. Samples from firearms and burglary points of entry were the least successful. Mixtures were recovered from 21% of samples, presenting a case for the collection of more elimination profiles to enable more samples to be used for database purposes. The research highlighted the difficulties in collecting data relating to the success rates of samples. Computerised automation of this process would be extremely beneficial in the assistance of policy development, method application, training, and investigative usefulness.     

The tendency of individuals to transfer DNA to handled items

This research investigates factors influencing the transfer of DNA to handled objects and the process known as 'shedding'. Volunteers were recruited to hold sterile plastic tubes using experiments originally designed by Lowe et al. [A. Lowe, C. Murray, J. Whitaker, G. Tully, P. Gill, The propensity of individuals to deposit DNA and secondary transfer of low level DNA from individuals to inert surfaces, Forensic Sci. Int. 129 (2002) 25-34]. Transferred cellular material was collected from the tubes and STR profiles generated using the AmpFlSTR SGM Plus multiplex with 28 and 34 PCR cycles. Volunteers were asked to hold the tubes with each hand, and to participate in a series of handwashing experiments. The DNA profiling results obtained from the transferred skin cells were compared. An attempt was made to characterize the volunteers as 'good' or 'bad' shedders and to establish which, if any, of the experimental variables were associated with 'good' shedding. Our results suggest that many factors significantly influence shedding, including which hand an individual touches an item with and the time that has elapsed since they last washed their hands. We have found that it may be more complicated than previously reported to categorise a person as being either a 'good' or a 'bad' shedder and that if truly 'good' shedders exist they may be significantly rarer than some have estimated. In the current research no 'good' shedders were observed in a group of 60 volunteers. Given these results, it seems that rather than being applied to individual forensic cases, knowledge of shedding characteristics will be most useful in providing general background data for the interpretation of trace DNA evidence.     

Assessing a novel room temperature DNA storage medium for forensic biological samples

The ability to properly collect, analyze and preserve biological stains is important to preserving the integrity of forensic evidence. Stabilization of intact biological evidence in cells and the DNA extracts from them is particularly important since testing is generally not performed immediately following collection. Furthermore, retesting of stored DNA samples may be needed in casework for replicate testing, confirmation of results, and to accommodate future testing with new technologies.A novel room temperature DNA storage medium, SampleMatrix™ (SM; Biomatrica, Inc., San Diego, CA), was evaluated for stabilizing and protecting samples. Human genomic DNA samples at varying amounts (0.0625-200 ng) were stored dry in SM for 1 day to 1 year under varying conditions that included a typical ambient laboratory environment and also through successive freeze-thaw cycles (3 cycles). In addition, spiking of 1-4× SM into samples prior to analysis was performed to determine any inhibitory effects of SM. Quantification of recovered DNA following storage was determined by quantitative PCR or by agarose gel electrophoresis, and evaluation of quantitative peak height results from multiplex short tandem repeat (STR) analyses were performed to assess the efficacy of SM for preserving DNA.Results indicate no substantial differences between the quality of samples stored frozen in liquid and those samples maintained dry at ambient temperatures protected in SM. For long-term storage and the storage of low concentration samples, SM provided a significant advantage over freezer storage through higher DNA recovery. No detectable inhibition of amplification was observed at the recommended SM concentration and complete profiles were obtained from genomic DNA samples even in the presence of higher than recommended concentrations of the SM storage medium. The ability to stabilize and protect DNA from degradation at ambient temperatures for extended time periods could have tremendous impact in simplifying and improving sample storage conditions and requirements. The current work focuses on forensics analysis; however this technology is applicable to all endeavors requiring storage of DNA.     

