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During the past few decades, Raman spectroscopy has progressed and captivated added attention in the field of science. However, the application of Raman spectroscopy is not limited to the field of forensic science and analytical chemistry; it is one of the emerging spectroscopic techniques, utilized in the field of forensic science which in turn could be a supporting tool in the law and justice system. The advantage of Raman spectroscopy over the other conventional techniques is that it is rapid, reliable, and non-destructive in nature with minimal or no sample preparation. The quantitative and qualitative analysis of evidence from biological and non-biological origins could easily be performed by using Raman spectroscopy. The forensic domain is highly complex with multidisciplinary branches, and therefore a plethora of techniques are utilized for the detection, identification, and differentiation of innumerable pieces of evidence for the purpose of law and justice. Herein, a systematic review is carried out on the application of Raman spectroscopy in the realm of forensic biology and serology considering its usefulness in practical perspectives. This review paper highlights the significance of modern techniques, including micro-Raman spectroscopy, confocal Raman spectroscopy, surface-enhanced Raman spectroscopy, and paper-based surface-enhanced Raman spectroscopy, in the field of Raman spectroscopy. These techniques have demonstrated notable advancements in terms of their applications and capabilities. Furthermore, to comprehensively capture the progress in the development of Raman spectroscopy, all the published papers which could be retrieved from the available databases from the year 2007 to 2022 were incorporated. [ABSTRACT FROM AUTHOR]

The use of PCR is widespread in biological fields. Some fields, such as forensic biology, push PCR to its limits as DNA profiling may be required in short timeframes, may be produced from minute amounts of starting material, and may be required to perform in the presence of inhibitory compounds. Due to the extreme high-throughput of samples using PCR in forensic science, any small improvement in the ability of PCR to address these challenges can have dramatic effects for the community. At least part of the improvement in PCR performance could potentially come by altering PCR cycling conditions. These alterations could be general, in that they are applied to all samples, or they could be tailored to individual samples for maximum targeted effect. Further to this, there may be the ability to respond in real time to the conditions of PCR for a sample and make cycling parameters change on the fly. Such a goal would require both a means to track the conditions of the PCR in real time, and the knowledge of how cycling parameters should be altered, given the current conditions. In Part 1 of our work, we carry out the theoretical groundwork for the ambitious goal of creating a smart PCR system that can respond appropriately to features within individual samples in real time. We approach this task using an open qPCR instrument to provide real-time feedback and machine learning to identify what a successful PCR 'looks like' at different stages of the process. We describe the fundamental steps to set up a real-time feedback system, devise a method of controlling PCR cycling conditions from cycle to cycle, and to develop a system of defining PCR goals, scoring the performance of the system towards achieving those goals. We then present three proof-of-concept studies that prove the feasibility of this overall method. In a later Part 2 of our work, we demonstrate the performance of the theory outlined in this paper on a large-scale PCR cycling condition alteration experiment. The aim is to utilise machine learning so that throughout the process of PCR automatic adjustments can be made to best alter cycling conditions towards a user-defined goal. The realisation of smart PCR systems will have large-scale ramifications for biological fields that utilise PCR. [ABSTRACT FROM AUTHOR]

The introduction of PCR into forensic science and the rapid increases in the sensitivity, specificity and discrimination power of DNA profiling that followed have been fundamental in shaping the field of forensic biology. Despite these developments, the challenges associated with the DNA profiling of trace, inhibited and degraded samples remain. Thus, any improvement to the performance of sub-optimal samples in DNA profiling would be of great value to the forensic community. The potential exists to optimise the PCR performance of samples by altering the cycling conditions used. If the effects of changing cycling conditions upon the quality of a DNA profile can be well understood, then the PCR process can be manipulated to achieve a specific goal. This work is a proof-of-concept study for the development of a smart PCR system, the theoretical foundations of which are outlined in part 1 of this publication. The first steps needed to demonstrate the performance of our smart PCR goal involved the manual alteration of cycling conditions and assessment of the DNA profiles produced. In this study, the timing and temperature of the denaturation and annealing stages of the PCR were manually altered to achieve the goal of reducing PCR runtime while maintaining an acceptable quality and quantity of DNA product. A real-time feedback system was also trialled using an STR PCR and qPCR reaction mix, and the DNA profiles generated were compared to profiles produced using the standard STR PCR kits. The aim of this work was to leverage machine learning to enable real-time adjustments during a PCR, allowing optimisation of cycling conditions towards predefined user goals. A set of parameters was found that yielded similar results to the standard endpoint PCR methodology but was completed 30 min faster. The development of an intelligent system would have significant implications for the various biological disciplines that are reliant on PCR technology. [ABSTRACT FROM AUTHOR]

