Blanka Borowiec, Tomasz Hadada and Magdalena Kosińska
For a very long time, scientists have been interested in preterm births, which are an extremely interesting object of research, and are worth investigating in context of a vast number of new clinical applications. A number of factors may contribute to the early termination of pregnancy, including groups related to the conditions of mother's organism. In this work, it was examined whether environmental, maternal and fetal factors could affect normal time of birth. Variables that were considered to influence the length of prenatal development were categorized as biological, describing the state of mother's health and pregnancy, and those describing the socio-economic status of parents. The research material was subjected to exploratory analysis. For categorized variables, numerical tables were constructed, and for quantitative variables the basic measures of position and variability were presented (mean, minimum, maximum, standard deviation, median, quartile distribution). To analyze the differences in the frequency distributions of the examined variables, the χ2 tests were used. To estimate the duration of the pregnancy of the tested mothers, the survival analysis (the product limit method - Kaplan-Meier method) was applied. The analysis showed a statistically significant effect on the duration of pregnancy and the nature of its course, indicating a shortening of pregnancy in the presence of risk factors (χ2=196,23; df = 4; p<0,001). The course of the curves shows an increase in the frequency of premature delivery in the case of clinically diagnosed risk factors. This will certainly provide a strong basis for improving the care of premature newborns and minimizing the risk for the mother.
Blanka Borowiec, Tomasz Hadada and Magdalena Kosińska
This study focused on the role of polymorphisms in prostaglandin expression regulating genes in the occurrence of early uterine contractions during pregnancy. The analyzed genes were: PLA2G4C, encoding calcium independent phospholipase A2, and PLA2G4C, encoding IVD phospholipase A2. It was examined if known reference polymorphisms in these genes (rs1366442, [A/C/T] for PLA2G4C; and rs4924618, [A/T] for PLA2G4D), have any influence on preterm birth. Additionally, other biological, genetic and socio-economic factors were taken into account and analyzed, based on their role in induction of early resolve of pregnancy in the study group. Blood samples were taken from 20 patients. 15 of them gave birth preterm, 5 gave birth at the predicted date of pregnancy resolve. DNA was isolated from the samples, and subjected to PCR, with obtained amplified samples separated using electrophoresis on 1,5% agarose gel. Resulting material was subjected to high-throughput sequencing. Statistical analysis was performed using Statistica 13 software. Analyses have shown that the discussed biological, genetic and societal-economic factors have statistically significant influence on preterm birth. Sequencing results presented the suspected presence of the analyzed SNPs in most women from the studied groups, while not showing their presence in any of the controls. The biological, genetic, and socio-economic factors analyzed have a significant influence on pre-term birth. Presence of SNPs in PLA2G4C and PLA2G4D genes may increase the risk of early resolve of pregnancy. However, as the control and study groups were relatively small, it is suggested to repeat the studies on bigger samples to validate the results.
Małgorzata Popis, Blanka Borowiec and Maurycy Jankowski
The common fruit fly, or Drosophila Melanogaster, has been used as an object of biomedicals studies for over a century. It has been mostly employed in genetic research, as it exhibits several advantages which make its use relatively easy and cheap, with the results widely translatable into further vertebrate studies. This model been the basis of the work of Christiane Nusslein-Volhard, who together with Eric Wieschaus unravelled much of the mystery surrounding early drosophila development in the 1970s-1980s, laying foundations for broader understanding of multicellular organism embryogenesis, which brought them a Nobel prize in Physiology and Medicine in 1995. The knowledge gained from drosophila studies improves the basic understanding of developmental processes, while the model itself is relatively easy to maintain, analyse and translate the results onto other species. While models such as Zebrafish present better with other vertebrates, drosophila remains a very important element of genetic research, finding even more applications with the development of current science and medicine. Hence, in this short review, the outline of the history, breakthroughs and perspectives of the drosophila research has been presented.
Blanka Borowiec, Małgorzata Popis and Maurycy Jankowski
A large amount of complex hormone associated processes occurring continuously in the human organism is necessary to maintain homeostasis in response to various internal and external conditions. In the same time, as the hormones use the bloodstream as their transmission medium, it is essential that their expression is strictly controlled to maintain their activity only when it is required. Because of that, the endocrine system evolved complex, self-regulating machinery that allows for precise signalling to the glands to initiate hormone expression, as well as equally quick negative feedback in the moment of reaching the optimal blood hormone concentration. The pituitary gland serves as the true endocrine part of that system, expressing a range of hormones that mostly serve as regulators of sub-systems serving different functions, scattered around organisms. The hypothalamus is the neuroendocrine part of the hypothalamic-pituitary axis, meaning it integrates the neuronal and hormonal signals, effectively linking the nervous and endocrine systems. The processes of hypothalamus and pituitary development share some significant similarities, which is unsurprising considering their close association and anatomical proximity at the base of the brain. Arising in highly overlapping developmental timeframes, they are both initially patterned by the gradients of extrinsic signalling molecules. After the initial lineage commitment, in both of those structures, intrinsic factors expressed by the distinct cell populations sustain the morphogenesis to result in a final complexly patterned structure. In this short review, the processes of the pituitary and hypothalamus development are described, with the most important factors driving them discussed.