The utility of whole genome amplification for typing compromised forensic samples

Biological evidence has become invaluable in the crime laboratory; however, it may exist in limited quantity and/or quality. Given this, the ability to amplify total DNA obtained from evidence, in an unbiased manner, would be highly advantageous. Methods for whole genome amplification (WGA) have the potential to fulfill this role, resulting in a virtually unlimited supply of DNA. In the research presented, two WGA methods, improved primer extension preamplification and multiple displacement amplification (MDA), were tested using commercial kits. Control DNA, artificially degraded DNA, and DNA from fresh blood, aged blood, hair shafts, and aged bones underwent WGA, followed by short tandem repeat and mitochondrial DNA analysis. The methods did amplify DNA, but performed poorly on forensically relevant samples; the maximum amplicon size was reduced, and MDA often resulted in extraneous bands following polymerase chain reaction. Taken together, WGA appears to be of limited forensic utility unless the samples are of a very high quality.     

Investigation into the usefulness of DNA profiling of earprints

DNA profiling of biological trace evidence has been used for many years. The application of this technique specifically to the DNA profiling of earprints has not to date been thoroughly investigated. This report presents the results of 60 earprints collected from three healthy adult volunteers under controlled laboratory conditions. DNA profile analysis revealed that high levels of non-donor alleles are observed when earprints are collected for DNA profiling. The source of these non-donor alleles is investigated and the impact that their presence within the profile may have on the use of this technique is discussed.     

The Influence of Evidence on Animal Cruelty Prosecution and Case Outcomes: Results of a Survey

Two hundred prosecuting attorneys completed a survey concerning priorities in taking on animal cruelty cases and the factors that help or hinder prosecuting such cases. Respondents commented on the priority given such cases. Questions also addressed specific kinds of evidence that had been used to decide whether to take on a cruelty case and were used in court. Results showed that prosecutors most frequently relied upon “traditional” sources of evidence, including detailed medical and crime scene reports and good quality photographic evidence. Other sources of forensic evidence such as DNA, computer forensics, forensic accounting, blood, and trace evidence were rarely employed. Veterinary forensic evidence, including forensic necropsies and detailed medical reports, was viewed as an important factor by a majority of prosecutors in deciding whether to accept a case for prosecution and in achieving a successful outcome, but a need for additional training for investigators was indicated.     

Female criminals—It's not always the offender!

Numerous crimes (including murder), all having a common denominator, occurred in Germany and Austria between 1993 and 2009. All of these cases presented with identical female DNA traces being found at the crime scene. The crimes committed differed markedly, as did the suspects involved, which were of varied origin. Many of these cases could be solved. However, none of the suspects could identify an involved female. Fourteen of these cases (including one murder) occurred in Upper Austria.A special task force of the Austrian police, together with the Institute of Legal Medicine in Salzburg, began systematically searching for errors in the investigative process after the cases became more and more incoherent and nebulous. In the end, the DNA trace evidence was shown to be contaminated. A woman involved in the manufacture of the cotton swabs turned out to be the source of the female DNA profile.Following this, several products of other manufacturers were tested for contamination with DNA. It was noted that cotton swabs which had been sterilised with radiation were often contaminated. As a result, it is recommended that the manufacturing process, as well as the products themselves used in collection of DNA trace evidence, should be re-evaluated with the emphasis on preventing contamination.     

The use of DNA stabilizing solution to enable room temperature storage and transportation of buccal and trace sample swabs

It is proposed that a DNA stabilizing solution (DNA Genotek Inc.) designed to preserve DNA in saliva samples at room temperature can be extrapolated to the storage of swab heads. The aim of this study was to evaluate the effectiveness of the solution for the preservation of reference swabs (buccal) and trace samples (facial swabs). To this end, the solution was used during a twin-site DNA transfer project assessing background levels of carer DNA present in children. Tubes containing 400 μl of solution were used to store and transport swab heads. At the laboratory, samples were extracted using the QIAamp DNA Mini Kit (Qiagen), quantified using the Quantifiler Duo Kit and profiled using the AmpF?STR^® SGM Plus^® PCR Amplification Kit (both Applied Biosystems). Twenty-eight PCR cycles were applied to all samples. Thirty-four cycles or a longer electrophoresis injection time was applied to trace samples where necessary. All Reference swabs produced high quantities of DNA and full DNA profiles after 28 cycles. Profile morphology indicated good quality DNA with no degradation. Of the trace samples, sufficient profiles were achieved to study the transfer of carer DNA making the solution fit for continued use in this project. DNA stabilizing solution enables the storage and transportation of swabs without freezing. This is convenient, reduces transportation costs and enables instant analysis of samples upon arrival at the laboratory. This is a useful alternative for a multi-site research project as well as a reliable storage tool for use in remote areas.     