Sarcophaga (=Parasarcophaga) (Liopygia) ruficornis (Fabricius, 1794) is a species of medical–veterinary and forensic importance, as its immatures cause myiasis in humans and animals and colonize carcasses and cadavers. Therefore, investigations into the biology and morphology of this species, with a particular focus on pupae that constitute ≥50% of the developmental period for the immatures, are pertinent. Although there are biological and morphological studies of pupae, the intrapuparial development at different temperatures has not yet been analysed. Therefore, the present study aimed to describe how temperature (22, 27 and 32 ± 1°C) affects the development and morphology of S. (L.) ruficornis pupae at 60 ± 10% relative humidity and a 12‐h photoperiod. Ten pupae were collected, euthanized and fixed every 4 h from pupariation until 24 h and every 8 h until the emergence of the first adult. Emergence occurred at 440, 272 and 232 h at 22, 27 and 32°C, with 590, 380 and 330 pupae, respectively. The highest mortality occurred at 32°C. Eight periods were defined, and sex was determined in pharate adult stage; in addition, 40 key morphological characteristics to estimate pupal age were presented. These findings can serve as a basis for studies on the biology and morphology of the pupa of S. (L.) ruficornis, particularly in bionomics, control and forensics, helping researchers and experts. [ABSTRACT FROM AUTHOR]

This article is a tribute to the late Prof. Simon R. Leather, a highly respected entomologist. It includes personal reflections from colleagues who worked closely with Prof. Leather, highlighting his dedication, expertise, and impact on the field of entomology. The article also mentions his role as an editor for scientific journals and his contributions to research and education. Overall, it pays homage to Prof. Leather's significant contributions and influence in the field of entomology. [Extracted from the article]

Molecular biology is a complex, abstract, subject that can be challenging for higher education students to comprehend. The current manuscript describes the design, implementation, and evaluation of two immersive VR simulations of a DNA lab and a crime scene investigation (CSI) for a forensic molecular biology course in the context of the "TESLA" Erasmus+ project. It illustrates the instructional design and technical aspects of the VR simulations' development. The experimental study employed a comparative quantitative research design. The guiding research questions examined how instructional modalities (online vs. face-to-face) affect learners' perceptions of VR-based training in higher education and the key factors influencing learners' intention for their adoption. Forty-six (n = 46) undergraduate students completed a 17-item questionnaire, which served as the main data collection instrument. Results demonstrate that both online and face-to-face VR-based instruction can effectively convey core concepts, thus challenging the traditional notion that face-to-face interaction is inherently superior. Its implications underscore the potential of VR simulations to supplement or even substitute traditional teaching methods, particularly for complex science subjects. [ABSTRACT FROM AUTHOR]

Background New methods for processing ‘touch’ or trace biological samples is an ongoing priority for forensic caseworking laboratories. These samples often contain materials from multiple individuals in varying quantities and/or degrees of degradation. Rapid characterization of cellular material before DNA profiling can allow laboratories to screen samples for the presence of multiple contributors or the amount of biological material present. Methods This dataset contains autofluorescence and morphological profiles of epidermal cell populations analyzed using Imaging Flow Cytometry. The epidermal samples were aged for varying amounts of time prior to analysis. Multiple samples from the same individual were also collected to assess profile variations within and across the contributors. Conclusions This data set may be used to investigate variability in epidermal cell populations from different individuals and potential forensic signatures contained within the non-genetic components that comprise touch biological evidence.

