24 research outputs found

    Whole genome amplification and real-time PCR in forensic casework

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    <p>Abstract</p> <p>Background</p> <p>WGA (Whole Genome Amplification) in forensic genetics can eliminate the technical limitations arising from low amounts of genomic DNA (gDNA). However, it has not been used to date because any amplification bias generated may complicate the interpretation of results. Our aim in this paper was to assess the applicability of MDA to forensic SNP genotyping by performing a comparative analysis of genomic and amplified DNA samples. A 26-SNPs TaqMan panel specifically designed for low copy number (LCN) and/or severely degraded genomic DNA was typed on 100 genomic as well as amplified DNA samples.</p> <p>Results</p> <p>Aliquots containing 1, 0.1 and 0.01 ng each of 100 DNA samples were typed for a 26-SNPs panel. Similar aliquots of the same DNA samples underwent multiple displacement amplification (MDA) before being typed for the same panel. Genomic DNA samples showed 0% PCR failure rate for all three dilutions, whilst the PCR failure rate of the amplified DNA samples was 0% for the 1 ng and 0.1 ng dilutions and 0.077% for the 0.01 ng dilution. The genotyping results of both the amplified and genomic DNA samples were also compared with reference genotypes of the same samples obtained by direct sequencing. The genomic DNA samples showed genotype concordance rates of 100% for all three dilutions while the concordance rates of the amplified DNA samples were 100% for the 1 ng and 0.1 ng dilutions and 99.923% for the 0.01 ng dilution. Moreover, ten artificially-degraded DNA samples, which gave no results when analyzed by current forensic methods, were also amplified by MDA and genotyped with 100% concordance.</p> <p>Conclusion</p> <p>We investigated the suitability of MDA material for forensic SNP typing. Comparative analysis of amplified and genomic DNA samples showed that a large number of SNPs could be accurately typed starting from just 0.01 ng of template. We found that the MDA genotyping call and accuracy rates were only slightly lower than those for genomic DNA. Indeed, when 10 pg of input DNA was used in MDA, we obtained 99.923% concordance, indicating a genotyping error rate of 1/1299 (7.7 × 10<sup>-4</sup>). This is quite similar to the genotyping error rate of STRs used in current forensic analysis. Such efficiency and accuracy of SNP typing of amplified DNA suggest that MDA can also generate large amounts of genome-equivalent DNA from a minimal amount of input DNA. These results show for the first time that MDA material is suitable for SNP-based forensic protocols and in general when samples fail to give interpretable STR results.</p

    Antimicrobial resistance among migrants in Europe: a systematic review and meta-analysis

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    BACKGROUND: Rates of antimicrobial resistance (AMR) are rising globally and there is concern that increased migration is contributing to the burden of antibiotic resistance in Europe. However, the effect of migration on the burden of AMR in Europe has not yet been comprehensively examined. Therefore, we did a systematic review and meta-analysis to identify and synthesise data for AMR carriage or infection in migrants to Europe to examine differences in patterns of AMR across migrant groups and in different settings. METHODS: For this systematic review and meta-analysis, we searched MEDLINE, Embase, PubMed, and Scopus with no language restrictions from Jan 1, 2000, to Jan 18, 2017, for primary data from observational studies reporting antibacterial resistance in common bacterial pathogens among migrants to 21 European Union-15 and European Economic Area countries. To be eligible for inclusion, studies had to report data on carriage or infection with laboratory-confirmed antibiotic-resistant organisms in migrant populations. We extracted data from eligible studies and assessed quality using piloted, standardised forms. We did not examine drug resistance in tuberculosis and excluded articles solely reporting on this parameter. We also excluded articles in which migrant status was determined by ethnicity, country of birth of participants' parents, or was not defined, and articles in which data were not disaggregated by migrant status. Outcomes were carriage of or infection with antibiotic-resistant organisms. We used random-effects models to calculate the pooled prevalence of each outcome. The study protocol is registered with PROSPERO, number CRD42016043681. FINDINGS: We identified 2274 articles, of which 23 observational studies reporting on antibiotic resistance in 2319 migrants were included. The pooled prevalence of any AMR carriage or AMR infection in migrants was 25·4% (95% CI 19·1-31·8; I2 =98%), including meticillin-resistant Staphylococcus aureus (7·8%, 4·8-10·7; I2 =92%) and antibiotic-resistant Gram-negative bacteria (27·2%, 17·6-36·8; I2 =94%). The pooled prevalence of any AMR carriage or infection was higher in refugees and asylum seekers (33·0%, 18·3-47·6; I2 =98%) than in other migrant groups (6·6%, 1·8-11·3; I2 =92%). The pooled prevalence of antibiotic-resistant organisms was slightly higher in high-migrant community settings (33·1%, 11·1-55·1; I2 =96%) than in migrants in hospitals (24·3%, 16·1-32·6; I2 =98%). We did not find evidence of high rates of transmission of AMR from migrant to host populations. INTERPRETATION: Migrants are exposed to conditions favouring the emergence of drug resistance during transit and in host countries in Europe. Increased antibiotic resistance among refugees and asylum seekers and in high-migrant community settings (such as refugee camps and detention facilities) highlights the need for improved living conditions, access to health care, and initiatives to facilitate detection of and appropriate high-quality treatment for antibiotic-resistant infections during transit and in host countries. Protocols for the prevention and control of infection and for antibiotic surveillance need to be integrated in all aspects of health care, which should be accessible for all migrant groups, and should target determinants of AMR before, during, and after migration. FUNDING: UK National Institute for Health Research Imperial Biomedical Research Centre, Imperial College Healthcare Charity, the Wellcome Trust, and UK National Institute for Health Research Health Protection Research Unit in Healthcare-associated Infections and Antimictobial Resistance at Imperial College London

