Real-time PCR designs to estimate nuclear and mitochondrial DNA copy number in forensic and ancient DNA studies

We explore different designs to estimate both nuclear and mitochondrial human DNA (mtDNA) content based on the detection of the 5' nuclease activity of the Taq DNA polymerase using fluorogenic probes and a real-time quantitative PCR detection system. Human mtDNA quantification was accomplished by monitoring the real-time progress of the PCR-amplification of two different fragment sizes (113 and 287 bp) within the hypervariable region I (HV1) of the mtDNA control region, using two fluorogenic probes to specifically determine the mtDNA copy of each fragment size category. This mtDNA real-time PCR design has been used to assess the mtDNA preservation (copy number and degradation state) of DNA samples retrieved from 500 to 1500 years old human remains that showed low copy number and highly degraded mtDNA. The quantification of nuclear DNA was achieved by real-time PCR of a segment of the X-Y homologous amelogenin (AMG) gene that allowed the simultaneous estimation of a Y-specific fragment (AMGY: 112 bp) and a X-specific fragment (AMGX: 106 bp) making possible not only haploid or diploid DNA quantitation but also sex determination. The AMG real-time PCR design has been used to quantify a set of 57 DNA samples from 4-5 years old forensic bone remains with improved sensitivity compared with the slot-blot hybridization method. The potential utility of this technology to improve the quality of some PCR-based forensic and ancient DNA studies (microsatellite typing and mtDNA sequencing) is discussed.     

A light stabilizer Tinuvin 770-induced toxic injury of adult rat cardiac myocytes

Tinuvin 770/bis(2,2,6,6-tetramethyl-4-piperidinyl)sebacate is a worldwide used light stabilizer for plastic materials like polyolefins. Tinuvin 770 is a biologically active component of polypropylene tubes. Glossmann and his study group managed to extract this compound by aqueous or organic solvents from laboratory plastic tubes, and propose that Tinuvin 770 is a potent blocker of L-type Ca(2+)-channel through the phenylalkylamine and benzothiazepine-selective drug binding domains of the alpha(1) subunit of the receptor [Proc. Natl. Acad. Sci. U.S.A. 90 (1993) 9523].We examined the direct morphological effect of Tinuvin 770 in give 25nmol, 0, 30, 60, 120 minute exposure time in isolated cardiomyocytes from adult rats. Incubation of myocytes with Tinuvin resulted in a progressive decline of rod-shaped and viable cells. It was accompanied by an increase in number of hypercontracted myocytes with microbleb formation compared to control and depletion of ATP level.In summary, our results demonstrate that plasma membrane damage and hypercontraction are manifestations of Tinuvin-induced injury of isolated cardiomyocytes.     

STR data for the AmpFlSTR Profiler Plus and COfiler loci from the Maghreb (North Africa)

Allele frequencies for 13 STRs (D3S1358, vWA, FGA, D8S1179, D21S11, D18S51, D5S818, D13S317, D7S820, THO1, TPOX, CSF1PO and D16S539) included in the AmpFlSTR Profiler Plus and COfiler kits were determined for a population sample from the Maghreb (Northern Africa).     

Recreating the past in the ornamentation of stately homes and the leisure activities conducted within them: the Duchess of Parcent as a case study for the Spanish aristocracy in the early twentieth century

This paper attempts to vindicate the part played by certain Spanish aristocratic women in the management, conservation, and transmission of their artistic patrimony in the decades around the turn of the twentieth century by creating art collections and displaying them in their own homes. The works were exhibited in rooms that were decorated and furnished to suggest historical ambiences in which French trends rubbed shoulders with styles that had been developed in Spain throughout its history, turning their homes into devices that recreated past times. Similarly, retrospective views of Spanish art were the focus of leisure and entertainment activities driven by ladies from the upper echelons of society through the performance of tableaux vivants. The focus of this study will be Trinidad Scholtz Hermensdorff, Duchess of Parcent.     

What Constitution? On Chile’s Constitutional Awakening

Law and Critique - This paper explores the political awakening of the Chilean people that began in October 2019. It puts forward an alternative reading of the people’s claim for a new...     

Age Estimation Based on DNA Methylation Using Blood Samples From Deceased Individuals

Age estimation using DNA methylation levels has been widely investigated in recent years because of its potential application in forensic genetics. The main aim of this study was to develop an age predictor model (APM) for blood samples of deceased individuals based in five age-correlated genes. Fifty-one samples were analyzed through the bisulfite polymerase chain reaction (PCR) sequencing method for DNA methylation evaluation in genes ELOVL2, FHL2, EDARADD, PDE4C, and C1orf132. Linear regression was used to analyze relationships between methylation levels and age. The model using the highest age-correlated CpG from each locus revealed a correlation coefficient of 0.888, explaining 76.3% of age variation, with a mean absolute deviation from the chronological age (MAD) of 6.08 years. The model was validated in an independent test set of 19 samples producing a MAD of 8.84 years. The developed APM seems to be informative and could have potential application in forensic analysis.     

Networks as ‘laboratories of experience’: exploring the life cycle of the suffrage movement and its aftermath in Ireland 1870–1937

ABSTRACT

Applying an innovative conceptual framework this article presents an interdisciplinary re-appraisal of the suffrage movement and its aftermath in Ireland throughout the years 1870–1937. New social movement theory is utilised to consider how, in the words of the Italian sociologist Alberto Melucci ‘the submerged networks of social movements are laboratories of experience’. Going beyond the previously published work of each author, this article uses the sociological lens of ‘laboratories of experience’ to re-analyse aspects of the suffrage movement, female activism and the wider women's movement in Ireland. This application of social movement theory to female networks, their origins, aims and strategies, along with their interconnectedness, provides a more nuanced and detailed understanding of the ‘life-cycle’ of this movement. The article aims to demonstrate how an analysis of network dynamics and application of the concept of ‘latency’ is useful in further understanding the significance, impact and longevity of the women's movement in Ireland.     

Detection of Cutting Agents in Drug‐Positive Seized Exhibits within the United States

The following report summarizes a study performed on seized drug exhibits collected in two U.S. states to evaluate the presence and identification of cutting agents. Aliquots of seized drug materials from Kentucky (n = 200) and Vermont (n = 315) were prepared using a dilute‐and‐shoot procedure. Initial analysis was performed using gas chromatography–mass spectrometry (GC‐MS) followed by analysis using liquid chromatography quadrupole time‐of‐flight mass spectrometry (LC‐QTOF). Active compounds detected overall included caffeine (31.0%), quinine/quinidine (24.7%), levamisole (11.6%), acetaminophen, (8.2%) and procaine (8.2%). These compounds were found with several drugs of abuse, such as heroin, fentanyl, methamphetamine, and cocaine. This novel information about cutting agents used to dilute or alter drugs of abuse is important to criminal investigations and in the management of acute intoxications at health centers. However, common methodologies for analysis and standard reporting practices frequently do not include cutting agents, resulting in lacking or inadequate information regarding prevalence of these substances.