Human remains have been recovered from a number of historic shipwrecks and their associated sites, often hundreds of years post-mortem. While remains of victims who have fled following wrecking can be subject to a range of environmental exposures, human remains in marine environments are subject to unique decomposition processes, faunal predation and impacts on DNA. Researchers in museums and academic institutions holding historic shipwreck remains have applied a plethora of scientific testing methods to extract information from artefacts and shipwreck remains. Specialist forensic DNA techniques, often adapted from ancient and archaeological DNA methods, are designed to maximize DNA recovery, and advances in technology and forensic biology have increased options for genotyping compromised human skeletal samples. A vast array of new genetic markers can now be targeted for interrogation to reveal externally visible characteristics, biogeographical ancestry or extended genetic relatives of victims. Some of these techniques have already been applied to historic shipwreck remains. This paper reviews current and emerging forensic DNA techniques available for recovering and revealing genetic information from historic shipwreck remains. It aims to direct investigators conducting genetic testing on historic shipwreck human remains to forensic DNA techniques as a possible approach for yielding further invaluable information. [ABSTRACT FROM AUTHOR]

Body fluids are one of the most encountered types of evidence in any crime and are commonly used for identifying a person's identity. In addition to these, they are also useful in ascertaining the nature of crime by determining the ty pe of fluid such as blood, semen, saliva, urine etc. Body fluids collected from crime scenes are mostly found in degraded, trace amounts and/or mixed with other fluids. However, the existing immunological and enzyme-based methods used for differentiating these fluids show limited specificity and sensitivity in such cases. To overcome these challenges, a new method utilizing microRNA expression of the body fluids has been proposed. This method is believed to be non-destructive as well as sensitive in nature and researches have shown promising results for highly degraded samples as well. This systematic review focuses on and explores the use and reliability of miRNAs in body fluid identification. It also summarizes the researches conducted on various aspects of miRNA in terms of body fluid examination in forensic investigations. [ABSTRACT FROM AUTHOR]

The study presents the importance of forensic bacteriology, its use in forensic cases, the methods for bacteriological sampling from corpses, the types and species of bacteria isolated from human and pig corpses, which are most commonly used in forensic biology. The microbial changes that occur after death remain unclear. Postmortem microbiology is a relatively new field of research. After death, the failure of the immune system and other physical barriers leads to the proliferation and spread of microbes. In order for bacteriological information to be accepted within the scope of forensic bacteriology, the court must find suspicion to be present about whether the bacteria seen on the body will contribute to solving the case. Experts must be appointed to examine the issue in line with this suspicion, and these experts must prepare and submit their reports to the court at the requested time. When considering the literature studies, forensic bacteriology has been suggested to be a scientific discipline in the developmental stage and to only be able to provide circumstantial evidence in forensic cases as opposed to primary evidence. According to the literature review, most bacterial studies isolated from corpses were conducted in Romania. Although bacterial samples were isolated from various parts of the corpses, bacteria were mostly isolated from their blood samples. According to literature searches from various scientific journal databases, no study has occurred with a list of the bacteria isolated from corpses. This study is thought to be able to fill this important gap. [ABSTRACT FROM AUTHOR]

Copyright of Forensic Science & Technology is the property of Institute of Forensic Science, Ministry of Public Security and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Parasites are ubiquitous, diverse, and have close interactions with humans and other animals. Despite this, they have not garnered significant interest from forensic scientists, and their utility as indicators in criminal investigations has been largely overlooked. To foster the development of forensic parasitology we explore the utility of parasites as forensic indicators in five broad areas: (i) wildlife trafficking and exploitation, (ii) biological attacks, (iii) sex crimes, (iv) criminal neglect of humans and other animals, and (v) indicators of movement and travel. To encourage the development and growth of forensic parasitology as a field, we lay out a four-step roadmap to increase the use and utility of parasites in criminal investigations. [ABSTRACT FROM AUTHOR]

