Postmortem genetic testing in sudden unexplained death: A public health laboratory experience

Sudden unexplained death in the young poses a diagnostically challenging situation for practicing autopsy pathologists, especially in the absence of anatomic and toxicological findings. Postmortem genetic testing may identify pathogenic variants in the deceased of such cases, including those associated with arrhythmogenic channelopathies and cardiomyopathies. The Wisconsin State Laboratory of Hygiene (WSLH) is a state-run public health laboratory which performs postmortem genetic testing at no cost to Wisconsin medical examiners and coroners. The current study examines sequencing data from 18 deceased patients (ages 2 months to 49 years, 5 females) submitted to WSLH, from 2016 to 2021. Panel-based analysis was performed on 10 cases, and whole exome sequencing was performed on the most recent 8 cases. Genetic variants were identified in 14 of 18 decedents (77.8%), including 7 with pathogenic or likely pathogenic variants (38.9%). Whole exome sequencing was more likely to yield a positive result, more variants per decedent, and a larger number of variants of uncertain significance. While panel-based testing may offer definitive pathogenic variants in some cases, less frequent variants may be excluded. Whole exome testing may identify rare variants missed by panels, but increased yield of variants of uncertain significance may be difficult to interpret. Postmortem genetic testing in young decedents of sudden unexplained death can provide invaluable information to autopsy pathologists to establish accurate cause and manner of death and to decedent's relatives to allow appropriate management. A public health laboratory model may be a financially advisable alternative to commercial laboratories for medical examiner's/coroner's offices.     

Identifying reliable ions for the statistical differentiation of structurally similar fentanyl analogs

Definitive identification of fentanyl analogs based on mass spectral comparison is challenging given the high degree of structural and, hence, spectral similarity. To address this, a statistical method was previously developed in which two electron-ionization (EI) mass spectra are compared using the unequal variance t-test. Normalized intensities of corresponding ions are compared, testing the null hypothesis (H₀) that the difference in intensity is equal to zero. If H₀ is accepted at all m/z values, the two spectra are statistically equivalent at the specified confidence level. If H₀ is not accepted at any m/z value, then there is a significant difference in intensity at that m/z value between the two spectra. In this work, the statistical comparison method is applied to distinguish EI spectra of valeryl fentanyl, isovaleryl fentanyl, and pivaloyl fentanyl. Spectra of the three analogs were collected over a 9-month period and at different concentrations. At the 99.9% confidence level, the spectra of corresponding isomers were statistically associated. Spectra of different isomers were statistically distinct, and ions responsible for discrimination were identified in each comparison. To account for inherent instrument variations, discriminating ions for each pairwise comparison were ranked based on the magnitude of the calculated t-statistic (t_calc) value. For a given comparison, ions with higher t_calc values are those with the greatest difference in intensity between the two spectra and, therefore, are considered more reliable for discrimination. Using these methods, objective discrimination among the spectra was achieved and ions considered most reliable for discrimination of these isomers were identified.     

平衡、融合与引导——试析新世界秩序构建中的中国作用

频繁的世界危机凸显出全球化时代下国际问题不可阻挡的跨国溢出与无序的世界治理秩序之间的深刻矛盾。从能源到气候变化问题,从大规模杀伤性武器的扩散到传染性疾病的防治,从国际金融系统的稳定到跨国宏观经济协调,政治、经济、安全问题复杂交织,世界越来越陷入动荡不安之中。当前处于世界新秩序重构的重要转型时期,中国不断增加的国家实力、声望和影响力赋予其在世界新秩序的构建中举足轻重的地位。本文旨在阐述中国在新秩序构建中的力量平衡、制度融合与理念引导作用。     

Heteroplasmy in hair: differences among hair and blood from the same individuals are still a matter of debate

The analysis of mitochondrial DNA (mtDNA) is a useful tool in forensic cases when sample contents too little or degraded nuclear DNA to genotype by autosomal short tandem repeat (STR) loci, but it is especially useful when the only forensic evidence is a hair shaft. Several authors have related differences in mtDNA from different tissues within the same individual, with high frequency of heteroplasmic variants in hair, as also in some other tissues. Is still a matter of debate how the differences influence the interpretation forensic protocols. One difference between two samples supposed to be originated from the same individual are related to an inconclusive result, but depending on the tissue and the position of the difference it should have a different interpretation, based on mutation-rate heterogeneity of mtDNA. In order to investigate it differences in the mtDNA control region from hair shafts and blood in our population, sequences from the hypervariable regions 1 and 2 (HV1 and HV2) from 100 Brazilian unrelated individuals were compared. The frequency of point heteroplasmy observed in hair was 10.5% by sequencing. Our study confirms the results related by other authors that concluded that small differences within tissues should be interpreted with caution especially when analyzing hair samples.     

