Introduction: This contribution presents the results of a research focused on speech therapy in kindergartens. This research was realized in Zlín Region. It explains how speech therapy prevention is realized in kindergartens, determines the educational qualifications of teachers for this activity and verifies the quality of the applied methodologies in the daily program of kindergartens. Methods: The empirical part of the study was conducted through a qualitative research. For data collection, we used participant observation. We analyzed the research data and presented them verbally, using frequency tables and graphs, which were subsequently interpreted. Results: In this research, 71% of the teachers completed a course of speech therapy prevention, 28% of the teachers received pedagogical training and just 1% of the teachers are clinical speech pathologists. In spite of this, the research data show that, in most of kindergartens, the aim of speech therapy prevention is performed in order to correct deficiencies in speech and voice. The content of speech therapy prevention is implemented in this direction. Discussion: Awareness of the teachers’/parents’ regarding speech therapy prevention in kindergartens. Limitations: This research was implemented in autumn of 2016 in Zlín Region. Research data cannot be generalized to the entire population. We have the ambition to expand this research to other regions next year. Conclusions: Results show that both forms of speech therapy prevention - individual and group - are used. It is also often a combination of both. The aim of the individual forms is, in most cases, to prepare a child for cooperation during voice correction. The research also confirmed that most teachers do not have sufficient education in speech therapy. Most of them completed a course of speech therapy as primary prevention educators. The results also show that teachers spend a lot of time by speech therapy prevention in kindergartens. Educators often develop the communication skills of children by interesting ways and methods.
Ješeta Michal, Chmelíková Eva, Crha Igor, Sedmíková Markéta, Žáková Jana and Ventruba Pavel
Endocrine disruptors (EDs) are chemical substances that affect physiological processes in the organism via hormonal regulation. The EDs are present in the environment and objects of everyday use. They are often detected in food, particularly released from packaging of canned food, but also from plastic water bottles, and they are also found in cosmetics and fertilizers. They are commonly detected in children's toys, banknotes, receipts and many more objects. Permanent and long-term utilization of EDs has harmful effects on human reproductive health mainly by interference with sex hormone synthesis and mechanism of action. The endocrine disruptors show many negative effects on male reproductive system. Any change during synthesis or activity of sex hormones can cause abnormal reproduction, including developmental anomalies of the sexual system, disruption of testicular development or deterioration of sperm quality. Mainly the impact on the development of testicles in prenatal and early postnatal period can be crucial for reproductive health in males. This review provides an overview of the EDs and their possible impact on reproductive health in males with focus on sperm quality and development of testicles.
Michal Ješeta, Eliska Boženková, Jana Žá Žáková, Pavel Ventruba, Igor Crha, Eva Lousová, Petra Coufalová and Bartosz Kempisty
Correct selection of spermatozoa before their using for an assisted reproductive techniques is one of the crucial step in therapy of human infertility. It was previously reported that male factor plays a major role in infertility. Basic semen analyses and standard methods for sperm selection in many cases does not eliminate sufficiently proportion of spermatozoa with genetics defects. Magnetic Activated Cell Sorting (MACS) is a selection method which reduce apoptotic sperm and improve sperm and embryo quality. The aim of our study was the comparison of swim-up method and MACS and their combination. We tested swim-up and MACS alone and treatment of spermatozoa in combination when was is first swim-up and second MACS and vice versa. In this study we evaluated sperm concentration, motility and their DNA integrity before and after separtion methods. On the basis of our results we recommend to use swim-up before MACS method. This approach brings better results in the sperm selection - lower proportion of spermatozoa with fragmented DNA and also it brings better gain of total spermatozoa usable for next IVF or ICSI methods.
Jana Žáková, Igor Crha, Ladislava Jelínková, Tonko Mardešic, Zlatko Pastor, Pavel Trávník, Bartosz Kempisty, Pavel Ventruba and Michal Ješeta
Assisted reproduction is a very dynamic part of reproductive gynaecology with fast changes in therapeutic approaches resulting from intensive research supported by private or public companies. This progress brings also new questions regarding ethical, legal or therapeutic issues. At the 26th Czech-Slovak symposium of assisted reproduction in Brno, several interesting topics of assisted reproduction were presented. During two days, more than 350 specialists discussed current topics related to gynaecology, embryology, andrology, genetics and sexology. From this fruitful discussion we chose the following topics that we consider to be the most important: (1) “Soft” versus “standard” stimulation protocols for IVF (2) Use of exogenous and endogenous progestins in IVF stimulation protocols (3) Cryopreservation and transplantation of ovarian tissue - state of the art and situation in the Czech Republic (4) Selection of spermatozoa with fragmented DNA: practical recommendations (5) Artificial activation of oocytes (8) Access of single and lesbian women to assisted reproduction technologies and possibility of their reproduction in the Czech Republic. Panel conclusions were presented at the end of the conference, which had great attendance, invoked lively commentaries and produced some definitive consensus. Certain issues remained inconclusive and these matters will be the subject of further discussion in future. This is a summary of the most important theses from this field in the Czech Republic in 2017.