    Suicidal Behavior and Alcohol Abuse

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    Suicide is an escalating public health problem, and alcohol use has consistently been implicated in the precipitation of suicidal behavior. Alcohol abuse may lead to suicidality through disinhibition, impulsiveness and impaired judgment, but it may also be used as a means to ease the distress associated with committing an act of suicide. We reviewed evidence of the relationship between alcohol use and suicide through a search of MedLine and PsychInfo electronic databases. Multiple genetically-related intermediate phenotypes might influence the relationship between alcohol and suicide. Psychiatric disorders, including psychosis, mood disorders and anxiety disorders, as well as susceptibility to stress, might increase the risk of suicidal behavior, but may also have reciprocal influences with alcohol drinking patterns. Increased suicide risk may be heralded by social withdrawal, breakdown of social bonds, and social marginalization, which are common outcomes of untreated alcohol abuse and dependence. People with alcohol dependence or depression should be screened for other psychiatric symptoms and for suicidality. Programs for suicide prevention must take into account drinking habits and should reinforce healthy behavioral patterns

    Surgical site infection after gastrointestinal surgery in high-income, middle-income, and low-income countries: a prospective, international, multicentre cohort study

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    Background: Surgical site infection (SSI) is one of the most common infections associated with health care, but its importance as a global health priority is not fully understood. We quantified the burden of SSI after gastrointestinal surgery in countries in all parts of the world. Methods: This international, prospective, multicentre cohort study included consecutive patients undergoing elective or emergency gastrointestinal resection within 2-week time periods at any health-care facility in any country. Countries with participating centres were stratified into high-income, middle-income, and low-income groups according to the UN's Human Development Index (HDI). Data variables from the GlobalSurg 1 study and other studies that have been found to affect the likelihood of SSI were entered into risk adjustment models. The primary outcome measure was the 30-day SSI incidence (defined by US Centers for Disease Control and Prevention criteria for superficial and deep incisional SSI). Relationships with explanatory variables were examined using Bayesian multilevel logistic regression models. This trial is registered with ClinicalTrials.gov, number NCT02662231. Findings: Between Jan 4, 2016, and July 31, 2016, 13 265 records were submitted for analysis. 12 539 patients from 343 hospitals in 66 countries were included. 7339 (58·5%) patient were from high-HDI countries (193 hospitals in 30 countries), 3918 (31·2%) patients were from middle-HDI countries (82 hospitals in 18 countries), and 1282 (10·2%) patients were from low-HDI countries (68 hospitals in 18 countries). In total, 1538 (12·3%) patients had SSI within 30 days of surgery. The incidence of SSI varied between countries with high (691 [9·4%] of 7339 patients), middle (549 [14·0%] of 3918 patients), and low (298 [23·2%] of 1282) HDI (p < 0·001). The highest SSI incidence in each HDI group was after dirty surgery (102 [17·8%] of 574 patients in high-HDI countries; 74 [31·4%] of 236 patients in middle-HDI countries; 72 [39·8%] of 181 patients in low-HDI countries). Following risk factor adjustment, patients in low-HDI countries were at greatest risk of SSI (adjusted odds ratio 1·60, 95% credible interval 1·05–2·37; p=0·030). 132 (21·6%) of 610 patients with an SSI and a microbiology culture result had an infection that was resistant to the prophylactic antibiotic used. Resistant infections were detected in 49 (16·6%) of 295 patients in high-HDI countries, in 37 (19·8%) of 187 patients in middle-HDI countries, and in 46 (35·9%) of 128 patients in low-HDI countries (p < 0·001). Interpretation: Countries with a low HDI carry a disproportionately greater burden of SSI than countries with a middle or high HDI and might have higher rates of antibiotic resistance. In view of WHO recommendations on SSI prevention that highlight the absence of high-quality interventional research, urgent, pragmatic, randomised trials based in LMICs are needed to assess measures aiming to reduce this preventable complication

