Expression and co-expression of surface markers of pluripotency on human amniotic cells cultured in different growth media

Objectives: Despite constant advances in the field of biology and medical application of human embryonic stem cells, the molecular mechanism of pluripotency remains largely unknown. So far, definitions of pluripotent stem cells (SC) have been based on a limited number of antigenic markers and have not allowed for unambiguous determination of the homogeneity of each subpopulation. Moreover, the use of some crucial pluripotency markers such as SSEA-3 and SSEA-4 has recently been questioned due to the possibility that the pattern of surface glycans may be changed depending on the content of the cell culture medium. Aim: Quantitative analysis of amniotic SC subpopulations cultured in different media, based on the following pluripotency surface markers: SSEA-3, SSEA-4, TRA-1-60 and TRA-1-81 expression and co-expression. Material and methods: Immunofluorescence and fluorescence microscopy were used to identify and localize S.C. within a normal human placenta at term. The number of SSEA-4+, SSEA-3+, TRA-1-60+ and TRA-1-81+ cells and cells with co-expression of the above mentioned markers, cultured in media containing different protein supplements of animal origin, was counted by flow cytometry. Results and conclusions: Cells with characteristics of embryonic SC were identified in the amniotic epithelium and the chorion, but not in the decidua basalis. Amniotic epithelium contained various types of SC, with SSEA-4+ as the most numerous. Disproportion in the number of SSEA-4+, SSEA-3+, TRA-1-60+ and TRA-1-81+ cells and cells characterized by co-expression of these antigens, as well as lack of quantitative differences between S.C. subpopulations cultured in different media, was observed. In conclusion, the amniotic epithelium is composed of SC at different stages of the development but human amnion might become an alternative source of SSEA-4+ embryonic-like SC. The composition of the evaluated media, characterized by different content of animal-derived proteins, does not influence the number of cells identified within the SC subpopulations

Long-term space missions’ effects on the human organism: what we do know and what requires further research

Space has always fascinated people. Many years have passed since the first spaceflight, and in addition to the enormous technological progress, the level of understanding of human physiology in space is also increasing. The presented paper aims to summarize the recent research findings on the influence of the space environment (microgravity, pressure differences, cosmic radiation, etc.) on the human body systems during short-term and long-term space missions. The review also presents the biggest challenges and problems that must be solved in order to extend safely the time of human stay in space. In the era of increasing engineering capabilities, plans to colonize other planets, and the growing interest in commercial space flights, the most topical issues of modern medicine seems to be understanding the effects of long-term stay in space, and finding solutions to minimize the harmful effects of the space environment on the human body

The smell of death. State-of-the-art and future research directions

The decomposition of a body is inseparably associated with the release of several types of odors. This phenomenon has been used in the training of sniffer dogs for decades. The odor profile associated with decomposition consists of a range of volatile organic compounds (VOCs), chemical composition of which varies over time, temperature, environmental conditions, and the type of microorganisms, and insects colonizing the carcass. Mercaptans are responsible for the bad smell associated with corpses; however, there are no unified recommendations for conducting forensic analysis based on the detectable odor of revealed corpses and previous research on VOCs shows differing results. The aim of this review is to systematize the current knowledge on the type of volatile organic compounds related to the decomposition process, depending on a few variables. This knowledge will improve the methods of VOCs detection and analysis to be used in modern forensic diagnostics and improve the methods of training dogs for forensic applications

Post-tracheostomy complications: respiratory failure caused by authologic foreign body – case report

Tracheostomy is performed frequently as a palliative treatment in patients with end-stage respiratory failure (RF). However, in patients requiring prolonged mechanical ventilation it may be difficult to recognize and can often lead to life-threatening RF. We present two cases of acute-on-chronic respiratory failure (ACRF) occurring in patients who had undergone tracheostomy [one with percutaneous dilatational tracheostomy (PDT) and the second with surgical tracheostomy (ST)]. The first case was admitted due to ACRF several months after previous successful decannulation and the second case after failure of several attempts of weaning from tracheal cannula. In both cases, noninvasive mechanical ventilation assisted flexible bronchoscopy (NIV-FB) was able to identify and solve the tracheal stenosis secondary to stiff bananashaped whitish foreign bodies. Histology sampling and genetic testing confirmed autologous foreign body formation—tracheal cartilage calcification. NIV-FB was found to be safe and effective in both diagnosis and treatment of the tracheal stenosis. Life-threatening RF connected with tracheal stenosis may be caused by rupture of tracheal cartilage ossification in patients with a history of ST and PDT. Bronchofiberoscopy performed with NIV will be a useful procedure to evaluate and treat the respiratory tract in patients with RF with suspected tracheal stenosis

