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Oxycodone is a strong opioid drug commonly used to treat acute, cancer, and chronic non-malignant pain. In this study, all oxycodone-related medico-legal cases where death had occurred in a hospital or nursing home in Finland were investigated to determine the range of post-mortem (PM) oxycodone blood concentrations in a therapeutic setting. All toxicology cases in which oxycodone was detected in PM femoral blood during the 4-year period of 2016–2019 in Finland were retrieved from the national PM toxicology database. In this material, the 365 deceased hospital patient cases that met the study inclusion criteria were divided into four groups according to the cause and manner of death. The reference group of 121 fatal oxycodone poisoning cases comprised two groups: those with verified associated drug abuse and those without drug abuse. The median oxycodone concentration in PM blood was significantly higher in cancer patients (0.10 mg/L) than in patients with recent surgery (0.07 mg/L) or other disease (0.06 mg/L) (p p

Informed consent is based on basic ethical principles that should be considered when conducting biomedical and behavioral research involving human subjects. These principles—respect, beneficence, and justice—form the foundations of informed consent which in itself is grounded on three fundamental elements: information, comprehension, and voluntary participation. While informed consent has focused on human subjects and research, the practice has been adopted willingly in the forensic science arena primarily to acquire reference samples from family members to assist in identifying missing persons. With advances in molecular biology technologies, data mining, and access to metadata, it is important to assess whether the past informed consent process and in particular associated risks are concomitant with these increased capabilities. Given the state-of-the-art, areas in which informed consent may need to be modified and augmented are as follows: reference samples from family members in missing persons or unidentified human remains cases; targeted analysis of an individual(s) during forensic genetic genealogy cases to reduce an investigative burden; donors who provide their samples for validation studies (to include population studies and entry into databases that would be applied to forensic statistical calculations) to support implementation of procedures and operations of the forensic laboratory; family members that may contribute samples or obtain genetic information from a molecular autopsy; and use of medical and other acquired samples that could be informative for identification purposes. The informed consent process should cover (1) purpose for collection of samples; (2) process to analyze the samples (to include type of data); (3) benefits (to donor, target, family, community, etc. as applicable); (4) risks (to donor, target, family, community, etc. as applicable); (5) access to data/reports by the donor; (6) sample disposition; (7) removal of data process (i.e., expungement); (8) process to ask questions/assessment of comprehension; (9) follow-up processes; and (10) voluntary, signed, and dated consent. Issues surrounding these topics are discussed with an emphasis on addressing risk factors. Addressing informed consent will allow human subjects to make decisions voluntarily and with autonomy as well as secure the use of samples for intended use.

Laboratories and their criminal justice systems are confronted with challenges for implementing new technologies, practices, and policies even when there appears to be demonstrative benefits to operational performance. Impacting decisions are the often higher costs associated with, for example, new technologies, limited current budgets, and making hard decisions on what to sacrifice to take on the seemingly better approach. A prospective cost–benefit analysis (CBA) could help an agency better formulate its strategies and plans and more importantly delineate how a relatively small increase to take on, for example, a new technology can have large impact on the system (e.g., the agency, other agencies, victims and families, and taxpayers). To demonstrate the process and potential value a CBA was performed on the use of an alternate and more expensive swab with reported better DNA yield and being certified human DNA free (i.e., nylon 4N6FLOQSwabs®), versus the traditional less costly swab (i.e., cotton swab). Assumptions are described, potential underestimates and overestimates noted, different values applied (for low and modest to high), and potential benefits (monetary and qualitative) presented. The overall outcome is that the cost of using the more expensive technology pales compared with the potential tangible and intangible benefits. This approach could be a guide for laboratories (and associated criminal justice systems) worldwide to support increased funding, although the costs and benefits may vary locally and for different technologies, practices, and policies. With well-developed CBAs, goals of providing the best services to support the criminal justice system and society can be attained.

Although knee measurements yield high classification rates in metric sex estimation, there is a paucity of studies exploring the knee in artificial intelligence-based sexing. This proof-of-concept study aimed to develop deep learning algorithms for sex estimation from radiographs of reconstructed cadaver knee joints belonging to the Terry Anatomical Collection. A total of 199 knee radiographs were obtained from 100 skeletons (46 male and 54 female cadavers; mean age at death 64.2 years, range 50-102 years) whose tibiofemoral joints were recon-structed in standard anatomical position. The AIDeveloper software was used to train, validate, and test neural network architectures in sex estimation based on image classification. Of the explored algorithms, an MhNet-based model reached the highest overall testing accuracy of 90.3%. The model was able to classify all females (100.0%) and most males (78.6%) correctly. These preliminary findings encourage further research on artificial intelligence-based methods in sex estimation from the knee joint. Combining radiographic data with automated and externally validated algorithms may establish valuable tools to be utilized in forensic anthropology.

Background: Quantitative changes in the cardiac autonomic nervous system may play an important role in the pathogenesis of various cardiovascular diseases. In the present morphological analysis, we aimed to study autonomic nerve density in myocardial sleeves and surrounding fibro-fatty tissue around caval veins. We correlated the nerve distribution with cardiovascular mortality and a history of atrial fibrillation. Materials and Methods: A total of 24 autopsied adult hearts were excised together with the superior and inferior vena cava and grouped according to the immediate and underlying causes of death (cardiovascular vs. non-cardiovascular), and documented heart rhythm history (atrial fibrillation vs. sinus rhythm). The density of autonomic nerves was quantified by measuring the area of immunohistochemical staining for sympathetic (tyrosine hydroxylase, TH) and parasympathetic (choline acetyltransferase, CHAT) nerves and ganglia. Growth-associated protein 43 (GAP43) was used as a neural growth marker. Results: The mean density of TH-positive nerves in the superior vena cava myocardial sleeves was significantly decreased between groups with documented underlying cardiovascular vs. non-cardiovascular cause of death (mean density ± standard deviation (SD): 704.81±1016.41 µm2/mm2 vs. 2391.01±1841.37 µm2/mm2; P =.008). Similarly, the nerve density of GAP43-positive nerves in the superior vena cava myocardial sleeves was significantly lower in subjects with documented underlying cardiovascular cause of death (mean density ± SD: 884.74±1240.16 µm2/mm2 vs. 2132.89±1845.89 µm2/mm2; P =.040). The mean age was significantly higher in subjects with documented underlying cardiovascular vs. non-cardiovascular cause of death (mean age ± SD: 69.2±11.9 years, vs. 57.5±11.2 years, P =.016). No differences were found in nerve densities of TH-positive (953.01±1042.93 µm2/mm2 vs. 919.26±1677.58 µm2/mm2), CHAT-positive (180.8±532.9 µm2/mm2 vs. 374.22±894.76 µm2/mm2), and GAP43-positive nerves (593.58±507.97 µm2/mm2 vs. 1337.34±1747.69 µm2/mm2) in myocardial sleeves around the inferior vena cava between groups with documented immediate cardiovascular vs. non-cardiovascular cause of death. Similarly, no differences were found between groups with documented underlying cardiovascular vs. non-cardiovascular cause of death (TH: 717.23±887.31 µm2/mm2 vs. 1365.51±2149.10 µm2/mm2; CHAT: 256.18±666.86 µm2/mm2 vs. 368.53±959.47 µm2/mm2; GAP43: 661.21±839.51 µm2/mm2 vs. 1759.90±2008.80 µm2/mm2). Moreover, there was no association found in nerve densities between subjects with documented atrial fibrillation vs. sinus rhythm (TH: 235.07±425.69 µm2/mm2 vs. 1166.08±1563.84 µm2/mm2; CHAT: 648.59±1017.33 µm2/mm2 vs. 175.31±641.65 µm2/mm2; GAP43: 990.17±1315.18 µm2/mm2 vs. 1039.86±1467.23 µm2/mm2). Conclusions: Decrease of superior vena cava myocardial sleeve sympathetic nerves may be associated with cardiovascular mortality and/or aging. No difference in autonomic innervation was found between subjects with documented atrial fibrillation vs. sinus rhythm. publishedVersion

The medical autopsy (also called hospital or clinical autopsy) is a highly specialised medical procedure, which requires professional expertise and suitably equipped facilities. To ensure high standards of performance, the Working Group of Autopsy Pathology of the European Society of Pathology (ESP) suggests a code of practice as a minimum standard for centres performing medical autopsies. The proposed standards exclusively address autopsies in adults, and not forensic autopsies, perinatal/or paediatric examinations. Minimum standards for organisation, standard of premises, and staffing conditions, as well as minimum requirements for level of expertise of the postmortem performing specialists, documentation, and turnaround times of the medical procedure, are presented. Medical autopsies should be performed by specialists in pathology, or by trainees under the supervision of such specialists. To maintain the required level of expertise, autopsies should be performed regularly and in a number that ensures the maintenance of good practice of all participating physicians. A minimum number of autopsies per dedicated pathologist in a centre should be at least 50, or as an average, at least one autopsy per working week. Forensic autopsies, but not paediatric/perinatal autopsies may be included in this number. Turnaround time for final reports should not exceed 3 weeks (14 working days) for autopsies without fixation of brain/spinal cord or other time-consuming additional examinations, and 6 weeks (30 working days) for those with fixation of brain/spinal cord or additional examinations.