Background: Different types of RNA take on multiple crucial functions in living cells and tissues. Messenger RNA (mRNA) is a temporary molecular carrier of genetic information. Analysis of the composition of all mRNA contained in a cell at a given moment, the so-called transcriptome, enables the determination of the type of cell and its condition, e.g., in pathologically altered states. Method: This review is based on pertinent publications retrieved by a selective literature search. Results: The analysis of differential gene expression has already been used in forensic molecular biology to determine the type of tissue contained in biological specimens. It is also being used in criminal investigations to determine the composition of mixed traces of various bodily fluids and/or organ tissues. The method is limited by degradation of the mRNA molecules through environmental influences. The use of newly developed molecular biological methods such as massive parallel sequencing can expand the information obtainable by this investigative method. Current research also addresses the forensic potential of deriving relevant information about the crime--e.g., its timing, or the condition of the involved persons--from the totality of mRNA species present in the specimens. Conclusion: Forensic RNA analysis can yield a great deal of relevant information. It is likely to be applicable in a much wider variety of forensic situations in the near future. [ABSTRACT FROM AUTHOR]

The use of messenger RNA for body fluid identification has been well documented. While reference genes are commonly used for normalizing gene expression, the use of reference genes in forensic science is not straightforward. Degradation of samples, mixed body fluids, and small volumes contribute to the complexity of normalizing gene expression. To demonstrate normalization of gene expression for forensic body fluid identification, a preliminary study involving two reference genes (beta-2-microglobulin, B2M and glucose-6-phosphate dehydrogenase, G6PD) was undertaken. Novel reverse-transcription quantitative PCR (RT-qPCR) assays for circulatory blood, buccal swabs, menstrual fluid, spermatozoa, seminal fluid, and vaginal material were designed. An appropriate PCR efficiency for the B2M and G6PD reference gene assays across the body fluid panel was confirmed prior to assessing their expression levels. B2M was found to be more highly expressed across the different body fluids than G6PD. The normalization of target assay expression using both genes was investigated. However, high variation and poor sensitivity, particularly in buccal and semen samples, were observed for both, which would limit sensitivity and mixture detection in forensic casework. If CT values are the intended output rather than comparative quantification, we propose that reference gene normalization is not required and could be detrimental to interpretation. [ABSTRACT FROM AUTHOR]

Badly burned skeletal remains are commonly submitted to forensic laboratories for victim identification via DNA analysis methods. Burned skeletal remains present many challenges for DNA analysis as they can contain low amounts of DNA which can also be damaged and degraded, resulting in partial or no STR profiles. Therefore, a simple, but effective screening method that identifies which samples may provide the most successful STR or mtDNA typing results for identification would enable forensic laboratories to save time, money, and resources. One metric that can be used and a screening method is the color of burned bone, as bone color changes with exposure to fire as temperature and length of exposure increase. This research developed a quantitative screening method based on the surface color of burned bone. The different visual bone colors (light brown, dark brown, black, gray, and white) were quantified using the Commission on Illumination L*a*b color space. These values were then compared to DNA yield, STR, and mtDNA profile completeness to identify whether the L*a*b values can predict genotyping success. A Bayesian network was constructed to determine the probability of STR typing success, given a set of L*a*b values. Results demonstrated that samples with an a* value greater than or equal to one and b* value greater than eight (light brown and dark brown burned samples) were the most predictive of STR typing success and mtDNA typing success. A decision tree for processing burned bones was constructed based on the color value thresholds. [ABSTRACT FROM AUTHOR]

Copyright of Forensic Science & Technology is the property of Institute of Forensic Science, Ministry of Public Security and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Identifying the body fluid source of stains can give an indication of activities, prioritize samples for DNA profiling, and provide probative evidence when the presence of DNA alone does not. Messenger RNA profiling assays using endpoint reverse-transcription polymerase-chain reaction (RT-PCR), were developed to address the issues with conventional body fluid identification and have since been used in forensic casework. However, endpoint RT-PCR has a narrow linear dynamic range (LDR), and there is currently no specific method for mRNA quantification to direct case strategy and optimize input template. Real-time reverse-transcription quantitative PCR (RT-qPCR) collects data during the amplification phase which reflects the starting quantity. The application of RT-qPCR assays as a quantification precursor to endpoint RT-PCR was investigated using previously described assays for circulatory blood, saliva, menstrual fluid, spermatozoa, seminal fluid, and vaginal material. While blood marker SLC4A1 and vaginal marker CYP2B7P showed a strong relationship, the remaining body fluid markers lacked a robust predictive relationship, likely due to differences in LDR. CYP2B7P and SLC4A1 showed high sensitivity when compared with other markers – resulting in a greater number of data points for estimation of the linear relationship. Compared with endpoint, the RT-qPCR assays had higher precision, LDR, and sensitivity. [ABSTRACT FROM AUTHOR]