Magdalena Kulus, Jakub Kulus, Maurycy Jankowski, Blanka Borowiec, Michal Jeseta, Dorota Bukowska, Klaus P. Brüssow, Bartosz Kempisty and Paweł Antosik
Constant advances in medicine, both human and veterinary, lead to continuous discovery of new drugs and treatments. Recently, the aspect of stem cell use in regenerative medicine has been very popular. There are still too few clinical trials on animals that could precisely estimate the therapeutic efficacy of cell therapy. However, stem cells are a source of extraordinary potential for multiplication and differentiation which, if used properly, can prove to be an effective mean of treatment of numerous diseases that are currently considered untreatable. The purpose of review is the characterization and clinical use of stem cells in mostly occurring diseases. Particular attention has been given to the issue of mesenchymal stromal cells, which so far have been most widely used in clinical practice. Current research into stem cells has allowed scientists to discover many different types of these cells, describe their characteristics and divide them into groups, with the most important being embryonic stem cells and somatic (adult) stem cells. Adult stem cells, due to their availability and lack of ethical problems, are used in veterinary practice. Different types of mesenchymal stem cells are distinguished, based on their origin. Adipose tissue derived stem cells and stromal vascular fraction find the widest clinical application. In veterinary medicine, stem cells therapies are most commonly used in the case of horse orthopedic injuries and in diseases of various origin in dogs and cats. While further research is needed to confirm the effectiveness of cell therapies, they have much potential to find plenty of potential applications in future medicine.
Marta Rybska, Sandra Knap, Maurycy Jankowski, Blanka Borowiec, Michal Jeseta, Dorota Bukowska, Paweł Antosik, Michał Nowicki, Maciej Zabel, Bartosz Kempisty and Jędrzej M. Jaśkowski
Ovarian cysts remain to be one of the most common and serious problems in reproduction of farm animals, as well as humans. Apart from causing the fall in reproductive potential of the ovaries, occupying the place in which folliculogenesis and oogenesis occur, they also cause hormone imbalances, by preventing corpus luteum formation, hence lowering the amount of steroid hormone production. While singular cysts rarely affect fertility, hormone fluctuations that are associated with their presence promotes their multiplication, which usually has more adverse effects. While the cysts are easily detectable in humans, possessing distinct echography while examined by ultrasound, multiple factors prevent widespread use of effective detection methods among large herds of farm animals. Because of lack of noticeable symptoms of early stages of such malignancies, they rarely get detected before the animal stops to exhibit symptoms of heat. That causes scientific research to be focused on not only methods of detection, but also the ways to negate the effects of ovarian cysts and bring the affected specimen back to reproductive potential. Despite that, high costs of diagnosis and treatment, cause them to be uncommon on commercial farms. As lack of fertility eliminates animals from breeding purposed herds, ovarian cysts persist as a cause of large losses of the animal husbandry business. Continuous research, focused on natural examples of ovarian cysts should be conducted, in order to improve methods of detection, prevention, treatment and recovery from the effects of ovarian cysts.
Artur Bryja, Marta Dyszkiewicz-Konwińska, Maurycy Jankowski, Piotr Celichowski, Katarzyna Stefańska, Agata Chamier-Gliszczyńska, Blanka Borowiec, Katarzyna Mehr, Dorota Bukowska, Paweł Antosik, Małgorzata Bruska, Maciej Zabel, Michał Nowicki and Bartosz Kempisty
The mucous membrane is composed of two layers. The layer of stratified squamous epithelium and the underlying layer of the connective tissue. The epithelium is composed of keratinocytes that are in different stages of differentiation, depending on their localization. In our research, after isolation of primary in vitro cultured buccal pouch mucosal cells, we observed keratinocytes in various stages of differentiation and fibroblasts. These cells, depending on the ionic dynamics, may be subject to different morphological and biochemical transformations. Understanding the expression profile of the normal oral mucosal tissue is important for further research into the effects of biomaterials on the mucosal cells, their growth, proliferation, and differentiation.
The porcine buccal pouch mucosal cells were used in this study. The oral mucosa was separated surgically and isolated enzymatically. The cells were in vitro cultured for 30 days, and after each step of in vitro culture (7 days, 15 days, 30 days), samples were collected for isolation of total RNA. The gene expression profile was measured using Affymetrix microarray assays.
