The hero of American and Polish nations: a molecular look at Thaddeus Kosciuszko's cause of death suggests a contribution of endocarditis caused by Cutibacterium acnes infection.

The remains of the heart tissue of Thaddeus Kosciuszko have been investigated as the possible cause of disease and death of the hero of Polish and American nations. Three specimens, DNA isolated from scrappings of wax surface, from the surface of a wooden plate, and from the linen cloth that have had contact with the object were subjected to nanosequencing. From the first two, among all reads identified, only one classified as Propionibacterium acnes (synonymous current name Cutibacterium acnes), had a purported clinical significance. The observed identity between the P. acnes sequences and reference was 89-90% consistent with the hypothesis that the identified reads represent the ancient P. acnes DNA (aDNA), which underwent fragmentation and sequence changes caused by its long-time presence in the environmental conditions conducive to degradation. We present a reasonable and entirely new hypothesis that the analyzed samples could reflect the presence of the bacteria in the original Kosciuszko's heart tissue and that the process of C. acnes infection was progressing inside the organ (endocarditis), not on its surface (pericarditis) leading to rapid deterioration of health and eventually death. We again point out that normal skin and mucosal membranes commensal, a causative agent of common skin acne, may be associated with various severe organ infections posing a threat to health and life.

Role of hypoxia on microRNA-dependant regulation of HGFA - HGF - c-Met signalling pathway in human progenitor and mature endothelial cells.

Hepatocyte growth factor (HGF) is considered to be one of the key pro-angiogenic cytokines that stimulates endothelial cells to proliferate and migrate. The activation of the precursor form of HGF is primarily undertaken by the serine protease HGFA. Research indicates that HIF-1α hypoxia stimulates the expression of HGFA, which is synthesized by a range of cells including fibroblasts, endothelium, and macrophages. To date, little is known about the potential role of epigenetic factors in the regulation of the HGFA - HGF - c-Met signalling pathway. The literature suggests that there are several microRNAs (miRNAs, miRs) directly affecting the expression of c-Met under normoxic conditions. The main objective of the research described was to explore the effect of chemically-induced hypoxia on the expression of miRNA molecules in human progenitor and mature endothelial cells, with particulate attention paid to those miRNAs that may specifically affect the HGFA - HGF - c-Met signalling pathway. This publication sheds new light on the role of miRNAs in hypoxia, as well as identifying several miRNAs directly involved in the regulation of HGFA, HGF and c-Met expression in hypoxic conditions. The results indicate that hsa-miR-335-5p, hsa-miR-425-5p and hsa-miR-101-3p are the major miRNAs that appear to play an important role in the regulation of the HGFA - HGF - c-Met signalling pathway.

SARS-CoV-2 Viral RNA Is Detected in the Bone Marrow in Post-Mortem Samples Using RT-LAMP.

Since the emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in late 2019, viral RNA has been detected in several different human tissues and organs. This study reports the detection of SARS-CoV-2 RNA in the bone marrow. Post-mortem samples were taken in a sterile manner during two forensic autopsies from the nasopharyngeal region, vitreous humor, cerebrospinal fluid, and bone marrow. SARS-CoV-2 was subsequently diagnosed via Genomtec® SARS-CoV-2 EvaGreen® RT-LAMP CE-IVD Duo Kit. In both postmortem patients, SARS-CoV-2 RNA was detected in bone marrow samples. However, both the vitreous humor and cerebrospinal fluid from the same patients gave negative results using the same test system. The evidence of viral RNA in the bone marrow, along with other reports supports the thesis that SARS-CoV-2 infections are systemic in nature, the consequences of which would profoundly influence both the testing and survival of patients.

Role of Laboratory Medicine in SARS-CoV-2 Diagnostics. Lessons Learned from a Pandemic.

Since the 2019 novel coronavirus outbreak began in Wuhan, China, diagnostic methods in the field of molecular biology have been developing faster than ever under the vigilant eye of world's research community. Unfortunately, the medical community was not prepared for testing such large volumes or ranges of biological materials, whether blood samples for antibody immunological testing, or salivary/swab samples for real-time PCR. For this reason, many medical diagnostic laboratories have made the switch to working in the field of molecular biology, and research undertaken to speed up the flow of samples through laboratory. The aim of this narrative review is to evaluate the current literature on laboratory techniques for the diagnosis of SARS-CoV-2 infection available on pubmed.gov, Google Scholar, and according to the writers' knowledge and experience of the laboratory medicine. It assesses the available information in the field of molecular biology by comparing real-time PCR, LAMP technique, RNA sequencing, and immunological diagnostics, and examines the newest techniques along with their limitations for use in SARS-CoV-2 diagnostics.