This document is an editorial from the International Journal of Molecular Sciences that discusses the importance of molecular biology in various fields, including forensic science. It highlights advancements in molecular biology techniques that have improved forensic investigations, paternity disputes, and the identification of missing persons. The editorial also mentions potential future developments in forensic molecular biology, such as next-generation sequencing and microRNA analysis. The Special Issue includes ten articles that cover topics such as post-mortem interval determination, sampling methods in forensic genetics, DNA methylation-based age prediction, and the use of animal genomics in forensic science. The articles provide new findings and contribute to the advancement of molecular biology applications in forensic science. The editorial concludes by emphasizing the need for future research efforts and standardized protocols and guidelines in the field. [Extracted from the article]

Mitochondrial (mt) DNA plays an important role in the fields of forensic and clinical genetics, molecular anthropology, and population genetics, with mixture interpretation being of particular interest in medical and forensic genetics. The high copy number, haploid state (only a single haplotype contributed per individual), high mutation rate, and well-known phylogeny of mtDNA, makes it an attractive marker for mixture deconvolution in damaged and low quantity samples of all types. Given the desire to deconvolute mtDNA mixtures, the goals of this study were to (1) create a new software, MixtureAceMT™, to deconvolute mtDNA mixtures by assessing and combining two existing software tools, MixtureAce™ and Mixemt, (2) create a dataset of in-silico MPS mixtures from whole mitogenome haplotypes representing a diverse set of population groups, and consisting of two and three contributors at different dilution ratios, and (3) since amplicon targeted sequencing is desirable, and is a commonly used approach in forensic laboratories, create biological mixture data associated with two amplification kits: PowerSeq™ Whole Genome Mito (Promega™, Madison, WI, USA) and Precision ID mtDNA Whole Genome Panel (Thermo Fisher Scientific by AB™, Waltham, MA, USA) to further validate the software for use in forensic laboratories. MixtureAceMT™ provides a user-friendly interface while reducing confounding features such as NUMTs and noise, reducing traditionally prohibitive processing times. The new software was able to detect the correct contributing haplogroups and closely estimate contributor proportions in sequencing data generated from small amplicons for mixtures with minor contributions of ≥5%. A challenge of mixture deconvolution using small amplicon sequencing is the potential generation of spurious haplogroups resulting from private mutations that differ from Phylotree. MixtureAceMT™ was able to resolve these additional haplogroups by including known haplotype/s in the evaluation. In addition, for some samples, the inclusion of known haplotypes was also able to resolve trace contributors (minor contribution 1–2%), which remain challenging to resolve even with deep sequencing. [ABSTRACT FROM AUTHOR]

Body fluid identification is an essential step in the forensic biology workflow that can assist DNA analysts in determining where to collect DNA evidence. Current presumptive tests lack the specificity that molecular techniques can achieve; therefore, molecular methods, including microRNA (miRNA) and microbial signature characterization, have been extensively researched in the forensic community. Limitations of each method suggest combining molecular markers to increase the discrimination efficiency of multiple body fluids from a single assay. While microbial signatures have been successful in identifying fluids with high bacterial abundances, microRNAs have shown promise in fluids with low microbial abundance (blood and semen). This project synergized the benefits of microRNAs and microbial DNA to identify multiple body fluids using DNA extracts. A reverse transcription (RT)‐qPCR duplex targeting miR‐891a and let‐7g was validated, and miR‐891a differential expression was significantly different between blood and semen. The miRNA duplex was incorporated into a previously reported qPCR multiplex targeting 16S rRNA genes of Lactobacillus crispatus, Bacteroides uniformis, and Streptococcus salivarius to presumptively identify vaginal/menstrual secretions, feces, and saliva, respectively. The combined classification regression tree model resulted in the presumptive classification of five body fluids with 94.6% overall accuracy, now including blood and semen identification. These results provide proof of concept that microRNAs and microbial DNA can classify multiple body fluids simultaneously at the quantification step of the current forensic DNA workflow. [ABSTRACT FROM AUTHOR]

