Exposure to air pollution and ovarian reserve parameters.

Exposure to air pollution is associated with many different health effects, especially cardiovascular and respiratory diseases. Additionally, highly significant links between exposure to air pollution and fertility, particularly male fertility was observed, however the studies regarding exposure to selected air pollutants and female fertility assessed by ovarian reserve are rare. Hence, the main aim of the study was to analyze relationship between exposure to ambient air pollution and ovarian reserve parameters among Polish women. The study population consisted of 511 women, who attended to infertility clinic because of diagnostic purposes. Participants filled in the questionnaire about social-demographic, lifestyle and health factors. Infertility specialists assessed ovarian parameters such as: antral follicle count (AFC) and concentration of hormones: Anti-Müllerian hormone (AMH), follicle stimulating hormone (FSH) and estradiol (E2). The air pollutants level (sulfur dioxide, nitrogen dioxide, carbon monoxide, ozone, particulate matters) were obtained via National Environmental Protection Inspectorate database. Significant negative association between PM2,5 and AHM (p = 0.032) as well as AFC (p = 0.044) was observed. Moreover, SO2 concentrations decrease AFC (p = 0.038). The results also suggest that PM10, PM2.5, SO2 exposure on antral follicle count may be more pronounced among women with a female factor infertility diagnosis. Additionally, exposure to PM2.5 and NOx on AFC and AMH was stronger among older women (> 35 years of age). To conclude, the present study found that air pollution could lead to decrease in follicle antral count and Anti-Müllerian hormone level, especially exposure to PM2,5 and SO2 thus the evidence suggest negative impact to ovarian reserve.

Sylvian fissure lipoma associated with fusiform aneurysm in the middle cerebral artery trifurcation: A case report and literature review.

Background: The intracranial lipomas are rare congenital malformations accounting for approximately 0.1-1.3% of all intracranial tumors, of which Sylvian fissure lipomas account for <5%. These lesions are frequently associated with dysgenesis of neuronal brain tissues and vascular malformations and in the majority are asymptomatic. Intracranial lipomas on magnetic resonance imaging (MRI) may mimic late subacute hemorrhage due to similar radiological features. Due to the tight adhesion of the lipoma to the surrounding nerve structures and vessels, complete removal is difficult and does not guarantee the disappearance of symptoms. Case Description: We present the case of a 42-year-old woman with chronic headaches and short-term memory impairment who was admitted to the emergency room after an out-of-hospital brain MRI with suspected ruptured right middle cerebral artery (MCA) aneurysm and late subacute intracranial hemorrhage. In the hospital, after clinical evaluation, emergency computed tomography (CT) angiography was performed, which revealed an unruptured fusiform aneurysm located in the right MCA trifurcation surrounded by an extremely hypodense lesion corresponding to fat in the right Sylvian fissure. No features of intracranial hemorrhage were present. The diagnosis of intracranial lipoma was finally confirmed after the MRI of the brain with a fat suppression sequence. Surgical treatment was not attempted, and the patient was treated conservatively with a satisfactory general outcome. Conclusion: A Sylvian fissure lipoma may be associated with a fusiform aneurysm in the MCA trifurcation. By modifying the standard MRI protocol and performing a CT scan, an intracranial lipoma can be detected and a late subacute intracranial hemorrhage can be excluded.

Comparison of SOX2 and POU5F1 Gene Expression in Leukapheresis-Derived CD34+ Cells before and during Cell Culture.

