Molecular mass screening of mosquitoes for filarial parasites in Germany – re-interpretation of PCR xenomonitoring results would be required

Abstract Comments concerning interpretation of the PCR xenomonitoring results in the article „Molecular detection of Setaria tundra (Nematoda: Filarioidea) and an unidentified filarial species in mosquitoes in Germany” Parasites & Vectors 2012, 5:14

Ligation mediated PCR performed at low denaturation temperatures—PCR melting profiles

We show that using low denaturation temperatures (80–88°C) during ligation mediated PCR (LM PCR) of bacterial DNA leads to the amplification of limited sets of the less stable DNA fragments. A set of electrophoretic patterns of such fragments obtained at different denaturation temperatures forms the PCR melting profile (PCR MP). A single pattern obtained for a given temperature and a set of patterns arising after application of several denaturation temperatures (PCR MP) are very specific for the given bacterial genome and may be used for strain characterisation and differentiation. The method may also be used for amplification and isolation of the less stable DNA fragments in a genome

Identification of monozygotic twins in the era of next‑generation sequencing

Bliźnięta monozygotyczne stanowią wyzwanie dla kryminalistyki ze względu na brak standaryzowanych narzędzi do ich różnicowania w obrębie pary, utrudniający lub uniemożliwiający sądowi ustalenie, które z bliźniąt jest sprawcą przestępstwa. Pojawienie się technologii sekwencjonowania nowej generacji, sekwencjonowania całogenomowego oraz analiz epigenomu ludzkiego otworzyło drogę do identyfikacji bliźniąt monozygotycznych w celach kryminalistycznych. Kwestią otwartą pozostaje odsetek bliźniąt monozygotycznych, który można różnicować, oraz poziom wiarygodności uzyskanych wyników. Dowody uzyskane za pomocą sekwencjonowania całogenomowego, w sprawach dotyczących identyfikacji bliźniąt monozygotycznych, budzą wątpliwości sądów. Wraz z nowymi technologiami pojawiał się szereg pytań odnośnie do ich parametrów: siły różnicowania, poziomu wiarygodności wyniku, sposobów walidacji metod oraz zagadnień natury prawnej. Nowe technologie identyfikacji bliźniąt wymagają dostosowania do wymagań stawianych metodom kryminalistycznym: weryfikacji, walidacji i standaryzacji tych metod oraz jednolitego podejścia do interpretacji wyników, aby mogły być powszechnie stosowane w kryminalistyce i uznawane przez sądy za wiarygodne dowody.Monozygotic twins pose a challenge to forensic science due to the lack of standardized tools to differentiate them within a pair, and thus making it difficult or impossible for a court to determine which of the twins is the perpetrator of the crime. The advent of next‑generation sequencing technologies, whole‑genome sequencing, and analyses of the human epigenome, has opened the way to the identification of monozygotic twins for forensic purposes. The percentage of monozygotic twins that can be differentiated and the level of reliability of results remain an issue open for investigation. Identification of monozygotic twins through whole‑genome sequencing raises doubts in the courts. New technologies raised a number of questions regarding the parameters: the power of differentiation, the level of reliability of the result, validation method and legal issues. New twin identification technologies require adapting to the requirements for forensic methods: verification, validation and standardization, as well as a uniform approach to the interpretation of results, so that they can be widely used in forensics and recognized by courts as reliable evidence

Integrons

Accumulation of antibiotic resistance among pathogenic bacteria has called attention to horizontal gene transfer that involves plasmids and transpozons. Integrons, usually placed on mobile genome elements, are very deeply engaged in the process of origin of multiple-drug-resistant strains. Integrons are genetic elements that contain determinants of the components of the site-specific recombination system that recognizes and captures mobile gene cassettes. More than 70 different antibiotic resistance genes covering most classes of antimicrobials presently in use have been detected in gene cassettes. Integrons are frequently found in clinical and environmental strains of gram-negative rods. The discovery of super-integrons, i.e. genetic structures gathering gene cassettes in a huge number, led to the conception of genome cassettes capture as an element of a broader phenomenon of bacterial genome modification in response to changing environmental conditions

Molecular Determinants of Drug Resistance and Mutation Patterns in Influenza Viruses Circulating in Poland Across Multiple Epidemic Seasons: Implications for Vaccination Strategies

Background: According to the WHO, up to 650 000 people die each year from seasonal flu-related respiratory illnesses. The most effective method of fighting the virus is seasonal vaccination. However, if an infection does occur, antiviral medications should be used as soon as possible. No studies of drug resistance in influenza viruses circulating in Poland have been systematically conducted. Therefore, the aim of the present study was to investigate the drug resistance and genetic diversity of influenza virus strains circulating in Poland by determining the presence of mutations in the neuraminidase gene. Material/Methods: A total of 258 clinical specimens were collected during the 2016-2017, 2017-2018, and 2018-2019 epidemic seasons. The samples containing influenza A and B were analyzed by RT-PCR and Sanger sequencing. Results: Differences were found between the influenza virus strains detected in different epidemic seasons, demonstrating the occurrence of mutations. Influenza A virus was found to be more genetically variable than influenza B virus (P<0.001, Kruskal-Wallis test). However, there was no significant difference in the resistance prevalence between the influenza A subtypes A/H1N1/pdm09 (4.8%) and A/H3N2/ (6.1%). In contrast, more mutations of drug-resistance genes were found in the influenza B virus (P<0.001, chi-square test). In addition, resistance mutations appeared en masse in vaccine strains circulating in unvaccinated populations. Conclusions: It seems important to determine whether the influenza virus strains tested for drug resistance as part of global influenza surveillance are equally representative of viruses circulating in populations with high and low vaccination rates, for all countries. Our results suggest that countries with low levels of influenza immunization may constitute reservoirs of drug-resistant influenza viruses.S

The role of powered support in ensuring the proper longwall working cross-section area in rockburst-prone seams

In Poland, for underground hard coal seam extraction, the longwall method is used. Retreat longwalls with natural roof caving in the gob are the most common. Currently it is estimated that about a half of the hard coal output in Poland originates from seams located in areas of rock burst hazard. The article shows information about 18 rock bursts, and 14 other seismic events which occurred between 2003 and 2012 in underground mines belonging to one of coal companies in Poland. In addition, negative consequences of those dynamic phenomena in the longwall workings are described. In the next part of paper, the most common types of damage to powered supports after the rock mass tremors and other dynamic phenomena in longwall workings are shown. In order to avoid damage of powered supports, in geo-mining conditions where dynamic phenomena occur, different types of protective means are applied. In the paper the methodology of assessing the powered support yield ability is described. At the end, an example of the assessed yield ability of a powered support in given geo-mining conditions of a longwall face affected by the seismic events is presented

The role of powered support in ensuring the proper longwall working cross-section area in rockburst-prone seams

In Poland, for underground hard coal seam extraction, the longwall method is used. Retreat longwalls with natural roof caving in the gob are the most common. Currently it is estimated that about a half of the hard coal output in Poland originates from seams located in areas of rock burst hazard. The article shows information about 18 rock bursts, and 14 other seismic events which occurred between 2003 and 2012 in underground mines belonging to one of coal companies in Poland. In addition, negative consequences of those dynamic phenomena in the longwall workings are described. In the next part of paper, the most common types of damage to powered supports after the rock mass tremors and other dynamic phenomena in longwall workings are shown. In order to avoid damage of powered supports, in geo-mining conditions where dynamic phenomena occur, different types of protective means are applied. In the paper the methodology of assessing the powered support yield ability is described. At the end, an example of the assessed yield ability of a powered support in given geo-mining conditions of a longwall face affected by the seismic events is presented