Scent identification evidence in jurisdiction (drawing on the example of judicial practice in Poland)

One of the most significant challenges for contemporary forensic science seems to be research of new sources of physical evidence. Particularly after successful implementation of revolutionary DNA identification law enforcement agencies look for other new and perhaps more efficient techniques.     

Decision support for using mobile Rapid DNA analysis at the crime scene

Mobile Rapid DNA technology is close to being incorporated into crime scene investigations, with the potential to identify a perpetrator within hours. However, the use of these techniques entails the risk of losing the sample and potential evidence, because the device not only consumes the inserted sample, it is also is less sensitive than traditional technologies used in forensic laboratories. Scene of Crime Officers (SoCOs) therefore will face a ‘time/success rate trade-off’ issue when making a decision to apply this technology.In this study we designed and experimentally tested a Decision Support System (DSS) for the use of Rapid DNA technologies based on Rational Decision Theory (RDT). In a vignette study, where SoCOs had to decide on the use of a Rapid DNA analysis device, participating SoCOs were assigned to either the control group (making decisions under standard conditions), the Success Rate (SR) group (making decisions with additional information on DNA success rates of traces), or the DSS group (making decisions supported by introduction to RDT, including information on DNA success rates of traces).This study provides positive evidence that a systematic approach for decision-making on using Rapid DNA analysis assists SoCOs in the decision to use the rapid device. The results demonstrated that participants using a DSS made different and more transparent decisions on the use of Rapid DNA analysis when different case characteristics were explicitly considered. In the DSS group the decision to apply Rapid DNA analysis was influenced by the factors “time pressure” and “trace characteristics” like DNA success rates. In the SR group, the decisions depended solely on the trace characteristics and in the control group the decisions did not show any systematic differences on crime type or trace characteristic.Guiding complex decisions on the use of Rapid DNA analyses with a DSS could be an important step towards the use of these devices at the crime scene.     

The reincorporation and redistribution of trace geoforensic particulates on clothing: An introductory study

Two experimental studies were undertaken to investigate the processes of reincorporation and redistribution of trace evidence on garments when worn by a suspect or a victim (reincorporation) or after the garments have been seized and packaged for subsequent forensic analysis (redistribution). The first experiment utilised UV powder, an established proxy for geoforensic trace particulates and the second experiment utilised daffodil pollen transferred onto garments under conditions that mimicked forensic reality. It was demonstrated that reincorporation of trace particulates occurs from upper to lower parts of the same garment and also from upper garments to lower garments. Reincorporation also occurred to all areas of the lower garments, however the highest concentration of particulates was found to be the lap area of the jeans. Particulates also tended to be preserved around technical details such as stitching or relief design features of the garments. Thus the decay of particulates after a contact has been made does not necessarily involve a loss of those particulates from the entire system. These findings have implications for the interpretation of trace evidence when seeking to establish the source of initial contacts or the chronology of pertinent events. The second study demonstrated that folding and packaging items of clothing leads to a redistribution of any trace particulate evidence that is present thereby eliciting an alteration in the spatial distribution of that evidence. There is therefore a necessity to take the context of trace evidence into account and also to follow protocols that are sensitive to these aspects of trace evidence behaviour as a failure to do so may have consequences for the correct interpretation of such evidence.     