Optical tweezers have a wide range of uses for mechanical manipulation of objects in the microscopic range. This includes both living and static cells in a variety of biomedical and research applications. Single‐focus optical tweezers, formed by focusing a laser beam through a high numerical aperture immersion objective, create a significant force, which enables controlled transport of a variety of different cell types and morphologies in three dimensions. Optical tweezers have been previously reported to capture and separate spermatozoa from a reconstituted simulated postcoital sample. We report herein the development of a simplified, more efficient cell transfer protocol that can separate and isolate both spermatozoa as well as leukocytes, with similar efficiencies as those previously reported. The new cell transfer method was used to separate sperm cells from a reconstituted mixture of spermatozoa and vaginal epithelial cells, with complete STR profiles developed from 50 cells with little evidence of contribution from the female contributor to the mixture. This modified protocol was then used to separate 21 samples of enriched leukocytes, with trapped cells ranging from 5 to 22 cells. Complete STR profiles were developed from as few as 10 leukocytes. Thus, with minimal sample preparation and a short trapping time, this method has the potential to provide an alternative to traditional differential extraction methods for separation of sperm:nonsperm mixtures while also providing versatility for separation of cells with differing morphologies. [ABSTRACT FROM AUTHOR]

DNA analysis obtained from forensic biological evidence provides strong evidence in identifying the perpetrator associated with the incident. The interindividual difference in the number of STR regions and the low mutation rate in these regions have made STRs preferred as genetic markers for donor identification. However, if amount of DNA in the forensic biological material is too low or the DNA is corrupted to such an extent that it doesn't allow analysis, short tandem repeat regions, which are frequently used in forensic genetic methods, cannot be determined. In the case of doubt, when there is no match to the STR profile, any information that can assist in identifying the sample's donor would be invaluable. Therefore, in the forensic genetics, in the diagnosis of identity of biological samples that are difficult to analyze, determining the additional information about the physical appearence of the donor such as age, hair and eyes colours with forensic DNA phenotyping, also such as body fluid and tissue type determination with mRNA and miRNA analyses, new current molecular methods have started to develop in the genetic and epigenetic field. In this review, for analysis of STRs and other markers we review recent advances in this field to maximize the analysis potential in identifying the phenotype of interest, largely through the application of MSP, developments in the interpretation of mixture DNA profiles containing genetic material of more than one person, RNA profiling for body fluid identification, and inclusion of epigenetic methods such as examination of methylation profiles.

With the continuous development and integration of molecular biology and forensic science, non-coding RNAs (ncRNAs), especially ncRNAs with regulatory functions such as microRNA, long non-coding RNA, and circular RNA, have recently been actively explored by forensic scholars. In this study, we review the literature on these ncRNAs in various fields of forensic science, including postmortem interval determination, wound age estimation, forensic age assessment, cause of death analysis, and body fluid identification, aiming to evaluate the current research and provide a perspective for future applications. [ABSTRACT FROM AUTHOR]

One of the most important pieces of evidence to find the suspect of a sexual assault are semen stains. It has been observed that suspects do not use condoms in many cases and transfer seminal fluids over the body of the victim, to their clothes, to bed linen, or to other objects in the environment. However, in many cases, it has been seen that the suspects or the victims tend to clean their bodies and clothes because of psychological conditions such as fear, panic, and anxiety. This study aims to determine whether human identification can be accomplished from washed semen stains by evaluating the quantity of extracted DNA obtained from washed clothes. In this study, both cotton- and nylon-blended fabrics were stained with semen samples taken from 10 volunteers and washed with detergent at 40 °C, 60 °C, and 90 °C. After stains were made re-visible using an ultraviolet (UV) light source, DNA was extracted using two different extraction methods (phenol–chloroform and spin-column). As a result, DNA extraction methods, washing conditions, and obtained DNA quantity from different washed fabrics were compared. It was shown that DNA could be obtained from all samples and that these DNA amounts could be used to perform identification. This study gives hope to the victims who have not been able to go to the judicial units immediately after the incident for fear or any other reasons and who think that they cannot get results because they have washed the laundry. [ABSTRACT FROM AUTHOR]