Katarzyna Stefańska, Małgorzata Józkowiak, Paweł Antosik, Dorota Bukowska, Piotr Celichowski, Małgorzata Bruska, Michał Nowicki, Bartosz Kempisty, Jana Zakova, Marie Machatkova and Michal Jeseta
Correct maturation of the oocyte is crucial for further fertilization and embryogenesis. It comprises of both nuclear and cytoplasmic maturation, during which the proteins, nutrients and mRNAs are assembled. Cumulus cells are connected with the oocyte via gap-junctions, which enable bi-directional transfer of molecules, forming cumulus-oocyte complex (COC). The expression pattern in CCs is thought to resemble the genes expressed in the oocyte. The CCs surrounding the gamete of high developmental competence have an increased expression of apoptotic markers. Therefore, our aim in this study was to determine whether any apoptosis-related genes are upregulated in porcine oocytes before or after IVM. We isolated COCs from 45 pubertal crossbred gilts, performed brilliant cresyl blue (BCB) staining and analyzed the gene expression pattern in oocytes before and after IVM with the use of microarray analysis. The results include 419 differentially expressed transcripts, 25 of which belong to „regulation of apoptosis” and „regulation of cell death” GO BP terms. This set of genes includes BCLAF1, EIF2AK3, KLF10, MIF, MAP3K1, NOTCH2, TXNIP and APP, all of which have been upregulated in immature porcine oocytes. Our results suggest that they play part in porcine oocyte maturation and could be used as potential markers of female gamete’s developmental competence. This knowledge could serve as a basis to improve ART in pigs.
Agata Chamier-Gliszczyńska, Sandra Kałużna, Katarzyna Stefańska, Piotr Celichowski, Paweł Antosik, Dorota Bukowska, Małgorzata Bruska, Jana Zakova, Marie Machatkova, Michal Jeseta and Michał Nowicki
The formation of mammalian oocytes begins in the ovary during fetal development. The proper development of oocytes requires close communication with surrounding somatic cells, the substances they emit allow proper maturation of oocytes. Somatic cumulus (CC) cells and oocytes form cumulus-oocyte (COC) complexes.
In this study, the Affymetrix microarray analysis was used to investigate changes in gene expression occurring in oocytes before and after in vitro maturation (IVM). The aim of the study was to examine oocyte genes involved in two ontological groups, “regulation of cell migration” and “regulation of cell proliferation” discovered by the microarray method.
We found a reduced expression of all 28 genes tested in the ontological groups: ID2, VEGFA, BTG2, CCND2, EDNRA, TGFBR3, GJA, LAMA2, RTN4, CDK6, IHH, MAGED1, INSR, CD9, PTGES, TXNIP, ITGB1, SMAD4, MAP3K1, NOTCH2 , IGFBP7, KLF10, KIT, TPM1, PLD1, BTG3, CD47 and MITF. We chose the most regulated genes down the IVM culture, and pointed out those belonging to two ontological groups.
Increased expression of the described genes before IVM maturation may indicate the important role of these genes in the process of ovum maturation. After the maturation process, the proteins produced by them did not play such an important role. In summary, the study provides us with many genes that can serve as molecular markers of oocyte processes associated with in vitro maturation. This knowledge can be used for detailed studies on the regulation of oocyte maturation processes.
Running title: Genes regulating cellular migration and proliferation in porcine oocytes
Katarzyna Stefańska, Agata Chamier-Gliszczyńska, Maurycy Jankowski, Piotr Celichowski, Magdalena Kulus, Magdalena Rojewska, Paweł Antosik, Dorota Bukowska, Małgorzata Bruska, Michał Nowicki, Bartosz Kempisty, Michal Jeseta and Jana Zakova
The correct oviductal development and morphogenesis of its epithelium are crucial factors influencing female fertility. Oviduct is involved in maintaining an optimal environment for gametes and preimplantation embryo development; secretory oviductal epithelial cells (OECs) synthesize components of oviductal fluid. Oviductal epithelium also participates in sperm binding and its hyperactivation. For better understanding of the genetic bases that underlay porcine oviductal development, OECs were isolated from porcine oviducts and established long-term primary culture. A microarray approach was utilized to determine the differentially expressed genes during specific time periods. Cells were harvested on day 7, 15 and 30 of in vitro primary culture and their RNA was isolated. Gene expression was analyzed and statistical analysis was performed. 48 differentially expressed genes belonging to “tube morphogenesis”, “tube development”, “morphogenesis of an epithelium”, “morphogenesis of branching structure” and “morphogenesis of branching epithelium” GO BP terms were selected, of which 10 most upregulated include BMP4, ARG1, SLIT2, FGFR1, DAB2, TNC, EPAS1, HHEX, ITGB3 and LOX. The results help to shed light on the porcine oviductal development and its epithelial morphogenesis, and show that after long-term culture the OECs still proliferate and maintain their tube forming properties.