Little is known about the potential of artificial intelligence in forensic shotgun pattern interpretation. As shooting distance is among the main factors behind shotgun patterning, this proof-of-concept study aimed to explore the potential of neural net architectures to correctly classify shotgun pattern images in terms of shooting distance. The study material comprised a total of 106 shotgun pattern images from two discrete shooting distances (n = 54 images from 10 m and n = 52 images from 17.5 m) recorded on blank white paper. The dataset was used to train, validate and test deep learning algorithms to correctly classify images in terms of shooting distance. The open source AIDeveloper software was used for the deep learning procedure. In this dataset, a TinyResNet-based algorithm reached the highest testing accuracy of 94%. Of the testing set, the algorithm classified all 10 m patterns correctly, and misclassified one 17.5 m pattern. On the basis of these preliminary data, it seems achievable to develop algorithms that would serve as a beneficial tool for forensic investigators when estimating shooting distances from shotgun patterns. In the future, studies with larger and more complex datasets are needed to develop robust and applicable algorithms for forensic shotgun pattern interpretation.

Sex estimation is an important part of osteological analysis of skeletons and forensic identification process. Traditionally cranial and pelvic traits are utilized for accurate sex estimation. However, post-cranial measurements have also been proven to accurately estimate sex especially from robust bones such as the femur. In this study, we investigated the potential of knee breadth dimensions in sex estimation in a Finnish population. To conduct this study we utilized a study sample (n=1654) belonging to the Northern Finland Birth Cohort 1966. All individuals were 46 years of age at the time of the examination. Three knee breadth dimensions were measured from subjects' knee posteroanterior radiographs: femoral biepicondylar breadth (FBEB), mediolateral breadth of the femoral condyles (FCML), and mediolateral breadth of the tibial plateau (TPML). Sex estimation was performed using logistic regression. The study clearly demonstrated that all three measurements were different between males and females. Sectioning points for individual knee breadth measurements were 82.9 mm for FBEB, 76.6 mm for FCML and 75.4 mm for TPML. The classification rates ranged from 90.9% to 93.6%. The less commonly used measurements of FCML and TPML showed higher accuracy than FBEB in sex estimation. Our study confirmed that knee breadths can be successfully utilized to improve sex estimation in cases where the skeleton is only partially preserved and other major components of sex estimation are absent. We can also provide new standards for sex estimation from the knee joint in a Finnish population. Sex estimation is an important part of osteological analysis of skeletons and forensic identification process. Traditionally cranial and pelvic traits are utilized for accurate sex estimation. However, post-cranial measurements have also been proven to accurately estimate sex especially from robust bones such as the femur. In this study, we investigated the potential of knee breadth dimensions in sex estimation in a Finnish population. To conduct this study we utilized a study sample (n=1654) belonging to the Northern Finland Birth Cohort 1966. All individuals were 46 years of age at the time of the examination. Three knee breadth dimensions were measured from subjects' knee posteroanterior radiographs: femoral biepicondylar breadth (FBEB), mediolateral breadth of the femoral condyles (FCML), and mediolateral breadth of the tibial plateau (TPML). Sex estimation was performed using logistic regression. The study clearly demonstrated that all three measurements were different between males and females. Sectioning points for individual knee breadth measurements were 82.9 mm for FBEB, 76.6 mm for FCML and 75.4 mm for TPML. The classification rates ranged from 90.9 % to 93.6 %. The less commonly used measurements of FCML and TPML showed higher accuracy than FBEB in sex estimation. Our study confirmed that knee breadths can be successfully utilized to improve sex estimation in cases where the skeleton is only partially preserved and other major components of sex estimation are absent. We can also provide new standards for sex estimation from the knee joint in a Finnish population.

Publisher Copyright: © 2021 The Author(s) Propranolol is a widely used beta-blocker mainly prescribed for the treatment of hypertension and other cardiac conditions. This medicine is also a frequent finding in drug screens, but little is known about its post-mortem toxicological profile. Our aim was to examine all post-mortem toxicology cases positive for propranolol in a three-year period, between 2016 and 2018 in Finland, and to compare these cases to those positive for metoprolol, another beta-blocker commonly used to treat cardiac diseases. There were 179 cases positive for propranolol and 416 for metoprolol in the study period. In the majority of propranolol cases (53%), the drug concentration in the blood was above the typical therapeutic range, but among the metoprolol cases this proportion was 18%. Propranolol was significantly more common than metoprolol in fatal poisonings, suicides and in cases with a history of drug abuse. Alcohol, benzodiazepines, antipsychotics and antidepressants were significantly more often detected in propranolol cases than in metoprolol cases. The deceased positive for propranolol were significantly younger than those positive for metoprolol. Cardiovascular diseases as the underlying cause of death were significantly more common among the metoprolol cases than among the propranolol cases. Our results showed significant differences between the propranolol group and the metoprolol group in post-mortem toxicology cases. The two drugs were used by two very different groups of people, with propranolol use being associated with psychiatric conditions.

We present a case of an accidental fatal fentanyl overdose caused by increased uptake of the drug from a transdermal patch while experiencing the heat of a sauna. The transdermal patch administers fentanyl at a relatively constant rate through the skin. However, in the subcutaneous tissue, blood circulation greatly influences the rate of this drug's systemic intake. In the present case, an elderly woman with multiple health conditions was prescribed fentanyl patches but was unaware of the risks associated with external heat sources when one wears the patch. She was found dead in the sauna with a postmortem femoral blood concentration of fentanyl that was elevated (15 mu g/L). The cause of death was determined to be fatal poisoning by fentanyl with the contributing factor of external heat from the sauna. Risks associated with transdermal administration of a potent opioid-like fentanyl are widely described in the scientific literature and described in the manufacturer's summary of product characteristics. Physicians and pharmacists should take particular care to ensure that patients understand these risks. Non

This study is the seventh report on fatal poisonings among drug addicts in the Nordic countries. In this report, we analyse data from the five Nordic countries: Denmark, Finland, Iceland, Norway and Sweden. Data on gender, number of deaths, places of deaths, age, main intoxicants and substances detected in blood were recorded to obtain national and comparable Nordic data, and to allow comparison with earlier studies conducted in 1984, 1991, 1997, 2002, 2007 and 2012. The death rate (number of deaths per 100,000 inhabitants) was highest in Iceland (6.58) followed closely by Sweden (6.46) and then lowest in Denmark (4.29). The death rate increased in Finland (5.84), Iceland and Sweden and decreased in Denmark compared to earlier studies. The death rate in Norway, which has decreased since 2002, has stabilised around 5.7 as of 2017. Women accounted for 7-23% of the fatal poisonings. The percentage was lowest in Iceland and highest in Finland and Norway. The age range was 14-70 years. The median age (41 years) was highest in Denmark and Norway. The other countries had a median age between 33 and 35 years. Opioids were the main cause of death. Methadone remained the main intoxicant in Denmark, while heroin/morphine was still the main intoxicant in Norway, as was buprenorphine in Finland. However, the picture has changed in Sweden compared to 2012, where heroin/morphine caused most deaths in 2017. Sweden also experienced the highest number of deaths from fentanyl analogues (67 deaths) and buprenorphine (61 deaths). Deaths from fentanyl analogues also occurred in Denmark, Finland and Norway, but to a smaller extent. Over the years, the proportion of opioid deaths has decreased in all countries except Sweden, which has experienced an increase. This decline has been replaced by deaths from CNS stimulants like cocaine, amphetamine and methylenedioxymethamphetamine (MDMA). Cocaine deaths have occurred in all countries but most frequently in Denmark. MDMA deaths have increased in all countries but mostly in Finland. Poly-drug use was widespread, as seen in the earlier studies. The median number of detected drugs per case varied from 4-6. Heroin/morphine, methadone, buprenorphine, cocaine, amphetamine, methamphetamine, MDMA, tetrahydrocannabinol (THC) and benzodiazepines were frequently detected. Pregabalin and gabapentin were detected in all countries, especially pregabalin, which was detected in 42% of the Finnish cases. New psychoactive substances (NPS) occurred in all countries except Iceland. (C) 2020 Elsevier B.V. All rights reserved.