Bone marrow is an abundant source of both hematopoietic as well as non-hematopoietic stem cells. Embryonic, fetal and stem cells located in tissues (adipose tissue, skin, myocardium and dental pulp) express core transcription factors, including the SOX2, POU5F1 and NANOG gene responsible for regeneration, proliferation and differentiation into daughter cells. The aim of the study was to examine the expression of SOX2 and POU5F1 genes in CD34-positive peripheral blood stem cells (CD34+ PBSCs) and to analyze the influence of cell culture on the expression of SOX2 and POU5F1 genes. The study material consisted of bone marrow-derived stem cells isolated by using leukapheresis from 40 hematooncology patients. Cells obtained in this process were subject to cytometric analysis to determine the content of CD34+ cells. CD34-positive cell separation was conducted using MACS separation. Cell cultures were set, and RNA was isolated. Real-time PCR was conducted in order to evaluate the expression of SOX2 and POU5F1 genes and the obtained data were subject to statistical analysis. We identified the expression of SOX2 and POU5F1 genes in the examined cells and demonstrated a statistically significant (p < 0.05) change in their expression in cell cultures. Short-term cell cultures (<6 days) were associated with an increase in the expression of SOX2 and POU5F1 genes. Thus, short-term cultivation of transplanted stem cells could be used to induce pluripotency, leading to better therapeutic effects.

The silencing of ets-4 mRNA relies on the functional cooperation between REGE-1/Regnase-1 and RLE-1/Roquin-1.

Regnase-1 is an evolutionarily conserved endoribonuclease. It degrades diverse mRNAs important for many biological processes including immune homeostasis, development and cancer. There are two competing models of Regnase-1-mediated mRNA silencing. One model postulates that Regnase-1 works together with another RNA-binding protein, Roquin-1, which recruits Regnase-1 to specific mRNAs. The other model proposes that the two proteins function separately. Studying REGE-1, the Caenorhabditis elegans ortholog of Regnase-1, we have uncovered its functional relationship with RLE-1, the nematode counterpart of Roquin-1. While both proteins are essential for mRNA silencing, REGE-1 and RLE-1 appear to associate with target mRNA independently of each other. Thus, although the functional interdependence between REGE-1/Regnase-1 and RLE-1/Roquin-1 is conserved, the underlying mechanisms may display species-specific variation, providing a rare perspective on the evolution of this important post-transcriptional regulatory mechanism.

RNA Binding Properties of the Ty1 LTR-Retrotransposon Gag Protein.

A universal feature of retroelement propagation is the formation of distinct nucleoprotein complexes mediated by the Gag capsid protein. The Ty1 retrotransposon Gag protein from Saccharomyces cerevisiae lacks sequence homology with retroviral Gag, but is functionally related. In addition to capsid assembly functions, Ty1 Gag promotes Ty1 RNA dimerization and cyclization and initiation of reverse transcription. Direct interactions between Gag and retrotransposon genomic RNA (gRNA) are needed for Ty1 replication, and mutations in the RNA-binding domain disrupt nucleation of retrosomes and assembly of functional virus-like particles (VLPs). Unlike retroviral Gag, the specificity of Ty1 Gag-RNA interactions remain poorly understood. Here we use microscale thermophoresis (MST) and electrophoretic mobility shift assays (EMSA) to analyze interactions of immature and mature Ty1 Gag with RNAs. The salt-dependent experiments showed that Ty1 Gag binds with high and similar affinity to different RNAs. However, we observed a preferential interaction between Ty1 Gag and Ty1 RNA containing a packaging signal (Psi) in RNA competition analyses. We also uncover a relationship between Ty1 RNA structure and Gag binding involving the pseudoknot present on Ty1 gRNA. In all likelihood, the differences in Gag binding affinity detected in vitro only partially explain selective Ty1 RNA packaging into VLPs in vivo.

Decontamination of microbiologically contaminated abiotic porous surfaces in an oral surgery clinic using vaporised hydrogen peroxide (VHP).