Preliminary analysis of polymorphism of STR markers in the population of domestic cats (Felis catus) in Lower Silesia.

AIM OF THE STUDY: Genetic tests play a crucial role in the crime investigation process and often provide the strongest evidence for case resolution. Although the majority of genetic analyses in the field of criminalistics focus on the human DNA, genetic identification of animals is becoming an increasingly common procedure. Domestic animals, which live around people, may be silent witnesses and even victims of criminal activity. Their typically limited value as evidence in such cases could radically change thanks to the possibility of using animal biological material present at the crime scene. In addition to forensic medicine, genetic identification methods of this type may also become a valuable tool in many other areas of life. Recently, there has been an increase in public interest in verifying the pedigree of animals, investigating poaching and illegal shooting of animals, e.g. protected wildcats and lynx, as well as illegal trade in animals. The main aims of the studies reported in this paper were to assess the degree of polymorphism of the analyzed STR markers in feline genetic material, and to perform a preliminary evaluation of their suitability for developing an original feline genetic identification test. MATERIAL AND METHODS: The studies involved an analysis of genetic material samples obtained from a population consisting of 123 unrelated cats representing various domestic cat breeds, living in the Lower Silesia region. The material collected from individual cats in the form of blood drops or buccal swabs was subjected to an analysis of five STR markers forming a single multiplex assay (FCA742, FCA744, F124, FCA732, FCA749). RESULTS: The results obtained for each marker separately were analyzed statistically and, using the 2 test, the concordance of the study population with the Hardy-Weinberg principle was evaluated. CONCLUSIONS: The findings demonstrate a significant potential of the analyzed markers for the development of genetic identification tests.

DNA profiling of oaks (Quercus spp.)

AIM OF THE STUDY: Forensic botany demands tools to verify the value of plant-origin evidence brought into the process of criminal investigation. Molecular biology provides techniques for comparing material from the crime scene with other biological material of evidence. In this paper, we propose a set of seven markers based on Short Tandem Repeats (STRs) loci for DNA profiling of Quercus spp. STR markers of the highest observed heterozygosity were selected, according to available literature. Oaks were chosen due to their wide dissemination in the northern hemisphere. They served as an object of study to develop a method for obtaining comparable genetic profiles of plant evidence material. MATERIAL AND METHODS: In the study, we verified the specificity of primers towards selected species of oaks. Twenty-three species, including most of those previously studied, were investigated for the presence of selected loci. After DNA extraction, STR sequences were amplified using multiplex PCR, and amplification products were then analysed with capillary electrophoresis. The frequency of genotypes was tested with the χ2 test. RESULTS: Out of 23 investigated species of oak, full genetic profiles were obtained for 13 species. CONCLUSIONS: An incomplete genetic profile may result from DNA degradation or lower homology in primer binding sites. Full profiles were successfully obtained for most of the species tested; however, deficient profiles were yielded in species for which a full profile was expected. This may be due to the loss in DNA quality caused by ageing processes of plant material and inappropriate storage conditions or method of preservation.

15th Anniversary of the Molecular Techniques Unit at the Department of Forensic Medicine at Wroclaw Medical University.

Molecular Techniques Unit at the Department of Forensic Medicine, Wroclaw Medical University has been operating since December 2003. Soon it will be 15 years since its establishment. This anniversary become an inspiration to write down the story of this institution whose origins illustrate the immense changes that have taken place in forensic genetics. The aim of our work was also to consolidate the professional achievements of Professor Tadeusz Dobosz, chief of the Unit, one of the pioneers of introducing DNA testing technology into Polish forensic medicine. The most important achievements of the Unit include participation in two EU research projects, the development of a non-destructive method of extraction of genetic material, research in field of gene therapy and certification of the Laboratory of the Molecular Techniques Unit by the Polish Accreditation Center (PCA) confirming compliance with the requirements of the PN-EN ISO/IEC 17025:2005 standard.