DNA transfer when using gloves in burglary simulations.

Several studies have demonstrated that DNA can be indirectly transferred from an individual onto a surface. Therefore, the presence of DNA that is compatible with a given person does not necessarily mean that this person has touched the surface on which the DNA was recovered. The present work simulates cases, where DNA is recovered on a door handle and compared to several reference DNA profiles. The DNA profile of the trace shares DNA components with a person of interest (POI). When asked about the DNA results, the POI says he has nothing to do with the incident and has never been at the scene. However, a possibility would be that the DNA came from his recently stolen gloves. Someone else, the alternative offender (AO), could have opened the door wearing his gloves (POI's gloves), and transferred his DNA (POI's DNA). Based on the above-mentioned scenario, 60 burglary simulations experiments were carried out to generate data to assess DNA results given these allegations. The quantity and quality of DNA profiles (NGM SElect) recovered when the POI opened/closed the door bare-handed or when someone else performed the same activity but using POI's gloves, were compared. The gloves were regularly worn during at least three months by their owner during the winter. On the contrary, the AO wore them only for two minutes. Among the traces collected on the door handles, less than 50% of the traces led to interpretable DNA profiles. In 30% of the cases (3/10), when the door was opened/closed with bare hands, the DNA found on the door handle led to a mixed DNA profile with the POI's DNA aligning with the major contributor. For the experiments where the AO opened/closed the door with the POI's gloves, the POI's DNA was compatible with 22% (11/50) of the mixed DNA profile, aligning with the major in 8% of the cases (4/50). The DNA profiles of the offices' occupants were observed on the door handles, but not the AO's. In addition to the results of the experiments, we show two examples of how one can assess results observed in casework. Given the possibility of indirect transfer of minute DNA quantities, this research emphasizes the need to evaluate DNA results given the activities when the POI has a legitimate reason that can explain the presence of their DNA.

A probabilistic approach to evaluate salivary microbiome in forensic science when the Defense says: `It is my twin brother'.

Salivary microbiota profiles may represent a valid contribution to forensic investigation when standard DNA genotyping methods fail. Starting from questioned and control materials in the form of saliva, the evidence can be expressed by means of a distance between those materials taking into account specific aspects of the microbiota composition. The value of the evidence for forensic discrimination purposes is quantified by means of a Bayes' factor, that allows one to overcome the major limitations and pitfalls of intuition connected to the use of cut-off values as a mean of decision.

Voiding cystourethrography and 99MTC-MAG3 renal scintigraphy in pediatric vesicoureteral reflux: what is the role of indirect cystography?

BACKGROUND: Primary vesicoureteral reflux (VUR) is the most common urological anomaly in children. Voiding cystourethrography (VCUG) is considered the reference standard for the diagnosis of VUR. Even if it is a secure and standardized technique, it is still an invasive method, hence, the effort to find an alternative method to diagnose VUR. The aim of the study is to evaluate the diagnostic accuracy of 99mTC-MAG3 scintigraphy with indirect cystography in detecting VUR and to estimate any interobserver variability in 99mTC-MAG3 scintigraphy interpretation. METHODS: The authors retrospectively reviewed all the pediatric patients who underwent both a VCUG and a 99mTC-MAG3 renal scintigraphy at the study institution between 2012 and 2016. RESULTS: A total of 86 children (and 168 renal units) were included. MAG3 scan revealed a sensitivity of 54% and a specificity of 90% with positive predictive value of 79% and negative predictive value of 73%. Each MAG3 scintigraphy was then independently and blindly evaluated by a pediatric urologist and two nuclear physicians. After revision, the concordance between VCUG and MAG3 in reflux cases dropped from 54% to 27% (on average), and the reviewers reclassified most examinations as non-conclusive. CONCLUSIONS: 99mTC-MAG3 renal scintigraphy with indirect cystography showed low sensitivity in detecting VUR of any grade and cannot, therefore, be proposed as completely alternative to VCUG in the diagnosis of VUR. Moreover, MAG3 scintigraphy interpretation for the diagnosis of VUR has a very high interobserver variability, mostly because of the lack of a correct and complete voiding phase.

