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The article presents a study of determining cannabinoids in whole blood using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS-MS). UPLC-MS-MS method was developed and formalized to identify the cap delta9-tetrahydrocannabinol (THC), 11-hydroxy-cap delta9-tetrahydrocannabinol (11-OH-THC-), and 11-nor-9-carboxy-cap delta9-tetrahydrocannabinol (THC-COOH) in whole blood. Samples acquired from protein precipitation were examined followed by a solid-phase extraction. Data were gathered using positive electrospray ionization and multiple reaction monitoring. Results show that UPLC-MS-MS method was sensitive, specific, and rapid because it does not need derivatization step.

In the present study, nuclear (autosomal and Y-chromosome short tandem repeats) and mitochondrial (hypervariable region I) ancient DNA data previously obtained from a 2,300-year-old Xiongnu population of the Egyin Gol Valley (south of Lake Baikal in northern Mongolia) (Keyser-Tracqui et al. [2003] Am. J. Hum. Genet. 73:247-260) were compared with data from two contemporary Mongolian populations: one from the same location (Egyin Gol Valley plus a perimeter of less than 100 km around the valley), and one from the whole of Mongolia. The principal objective of this comparative analysis was to assess the likelihood that genetic continuity exists between ancient and present-day Mongolian populations. Since the ancient Xiongnu sample might have been composed of some of the ancestors of the present-day Yakuts, data from a present-day Yakut population, as well as published data from Turkish populations, were also included in the comparative analysis. The main result of our study was the genetic similarity observed among Mongolian samples from different periods and geographic areas. This result supports the hypothesis that the succession over time of different Turkic and Mongolian tribes in the current territory of Mongolia resulted in cultural rather than genetic exchanges. Furthermore, it appears that the Yakuts probably did not find their origin among the Xiongnu tribes, as we previously hypothesized. [ABSTRACT FROM AUTHOR]

Mutation analysis in the mitochondrial DNA (mtDNA) control region is widely used in population genetic studies as well as in forensic medicine. Among the difficulties linked to the mtDNA analysis, one can find the detection of heteroplasmy, which can be inherited or somatic. Recently, age-related point mutation A189G was described in mtDNA and shown to accumulate with age in muscles. We carried out the detection of this 189 heteroplasmic point mutation using three technologies: automated DNA sequencing, Southern blot hybridization using a digoxigenin-labeled oligonucleotide probe, and peptide nucleic acid (PNA)/real-time PCR combined method on different biological samples. Our results give additional information on the increase in mutation frequency with age in muscle tissue and revealed that the PNA/real-time PCR is a largely more sensitive method than DNA sequencing for heteroplasmy detection. These investigations could be of interest in the detection and interpretation of mtDNA heteroplasmy in anthropological and forensic studies. [ABSTRACT FROM AUTHOR]

The excavation of five frozen graves at the Sytygane Syhe and Istekh-Myrane burial sites (dated at 400 years old) in central Yakutia revealed five human skeletons belonging to the Yakut population. To investigate the origin and evolution of the Yakut population as well as the kinship system between individuals buried in these two sites, DNA was extracted from bone samples and analyzed by autosomal short tandem repeats (STRs) and by sequencing hypervariable region I (HV1) of the mitochondrial DNA (mtDNA) control region. The results showed a diversity of sepulchral organizations linked probably to the social or genetic background of the subjects. Comparison of STR profiles, mitochondrial haplotypes, and haplogroups with data from Eurasian populations indicated affinities with Asian populations and suggested a relative specificity and continuity of part of the Yakut mitochondrial gene pool during the last five centuries. Moreover, our results did not support a Central Asian (with the exception of maternal lineage of West Eurasian origin) or Siberian origin of the maternal lineages of these ancient Yakut subjects, implying an ethnogenesis of the Yakut population probably more complex than previously proposed. [ABSTRACT FROM AUTHOR]

Abstract: In a previous study, we extracted, amplified and sequenced a DNA fragment from bone lesions similar to those of bone tuberculosis in a predynastic skeleton (Egypt, around 3400 bc). This 65kDa gene fragment encodes the surface of the mycobacterium responsible for the lesions. In the present study, we reconstructed the phylogenetic tree of the Mycobacterium family using this fragment and 51 sequences of pathogens and environmental mycobacteria. This reconstruction enabled us to polarise the phylogenic tree and to confirm the originality of the sequence and its ancestral character. According to a recent evolutionary scenario for the Mycobacterium tuberculosis complex, these data suggest the pathogenicity of an archaic mycobacterium at the emergence of urban life. This result could lead to a better understanding of present day evolutionary processes, especially the emergence or re-emergence of non-tuberculosis mycobacteria infection and disease. [Copyright &y& Elsevier]