PURPOSE: The aims of the study were to identify microorganisms, including those in the VBNC state, inhabiting porous surfaces in oral surgery offices and to assess the biocidal effectiveness and impact of 300 ppm vaporised hydrogen peroxide (VHP) for 20 min on decontaminated materials. METHODS: From the surfaces of textured armrests of dental chairs, pinewood doors and window frames and cotton medical aprons, 30 swabs were taken with moistened sponges. The identification of isolated microorganisms was performed using molecular methods with MALDI-TOF MS, DNA Sanger sequencer and Illumina MiSeq. To evaluate the impact of VHP decontamination (independent variable) on the number of microorganisms (response variable) ANOVA and LSD tests were used. After application of 10 processes of VHP decontamination, changes in the properties of the materials were assessed using FTIR spectroscopy, SEM microscopy and XPS spectrometry. RESULTS: The concentration of microorganisms was 101-104 CFU/100 cm2 on the tested surfaces and 102 CFU/m3 in the air. Twenty species of bacteria, one yeast and 16 filamentous fungi were identified, with the predominance of Bacillus, Staphylococcus, Alternaria, Aspergillus and Penicillium. Moreover, Janthinobacterium, Acremonium, Aureobasidium, Coprinellus and Cosmospora in the VBNC state were metagenomically detected. VHP decontamination resulted in a reduction in the majority of tested microbial strains by a minimum of 3 log, and all tested mixed cultures inhabiting porous surfaces were above 98% and in the air, 100%. VHP decontamination did not affect the structural and morphological properties of cotton fibres, wood or stainless steel. CONCLUSIONS: VHP decontamination at a concentration of 300 ppm for 20 min can be used for the holistic disinfection of air, surfaces and equipment in oral surgery offices.

For good and for bad: the role of endogenous retroelements in humans / Na dobre i na zle: rola endogennych retroelementów u czlowieka.

Transposable elements (TEs) are the sequences that are able to "jump" across the genome. They are found in virtually all organisms including human. Although in human, the majority of TEs lost their ability to autonomous transposition, they make up almost half of our genome, and played important roles in genome evolution. Fast progress in deep sequencing and functional analysis has revealed the importance of domes­ticated copies of transposable elements, including their regulatory sequences, transcripts and proteins in normal cells functioning. However, a growing numer of evidence suggest the involvment of TEs in development and progression of autoimmune and neurodegenerative disaeses as well as in many types of cancer. In this review we summarize the current state of knowledge about the LTR retroelements: endogenous retroviruses (ERVs) and Ty3/Gypsy retrotransposons, and their role in human organism.

Retroviral-like determinants and functions required for dimerization of Ty1 retrotransposon RNA.

During replication of long terminal repeat (LTR)-retrotransposons, their proteins and genome (g) RNA assemble into virus-like particles (VLPs) that are not infectious but functionally related to retroviral virions. Both virions and VLPs contain gRNA in a dimeric form, but contrary to retroviruses, little is known about how gRNA dimerization and packaging occurs in LTR-retrotransposons. The LTR-retrotransposon Ty1 from Saccharomyces cerevisiae is an informative model for studying LTR-retrotransposon and retrovirus replication. Using structural, mutational and functional analyses, we explored dimerization of Ty1 genomic RNA. We provide direct evidence that interactions of self-complementary PAL1 and PAL2 palindromic sequences localized within the 5'UTR are essential for Ty1 gRNA dimer formation. Mutations disrupting PAL1-PAL2 complementarity restricted RNA dimerization in vitro and Ty1 mobility in vivo. Although dimer formation and mobility of these mutants was inhibited, our work suggests that Ty1 RNA can dimerize via alternative contact points. In contrast to previous studies, we cannot confirm a role for PAL3, tRNAiMet as well as recently proposed initial kissing-loop interactions in dimer formation. Our data also supports the critical role of Ty1 Gag in RNA dimerization. Mature Ty1 Gag binds in the proximity of sequences involved in RNA dimerization and tRNAiMet annealing, but the 5' pseudoknot in Ty1 RNA may constitute a preferred Gag-binding site. Taken together, these results expand our understanding of genome dimerization and packaging strategies utilized by LTR-retroelements.

Filamin A upregulation correlates with Snail-induced epithelial to mesenchymal transition (EMT) and cell adhesion but its inhibition increases the migration of colon adenocarcinoma HT29 cells.