More on the question 'When does absence of evidence constitute evidence of absence?' How Bayesian confirmation theory can logically support the answer.

In forensic science it is not rare that common sayings are used to support particular inferences. A typical example is the adage 'The absence of evidence is not evidence of absence'. This paper analyzes the rationale hidden behind such statement and it offers a structural way to approach the analysis of this particular adage throughout a careful analysis of four different scenarios.

Critical analysis of forensic cut-offs and legal thresholds: A coherent approach to inference and decision.

In this paper we critically discuss the definition and use of cut-off values by forensic scientists, for example in forensic toxicology, and point out when and why such values - and ensuing categorical conclusions - are inappropriate concepts for helping recipients of expert information with their questions of interest. Broadly speaking, a cut-off is a particular value of results of analyses of a target substance (e.g., a toxic substance or one of its metabolites in biological sample from a person of interest), defined in a way such as to enable scientists to suggest conclusions regarding the condition of the person of interest. The extent to which cut-offs can be reliably defined and used is not unanimously agreed within the forensic science community, though many practitioners - especially in operational laboratories - rely on cut-offs for reasons such as ease of use and simplicity. In our analysis, we challenge this practice by arguing that choices made for convenience should not be to the detriment of balance and coherence. To illustrate our discussion, we will choose the example of alcohol markers in hair, used widely by forensic toxicologists to reach conclusions regarding the drinking behaviour of individuals. Using real data from one of the co-authors' own work and recommendations of cut-offs published by relevant professional organisations, we will point out in what sense cut-offs are incompatible with current evaluative guidelines (e.g., [31]) and show how to proceed logically without cut-offs by using a standard measure for evidential value. Our conclusions run counter to much current practice, but are inevitable given the inherent definitional and conceptual shortcomings of scientific cut-offs. We will also point out the difference between scientific cut-offs and legal thresholds and argue that the latter - but not the former - are justifiable and can be dealt with in logical evaluative procedures.

Analysing and exemplifying forensic conclusion criteria in terms of Bayesian decision theory.

There is ongoing discussion in forensic science and the law about the nature of the conclusions reached based on scientific evidence, and on how such conclusions - and conclusion criteria - may be justified by rational argument. Examples, among others, are encountered in fields such as fingermarks (e.g., 'this fingermark comes from Mr. A's left thumb'), handwriting examinations (e.g., 'the questioned signature is that of Mr. A'), kinship analyses (e.g., 'Mr. A is the father of child C') or anthropology (e.g., 'these are human remains'). Considerable developments using formal methods of reasoning based on, for example (Bayesian) decision theory, are available in literature, but currently such reference principles are not explicitly used in operational forensic reporting and ensuing decision-making. Moreover, applied examples, illustrating the principles, are scarce. A potential consequence of this in practical proceedings, and hence a cause of concern, is that underlying ingredients of decision criteria (such as losses quantifying the undesirability of adverse decision consequences), are not properly dealt with. There is merit, thus, in pursuing the study and discussion of practical examples, demonstrating that formal decision-theoretic principles are not merely conceptual considerations. Actually, these principles can be shown to underpin practical decision-making procedures and existing legal decision criteria, though often not explicitly apparent as such. In this paper, we will present such examples and discuss their properties from a Bayesian decision-theoretic perspective. We will argue that these are essential concepts for an informed discourse on decision-making across forensic disciplines and the development of a coherent view on this topic. We will also emphasize that these principles are of normative nature in the sense that they provide standards against which actual judgment and decision-making may be compared. Most importantly, these standards are justified independently of peoples' observable decision behaviour, and of whether or not one endorses these formal methods of reasoning.

A solution for the rare type match problem when using the DIP-STR marker system.