Abstract: A collaborative study was carried out by the European DNA Profiling Group (EDNAP) in order to evaluate the performance of Y-chromosome binary polymorphism analysis in different European laboratories. Four blood samples were sent to the laboratories, to be analysed for 11 Y-chromosome single nucleotide polymorphisms (SNPs): SRY-1532, M40, M35, M213, M9, 92R7, M17, P25, M18, M153 and M167. All the labs were also asked to submit a population study including these markers. All participating laboratories reported the same results, indicating the reproducibility and robustness of Y-chromosome SNP typing. A total of 535 samples from six different European populations were also analysed. In Galicia (NW Spain) and Belgium, the most frequent haplogroup was R1b*(xR1b1,R1b3df). Haplogroup F*(xK) is one of the most frequent in Austria and Denmark, while the lowest frequency appear in Belgium. Haplogroup frequencies found in this collaborative study were compared with previously published European Y-chromosome haplogroup data. [Copyright &y& Elsevier]

Abstract: We extracted the DNA contained in samples of bones and teeth from 10 skeletons excavated from the Gravette site (400–1000 AD, south of France). Ancient DNA was analysed by autosomal short tandem repeats (STRs). The DNA present in these ancient remains appeared very degraded, but nevertheless, better conserved in tooth than in bone samples. Moreover, we showed that the DNA extracted from ancient dental pulp was not exempt from polymerase chain reaction (PCR) inhibitors, which could result from extreme DNA fragmentation. An adapted protocol with a supplementary step of purification removed this inhibition. [Copyright &y& Elsevier]

Evaluates a single nucleotide polymorphism panel for both forensic and anthropological studies. Autosomal and X-chromosome informative biallelic markers analyzed in order to establish individual specific profiles; Intronic position allied to stability against mutation; Representation at high frequencies among populations; Lack of linkage disequilibria between markers located on the same chromosome confirmed by Genepop calculations using the obtained genotyping data.

We successfully extracted DNA from a bone sample of a Neolithic skeleton (dated 3,600 ± 60 years BP) excavated in northeastern Yakutia (east Sibe- ria). Ancient DNA was analyzed by autosomal STRs (short tandem repeats) and by sequencing of the hypervariable region I (HV1) of the mitochondrial DNA (mtDNA) control region. The STR profile, the mitochondrial haplotype, and the haplogroup determined were compared with those of modern Eurasian and Native American populations. The results showed the affinity of this ancient skeleton with both east Siberian/Asian and Native American populations. [ABSTRACT FROM AUTHOR]

The purported lowering of sex opposition, coupled with a possible abrupt unconsciousness-inducing effect and ease of administration in spiked drinks have resulted in the use of hypnotics in cases of drug-facilitated offense. Among these compounds, lorazepam possesses amnesic properties and can impair an individual rapidly.The chances to detect this substance increase if the most sensitive methods are used and if the biological fluid which allows the longest possible detection time is available.In order to document the window of detection of lorazepam, we have orally administered 2.5 mg of the drug to three volunteers and collected oral fluid (n=l) over 8 h, urine (n=2) over 144 h and hair (n=3) 4 weeks after exposure. Lorazepam was analyzed by LC-MS/MS after alkalinisation (to pH 8.4 with phosphate buffer) and extraction by dichloromethane/diethyl ether in presence of diazepam-d5, used as internal standard. Reversed-phase separation on a XTerra C18 column was achieved in 12 min, under gradient conditions. Molecular ions (m/z 321 and 290 for lorazepam and the IS, respectively) were selected in Ql and the corresponding daughter ions (m/z 303 and 275 for lorazepam and m/z 154 and 198 for the IS) were detected in Q3 after collision with argon.Urine tested positive for lorazepam over 144 h (2–4 ng/ml), with a peak detected after 24 h exposure (411–880 ng/ml). Oral fluid tested positive for lorazepam over 8 h (0.7 ng/ml). Despite a limit of quantitation at 1 pg/mg, we were unable to detect a single lorazepam dose in hair, contrarily to most other benzodiazepines that are detectable.Therefore, in case of drug-facilitated crimes involving lorazepam, urine appears as the best specimen to document exposure, particularly if LC-MS/MS is used. [Copyright &y& Elsevier]

Reports on the use of drug to modify a person's behavior for criminal gain in France. Concerns regarding a recent increase in reports of drug-facilitated crimes in the general public; Difficulties faced by medical examiners on the detection of drugs used by these persons; Efforts of the researchers to develop new techniques for the collection and screening methods used to detect different compounds.