Synchrotron X‐ray phase‐contrast microtomography (SR‐PhC micro‐CT) is well established, fast and non‐destructive imaging technique for data acquisition that is currently being used to obtain new insights into insect anatomy and function in physiological, morphological and phylogenetic studies. In this study, we described in situ the internal organs of the red flour beetle Tribolium castaneum Herbst 1797, a widespread pest of cereals and stored food causing serious damage to the human economy. Two‐dimensional virtual sections and volumetric reconstructions of the nervous, alimentary and reproductive systems were carried out in both sexes. The results provided a comprehensive overview of the morphological characteristics of this species, such as the different maturation stages of ovarioles and the realistic location, size and shape of internal organs. Given the great interest in this model species in experimental biology and forensic entomology, complete knowledge of the general anatomy is required for future functional applications in pest control and experimental studies. In addition, this study confirms SR‐PhC micro‐CT as a powerful and innovative tool in entomology, particularly suitable for small species and chitinized structures that are difficult to analyse using conventional dissection and histological methods. [ABSTRACT FROM AUTHOR]

Copyright of Forensic Science & Technology is the property of Institute of Forensic Science, Ministry of Public Security and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Forensic Science & Technology is the property of Institute of Forensic Science, Ministry of Public Security and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Considering activity level propositions in the evaluation of forensic biology findings is becoming more common place. There are increasing numbers of publications demonstrating different transfer mechanisms that can occur under a variety of circumstances. Some of these publications have shown the possibility of DNA transfer from site to site on an exhibit, for instance as a result of packaging and transport. If such a possibility exists, and the case circumstances are such that the area on an exhibit where DNA is present or absent is an observation that is an important diagnostic characteristic given the propositions, then site to site transfer should be taken into account during the evaluation of observations. In this work we demonstrate the ways in which site to site transfer can be built into Bayesian networks when carrying out activity level evaluations of forensic biology findings. We explore the effects of considering qualitative vs quantitative categorisation of DNA results. We also show the importance of taking into account multiple individual's DNA being transferred (such as unknown or wearer DNA), even if the main focus of the evaluation is the activity of one individual. • Site-to-site DNA transfer on an exhibit can be important to consider in an evaluation. • We show several methods of incorporating site-to-site transfer in a Bayesian network. • Modelling DNA amounts vs presence/absence has an effect on evaluation outcomes. • Modelling multiple people's DNA transfer within a trace effects evaluation outcome [ABSTRACT FROM AUTHOR]

Forensic Biology is contingent upon matching DNA profiles between a crime sample and a reference sample. There are several capillary electrophoresis kits available to generate a short tandem repeat (STR) profile from DNA samples, while newer methods using massively parallel sequencing are slowly being implemented in forensic laboratories worldwide. During evaluation of a newer capillary electrophoresis kit, Applied Biosystems™ VeriFiler™ Plus, a discordance was observed in the Penta D locus. The previous kit, Promega PowerPlex 21® System produced a 13.4,14 genotype, whilst VeriFiler™ Plus produced a 14,14 genotype. An expanded investigation into Penta D microvariant alleles revealed that multiple discordances were observed for DNA profiles containing larger x.4 variants. There was full concordance between PowerPlex® 21 and QIAGEN Investigator® 26plex, however discordances were observed between VeriFiler™ Plus and the other three kits tested, including the massively parallel sequencing kit, Verogen ForenSeq® MainstAY. Notably, four of these discordances resulted in null alleles with the VeriFiler™ Plus kit. A review of the Penta D DNA sequences in MainstAY revealed fully concordant microvariant alleles involved deletions within the repeat region, whilst variability in the discordances observed were dependent on the location of the variation outside the repeat region and the analysis method used. Variations observed within the 5' flanking region produced the same allele designation across all capillary electrophoresis kits. However, deletions within the 3' region either produced a null allele for VeriFiler™ Plus where the deletion is thought to overlap the primer binding site, or microvariant alleles for the PowerPlex® 21 and Investigator 26plex kits, which produced longer Penta D amplicons. The discovery of these variations in the Penta D flanking sequences is informative as it increases the awareness of Penta D discordances between different kit chemistries in nominated reference DNA profile comparisons and DNA database searching and matching alike, and provides support for this phenomenon when providing evidence as to the admissibility of such results in trial proceedings. • Discordances in allele calls can be observed between different forensic kits. • Sequencing identified several deletions in the Penta D flanking sequence. • Kit specific Penta D allele designations was dependent upon deletion position. • Scientists should consider discordances when reporting results from different kits. [ABSTRACT FROM AUTHOR]