The rare type match problem is an evaluative challenging situation in which the analysis of a DNA profile reveals the presence of (at least) one allele which is not contained in the reference database. This situation is challenging because an estimate for the frequency of occurrence of the profile in a given population needs sophisticated evaluative procedures. The rare type match problem is very common when the DIP-STR marker system, which has proven itself very useful for dealing with unbalanced DNA mixtures, is used, essentially due to the limited size of the available database. The object-oriented Bayesian network proposed in Cereda et al. [7] to assess the value of the evidence for general scenarios, was not designed to deal with this particular situation. In this paper, the model is extended and partially modified to be able to calculate the full Bayesian likelihood ratio in presence of any (observed and not yet observed) allele of a given profile. The method is based on the approach developed in Cereda [5] for Y-STR data. Alternative solutions, such as the plug-in approximation and an empirical Bayesian methodology are also proposed and compared with the results obtained with the full Bayesian approach.

Altered TDP-43-dependent splicing in HSPB8-related distal hereditary motor neuropathy and myofibrillar myopathy.

BACKGROUND AND PURPOSE: Mutations in the small heat-shock protein 22 gene (HSPB8) have been associated with Charcot-Marie-Tooth disease type 2L, distal hereditary motor neuropathy (dHMN) type IIa and, more recently, distal myopathy/myofibrillar myopathy (MFM) with protein aggregates and TDP-43 inclusions. The aim was to report a novel family with HSPB8K141E -related dHMN/MFM and to investigate, in a patient muscle biopsy, whether the presence of protein aggregates was paralleled by altered TDP-43 function. METHODS: We reviewed clinical and genetic data. We assessed TDP-43 expression by qPCR and alternative splicing of four previously validated direct TDP-43 target exons in four genes by reverse transcriptase-polymerase chain reaction. RESULTS: The triplets and their mother presented in the second to third decade of life with progressive weakness affecting distal and proximal lower limb and truncal muscles. Nerve conduction study showed a motor axonal neuropathy. The clinical features, moderately raised creatin kinase levels, selective pattern of muscle involvement on magnetic resonance imaging and pathological changes on muscle biopsy, including the presence of protein aggregates, supported the diagnosis of a contemporary primary muscle involvement. In affected muscle tissue we observed a consistent alteration of TDP-43-dependent splicing in three out of four TDP-43-target transcripts (POLDIP3, FNIP1 and BRD8), as well as a significant decrease of TDP-43 mRNA levels. CONCLUSIONS: Our study confirmed the role of mutated HSPB8 as a cause of a combined neuromuscular disorder encompassing dHMN and MFM with protein aggregates. We identified impaired RNA metabolism, secondary to TDP-43 loss of function, as a possible pathological mechanism of HSPB8K141E toxicity, leading to muscle and nerve degeneration.

The meaning of justified subjectivism and its role in the reconciliation of recent disagreements over forensic probabilism.

In this paper we reply to recent comments in this Special Issue according to which subjective probability is not considered to be a concept fit for use in forensic evaluation and expert reporting. We identify the source of these criticisms to lie in a misunderstanding of subjective probability as unconstrained subjective probability; a lack of constraint that neither corresponds to the way in which we referred to subjective probability in our previous contributions, nor to the way in which probability assignment is understood by current evaluative guidelines (e.g., of ENFSI). Specifically, we explain that we understand subjective probability as a justified assertion, i.e. a conditional assessment based on task-relevant data and information, that may be thought of as a constrained subjective probability. This leads us to emphasise again the general conclusion that there is no gap between justified (or, reasonable) subjective probability and other concepts of probability in terms of its ability to provide assessments that are soundly based on whatever relevant information available. We also note that the challenges an expert faces in reporting probabilities apply equally to all interpretations of probability, not only to subjective probability.

The consequences of understanding expert probability reporting as a decision.