Nucleic Acids Persistence—Benefits and Limitations in Forensic Genetics

The analysis of genetic material may be the only way to identify an unknown person or solve a criminal case. Often, the conditions in which the genetic material was found determine the choice of the analytical method. Hence, it is extremely important to understand the influence of various factors, both external and internal, on genetic material. The review presents information on DNA and RNA persistence, depending on the chemical and physical factors affecting the genetic material integrity. One of the factors taken into account is the time elapsing to genetic material recovery. Temperature can both preserve the genetic material or lead to its rapid degradation. Radiation, aquatic environments, and various types of chemical and physical factors also affect the genetic material quality. The substances used during the forensic process, i.e., for biological trace visualization or maceration, are also discussed. Proper analysis of genetic material degradation can help determine the post-mortem interval (PMI) or time since deposition (TsD), which may play a key role in criminal cases

Pośmiertna detekcja etanolu w zwłokach poddanych balsamacji – badania wstępne

Nowadays, embalming has become more popular as it is often performed due to sanitary, hygienic, or psychological reasons. In special circumstances, like transporting a corpse from abroad, embalming is an administrative requirement. Embalming is performed using chemical substances that inhibit enzymes, bacteriological activity, and therefore cease decomposition processes. The study shows the toxicological analyses of sampled materials collected during post mortem examinations performed on six corpses subjected to the embalming process. It also presents the concentration of used preservatives in the tissues and body fluids. Analysis performed using gas chromatography with a flame ionization detector (GC-FID) revealed that in five cases, the deceased were intoxicated with ethanol during the time of death, and liquid chromatography-mass spectrometry (LC-MS) excluded the presence of psychoactive substances in all six cases. The results showed that in cases of toxicological analyses of embalmed corpses, it is crucial to secure isolated body fluids, such as cerebrospinal fluid, vitreous humor, and a wide range of reference material.Balsamacja zwłok staje się coraz bardziej popularna ze względów sanitarnych, higienicznych lub psychologicznych. W szczególnych okolicznościach, jak transport zwłok z zagranicy, balsamowanie jest wymogiem administracyjnym. Balsamacja odbywa się za pomocą substancji chemicznych, które hamują aktywność enzymatyczną i bakteriologiczną, wstrzymując tym samym procesy rozkładu. Prezentowana praca przedstawia analizy toksykologiczne materiałów po-branych podczas sekcji sześciu zwłok poddanych procesom balsamowania oraz wyniki pomiarów stężenia użytych konserwantów w tkankach i płynach ustrojowych badanych zwłok. Analiza przeprowadzona za pomocą chromatografii gazowej z detektorem płomieniowo-jonizacyjnym (gas chromatography with a flame ionization detector – GC-FID) udowodniła obecność etanolu w pięciu przypadkach. Chromatografia cieczowa połączona ze spektrometrią mas (liquid chromatography-mass spectrometry – LC-MS) wykluczyła obecność substancji psychoaktywnych we wszystkich sze-ściu przypadkach. Wyniki wykazały, że w przypadku analiz toksykologicznych zabalsamowanych zwłok kluczowe znaczenie ma zabezpieczenie izolowanych płynów ustrojowych, takich jak płyn mózgowo-rdzeniowy, płyn gałki ocznej, oraz wielu materiałów referencyjnych

Insects feeding on cadavers as an alternative source of human genetic material

In some criminal cases, the use of classical sources of human genetic material is difficult or even impossible. One solution may be the use of insects, especially blowfly larvae which feed on corpses. A recent review of case reports and experimental studies available in biomedical databases has shown that insects can be a valuable source of human mitochondrial and genomic deoxyribonucleic acid (DNA), allowing for an effective analysis of hypervariable region (HVR) sequences and short tandem repeat (STR) profiles, respectively. The optimal source of human DNA is the crop (a part of the gut) of active third-instar blowfly larvae. Pupae and insect faeces can be also used in forensic genetic practice instead of the contents of the alimentary tract

Polymorphism of SE 33 locus in a population sample from Upper Silesia (southern Poland)

Praca przedstawia wyniki badań populacyjnych w obrębie locus ACTBP2 (human beta-actinrelatedpseudogene (SE33) w populacji Górnego Śląska. Locus SE33 jest najbardziej polimorficznym ze wszystkich markerów stosowanych dotychczas w genetyce sądowej, zarówno w badaniach dotyczących ustalania sporności ojcostwa, pokrewieństwa i w identyfikacji śladów biologicznych. Badania przeprowadzono w grupie 1315 osobników dorosłych, niespokrewnionych, płci męskiej i żeńskiej. Celem niniejszej pracy było określenie częstości występowania poszczególnych alleli locus SE33 w badanej populacji, ocena zgodności rozkładu alleli z prawem Hardy-Weinberga, obliczenie parametrów oceniających przydatność markera w medycynie sądowej. Zbadanie homogenności rozkładu alleli SE33 pomiędzy badaną populacją a populacjami z innych obszarów Polski.In the paper the authors show the results of the population research on locus SE 33 in a representative sample of 1315 unrelated individuals (males and females) of European origin living in a southern part of Poland (Upper Silesia)