Filamin A (FLNA) is actin filament cross-linking protein involved in cancer progression. Its importance in regulating cell motility is directly related to the epithelial to mesenchymal transition (EMT) of tumor cells. However, little is known about the mechanism of action of FLNA at this early stage of cancer invasion. Using immunochemical methods, we evaluated the levels and localization of FLNA, pFLNA[Ser2152], ß1 integrin, pß1 integrin[Thr788/9], FAK, pFAK[Y379], and talin in stably transfected HT29 adenocarcinoma cells overexpressing Snail and looked for the effect of Snail in adhesion and migration assays on fibronectin-coated surfaces before and after FLNA silencing. Our findings indicate that FLNA upregulation correlates with Snail-induced EMT in colorectal carcinoma. FLNA localizes in the cytoplasm and at the sites of focal adhesion (FA) of invasive cells. Silencing of FLNA inhibits Snail-induced cell adhesion, reduces the size of FA sites, induces the relocalization of talin from the cytoplasm to the membrane area and augments cell migratory properties. Our findings suggest that FLNA may not act as a classic integrin inhibitor in invasive carcinoma cells, but is involved in other pro-invasive pathways. FLNA upregulation, which correlates with cell metastatic properties, maybe an additional target for combination therapy in colorectal carcinoma tumor progression.

[Modern methods in analysis of microtubules dynamic in vivo]. / Nowoczesne metody przyzyciowej analizy polimeryzacji mikrotubul.

Microtubules are involved in any vital cellular activities, including the maintenance of cell shape, division, migration and intracellular transport. Microtubule dynamics is regulated by the balance between their polymerization and depolymerization. Microtubule stability is dependent on their alpha and beta subunits composition, tubulin post-translational modifications and interaction of microtubules with microtubule-associated proteins (MAPs). Disruption of these processes can lead to a number of pathological conditions such as cancer, cardiovascular disease, or the fibrosis development. This review summarizes the current knowledge of the modern methods of microtubule polymerization analysis. This allows a better understanding of the structure and mechanisms played by microtubules in their physiological functions and the development of pathological conditions resulting from their disorder.

RNA-Puzzles Round III: 3D RNA structure prediction of five riboswitches and one ribozyme.

RNA-Puzzles is a collective experiment in blind 3D RNA structure prediction. We report here a third round of RNA-Puzzles. Five puzzles, 4, 8, 12, 13, 14, all structures of riboswitch aptamers and puzzle 7, a ribozyme structure, are included in this round of the experiment. The riboswitch structures include biological binding sites for small molecules (S-adenosyl methionine, cyclic diadenosine monophosphate, 5-amino 4-imidazole carboxamide riboside 5'-triphosphate, glutamine) and proteins (YbxF), and one set describes large conformational changes between ligand-free and ligand-bound states. The Varkud satellite ribozyme is the most recently solved structure of a known large ribozyme. All puzzles have established biological functions and require structural understanding to appreciate their molecular mechanisms. Through the use of fast-track experimental data, including multidimensional chemical mapping, and accurate prediction of RNA secondary structure, a large portion of the contacts in 3D have been predicted correctly leading to similar topologies for the top ranking predictions. Template-based and homology-derived predictions could predict structures to particularly high accuracies. However, achieving biological insights from de novo prediction of RNA 3D structures still depends on the size and complexity of the RNA. Blind computational predictions of RNA structures already appear to provide useful structural information in many cases. Similar to the previous RNA-Puzzles Round II experiment, the prediction of non-Watson-Crick interactions and the observed high atomic clash scores reveal a notable need for an algorithm of improvement. All prediction models and assessment results are available at http://ahsoka.u-strasbg.fr/rnapuzzles/.

ß-III tubulin modulates the behavior of Snail overexpressed during the epithelial-to-mesenchymal transition in colon cancer cells.