The imprints left by persistent DNA viruses in the tissues can testify to the changes driving virus evolution as well as provide clues on the provenance of modern and ancient humans. However, the history hidden in skeletal remains is practically unknown, as only parvovirus B19 and hepatitis B virus DNA have been detected in hard tissues so far. Here, we investigated the DNA prevalences of 38 viruses in femoral bone of recently deceased individuals. To this end, we used quantitative PCRs and a custom viral targeted enrichment followed by next generation sequencing. The data was analyzed with a tailor-made bioinformatics pipeline. Our findings revealed bone to be a much richer source of persistent DNA viruses than earlier perceived, discovering ten additional ones, including several members of the herpesand polyomavirus families, as well as human papillomavirus 31 and torque teno virus. Remarkably, many of the viruses found have oncogenic potential and/or may reactivate in the elderly and immunosuppressed individuals. Thus, their persistence warrants careful evaluation of their clinical significance and impact on bone biology. Our findings open new frontiers for the study of virus evolution from ancient relics as well as provide new tools for the investigation of human skeletal remains in forensic and archaeological contexts.

Flualprazolam is a novel designer benzodiazepine, structurally related to alprazolam, flubromazolam and triazolam. In the last couple of years, it has been frequently detected in seizures and in forensic cases in Sweden and Finland. However, there is a lack of published blood concentrations for the drug, which presents difficulties when assessing its relevance for the cause of death. A quantitative method for the determination of flualprazolam in post-mortem blood was developed and validated, and subsequently used to analyse samples from 33 deaths previously screened as testing positive for flualprazolam in Sweden and Finland. Most of the cases in the study were accidental deaths (61 %) or suicides (18 %). The median (range) flualprazolam concentration was 18.0 (3.0-68) ng/g. The majority of the deceased were male (82 %) and the median age was 30 years. The median age in the Swedish cases was significantly higher (35 years) than in the Finnish cases (23 years) (p

Purity assessment of seized material containing new psychoactive substances (NPS) is complicated without appropriate primary reference standards. Here we present a method for fast quantitative estimation of stimulant-type NPS with use of secondary reference standards, based on gas chromatography nitrogen chemiluminescence detection coupled with atmospheric pressure chemical ionization quadrupole time-of-flight mass spectrometry (GC-NCD-APCI-QTOFMS). Quantification was based on the detector’s N-equimolar response to nitrogen and using two external nitrogen-containing calibrators, MDMA for prim- and sec- amines and α-PVP for tert- amines. Sample preparation involved dissolving the seized powdery material in an organic solvent mixture followed by acylation with N-methyl-bis-trifluoroacetamide (MBTFA). The method’s between-day accuracy and precision over a five-day period was measured for twenty-eight stimulants: the grand mean equimolarity was 91.9% (CV 5.5%), as compared with primary reference standards. The GC-NCD-APCI-QTOFMS method was applied to the purity estimation of forty-two seized powder samples previously found to contain stimulant-type NPS by appropriate methods. The quantitative results were compared to those obtained by an established method relying on liquid chromatography chemiluminescence detection (LC-CLND), the latter using caffeine as an external calibrator. The mean difference of purity values between the methods was 8.1% (range 0.4 - 26.7%). The presented method might find use as a tool for instant purity assessment in forensic laboratories.

We describe the sudden death of a middle-aged man while having a sauna under the influence of α-pyrrolidinovalerophenone (α-PVP) (PM blood concentration: 0.8 mg/L), amphetamine (0.34 mg/L), and other drugs (buprenorphine, benzodiazepines), and engaging in solitary sexual activities. The drugs’ effects on the cardio-circulatory system and on body thermoregulation combined with the high temperatures are likely to have been central mechanisms leading to death. The high levels of adrenaline triggered by sexual arousal and the respiratory depression caused by buprenorphine, in association with benzodiazepines, may have also contributed to his death. This previously unreported type of accidental autoerotic death illustrates the risk of using amphetamine-like sympathomimetic drugs (e.g. cathinone derivates) in hot environments such as a sauna, and during sexual activities therein.

Diabetes mellitus (DM) could cause pilot incapacitation and result in aviation fatalities. The mechanisms could be directly as a consequence of acute hypoglycemia/subacute diabetic ketoacidosis (DKA) or indirectly as an acute cardiovascular event by contributing to the development of atherosclerosis in coronary or carotid and cerebral arteries. In this study, DM-related fatal flight accidents in the US National Transport Bureau's database between years 2011-2016 were analyzed with special emphasis on postmortem (PM) glucose levels and correlation of toxicological reports with anamnestic information on DM. Additionally, autopsy results on coronary arteries were reviewed. In 43 out of 1491 (similar to 3%) fatal accidents pilots had DM. Postmortem glucose or glycated hemoglobin percentage (Hb1Ac) was measured in 12 of the 43 cases; while antidiabetic medication was found in 14 of the cases (only two of the cases had both glucose measurements and medication). With the increasing prevalence of DM, a possibility of pilot incapacitation due to DM or complications of DM should be actively studied, even if no anamnestic information of DM was available. While PM hypoglycemia is difficult to assess, we propose a systematic investigation based on measurement of glucose, Hb1Ac%, and ketone bodies, and documentation of atherosclerotic lesions in major arteries to identify or rule out DM as a cause of pilot incapacitation.

Autosomal DNA data from Peru for human identity testing purposes are scarce in the scientific literature, which hinders obtaining an appropriate portrait of the genetic variation of the resident populations. In this study we genetically characterize five populations from the Northeastern Peruvian Andes (Chachapoyas, Awajun, Wampis, Huancas and Cajamarca). Autosomal short tandem repeat (aSTR) and identity informative single nucleotide polymorphism (iiSNP) data from a total of 233 unrelated individuals are provided, and forensic genetic parameters are calculated for each population and for the combined set Northeastern Peruvian Andes. After correction for multiple testing in the whole dataset of the Northeastern Peruvian Andes, the only departure from Hardy-Weinberg equilibrium was observed in locus rs2111980. Twenty one out of 27 aSTR loci exhibited an increased number of alleles due to sequence variation in the repeat motif and flanking regions. For iiSNPs 33% of the loci displayed flanking region variation. The combined random match probability (RMP), assuming independence of all loci (aSTRs and iiSNPs), in the Chachapoyas, the population with the largest samples size (N = 172), was 8.14 x 10(-62) for length-based data while for sequence-based was 4.15 x 10(-67). In the merged dataset (Northeastern Peruvian Andes; N = 233), the combined RMP when including all markers were 2.96 x 10(-61) (length-based) and 3.21 x 10(-66) (sequence-based). These new data help to fill up some of the gaps in the genetic canvas of South America and provide essential length- and sequence-based background information for other forensic genetic studies in Peru.

It is not known how mortality differs between psychopathic and nonpsychopathic individuals. We linked data from subjects having been in forensic mental examinations at Niuvanniemi Hospital during 1984–1993 to the data from the National Death Registry to estimate the association between psychopathy and mortality. One hundred psychopathic individuals scoring 25 or higher in the PCL-R scale were followed up for 20–30 years. Two control groups were used as follows: 178 offenders scoring less than 25 on the PCL-R, and sample of general population drawn from the Finnish National Statistics database. Results reveal that psychopaths die younger than the general population, and the causes of death are more violent than in the nonpsychopath control group. There was a significant positive correlation between PCL-R score and mortality, and the mortality among psychopaths was about fivefold when compared with general population.