In this paper we reiterate that the personalist interpretation of probability is inevitable and as least as informed as any other allegedly more 'objective' definition of probability. We also argue that the problem faced by forensic scientists, the reporting on imperfect personal knowledge, in terms of probabilities, can be reconstructed as a decision problem. Tackling this problem through a rigorous decision theoretic analysis provides further argument in support of the view that optimal probability reporting is in terms of single numbers, not intervals.

Reframing the debate: A question of probability, not of likelihood ratio.

Evidential value is measured by a likelihood ratio. This ratio has two components, the probability, or probability density, of the evidence if the prosecution proposition is true and the probability (density) of the evidence if the defence proposition is true. It takes the form of a single value, even if these probabilities are subjective measures of belief of the reporting forensic scientist.

Analysis and evaluation of magnetism of black toners on documents printed by electrophotographic systems.

This paper reports on a study to assess the potential of measurements of magnetism, using a proprietary magnetic analysis system, for the routine analysis of toners on documents printed by black and white electrophotographic systems. Magnetic properties of black toners on documents printed by a number of different devices were measured and compared. Our results indicate that the analysis of magnetism is complementary to traditional methods for analysing black toners, such as FTIR. Further, we find that the analysis of magnetism is realistically applicable in closed set cases, that is when the number of potential printing devices can be clearly defined.

Probabilistic evaluation of n traces with no putative source: A likelihood ratio based approach in an investigative framework.

Analysis of marks recovered from different crime scenes can be useful to detect a linkage between criminal cases, even though a putative source for the recovered traces is not available. This particular circumstance is often encountered in the early stage of investigations and thus, the evaluation of evidence association may provide useful information for the investigators. This association is evaluated here from a probabilistic point of view: a likelihood ratio based approach is suggested in order to quantify the strength of the evidence of trace association in the light of two mutually exclusive propositions, namely that the n traces come from a common source or from an unspecified number of sources. To deal with this kind of problem, probabilistic graphical models are used, in form of Bayesian networks and object-oriented Bayesian networks, allowing users to intuitively handle with uncertainty related to the inferential problem.

A novel synonymous mutation in the MPZ gene causing an aberrant splicing pattern and Charcot-Marie-Tooth disease type 1b.

Charcot-Marie-Tooth disease (CMT) is an inherited peripheral neuropathy with a heterogeneous genetic background. Here, we describe two CMT1B families with a mild sensory-motor neuropathy and a novel synonymous variant (c.309G > T, p.G103G) in exon 3 of the MPZ gene. Next generation sequencing analysis on a 94 CMT gene panel showed no mutations in other disease genes. In vitro splicing assay and mRNA expression analysis indicated that the c.309T variant enhances a cryptic donor splice site at position c.304 resulting in the markedly increased expression of the r.304_448del alternative transcript in patients' cells. This transcript is predicted to encode a truncated P0 protein (p.V102Cfs11*) lacking the transmembrane domain, thus suggesting a possible haploinsufficiency mechanism for this mutation. This is the third reported synonymous MPZ variant associated with CMT1 and affecting splicing. These data confirm the functional impact of synonymous variants on MPZ splicing and their possible role as disease-causing mutations rather than silent polymorphisms.

The decisionalization of individualization.

Throughout forensic science and adjacent branches, academic researchers and practitioners continue to diverge in their perception and understanding of the notion of 'individualization', that is the claim to reduce a pool of potential donors of a forensic trace to a single source. In particular, recent shifts to refer to the practice of individualization as a decision have been revealed as being a mere change of label [1], leaving fundamental changes in thought and understanding still pending. What is more, professional associations and practitioners shy away from embracing the notion of decision in terms of the formal theory of decision in which individualization may be framed, mainly because of difficulties to deal with the measurement of desirability or undesirability of the consequences of decisions (e.g., using utility functions). Building on existing research in the area, this paper presents and discusses fundamental concepts of utilities and losses with particular reference to their application to forensic individualization. The paper emphasizes that a proper appreciation of decision tools not only reduces the number of individual assignments that the application of decision theory requires, but also shows how such assignments can be meaningfully related to constituting features of the real-world decision problem to which the theory is applied. It is argued that the decisonalization of individualization requires such fundamental insight to initiate changes in the fields' underlying understandings, not merely in their label.