《正当防卫指导意见》中的程序问题评析

《正当防卫指导意见》中关于程序问题的若干规范对于正确办理涉正当防卫案件意义重大。侦查机关在取证工作中,应当着重注意对于能够证明不法侵害行为存在证据的及时收集。公检法机关在依法公正处理涉正当防卫案件时,尤其应当认真听取辩护方意见,及时披露案件信息,回应社会关切。对属于正当防卫的案件应当及时作出不予追究决定。但对于防卫过当案件均应适用认罪认罚从宽制度的要求似乎不妥。对于陪审制的运用应扬长避短。在案件办理的不同环节均应注意释法析理,尤其应当注意在认定为正当防卫的案件中对被害人及其家属进行释法析理。在做好法治宣传教育工作中,既要教育公民自觉遵守法律,更要教育公民敢于拿起法律的武器,积极行使防卫权利。     

Training of legal professionals in DNA evidence

The South African Criminal Legal System is based on Roman Dutch law. Court proceedings are led by a single presiding officer of the court. Prosecutors and defence advocates present the court with evidence in an adversarial manner. This system has inherent advantages and disadvantages and therefore the training of legal professionals in handling DNA evidence in court is important. The prosecutors resort under the National Prosecuting Authority and the defence advocates act independently or e.g. under the auspices of Legal Aid South Africa.Education curricula of legal professional do not include forensic science evidence. Principles such as evidential value in the forensic context are not addressed. Training of legal professionals with our Essential DNA Evidence™ Course has been a multiplier of forensic science knowledge in the legal profession in South Africa. We present prosecution and defence perspectives in an unbiased manner, compensating for the possible subjective interpretations of evidence that may be presented in court. Forensic evidence is subsequently carefully evaluated prior to being court presentation thus improving court efficiency, and allowing for a more focussed approach to the presentation of evidence. Approaches to the customisation of course content that adds value has been identified via evaluation of training programmes.Experience has shown that legal professionals have the ability to incorporate relatively complex scientific concepts into their legal arguments if provided with the appropriate training opportunity. Appropriate training in DNA evidence has made the court process more effective, both in terms of time and costs, and ultimately serves justice.     

Touch DNA of Shed Skin Cells from the Deployed Airbag to Address Drunken Driving Crimes

In the criminal cases of driving under the influence （DUI）, DNA evidence can be collectedfrom the deployed airbag of the motor vehicle and submitted to the crime lab for touch DNA analysis.The evidence can be acquired when the skin cells are observed on the surface of the airbag in a trafficaccident. However, the low quantity or quality of the evidence collected from a crime scene preventsfurther identification analysis in many cases. In the current study, we reported a case of identifyingtouch DNA extraction from the shed skin cells from the deployed airbag of a motor vehicle. We man-aged to collect DNA evidence from the shed skin cells in an airbag using a proper approach of collec-tion and extraction. The 5.87 ng of extracted DNA was sufficient for genotyping and forensic identifica-tion, which helped to identify the driver of the car in collision with a pier in the street. In DUI casesand other traffic accidents, therefore, the amount of touch DNA extracted from the deployed airbag canbe sufficient for DNA marker genotyping and further analysis.     

Knowledge on DNA Success Rates to Optimize the DNA Analysis Process: From Crime Scene to Laboratory

DNA analysis has become an essential intelligence tool in the criminal justice system for the identification of possible offenders. However, it appears that about half of the processed DNA samples contains too little DNA for analysis. This study looks at DNA success rates within 28 different categories of trace exhibits and relates the DNA concentration to the characteristics of the DNA profile. Data from 2260 analyzed crime samples show that cigarettes, bloodstains, and headwear have relatively high success rates. Cartridge cases, crowbars, and tie‐wraps are on the other end of the spectrum. These objective data can assist forensics in their selection process.The DNA success probability shows a positive relation with the DNA concentration. This finding enables the laboratory to set an evidence‐based threshold value in the DNA analysis process. For instance, 958 DNA extracts had a concentration value of 6 pg/μL or less. Only 46 of the 958 low‐level extracts provided meaningful DNA profiling data.