Since gamma-hydroxybutyrate (GHB) is present in both blood and urine of the general population as an endogenous compound, toxicologists must be able to discriminate between these endogenous levels and a concentration resulting from exogenous exposure. The implementation of a cut-off concentration must be done cautiously, due to the wide distribution of endogenous concentrations.To verify the accuracy of a proposed 50 mg/l postmortem blood cut-off, we tested 71 autopsy cases of subjects where the cause of death could exclude GHB exposure. The delay between death and autopsy ranged from 12 to 72 h. GHB was tested by gas chromatography–mass spectrometry (GC–MS) after precipitation. Briefly, 20 μl of blood, bile, or vitreous humor was pipetted in a glass tube, followed by 20 μl of GHB-d6 and 45 μl of acetonitrile. After vortexing and centrifugation, the supernatant was collected and evaporated to dryness. The residue was derivatized with bis(trimethylsilyl)trifluoroacetamide (BSTFA) with 1% trimethylchlorosilane (TMCS) for 20 min at 70 °C. After injection on a 30 m HP5 MS capillary column, GHB (m/z 233, 204, and 147) and GHB-d6 (m/z 239) were identified by MS.GHB tested positive in all the 71 whole blood (cardiac) specimens, with concentrations in the range 0.4–409 mg/l, with a major distribution in the range 10–40 mg/l. A concentration >50 mg/l was observed in 14 cases. As there was no data to support GHB exposure, this was considered as postmortem formation. In order to discriminate this contamination, when available, femoral blood, bile or/and, vitreous humor were tested. The following results were obtained: cardiac blood (55–409 mg/l) versus bile (6.1–238 mg/l) in seven cases; cardiac blood (51–409 mg/l) versus femoral blood (17–44 mg/l) in five cases, and cardiac blood (51–409 mg/l) versus vitreous humor (3.9–2 mg/l) in six cases. It is obvious that bile does not fit the requirements for discrimination and that femoral blood and mostly vitreous humor can be of particular interest. These results demonstrate that a positive (>50 mg/l) postmortem blood GHB concentration cannot support alone drug exposure and that it is essential to document the case with other specimens, including peripheral blood and vitreous humor. [Copyright &y& Elsevier]

We extracted DNA from two skeletons belonging to the Sytho-Siberian population, which were excavated from the Sebÿstei site (dating back 2,500 years) in the Altai Republic (Central Asia). Ancient DNA was analyzed by autosomal short tandem repeats (STRs) and by the sequencing of the hypervariable region 1 (HV1) of the mitochondrial DNA (mtDNA) control region. The results showed that these two skeletons were not close relatives. Moreover, their haplogroups were characteristic of Asian populations. Comparison with the haplogroup of 3,523 Asian and American individuals linked one skeleton with a putative ancestral paleo-Asiatic population and the other with Chinese populations. It appears that the genetic study of ancient populations of Central Asia brings important elements to the understanding of human population movements in Asia. [ABSTRACT FROM AUTHOR]

Objectives. To investigate the epidemiology of bone changes related to degenerative joint disease and enthesopathies in the earliest Neolithic population identified in Europe (dated back 7700 years), to compare the findings with those in medieval and contemporary populations, and to draw etiological inferences. Methods. Examination of the skeletal remains of 119 individuals older than 20 years of age at death and buried in two Central European cemeteries dated as far back as 7700 years (5700 BC). Results. The propensity for developing degenerative disease seemed to vary among individuals, as it does today. The hands, shoulder, and hip were common targets of osteoarthritis, as they are today, and the elbow was more likely to be affected in the Neolithic and medieval populations than in contemporary populations. When age of the individuals is taken into account, the prevalence of knee osteoarthritis does not seem to have changed substantially over time. Conclusions. These results attest to the major role of microtrauma in osteoarthritis of the upper limb joints. At the other joints, after correction for age, the prevalence of degenerative disease and the combinations of joint involvements have remained remarkably constant over time, suggesting a major role for genetic factors (bone formers) already present in European populations in the early Neolithic period. [Copyright &y& Elsevier]

Objectives. To investigate the epidemiology of bone changes related to degenerative joint disease and enthesopathies in the earliest Neolithic population identified in Europe (dated back 7 700 years), to compare the findings with those in medieval and contemporary populations, and to draw etiological inferences. Methods. Examination of the skeletal remains of 119 individuals older than 20 years of age at death and buried in two Central European cemeteries dated as far back as 7 700 years (5 700 BC). Results. The propensity for developing degenerative disease seemed to vary among individuals, as it does today. The hands, shoulder, and hip were common targets of osteoarthritis, as they are today, and the elbow was more likely to be affected in the Neolithic and medieval populations than in contemporary populations. When age of the individuals is taken into account, the prevalence of knee osteoarthritis does not seem to have changed substantially over time. Conclusions. These results attest to the major role of microtrauma in osteoarthritis of the upper limb joints. At the other joints, after correction for age, the prevalence of degenerative disease and the combinations of joint involvements have remained remarkably constant over time, suggesting a major role for genetic factors (bone formers) already present in European populations in the early Neolithic period. [Copyright &y& Elsevier]