U-47,700 is a synthetic opioid that emerged on the novel psychoactive substance market a few years ago. After incorporating the substance into the urine UPLC-TOF-MS screening used in post-mortem toxicology, the drug was detected in 10 autopsy cases within routine case work. In all cases, the cause of death was accidental poisoning by U-47,700 alone or in combination with other psychoactive substances. The concentration of U-47,700 in the blood samples ranged between 0.15-2.0 mg/L with a median of 0.30 mg/L. In one of the cases with a U-47,700 concentration of 0.27 mg/L, no other psychoactive substances were detected. The stored TOF-MS analytical data from the year preceding the incorporation of U-47,700 into the screening was reprocessed in order to search for more positive cases. The data-independent acquisition of the original screening allowed for retrospective re-analysis of the full-scan data without additional experiments on the actual sample. The retrospective data-analysis revealed two additional cases positive for U-47,700. The first mention of U-47,700 on a Finnish internet discussion forum was in March 2015. After having been detected in several death cases, the drug was put under national control in November 2016 and the last fatality occurred in 2017. The toxic lifespan of U-47,700 thus lasted for approximately 2 years in Finland. Forensic and clinical laboratories need to rapidly adjust their screening procedures in order to adapt to the continuously expanding field of novel psychoactive substances. Retrospective data-analysis is a practical tool for monitoring the emergence of new substances onto the market. (C) 2019 Elsevier B.V. All rights reserved.

Estimation of an individual’s age has important applications in forensics. In young individuals, it often relies on separate evaluations of permanent teeth (PT) and third molars (TM) development. Here we analysed the age prediction performance of combined information from PT and TM in an unusual sample of healthy Somalis, born and living in Finland. PT development was staged according to Demirjian et al. (Hum Biol, 1973) and TM development according to Köhler et al. (Ann Anat, 1994), using panoramic radiographs from 803 subjects (397 males, 406 females) aged 3-23 years. A sex-specific Bayesian age-estimation model for the multivariate distribution of the stages conditional on age was fitted on PT, TM, and PT and TM combined. The age-estimation performances were validated and quantified. The approach combining PT and TM only overestimated age with an ME of -0.031 years in males and -0.011 years in females, indicating the best age prediction performance. ispartof: INTERNATIONAL JOURNAL OF LEGAL MEDICINE vol:133 issue:4 pages:1207-1215 ispartof: location:Germany status: Published online

AIM: The aim of the current study was to retrospectively collect dental panoramic radiographs from Somali children living in Finland, to use the radiographic data to develop a new age estimation model based on the model established by Willems et al. (J Forensic Sci 46(4):893-895, 2001), and to compare the age prediction performances of the Willems et al. model (WM) and the newly developed model. MATERIAL AND METHODS: Dental panoramic radiographs from 808 healthy Somalis born in Finland were selected. The development of the seven left mandibular permanent teeth, from the central incisor to the second molar, was staged according to Demirjian et al. (Hum Biol 45(2):211-227, 1973). Radiographs with all listed permanent teeth completely developed were excluded. The studied sample consisted of 635 subjects (311 females, 324 males) ranging in age from 4 to 18 years. Kappa and weighted Kappa statistics were used to quantify intra- and inter-observer agreement in stage allocation. The collected dataset was used to validate the WM, constructed on a Belgian Caucasian reference sample, and to establish a Somali-specific age estimation model (SM) based on the WM. Both models were validated and their age prediction performances quantified using mean error (ME), mean absolute error (MAE) and root mean squared error (RMSE). RESULTS: The SM resulted in a slight underestimation of age when the sex groups were analysed separately or combined, with ME varying between 0.04 (standard deviation (SD) 1.01) and 0.05 (SD 1.04) years, MAE between 0.77 and 0.80 years and RMSE between 1.01 and 1.04 years. The WM statistically significantly underestimated the age of females, with an ME of 0.20 (SD 1.01) years (p = 0.0006). For males, and for females and males combined, no statistically significant ME was observed. CONCLUSION: The WM and SM were similar in their age prediction performances, and the use of the WM in dental age assessment in the Somali population is justified. ispartof: INTERNATIONAL JOURNAL OF LEGAL MEDICINE vol:132 issue:6 pages:1-8 ispartof: location:Germany status: published

Various psychiatric problems in adolescence and early adulthood have been shown to associate with criminal behaviour. In this study the association of personality disorders (PDs) with criminal behaviour was examined in adolescents treated in psychiatric hospitals. The study sample consisted of 508 adolescents (age 13-17) admitted to acute psychiatric impatient care between April 2001 and March 2006. Crime data was obtained from the Finnish Legal Register Centre on September 2013. The Schedule for Affective Disorder and Schizophrenia for School-Age Children Present and Lifetime (K-SADS-PL) was used to assess psychiatric diagnoses in adolescence. The information on PDs in early adulthood was based on follow-up information on psychiatric treatments in either out-or inpatient settings until the end of 2012, and was extracted from the National Care Register for Health Care provided by the Finnish National Institute for Health and Welfare. A total of 22 (39%) of the 57 subjects with PD had committed a crime. In women, the likelihood for violent criminality was significantly increased in those with Borderline PD (OR 6.09, CI 1.24-29.84, p = 0.009) and was also associated with conduct disorder (OR 4.26, CI 1.38-13.19, p = 0.012), child welfare placement (OR 11.82, CI 3.61-38.76, p

We reviewed the 33 727 postmortem toxicology investigations performed in Finland over a period of 5 years (2009-2013) and identified those in which the antidepressant bupropion was detected. Cases positive for other antidepressant drugs were reviewed for comparison. The postmortem toxicological examination included, in all cases, the routine screening and quantification of hundreds of drugs and poisons using quality-controlled methods. Bupropion was detected in 65 cases. A large proportion of the bupropion-positive deaths resulted from suicide (55%). In fatal poisoning cases found positive for bupropion, the proportion of suicide was even higher (77%). The measured median bupropion postmortem blood concentration (0.69 mg/L) was markedly higher than the normal therapeutic range in plasma in the treatment of depression (up to 0.1 mg/L) and even higher in fatal bupropion poisonings (13 mg/L). Only 14% of the deceased positive for bupropion were estimated to be drug abusers. However, nearly all of the drug abuse cases were from the last year of the study (2013), indicating a recent increase of the use of bupropion among drug abusers and possibly even abuse of bupropion itself. Suicide victims positive for bupropion were younger than those who died with other antidepressant drugs in their blood. In addition, the percentage of fatal poisonings among bupropion-positive postmortem cases was higher than among the users of other antidepressant drugs. Suicide was significantly more common among the deceased positive for bupropion than among users of other antidepressant drugs. An unknown degree of bupropion degradation before the assay and post-mortem redistribution of bupropion may have impacted the measured levels. Nonetheless, all post-mortem concentrations of bupropion were elevated and especially high concentrations were detected in suicides. (C) 2016 Elsevier Ireland Ltd. All rights reserved.

A liquid chromatography-tandem mass spectrometric (LC-MS/MS) method was developed and validated for the analysis of seven common sartans (candesartan, eprosartan, losartan, olmesartan, telmisartan, valsartan, and losartan carboxylic acid (EXP3174), the active metabolite of losartan) in postmortem blood samples with liquid-liquid extraction. Detection was accomplished by triple-quadrupole MS in the selected reaction monitoring mode. The validation procedure showed low limits of quantitation of 5 ng/mL for all compounds with good accuracy and precision. The matrix effect was evaluated from the variation of peak areas and concentrations. The method shows some matrix effect but the effect was efficiently compensated by using an internal standard method. The method was applied to the analysis of sartans in postmortem blood samples, and produced 534 positive findings during 2014-2015. Nine deaths were encountered where the concentration of a sartan was very high, indicating intoxication together with other causes of death or multi-drug intoxication. The quantitative results indicate that sartans do not show significant postmortem redistribution; thus concentrations higher than the therapeutic range can be considered as being toxic or fatal.

Forensic and ancient DNA samples often are damaged and in limited quantity as a result of exposure to harsh environments and the passage of time. Several strategies have been proposed to address the challenges posed by degraded and low copy templates, including a PCR based whole genome amplification method called degenerate oligonucleotide-primed PCR (DOP-PCR). This study assessed the efficacy of four modified versions of the original DOP-PCR primer that retain at least a portion of the 5' defined sequence and alter the number of bases on the 3' end. The use of each of the four modified primers resulted in improved STR profiles from environmentally-damaged bloodstains, contemporary human skeletal remains, American Civil War era bone samples, and skeletal remains of WWII soldiers over those obtained by previously described DOP-PCR methods and routine STR typing. Additionally, the modified DOP-PCR procedure allows for a larger volume of DNA extract to be used, reducing the need to concentrate the sample and thus mitigating the effects of concurrent concentration of inhibitors. Published by Elsevier Ireland Ltd.