This review paper covers the forensic-relevant literature in biological sciences from 2019 to 2022 as a part of the 20th INTERPOL International Forensic Science Managers Symposium. Topics reviewed include rapid DNA testing, using law enforcement DNA databases plus investigative genetic genealogy DNA databases along with privacy/ethical issues, forensic biology and body fluid identification, DNA extraction and typing methods, mixture interpretation involving probabilistic genotyping software (PGS), DNA transfer and activity-level evaluations, next-generation sequencing (NGS), DNA phenotyping, lineage markers (Y-chromosome, mitochondrial DNA, X-chromosome), new markers and approaches (microhaplotypes, proteomics, and microbial DNA), kinship analysis and human identification with disaster victim identification (DVI), and non-human DNA testing including wildlife forensics. Available books and review articles are summarized as well as 70 guidance documents to assist in quality control that were published in the past three years by various groups within the United States and around the world.

While often necessary in sexual assault cases, confirmatory identification of body fluids can be a lengthy and/or costly process. In particular, the detection of vaginal fluid and menstrual fluid in forensic casework is limited to endpoint reverse-transcription PCR to detect fluid-specific messenger RNA (mRNA) markers as there are no robust chemical or enzymatic techniques available for these fluids. Similarly, testing for rectal mucosa is not possible with standard methods, the presence of which would provide probative value in cases of alleged anal penetration, although mRNA-based markers have recently been described. Reverse-transcription loop-mediated isothermal amplification (RT-LAMP) is an alternative technique that enables detection of mRNA at a single temperature (usually 60-65℃) for 10-30 minutes and has comparable sensitivity to PCR. We describe the coupling of RT-LAMP amplification (60℃ for 30 minutes) with CRISPR-mediated fluorescent detection of the body fluid specific mRNA markers MMP3 (menstrual fluid), CYP2B7P (vaginal material), TNP1 (spermatozoa), KLK2 (semen), and MUC12 (rectal mucosa). Following temperature optimization and final selection of RT-LAMP-CRISPR assays, their specificity across circulatory blood, buccal, menstrual fluid, vaginal material, semen, and rectal mucosa was assessed. Most assays were specific for their intended target body fluid, although MMP3 and CYP2B7P were detected in some rectal mucosa samples, the latter of which has been observed previously in the literature. A preliminary sensitivity assessment in target fluids was determined by a dilution series over six logs of RNA input. A range of assay approaches were investigated to develop a protocol suitable for use in a forensic screening laboratory. This included the determination of fluorescent assay results by eye, use of lyophilised reagents, and RT-LAMP and CRISPR reactions undertaken in one-tube in a lower resource setting., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Copyright of Forensic Science & Technology is the property of Institute of Forensic Science, Ministry of Public Security and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Forensic Science & Technology is the property of Institute of Forensic Science, Ministry of Public Security and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Forensic Science & Technology is the property of Institute of Forensic Science, Ministry of Public Security and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Forensic Science & Technology is the property of Institute of Forensic Science, Ministry of Public Security and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Forensic Science & Technology is the property of Institute of Forensic Science, Ministry of Public Security and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Forensic Science & Technology is the property of Institute of Forensic Science, Ministry of Public Security and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)