In results, we observed genes belonging to two ontology groups: cation homeostasis and cation transport. These genes were up-regulated after 7 days of in vitro culture as compared to down-regulation after 15 and 30 days of in vitro culture. These results suggested that dynamic growth, proliferation and cell adhesion are more intense in the first 7 days of in vitro culture. We also observed, for the first time, the expression of ATP13A3 in porcine oral mucosal cells.
Edyta Porowska, Magdalena Kulus, Maurycy Jankowski, Ievgenia Kocherova, Michal Jeseta, Agata Chamier-Gliszczyńska, Katarzyna Stefańska, Blanka Borowiec, Dorota Bukowska, Klaus P. Brüssow, Bartosz Kempisty and Paweł Antosik
Pyometra is the most common gynecological disease in female dogs. It usually occurs in middle age female dogs, usually about two months after the completion of heat. This disease is the accumulation of purulent fluid inside the uterus. Etiology of pyometra is not fully understood. It is assumed, that pyometra is a result of hormonal disorders in the endometrium combined with bacterial superinfection. The diagnosis is based on the interview, clinical examination, additional laboratory tests and ultrasound or x-ray of the abdomen. There are two treatments: ovariohysterectomy and conservative treatment with pharmacological agents for example prostaglandin, aglepriston, antibiotics with a broad spectrum of action. Currently conducted molecular studies have a large influence on the development of the present knowledge on the pathogenesis and course of pyometra, whose conclusions may be used to change the current therapeutic protocols.
Magdalena Kulus, Małgorzata Józkowiak, Jakub Kulus, Małgorzata Popis, Blanka Borowiec, Katarzyna Stefańska, Piotr Celichowski, Mariusz J. Nawrocki, Dorota Bukowska, Klaus P. Brüssow, Bartosz Kempisty, Michal Jeseta and Paweł Antosik
Morphological and biochemical changes in the cells surrounding the oocyte seem to be extremely important in an effective fertilization process. Thanks to advanced cell culture techniques, as well as biochemical and bioinformatics analyses, we can partly imitate the phenomena occurring in the living organism. Previous studies showed a possibility of short – and long – term OEC in vitro cultivation, during which these cells have shown to have significant proliferation and expression of genes responsible for differentiation. Our research was aimed at maintaining a culture of porcine oviduct epithelial cells and analyzing their gene expression profile. The study employed cross-bred gilts at the age of about 9 months, obtained from commercial herds. With the use of Affymetrix® Porcine Gene 1.1 ST Array Strip, we have examined the expression of 12257 transcripts. Genes with fold change higher than abs (2) and with corrected p-value lower than 0.05 were considered as differentially expressed. We chose 20 genes with the most marked expression (10 up – regulated, 10 down – regulated) for further investigation in the context of literature sources. These genes belonged to three ontological groups: “cell cycle process”, “cell division” and “cell proliferation”. The results obtained from these studies may be the basis for further molecular analyses.
Magdalena Kulus, Blanka Borowiec, Małgorzata Popis, Piotr Celichowski, Michal Jeseta, Dorota Bukowska, Hanna Piotrowska-Kempisty, Małgorzata Bruska, Maciej Zabel, Michał Nowicki, Bartosz Kempisty and Paweł Antosik
Among many factors, the epithelium lining the oviductal lumenis very important for the development of the oocyte and its subsequent fertilization. The oviductal epithelium is characterized by the presence of ciliary cells, supporting the movement of cumulus-oocyte complexes towards the uterus. By interacting with the semen, the epithelium of the fallopian tube makes the sperm acquire the ability to fertilize. So far, the exact molecular mechanisms of these changes have not been known. Hence, understanding the metabolism of oviduct epithelial cells and the level of expression of individual groups of genes seems to be a way to deepen the knowledge about the broadly understood reproduction.
In our research, we decided to culture oviductal epithelial cells (OECs) in vitro for a long period of time. After 24h, 7, 15 and 30 days, the OECs were harvested, with their RNA isolated. Transcriptomic changes were analyzed using microarrays. The “cellular response to lipid” group was represented by the following genes: MUC1, CYP24A1, KLF4, IL24, SNAI2, CXCL10, PPARD, TNC, ABCA10, while the genes belonging to the “cellular lipid metabolic processes” were: LIPG, ARSK, ACADL, FADS3, P2RX7, ACSS2, PPARD, KITLG, SPTLC3, ERBB3, KLF4, CRABP2. Additionally, PPARD and ACADL were members of the “fatty acid beta-oxidation” ontology group. Our study describes genes that are not directly related to fertility processes. However, significant changes in their expression in in vitro cultured OECs may indicate their usefulness as markers of OECs’ physiological processes.
Running title: Fatty acids changes in porcine oviductal epithelial cells in in vitro cultivation