Sublingual buprenorphine is used in opioid maintenance treatment but buprenorphine is also widely abused and causes fatal poisonings. The aim of this study was to investigate buprenorphine-positive fatalities in order to gain novel information on the magnitude and nature of buprenorphine abuse. All post-mortem toxicology cases positive for urinary buprenorphine, including fatal poisonings caused by buprenorphine and fatalities in which the cause of death was unrelated to buprenorphine, in the five year period of 2010-2014 in Finland were characterized according to urine buprenorphine and naloxone concentrations (n = 775). Urine concentrations were used to assess which buprenorphine preparation had been used; mono-buprenorphine or a buprenorphine-naloxone combination, and whether they had been administered parenterally. In at least 28.8% of the buprenorphine-positive cases the drug had been administered parenterally. The majority of the parenteral users (68.6%) had taken mono-buprenorphine. Fatal poisoning was significantly more common among the identified parenteral users (65.5%) than among other users of buprenorphine products (45.3%). The proportion of buprenorphine-related poisoning was similar in identified parenteral users of mono-buprenorphine (68.6%) and buprenorphine-naloxone (64.1%). In nearly all of the fatal poisoningss the deceased had used other drugs and/or alcohol along with buprenorphine (98.7%). The median age of the deceased increased significantly over the study period, from 32 to 38 years. Our results show that there is ongoing parenteral abuse of both mono-buprenorphine and buprenorphine-naloxone combination. Parenteral users of buprenorphine put themselves into a great risk of fatal poisoning or other accidental injury death which is further exacerbated by the frequent polydrug use. (c) 2018 Elsevier B.V. All rights reserved.

Drug seizures involving a wide variety of emerging new psychoactive substances (NPS) call for new approaches for instant quantification and valuation. Liquid chromatography-chemiluminescence nitrogen detection (LC-CLND) was used in the quantification of opioids with a single secondary standard (caffeine), utilizing the detector's equimolar response to nitrogen. The mean N-equimolarity of CLND for ten fentanyl derivatives and U-47700 by the present LC-CLND method was 96.4% (range 91-101%). The furanylfentanyl samples consisted of 112 powdery samples with a mean (median, range) hydrochloride purity of 13% (4.9%, 0.08-100%). The purity distribution of the furanylfentanyl samples was distinctly bipartite, showing only lower than 9% (NI = 98) and higher than 60% (N=14) purities. The carfentanil samples consisted of eight brownish sticky samples with a mean (median, range) hydrochloride purity of 0.064% (0.063%, 0.052-0.092%). The U-47700 samples consisted of seven powdery samples with a mean (median, range) hydrochloride purity of 89.0% (100%, 51-100%). The present application to synthetic opioid analysis widens the scope of the established LC-CLND method, previously found useful for single-calibrant quantification of stimulant/hallucinogenic and cannabinoid type of NPS. (C) 2019 Elsevier B.V. All rights reserved.

The pharmacogene, CYP2D6, is commonly used to infer metabolizer phenotype of many marketed drugs and endogenous toxins in ante- and post-mortem patients but only represents the efficiency of phase 1 metabolism. Downstream metabolic enzymes encoded by UGT2B7, ABCB1, OPRM1, and COMT also have been implicated in variable individual response to drugs due to their activity at different stages of the tramadol ADME (absorption, distribution, metabolism, and excretion) process. While commonly studied as single genes using targeted genotyping approaches, a more comprehensive tramadol metabolism profile has not been evaluated. 1000 Genomes Project data for UGT2B7, ABCB1, OPRM1, and COMT were used to characterize full-gene haplotypes and their effect on protein function using in-house excel-based workbooks, PopART, and TreeView. Population genetic summary statistics and intergenic analyses associated these haplotypes with full-gene CYP2D6-inferred metabolizer phenotype. The findings suggest that UGT2B7, ABCB1, OPRM1, and COMT may contribute to predicted metabolizer phenotype as opposed to relying solely on CYP2D6.

CYP2D6 is a critical pharmacogenetic target, and polymorphisms in the gene region are commonly used to infer enzyme activity score and predict resulting metabolizer phenotype: poor, intermediate, extensive/normal, or ultrarapid which can be useful in determining cause and/or manner of death in some autopsies. Current genotyping approaches are incapable of identifying novel and/or rare variants, so CYP2D6 star allele definitions are limited to polymorphisms known a priori. While useful for most predictions, recent studies using massively parallel sequencing data have identified additional polymorphisms in CYP2D6 that are predicted to alter enzyme function but are not considered in current star allele nomenclature. The 1000 Genomes Project data were used to produce full-gene haplotypes, describe their distribution in super-populations, and predict enzyme activity scores. Full-gene haplotypes generated lower activity scores than current approaches due to inclusion of additional damaging polymorphisms in the star allele. These findings are critical for clinical implementation of metabolizer phenotype prediction because a fraction of the population may be incorrectly considered normal metabolizers but actually may be poor or intermediate metabolizers.

The aim was to evaluate the performance of point-of-care (POC) tests in detecting glucose and ketone bodies in postmortem (PM) samples and to assess the usefulness of POC tests in sample screening for more precise analyses. Glucose and ketone body, beta-hydroxybutyrate (BHB), were measured from vitreous humor (VH) in 52 autopsy cases with a POC blood glucose monitoring device (BGMD). In addition glucose and ketone bodies, acetone (Ac) and acetoacetate (AcAc), were measured from urine samples in another set of 59 cases with semi-quantitative stick tests. The results were compared to the concentration in VH measured with validated methods (values > 7 mmol/l indicate possible hyperglycemia and total ketone body levels >= 3 mmol/l ketoacidosis). The sensitivity for glucose with the BGMD was 1.0 and specificity 0.94 when the threshold value for the meter to predict elevated glucose was set to >= 10 mmol/l. The correlation between the BGMD and the validated method was strong (R-2 = 0.89). For detecting ketoacidosis, the BGMD had a sensitivity of 1.0 and specificity of 0.73, when the threshold value was set to 2.5 mmol/l. The urine stick test presented a sensitivity of 0.89 and specificity of 0.90 for detecting elevated VH glucose concentration. The sensitivity and specificity for the stick test to detect cases with possible ketoacidosis were 0.84 and 0.68, respectively. According to the results, BGMD can be reliably applied for sample screening, although more samples need to be analyzed for delineating the correct threshold values. In the case of glucose, the urine stick tests could be indicative in detecting cases with VH glucose >= 10 mmol/l. For predicting possible ketoacidosis with elevated VH total ketone bodies, the stick test is not reliable as the test presented both false-positive and -negative results. (C) 2016 Elsevier Ireland Ltd. All rights reserved.

In Finland, forensic age assessment is strictly regulated by legislation. According to the Aliens Act (301/2004) and the amendment of the Act (549/2010), the police authorities, the frontier guard authorities, and the immigration authorities have the right to refer asylum seekers to the University of Helsinki, Department of Forensic Medicine, for age assessment. These assessments are especially performed to solve if the person is of major age, the cutoff being 18 completed years. The forensic age assessment is largely based on dental development, since the special permit of the Radiation and Nuclear Safety Authority (STUK) to the Department of Forensic Medicine of the University of Helsinki, allowing the use of ionizing radiation for non-medical purposes, includes dental and hand X-rays. Forensic age assessment is always performed by two forensic odontologists. In 2015, the total number of forensic age assessment examinations was 149, and the countries of origin of the asylum seekers were most commonly Iraq, Afghanistan, and Somalia. The current legislation on forensic age assessment has been well received and approved. Radiological and other examinations can be performed in different parts of Finland, but the forensic odontologist at the University of Helsinki is always involved in the process and ensures joint quality standards for the forensic age assessment.