Class III ß-tubulin (TUBB3) is a marker of drug resistance expressed in a variety of solid tumors. Originally, it was described as an important element of chemoresistance to taxanes. Recent studies have revealed that TUBB3 is also involved in an adaptive response to a microenvironmental stressor, e.g. low oxygen levels and poor nutrient supply in some solid tumors, independently of the microtubule targeting agent. Furthermore, it has been demonstrated that TUBB3 is a marker of biological aggressiveness associated with modulation of metastatic abilities in colon cancer. The epithelial-to-mesenchymal transition (EMT) is a basic cellular process by which epithelial cells lose their epithelial behavior and become invasive cells involved in cancer metastasis. Snail is a zinc-finger transcription factor which is able to induce EMT through the repression of E-cadherin expression. In the presented studies we focused on the analysis of the TUBB3 role in EMT-induced colon adenocarcinoma cell lines HT-29 and LS180. We observed a positive correlation between Snail presence and TUBB3 upregulation in tested adenocarcinoma cell lines. The cellular and behavioral analysis revealed for the first time that elevated TUBB3 level is functionally linked to increased cell migration and invasive capability of EMT induced cells. Additionally, the post-transcriptional modifications (phosphorylation, glycosylation) appear to regulate the cellular localization of TUBB3 and its phosphorylation, observed in cytoskeleton, is probably involved in cell motility modulation.

The matrix domain contributes to the nucleic acid chaperone activity of HIV-2 Gag.

BACKGROUND: The Gag polyprotein is a multifunctional regulator of retroviral replication and major structural component of immature virions. The nucleic acid chaperone (NAC) activity is considered necessary to retroviral Gag functions, but so far, NAC activity has only been confirmed for HIV-1 and RSV Gag polyproteins. The nucleocapsid (NC) domain of Gag is proposed to be crucial for interactions with nucleic acids and NAC activity. The major function of matrix (MA) domain is targeting and binding of Gag to the plasma membrane but MA can also interact with RNA and influence NAC activity of Gag. Here, we characterize RNA binding properties and NAC activity of HIV-2 MA and Gag, lacking p6 domain (GagΔp6) and discuss potential contribution of NC and MA domains to HIV-2 GagΔp6 functions and interactions with RNA. RESULTS: We found that HIV-2 GagΔp6 is a robust nucleic acid chaperone. HIV-2 MA protein promotes nucleic acids aggregation and tRNA(Lys3) annealing in vitro. The NAC activity of HIV-2 NC is affected by salt which is in contrast to HIV-2 GagΔp6 and MA. At a physiological NaCl concentration the tRNA(Lys3) annealing activity of HIV-2 GagΔp6 or MA is higher than HIV-2 NC. The HIV-2 NC and GagΔp6 show strong binding to the packaging signal (Ψ) of HIV-2 RNA and preference for the purine-rich sequences, while MA protein binds mainly to G residues without favouring Ψ RNA. Moreover, HIV-2 GagΔp6 and NC promote HIV-2 RNA dimerization while our data do not support MA domain participation in this process in vitro. CONCLUSIONS: We present that contrary to HIV-1 MA, HIV-2 MA displays NAC activity and we propose that MA domain may enhance the activity of HIV-2 GagΔp6. The role of the MA domain in the NAC activity of Gag may differ significantly between HIV-1 and HIV-2. The HIV-2 NC and MA interactions with RNA are not equivalent. Even though both NC and MA can facilitate tRNA(Lys3) annealing, MA does not participate in RNA dimerization in vitro. Our data on HIV-2 indicate that the role of the MA domain in the NAC activity of Gag differs not only between, but also within, retroviral genera.

[MRTFs protein family and its role in pathological stages]. / Bialka rodziny MRTF i ich udzial w patogenezie.

The MRTFs (myocardin-releated transcription factors) protein family consists of myocardin, MRTF-A (MKL1, MAL) and MRTF-B (MKL2). These proteins are included in the common family due to the presence of evolutionarily conserved domains which are responsible for homo- and heterodimerization with other members of the family as well as actin binding and transcription activation. Despite high structural homology, these factors present different characteristics in terms of localization. The expression of myocardin is limited to the myocardial cells and smooth muscle cells, exclusively, whereas MRTF-A and MRTF-B are commonly found in various cells and tissues. These proteins interact with MADS box transcription factors as well as serum response factors (SRF) thus being engaged into signal transduction, which results from cytoskeleton reorganisation, from cytoplasm to the nucleus. It has been concluded that these proteins both take part in muscle tissue differentiation as well as mediate the development of pathological conditions (vascular diseases, cancer and fibrosis).