Seventeen years of archaeological and anthropological expeditions in North-Eastern Siberia (in the Sakha Republic, Yakutia) have permitted the genetic analysis of 150 ancient (15th-19th century) and 510 modern individuals. Almost all males were successfully analysed (Y-STR) and this allowed us to identify paternal lineages and their geographical expansion through time. This genetic data was confronted with mythological, historical and material evidence to establish the sequence of events that built the modern Yakut genetic diversity. We show that the ancient Yakuts recovered from this large collection of graves are not representative of an ancient population. Uncommonly, we were also able to demonstrate that the funerary preference observed here involved three specific male lineages, especially in the 18th century. Moreover, this dominance was likely caused by the Russian conquest of Siberia which allowed some male clans to rise to new levels of power. Finally, we give indications that some mythical and historical figures might have been the actors of those genetic changes. These results help us reconsider the genetic dynamics of colonization in some regions, question the distinction between fact and myth in national histories and provide a rare insight into a funerary ensemble by revealing the biased process of its composition. Analysis of human remains from 15th-19th Century North-Eastern Siberia, in combination with modern samples, indicate the drivers of present day Yakut genetic diversity, and enable archaeological interpretations of population dynamics at this time. [ABSTRACT FROM AUTHOR]

A letter to the editor is presented about the identification of variola virus in a 300-year-old Siberian mummy.

Discusses the death of a 14-year-old boy due to repetitive exposure to buprenorphine. Pulmonary and visceral congestion; Toxicological analysis; Accidental asphyxia.

Presents a case report of an attack using a Japanese samurai sword. Claims of the attacker and victim; Evidence discussion; Suitability of the sword for stabbing and cutting; Potential of stabbing wounds to give fatal injuries; Difference between stab and incise wounds.

Abstract: Since 2004, a multidisciplinary Franco-Russian expedition discovered in the Sakha Republic (Yakutiya) more than 60 tombs preserved by the permafrost. In July 2006, an exceptionally well-preserved mummy was unearthed. The coffin, burial furniture and clothes suggested a shaman''s tomb. Multislice computed tomography (MSCT) was performed before autopsy with forensic and anthropological aims. Forensic study aimed to detect any lesions and determine the manner of death. Anthropological study aimed to determine the mummy''s gender, age at death, morphological affinity, stature and body mass. She was female and virginity status was assessed. The radiological and forensic conclusions were compared. Imaging confirmed most autopsy findings, suggesting that death followed disseminated infection. MSCT could not formally exclude a traumatic death because close examination of the skin was difficult, but was superior to conventional autopsy in diagnosis of infectious lesions of the left sacroiliac joint and one pelvic lesion. Autopsy detected a post-infectious spinal lesion, misinterpreted on MSCT as a Schmorl''s node. However, most conclusions of virtual and conventional anthropological studies agreed. Age at death was estimated around 19 years old. The morphology of the mummy was mongoloid. MSCT identified the craniometric characteristics as similar to those of the Buryat population. The deceased''s stature was 146cm and estimated body mass was 49kg. MSCT demonstrated its great potential and complementarity with conventional autopsy and anthropological techniques in the study of this natural female mummy buried in 1728. [Copyright &y& Elsevier]

The article reports on findings of hookworm infection in corpses found in a frozen burial site in the Altai mountains of Berel, Kazakhstan, in 1999. Two partly decomposed bodies were found in the burial chamber, a man and a woman. The woman was buried several years later. Samples from both bodies were investigated by direct examination and other methods, and found to contain hookworm eggs. The closest zone for hookworm infection is found in the shores of the Aral or Caspian sea, or in Iran. These findings confirm that the Scythe people covered large areas in their nomadic travels.

Fifteen somatic (D8S1179, D21S11, D7S820, CSF1PO, D3S1358, TH01, D13S317, D16S539, D2S1338, D19S433, VWA, TPOX, D18S51, D5S818, FGA) and five Y-chromosome (DYS19, DYS390, DYS391, DYS392, DYS393) short tandem repeat (STR) system analyses were carried out in a Mongolian population in order to define the possible relationship between Mongolians and old Hungarian population. For STR data analysis the Microsoft Excel-PowerStats program was used. Inter-population data analysis was performed with Arlequin Software ver. 2.000. The somatic markers showed meaningful difference between Mongolians and old Hungarians but the distribution of the Y-chromosome STR systems refers to a closer relationship between the old Hungarian and oriental populations. [Copyright &y& Elsevier]