There is a constant demand for the quantification of drug metabolites within post-mortem toxicology. Especially electrospray ionization-mass spectrometry techniques necessitate that calibration is carried out using primary reference standards due to the non-uniform ionization efficiency between parent drugs and their metabolites. As reference standards for metabolites are not readily available and their costs are high, alternative methods for immediate quantification are required. In this study, ultra-high performance liquid chromatography coupled with photodiode array detection and corona charged aerosol detection was utilized for the concurrent quantification of 23 drug metabolites using the corresponding parent drug for calibration. Based on this secondary calibration, the quantitative results for the N-demethylated metabolites by each detector were similar to those obtained by the ordinary calibration using reference standards. For O-demethylated metabolites, the differences in detector response caused somewhat larger biases using the secondary calibration. Using the validated secondary calibration, the blood sample data gathered from 633 post-mortem cases was retrospectively reprocessed to discover the combined metabolite-parent concentrations and metabolite to parent ratios for six toxicologically relevant drugs. These results, representing all causes of death, were compared to published data from therapeutic drug monitoring and post-mortem toxicology. (C) 2016 Elsevier Ireland Ltd. All rights reserved.

Blood samples preserved on FTA cards offer unique opportunities for genetic research. DNA recovered from these cards should be stable for long periods of time. However, it is not well established as how well the DNA stored on FTA card for substantial time periods meets the demands of forensic or genomic DNA analyses and especially so for from post-mortem (PM) samples in which the quality can vary upon initial collection. The aim of this study was to evaluate the time-dependent degradation on DNA quality and quantity extracted from up to 16 years old post-mortem bloodstained FTA cards. Four random FTA samples from eight time points spanning 1998 to 2013 (n = 32) were collected and extracted in triplicate. The quantity and quality of the extracted DNA samples were determined with Quantifiler (R) Human Plus (HP) Quantification kit. Internal sample and sample-to-sample variation were evaluated by comparing recovered DNA yields. The DNA from the triplicate samplings were subsequently combined and normalized for further analysis. The practical effect of degradation on DNA quality was evaluated from normalized samples both with forensic and pharmacogenetic target markers. Our results suggest that (1) a PM change, e.g. blood clotting prior to sampling, affects the recovered DNA yield, creating both internal and sample-to-sample variation; (2) a negative correlation between the FTA card storage time and DNA quantity (r = -0.836 at the 0.01 level) was observed; (3) a positive correlation (r = 0.738 at the level 0.01) was found between FTA card storage time and degradation levels. However, no inhibition was observed with the method used. The effect of degradation was manifested clearly with functional applications. Although complete STR-profiles were obtained for all samples, there was evidence of degradation manifested as decreased peak heights in the larger-sized amplicons. Lower amplification success was notable with the large 5.1 kb CYP2D6 gene fragment which strongly supports degradation of the stored samples. According to our results, DNA stored on FTA cards is rather stable over a long time period. DNA extracted from this storage medium can be used as human identification purposes as the method used is sufficiently sensitive and amplicon sizes tend to be

Despite abounding evidence for the harmful effects of synthetic anabolic-androgenic steroids, they are commonly misused for competitive and body-image reasons. Steroids are often used in the context of poly-drug misuse, which may mask their specific effects on behaviour, such as increasing aggression. We present our case report as a concrete example of current steroid-related substance-misuse trends. A 25-year old Finnish male with various psychiatric and drug-related symptomology, but almost no previous history of aggressive behaviour, battered his wife to death and mutilated her body after a five-week steroid regimen.

The fatal toxicity index (FTI) is the absolute number of fatal poisonings caused by a particular drug divided by its consumption figure. Consequently, it is a useful measure in evaluating toxicity of the drug and its relevance in fatal poisonings. In this study, we assessed the FTI of medicinal drugs in 3 years (2005, 2009, and 2013) in Finland. As the measure of drug consumption, we used the number of defined daily doses (DDD) per population in each year. There were 70 medicinal drugs in Finland for which the mean FTI expressed as the number of deaths per million DDD over the three study years was higher or equal to 0.1. The Anatomical Therapeutic Chemical (ATC) classification system was used for the classification of the active ingredients of medicinal drugs according to the organ or system which they act on. Of these 70 drugs, 55 drugs (78.6 %) acted on the nervous system (denoted by ATC code N), 11 (15.7 %) on the cardiovascular system (C), three (4.3 %) on the alimentary tract and metabolism (A), and one (1.4 %) on the musculoskeletal system (M). The nervous system drugs consisted of 20 psycholeptics, (ATC code N05), 20 psychoanaleptics (N06), eight analgesics (N02), six antiepileptics (N03), and one other nervous system drug (N07). The highest individual FTIs were associated with the opioids methadone, dextropropoxyphene, oxycodone, tramadol, and morphine; the antipsychotics levomepromazine and chlorprothixene; and the antidepressants doxepin, amitriptyline, trimipramine, and bupropion. Buprenorphine was not included in the study, because most of the fatal buprenorphine poisonings were due to smuggled tablets. A clearly increasing trend in FTI was observed with pregabalin and possibly with bupropion, both drugs emerging as abused substances.

In the middle of the 1980?s, I heard a highly profiled professor making a comment after a lecture about a brand new technique, the polymerase chain reaction (PCR). His comment was full of doubt about this novel technology, and the message was something like: ? it [PCR] can never become a widely used diagnostic tool due to the unavoidable contamination? . However, the PCR revolutionized life sciences from medicine to conservation genetics, the inventor was awarded with the Noble Prize, and many of us made careers using the very technique. Scientists, clinical diagnostics and forensic laboratories and others using PCR quickly learned to deal with contamination and build mechanisms to monitor for it. The contamination was there, but, it could be managed with the right laboratory environment, sample flow, and careful experimental design including proper sample handling and a set of controls.

High throughput sequencing (HTS) generates large amounts of high quality sequence data for microbial genomics. The value of HTS for microbial forensics is the speed at which evidence can be collected and the power to characterize microbial-related evidence to solve biocrimes and bioterrorist events. As HTS technologies continue to improve, they provide increasingly powerful sets of tools to support the entire field of microbial forensics. Accurate, credible results allow analysis and interpretation, significantly influencing the course and/or focus of an investigation, and can impact the response of the government to an attack having individual, political, economic or military consequences. Interpretation of the results of microbial forensic analyses relies on understanding the performance and limitations of HTS methods, including analytical processes, assays and data interpretation. The utility of HTS must be defined carefully within established operating conditions and tolerances. Validation is essential in the development and implementation of microbial forensics methods used for formulating investigative leads attribution. HTS strategies vary, requiring guiding principles for HTS system validation. Three initial aspects of HTS, irrespective of chemistry, instrumentation or software are: 1) sample preparation, 2) sequencing, and 3) data analysis. Criteria that should be considered for HTS validation for microbial forensics are presented here. Validation should be defined in terms of specific application and the criteria described here comprise a foundation for investigators to establish, validate and implement HTS as a tool in microbial forensics, enhancing public safety and national security. Published version