The effect of household corrosive substances on latent fingermark development in the context of deliberate corrosive substance attacks

Corrosive substance attacks (CSA) are a prevalent issue in the UK with 525 offenses involving a corrosive substance reported to the police in the year ending March 2022. Easy availability, low cost, and concealability in public are common reasons for choosing a corrosive substance as a weapon. The Metropolitan Police revealed that 68% of 1849 CSA cases resulted in no suspect identified or evidential difficulties. There is limited research into the effect of corrosive substances on latent fingermarks. This study aimed to determine the potential for fingermarks to be recovered from surfaces exposed to a household corrosive substance within the context of a deliberate CSA. Natural and sebaceous-loaded fingermarks were exposed to Domestos bleach, Harpic limescale remover (hydrochloric acid-based) and lemon juice. Harpic limescale remover had the most detrimental effect, with only 7.1% of fingermarks (n = 378) exposed being identifiable (defined as sufficient clear ridge detail for identification) after enhancement, followed by bleach with only 10.3% of fingermarks (n = 378) identifiable. Lemon juice had the least detrimental effect on fingermarks, with 40.5% fingermarks (n = 378) identifiable compared to 53.4% for the controls (not exposed to any substance; n = 378). Throughout the study, fewer natural fingermarks were identifiable after exposure to corrosive substances compared to sebaceous fingermarks which was as expected. Overall, this study demonstrated that there is potential to recover latent fingermarks, depending on their composition, following exposure to a household corrosive substance. This area warrants further research to establish best practice to maximize the potential to recover identifiable fingermarks.     

Examining Ethnic Identity and Self-Esteem Among Biracial and Monoracial Adolescents

The psychological well-being and ethnic identity of biracial adolescents are largely underrepresented topics in current scholarly literature, despite the growing population of biracial and multiracial individuals in the United States. This study examined self-esteem, ethnic identity, and the relationship between these constructs among biracial and monoracial adolescents (n = 3282). Using analysis of covariance, significant differences emerged between biracial and monoracial adolescents on both a measure of self-esteem and a measure of ethnic identity. Specifically, biracial adolescents showed significantly higher levels of self-esteem than their Asian counterparts, but significantly lower self-esteem than Black adolescents. Furthermore, biracial adolescents scored significantly higher than Whites on a measure of ethnic identity, but scored lower than their Black, Asian, and Latino peers on the same measure. Finally, correlational analyses revealed a significant and positive relationship between ethnic identity and self-esteem for all groups.     

An ocean for the Global South: Brazil and the zone of peace and cooperation in the South Atlantic

In this article, we analyse an instance of revitalisation of a dormant interregional organisation dating back to the Cold War: the Zone of Peace and Cooperation of the South Atlantic (ZOPACAS), initially launched by South American and African states in 1986 through the UN General Assembly. Drawing on the concepts of “consensual hegemony” we argue that the current phase of ZOPACAS’ existence is characterised by Brazil's efforts to rekindle it, thus reflecting its aspiration to create a new space of influence. Rather than pursuing more traditional forms of regional leadership, Brazil uses ZOPACAS as part of a persuasion-based strategy based on regional multilateralism that is designed in antagonism to other international organisations and Western powers. However, this strategy also faces important limitations resulting from resource constraints, lack of institutionalisation and an excessive exclusionary focus on minimising the role of global powers with interests in the region.     

Practical benzylation of N,N-substituted ethanolamines related to chemical warfare agents for analysis and detection by electron ionization gas chromatography–mass spectrometry

The benzylation of three low molecular weight N,N-disubstituted ethanolamines related to chemical warfare agents (CWAs) to furnish derivatives with improved gas chromatography-mass spectrometry (GC-MS) profiles is described. Due to their low molecular weight and polar nature, N,N-disubstituted ethanolamines are notoriously difficult to detect by routine GC-MS analyses during Organisation for the Prohibition of Chemical Weapons (OPCW) proficiency tests (PTs), particularly in scenarios when they are present at low levels (~1–10 ppm) amidst more abundant interferences. Our studies revealed that the optimal derivatization conditions involved the treatment of the ethanolamine with benzyl bromide in the presence of an inorganic base (e.g., Na₂CO₃) in dichloromethane at 55°C for 2 h. This optimized set of conditions was then successfully applied to the derivatization of N,N-dimethylethanolamine, N,N-diethylethanolamine and N,N-diisopropylethanolamine present separately at 1 and 10 μg/mL concentrations in a glycerol-rich matrix sample featured in the 48th OPCW PT. The benzylated derivatives of the three ethanolamines possessed retention times long enough to clear the massive glycerol-containing matrix interferences. The protocol herein is introduced as an alternative method for derivatization of these CWA and pharmaceutically important species and should find broad applicability in laboratories where routine forensic analysis is carried out.     

A statistical approach to aid examiners in the forensic analysis of handwriting

We develop a statistical approach to model handwriting that accommodates all styles of writing (cursive, print, connected print). The goal is to compute a posterior probability of writership of a questioned document given a closed set of candidate writers. Such probabilistic statements can support examiner conclusions and enable a quantitative forensic evaluation of handwritten documents. Writing is treated as a sequence of disjoint graphical structures, which are extracted using an automated and open-source process. The graphs are grouped based on the similarity of their shapes through a K-means clustering template. A person's writing pattern can be characterized by the rate at which graphs are emitted to each cluster. The cluster memberships serve as data for a Bayesian hierarchical model with a mixture component. The rate of mixing between two parameters in the hierarchy indicates writing style.