    La sécurité entre permanence et changement

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    The end of the bipolar era should have inevitably led to a new approach in security conceptions. But in what direction? Linking contributions from Barry Buzan, Ole Wæver, and Roxanne Lynn Doty, briefly retracing the conceptual evolution from state security to human security passing through societal security and proposing a transnational reading of security stakes and approaches, the author offers a reflection on the ways to understand security as an object of study and as a translation of the security stakes and issues which may concern the individual, society, and the State. Being in a constant flux, one will understand that security stakes are just the ultimate result of a combination of the natural and the rational

    The European Gendarmerie Force: a solution in search of problems?

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    By creating the European Gendarmerie Force (EGF), some European Union (EU) member states have devised a ‘structured solution’ to provide international civilian police capabilities. In this article, we undertake a critical examination of the EGF by first arguing that the EGF has been widely misrepresented, notably with regard to its general purpose and specific relationship to the EU. Next, we examine a range of security problems used to justify the EGF, arguing that its potential role in handling certain tasks has not been very carefully considered. Finally, we suggest that a major rationale behind the EGF was the shared desire among its members to draw attention to a policing model that is not universally appreciated, and to promote this model by offering its ‘third-type’ capabilities while keeping the EGF outside of EU institutional constraints. In the conclusion, we identify some crucial questions related to the EGF–EU relations, notably in terms of non-optimisation of EU resources and possible incoherence in EU/Common Security and Defence Policy efforts

    La diagnosi di annegamento: un'autopsia ancora problematica

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    L’annegamento è un fenomeno a bassa incidenza ma con elevata mortalità, risultando essere nel mondo una tra le principali cause di morte accidentale. Inoltre, non raramente, l’annegamento viene usato con finalità suicidiaria od omicidiaria, assumendo quindi una corretta diagnosi medico-legale un ruolo risolutivo, soprattutto in caso di permanenza in acqua del cadavere per periodi di tempo anche lunghi. L’indagine su corpi recuperati in acqua dunque comprende una quota importante dei compiti del medico-legale che con l’avanzare delle conoscenze scientifiche, specialmente nel campo della biologia molecolare e dell’immunoistochimica, può avvalersi sempre di nuove metodiche di indagine senza dover abbandonare le precedenti: un’attenta ispezione della scena del ritrovamento del cadavere ed un accurato esame autoptico devono essere completati da indagini tossicologiche e di laboratori

    Il tessuto ischemico miocardico in stato avanzato di putrefazione: uno studio istochimico

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    La diagnosi istologica in patologia forense è spesso resa difficoltosa dai fenomeni trasformativi, quali l'autolisi e la putrefazione. Frequentemente infatti il materiale biologico viene prelevato da cadaveri rinvenuti molto tempo dopo la morte o riesumati: questo comporta una serie di difficoltà sia nell'allestimento dei preparati che nell'interpretazione delle immagini microscopiche. Occorre considerare anche la differente struttura dei tessuti e degli organi che condiziona il diverso andamento dei fenomeni trasformativi. Usualmente, infatti, la putrefazione altera precocemente la struttura di organi quali il fegato, il rene e l'encefalo: maggiore invece è la resistenza alla putrefazione di altri tessuti, alcuni molto importanti per la diagnosi istologica, quali il polmone ed il miocardio.The histological diagnosis in forensic pathology is often made difficult by the transformative phenomena, such as autolysis and putrefaction. Frequently, in fact, the biological material is taken from corpses unearthed long after the death or exhumed: this involves a number of difficulties in setting both of the preparations in the interpretation of microscopic images. It should also consider the different structure of tissues and organs that affects the different performance of transformative phenomena. Usually, in fact, the putrefaction early alters the structure of organs such as the liver, the kidney and the brain: instead greater is the resistance to rot of other tissues, some very important for histological diagnosis, such as lung and myocardium
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