Molecular phylogenies have been used in forensic molecular epidemiological investigations of transmission events of human immunodeficiency virus (HIV) in dental and medical practices and in rape cases [1-7]. Some of these investigations have resulted in criminal charges [4-7] and typically have involved only a few transmission events in a short period of time. Gonzales-Candelas et al. report in BMC Biology[8] a case of an anesthetist from Valencia, Spain, who was convicted of professional malpractice by infecting over 270 of his patients with hepatitis C virus (HCV). This case differs from most other viral infection cases in that the origin of transmissions date back approximately 25 years and it involved a large number of patients. Greater than 300 candidates, who had undergone a minor surgery in one of two hospitals, were tested, and for the first time a molecular clock was used to estimate the date of various transmission events. HCV is a single-stranded RNA virus and member of Flaviviridae family. It is transmitted by blood-to-blood contact, e.g. in intravenous drug use or unprotected sexual intercourse, but also during medical procedures under poor-quality conditions (e.g. non-screened blood transfusion and re-use of syringes or needles). Up to 20% of infected individuals develop severe medical complications, including liver cancer, cirrhosis with a consequent liver failure. An HCV infection is extremely devastating to an infected individual and an epidemic is costly for the society. As the first cases were detected in 1998, and an epidemiological investigation that there was a large outbreak with possibly hundreds of patients infected, public health officials suspected that an anesthetist, working in a public and private hospital, was the likely source of the HCV transmissions. The epidemiologists investigated data of over 66,000 individuals and common risk factors for infection in surgery, e.g., surgeon, surgery room, type of surgery, anesthesiologist, and type of anesthesia. The only significant factor (adjusted Odds Ratio 28.5, 95% CI 9.83 to 82.59) was this anesthetist. This observation was supported by the fact that of the initial 197 cases considered in the outbreak, 184 had been anesthetized by the suspected individual. Since the suspected epidemic involved an exceedingly large number of patients and the overall period or transmission events were unknown, the authorities, including the judge, tasked experts in evolutionary biology and viral epidemiology to address the following questions: 1) was the suspect the source responsible for the outbreak?; 2) Could they ascertain which patients had been infected from a common source and thus could be included in the outbreak?; 3) Alternatively, which patients could have been infected from other sources?; 4) Could they exclude these alternative sources or the existence of different but simultaneous outbreaks?; 5) Could they determine the duration of the outbreak?; 6) Could they date the time of infection for each patient in the outbreak?; and 7) Could they determine the date of infection of the anesthetist? Gonzales-Candelas et al. [8] tackled the questions by sequencing 229-nucleotides (nts) of a conservative non-structural NS5B gene by reverse transcription of RNA to DNA, followed by Sanger sequencing of the DNA in all patients, the anesthetist, and a number of control samples. The difficulty with genetic analysis of HCV is, like HIV, it is a very fast-evolving virus, and thus, even individuals harboring the virus with the same origin will have differences in their viral RNA sequences. However, those sequences from the same source are likely to be more similar than those of epidemiologically unassociated patients. Therefore, phylogenetic approaches could prove informative, as was practiced in some previous cases analyzing HIV sequences [1-6]. Gonzales-Candelas et al. [8] typed patients positive for the 320 HCV-1a and 290 HCV-1b strains and 44 HCV-1a positive patients serving as geographically local controls. The HCV from the anesthetist was genotypically consistent with HCV-1a, and a dual (HCV-1A and -1b) origin was excluded. The authors then analyzed only the HCV-1a positive patients, and compared them with the controls. The sequence differences along the targeted 229 nts of the NS5B gene among the patients vs among the controls compared to the anesthetist were clear: there were 0–19 differences among the patients of which half had exactly the same sequence as the anesthetist and three out of four had two or less differences. The challenge of determining the direction and timing of the infection still remained, since phylogenetic trees reconstructed from the relatively short and slowly evolving NS5B gene did not have sufficient resolving power to separate the branches of the patients and local controls. Analysis then was performed on a faster evolving 406-nt region in the structural envelope coding E1-E2 region. The samples from the anesthetist were taken on 12th Feb 1998, and no further sampling was possible under Spanish law. Altogether over 4000 cloned sequences were used for the phylogenetic analysis, and 134 of them represented the anesthetist with 28 different haplotypes, which clustered in two groups. Neighbor-joining (NJ) and maximum-likelihood (ML) based phylogenetic trees showed highly supported internal branching (100% and 96% bootstraps, respectively). These trees were used to define which patients were included in the outbreak. This analysis divided the group into 274 patients considered associated with the outbreak (victims), 47 patients initially thought to be in the outbreak that were excluded, and 47 controls. In order to assess the support for this grouping the authors calculated the likelihood ratio (LR) for alternating hypotheses that the sequences of the patients group with the outbreak vs they group with the controls. The LRs tended to favor the hypothesis that they fall within the outbreak group and ranged from 1.051 to 6.622 × 1095. Similarly, for those excluded from the outbreak the LRs ranged from 1.330 to 4.408 × 1084 supporting the alternate hypothesis. Both of the figures provided a very strong support for the conclusions. For estimating the timing of the transmission events Gonzales-Candelas et al.[8] used a Bayesian method implemented in the program BEAST (Bayesian Evolutionary Analysis by Sampling Trees). For each of the 274 patient they used the E1-E2 region sequences to estimate the time to the most recent common ancestor (MRCA). For the analysis they incorporated the dates of the sampling, and the dates of the infection of 24 patients, who visited the anesthetist only once. Sequences from these 24 patients were used to calibrate molecular clock estimates for the MRCA. The authors concluded that the estimated transmission events of the patients was between Jan 1987 and April 1998, compared to the one obtained for the anesthetist August 1984 to October 1991). In the court these data were compared with those obtained from the non-molecular investigation (documents and testimonies), and in two out of three cases the molecular data estimates were congruent with other data. The analysis in this case was systematic and rigorous. However, it is important to note that the data presented by Gonzales-Candelas et al.[8], like in many other DNA investigations presented in the court, were not used to prove if the anesthetist was guilty, but rather in concert with other evidence. Even though only a piece of the puzzle, analyses, such as carried out by Gonzales-Candelas et al., will likely be used increasingly to assist epidemiologic and criminal (also civil) investigations.

Lactose persistence (LP), the ability to digest milk sugar (lactose), is one of the best examples of selection-based evolutionary change in humans from milk-drinking cultures. LP has been documented in populations with northern European ancestry and those originating in Central Asia, Middle East, the Arabian Peninsula, and Africa. Based on molecular genetic studies, it has been estimated that the selective change in the ability to digest lactose beyond childhood occurred within approximately the past 5,000-10,000 years [1]. The estimation is consistent with an advantage to LP in dairy farming-based subsistence, which came to prominence in certain geographical areas around the same time. The important enzyme in the breakdown of milk sugar, lactase (or more specifically lactase-phlorizin hydrolase (LPH)), is coded by the LCT gene. The molecular genetic data reported in recent years show several interesting aspects of LP. The first genetic variation described for European LP was not in the LCT gene, but in the in the intron 13 of the MCM6 gene, residing ~14 kb upstream of MCM6 [2]. However, it soon was realized that the MCM6 intron 13 T-13910 allele, which explained most LP in Europe, was absent in Africa [3,4]. Instead, three other SNPs in the same intron, C-14010, G-13907 and G-13915, were associated with lactase persistence in Africa. Also, these SNPs originated on different haplotype backgrounds from the European T-13910 allele and from each other providing a major model of convergent evolution in LP due to strong selective pressure [3,5]. To date, several SNPs associated with LP are known. In Europe, in addition to MCM6 intron 13 T-13910, intron 9 G-22018 is significantly associated with the LP trait. In Africa and the Arabian Peninsula, four SNPs (those above and G-14009), which are all located within 100 bp of T-13910 have been shown to be associated with the LP predominantly in pastoralist populations. Those variants do not, however, account for all the phenotypic variance of LP in Africa, implying that additional genetic variation may play a role. A new study by Ranciaro et al. in American Journal of Human Genetics[6] sought out new genetic variants associated with LP in Africa. They studied 819 individuals from 63 African populations, and 154 individuals from non-African populations from Europe, the Middle East and Asia. The authors sequenced MCM6 gene introns 9 and 13 and ~2 kb of the LCT gene promoter region in these samples. Additionally, in an effort to reconstruct the origin and spread of LP-associated alleles in Africa, four microsatellites were genotyped in a ~198 kb region in a subset of 252 individuals. The participants’ LP status was tested using a standardized blood test that tracks lactose digestion. With the new larger data set, and sampling from outside Africa, the authors confirmed the earlier observed association between the LP trait in Africa and three variants in intron 13, and they also found two additional LP-associated SNPs, one in intron 13 (G-12962) and one in the LCT promoter region (T-956). Further functional studies are naturally needed to prove whether these new SNPs are causative or just merely in linkage disequilibrium with the earlier identified variants. Ranciaro et al. [6] then used allele frequency and long-range linkage disequilibrium based neutrality tests to detect any signatures of selection. They found evidence for recent positive selection in eastern African populations and the Fulani from central Africa. While looking for the new variants for LP within Africa, the study by Ranciaro et al. [6] also considered that if LP patterns coincide with the rise in pastoralism, the LP-related alleles found in present-day populations should reveal past migrations of those populations. In order to do that, they analyzed a subset of 252 study participants using four microsatellites and combined these with the SNPs to create haplotypes. The haplotype analysis has some interesting insights about the migration of pastoralism groups in Africa. For example, the haplotype analysis supported an eastern African origin (Kenya and Tanzania) of the C-14010 LP-associated mutation now found in southern Africa in Bantu-speaking Xhosa and the hunter-gatherer San population. Also, the presence of Middle Eastern origin G-13915 allele in North African populations suggests migration between these regions. Interestingly, the data are supported by known historical interactions between these populations. Some of the data in Ranciaro et al. [6] is in agreement with earlier findings of distinct genetic LP variants that have arisen in various parts of Africa through convergent evolution. For example, the G-13907 was almost exclusively restricted to northern Kenya, Ethiopia and northern Sudan, as reported earlier. Last but not least, the authors also detected the traditionally European LP haplotype background in some West-and North-Central African populations. But, even with these new data from Ranciaro et al. [6], the genetic variants found and analyzed in various populations to date, are far from fully explaining LP in Africa.