Ty1 retrovirus-like element Gag contains overlapping restriction factor and nucleic acid chaperone functions.

Ty1 Gag comprises the capsid of virus-like particles and provides nucleic acid chaperone (NAC) functions during retrotransposition in budding yeast. A subgenomic Ty1 mRNA encodes a truncated Gag protein (p22) that is cleaved by Ty1 protease to form p18. p22/p18 strongly inhibits transposition and can be considered an element-encoded restriction factor. Here, we show that only p22 and its short derivatives restrict Ty1 mobility whereas other regions of GAG inhibit mobility weakly if at all. Mutational analyses suggest that p22/p18 is synthesized from either of two closely spaced AUG codons. Interestingly, AUG1p18 and AUG2p18 proteins display different properties, even though both contain a region crucial for RNA binding and NAC activity. AUG1p18 shows highly reduced NAC activity but specific binding to Ty1 RNA, whereas AUG2p18 shows the converse behavior. p22/p18 affects RNA encapsidation and a mutant derivative defective for RNA binding inhibits the RNA chaperone activity of the C-terminal region (CTR) of Gag-p45. Moreover, affinity pulldowns show that p18 and the CTR interact. These results support the idea that one aspect of Ty1 restriction involves inhibition of Gag-p45 NAC functions by p22/p18-Gag interactions.

[Filamin A as a mediator of alterations in cancer cells]. / Filamina A mediatorem zmian zachodzacych w komórkach nowotworowych.

Filamin A (FLNA, filamin-1) is a homodimeric protein, commonly expressed in animal organisms. Its basic function in the cell is actin crosslinking and forming 3D cytoskeleton structure. Filamin-1 interacts with more than 60 different proteins with various functions such as: cell membrane and cytoskeleton formation, maintaining cell shape, intracellular signaling, nuclear functions or GTP-binding proteins regulation. FLNA interactions with oncogenesis- and metastasis-related proteins, such as K-RAS, TRAF2 or NIK indicate its crucial role in cancer progression. Filamin-1 undergoes proteolytic fragmentation producing products, translocation of which to the nucleus may be related to alterations in the cell metastatic ability. It was also demonstrated that FLNA dysfunctions can lead to sensitization of cells to ionizing irradiation or common chemotherapeutics: bleomycin and cisplatin. These findings indicate that FLNA can be considered as a novel target in anti-cancer therapy.

[Cancer stem cells]. / Nowotworowe komórki macierzyste.

Cancer stem cell theory gains increasingly greater significance in the world of medicine. Numerous findings of scientific research in vivo and in vitro indicate that it is the population of undifferentiated, self-renewing cells which is responsible for recurrence of cancer and metastasis. Similarly to normal stem cells, cancer stem cells (CSC) function in the environment of the other cells of the organism, called the niche, where they receive signals for differentiation and proliferation processes. Disorders in the signaling pathways between CSC and the niche that result from e.g. acquired oncogenic mutations may lead to uncontrolled proliferation of stem cells, gaining independence from the primary niche or settling a new microenvironment. CSC are identified on the basis of specific markers - membrane proteins or cell enzymes. Methods based on the measurement of dye fluorescence (obtaining side population, SP) or fluorescence of the fluorophore conjugated with a monoclonal antibody directed against the specific CSC marker are used for isolation. A different method obtains morphologically miscellaneous clones by single cell cloning: holo-, mero- and paraclones. Tumor forming assay in NOD/SCID mice is a standard in vivo test that confirms the stem character of isolated cells. However, this model may not fully reflect the complexity of cancer illnesses in human beings. Solving the mystery of oncogenesis, including the existence of cancer stem cells, is undoubtedly one of the priorities of contemporary medicine that should contribute to the improvement of cancer therapy.