Early white matter involvement in an infant carrying a novel mutation in ACOX1.

We describe the clinical findings and MRI features observed in a child who presented a two-step disease course: he was hypotonic at birth and soon afterwards developed seizures, which were partially responsive to treatment; he subsequently showed developmental delay and a progressive neurological deterioration with the onset of severe seizures at around three years of age. Head MRI at age 20 days was unremarkable, whereas at 25 months it showed bilateral hyperintensity of the deep cerebellar nuclei; five months later, the signal hyperintensity was also present in the cerebellar white matter and ventral pontine fibre tracts. Molecular analysis revealed a novel ACOX1 mutation, predicting a largely truncated protein. The white matter involvement, which followed an ascending trajectory from cerebellar and brainstem structures to the cerebral hemispheres, seemed to originate from the perinuclear white matter of the deep cerebellar nuclei.

Statistical hypothesis testing and common misinterpretations: Should we abandon p-value in forensic science applications?

Many people regard the concept of hypothesis testing as fundamental to inferential statistics. Various schools of thought, in particular frequentist and Bayesian, have promoted radically different solutions for taking a decision about the plausibility of competing hypotheses. Comprehensive philosophical comparisons about their advantages and drawbacks are widely available and continue to span over large debates in the literature. More recently, controversial discussion was initiated by an editorial decision of a scientific journal [1] to refuse any paper submitted for publication containing null hypothesis testing procedures. Since the large majority of papers published in forensic journals propose the evaluation of statistical evidence based on the so called p-values, it is of interest to expose the discussion of this journal's decision within the forensic science community. This paper aims to provide forensic science researchers with a primer on the main concepts and their implications for making informed methodological choices.

The importance of distinguishing information from evidence/observations when formulating propositions.

The value of forensic results crucially depends on the propositions and the information under which they are evaluated. For example, if a full single DNA profile for a contemporary marker system matching the profile of Mr A is assessed, given the propositions that the DNA came from Mr A and given it came from an unknown person, the strength of evidence can be overwhelming (e.g., in the order of a billion). In contrast, if we assess the same result given that the DNA came from Mr A and given it came from his twin brother (i.e., a person with the same DNA profile), the strength of evidence will be 1, and therefore neutral, unhelpful and irrelevant(1) to the case at hand. While this understanding is probably uncontroversial and obvious to most, if not all practitioners dealing with DNA evidence, the practical precept of not specifying an alternative source with the same characteristics as the one considered under the first proposition may be much less clear in other circumstances. During discussions with colleagues and trainees, cases have come to our attention where forensic scientists have difficulty with the formulation of propositions. It is particularly common to observe that results (e.g., observations) are included in the propositions, whereas-as argued throughout this note-they should not be. A typical example could be a case where a shoe-mark with a logo and the general pattern characteristics of a Nike Air Jordan shoe is found at the scene of a crime. A Nike Air Jordan shoe is then seized at Mr A's house and control prints of this shoe compared to the mark. The results (e.g., a trace with this general pattern and acquired characteristics corresponding to the sole of Mr A's shoe) are then evaluated given the propositions 'The mark was left by Mr A's Nike Air Jordan shoe-sole' and 'The mark was left by an unknown Nike Air Jordan shoe'. As a consequence, the footwear examiner will not evaluate part of the observations (i.e., the mark presents the general pattern of a Nike Air Jordan) whereas they can be highly informative. Such examples can be found in all forensic disciplines. In this article, we present a few such examples and discuss aspects that will help forensic scientists with the formulation of propositions. In particular, we emphasise on the usefulness of notation to distinguish results that forensic scientists should evaluate from case information that the Court will evaluate.