The post-industrial lifestyle has many disadvantageous effects on our health. One of the factors is modern nutrition, which has been associated with epidemic burdens, such as obesity and cardiovascular diseases. At least two major shifts have occurred in the nutritional history of humans: the use of carbohydrate-rich diets which were adopted around 10,000 years BP due to Neolithic farming, and later the influence of industrially processed flour and white sugar after the industrial revolution in the 1850s. In a recent paper in Nature Genetics Adler et al. used a novel approach to see how these dietary changes affected the oral microbiome by analyzing the ancient microbial DNA in the calcified dental plaque from 34 early European skeletons.

September 2012 marked the 28th anniversary of an experiment that translated into the forerunner of a technology that impacts society worldwide till today, as well as the official retirement of the remarkable scientist who is widely known for this milestone discovery. That is, DNA fingerprinting invented, coined, further developed, and for the first time practically applied by Sir Alec Jeffreys, which changed forensic science for the better and forever. Sir Alec’s fame transcends forensic science, other scientific disciplines, and just as significant the lay public. His invention has promulgated one of the most successful scientific tools in solving and fighting crime, identifying missing persons and victims of mass disasters, and - just as important - in helping exonerate those innocent people who have been wrongly associated with a crime. Although most of his research career was outstanding beyond forensic DNA analysis, Sir Alec is widely considered by the apt moniker “The Founding Father of Forensic DNA Identification”. To contribute to the affirmation and acknowledgement that Sir Alec’s discovery of DNA fingerprinting has had on a wide range of scientific disciplines, we the Editors-in-Chief, thought it would be important and timely to devote a special series of review articles in Investigative Genetics on DNA fingerprinting, its past, present and future. We are pleased to have distinguished experts in forensics, anthropology, zoology, botany, and microbiology who contributed treatises on how Sir Alec’s discovery and subsequent developments of DNA fingerprinting have impacted - and continue to influence - their fields. All these scholars witnessed the nascent days of DNA fingerprinting and used – together with numerous colleagues worldwide - various similarly-based molecular genetic strategies to address a wide range of research questions in their respective fields. Mark Jobling, Investigative Genetics’ regular columnist and professor at the same University of Leicester’s Department of Genetics that Sir Alec resided for 35 years, contributed a Guest Editorial to summarize the story of DNA fingerprinting in greater detail. We remember well the original International DNA fingerprinting Conferences held on five continents from 1990 to 1999 (Bern, Switzerland in 1990; Belo Horizonte, Brazil in 1992; Hyderabad, India in 1994; Melbourne, Australia in 1996; and Port Elizabeth, South Africa in 1999). These conferences, initially organized by Sir Alec and colleagues, were unique and extremely valuable because scientists from different scientific backgrounds were brought together with a common interest in genetically characterizing and differentiating organisms, strains, or species by means of DNA fingerprinting. We recall the lively interactions and how it helped us and many others to gain a foothold in molecular genetics, and at the same time to learn how useful DNA fingerprinting is to address various open questions in the life sciences. Although this International DNA fingerprinting Conference series came to an end within a decade, the science and technology of DNA fingerprinting continued and was rapidly and fully absorbed by various life science disciplines. Predictably there was the necessary end of this exciting conference series; it had served its purpose of establishing DNA fingerprinting as a routine tool across life sciences. Yet, the end was unfortunate because this particular venue for vibrant exchange of science and technology within and across disciplines was gone. Especially in a scientific world where inter-disciplinary work has become increasingly important and at the same time more challenging because of the necessary drive for levels of specialization, it can be difficult to find a common language even among biological scientists. Sir Alec’s scientific interest in human genomic dynamics was much beyond DNA fingerprinting. In fact, he made numerous groundbreaking discoveries on the processes of mutation and recombination - two important pathways of creating and maintaining genetic diversity - and regularly published them in the highest ranked scientific journals. To keep focus, these notable scientific achievements that likely will provide the fodder for generations of scientists cannot be the target of this special article series; perhaps future ones in this or more specialized journals will be devoted to that part of his enormous research achievements. We hope that with this special article series on DNA fingerprinting Investigative Genetics will attract many readers, some who may read these articles with a nostalgic touch, others to gain a perspective on how a single discovery could have such a strong impact on science and society so quickly as DNA fingerprinting did, and those who seek the motivation to commit themselves to science with the same verve as Sir Alec Jeffreys has.

In a worldwide collaborative effort, 19,630 Y-chromosomes were sampled from 129 different populations in 51 countries. These chromosomes were typed for 23 short-tandem repeat (STR) loci (DYS19, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, DYS385ab, DYS437, DYS438, DYS439, DYS448, DYS456, DYS458, DYS635, GATAH4, DYS481, DYS533, DYS549, DYS570, DYS576, and DYS643) and using the PowerPlex Y23 System (PPY23, Promega Corporation, Madison, WI). Locus-specific allelic spectra of these markers were determined and a consistently high level of allelic diversity was observed. A considerable number of null, duplicate and off-ladder alleles were revealed. Standard single-locus and haplotype-based parameters were calculated and compared between subsets of Y-STR markers established for forensic casework. The PPY23 marker set provides substantially stronger discriminatory power than other available kits but at the same time reveals the same general patterns of population structure as other marker sets. A strong correlation was observed between the number of Y-STRs included in a marker set and some of the forensic parameters under study. Interestingly a weak but consistent trend toward smaller genetic distances resulting from larger numbers of markers became apparent. Fil: Corach, Daniel. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Servicio de Huellas Digitales Genéticas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Caputo, Mariela. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Servicio de Huellas Digitales Genéticas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Marino, Miguel Eduardo. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Laboratorio de Analisis de ADN; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Purps, Josephine. Charité-Universitätsmedizin; Alemania Fil: Siegert, Sabine. University of Cologne; Alemania Fil: Willuweit, Sascha. Charité-Universitätsmedizin; Alemania Fil: Nagy, Marion. Charité-Universitätsmedizin; Alemania Fil: Alves, Cíntia. Universidad de Porto; Portugal Fil: Salazar, Renato. Universidad de Porto; Portugal Fil: Angustia, Sheila M. T.. Philippine National Police Crime Laboratory; Filipinas Fil: Santos, Lorna H.. Philippine National Police Crime Laboratory; Filipinas Fil: Anslinger, Katja. Universitat Genzentrum Der Ludwing-maximilians; Alemania Fil: Bayer, Birgit. Universitat Genzentrum Der Ludwing-maximilians; Alemania Fil: Ayub, Qasim. The Wellcome Trust Sanger Institute; Reino Unido Fil: Wei, Wei. The Wellcome Trust Sanger Institute; Reino Unido Fil: Xue, Yali. The Wellcome Trust Sanger Institute; Reino Unido Fil: Tyler Smith, Chris. The Wellcome Trust Sanger Institute; Reino Unido Fil: Baeta Bafalluy, Miriam. Universidad de Zaragoza; España Fil: Martínez Jarreta, Begoña. Universidad de Zaragoza; España Fil: Egyed, Balazs. Eotvos University, Budapest; Argentina Fil: Balitzki, Beate. Universidad de Basilea; Suiza Fil: Tschumi, Sibylle. Universidad de Basilea; Suiza Fil: Ballard, David. King; Reino Unido Fil: Syndercombe Court, Denise. King; Reino Unido Fil: Barrantes, Xinia. Poder Judicial, Forensic Sciences Department; Costa Rica Fil: Bäßler, Gerhard. Landeskriminalamt Baden-Württemberg; Alemania Fil: Berger, Burkhard. Universidad de Innsbruck; Austria Fil: Niederstätter, Haral. Universidad de Innsbruck; Austria Fil: Parson, Walther. Universidad de Innsbruck; Austria. University Park; Estados Unidos Fil: Davis, Carey. Department of Molecular and Medical Genetics; Estados Unidos. Institute of Applied Genetics; Estados Unidos Fil: Furfuro, Sandra. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Laboratorio de Análisis de ADN; Argentina Fil: Locarno, Laura. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Laboratorio de Análisis de ADN; Argentina