Ceramide Synthase 6 Is a Novel Target of Methotrexate Mediating Its Antiproliferative Effect in a p53-Dependent Manner

We previously reported that ceramide synthase 6 (CerS6) is elevated in response to folate stress in cancer cells, leading to enhanced production of C16-ceramide and apoptosis. Antifolate methotrexate (MTX), a drug commonly used in chemotherapy of several types of cancer, is a strong inhibitor of folate metabolism. Here we investigated whether this drug targets CerS6. We observed that CerS6 protein was markedly elevated in several cancer cell lines treated with MTX. In agreement with the enzyme elevation, its product C16-ceramide was also strongly elevated, so as several other ceramide species. The increase in C16-ceramide, however, was eliminated in MTX-treated cells lacking CerS6 through siRNA silencing, while the increase in other ceramides sustained. Furthermore, the siRNA silencing of CerS6 robustly protected A549 lung adenocarcinoma cells from MTX toxicity, while the silencing of another ceramide synthase, CerS4, which was also responsive to folate stress in our previous study, did not interfere with the MTX effect. The rescue effect of CerS6 silencing upon MTX treatment was further confirmed in HCT116 and HepG2 cell lines. Interestingly, CerS6 itself, but not CerS4, induced strong antiproliferative effect in several cancer cell lines if elevated by transient transfection. The effect of MTX on CerS6 elevation was likely p53 dependent, which is in agreement with the hypothesis that the protein is a transcriptional target of p53. In line with this notion, lometrexol, the antifolate inducing cytotoxicity through the p53-independent mechanism, did not affect CerS6 levels. We have also found that MTX induces the formation of ER aggregates, enriched with CerS6 protein. We further demonstrated that such aggregation requires CerS6 and suggests that it is an indication of ER stress. Overall, our study identified CerS6 and ceramide pathways as a novel MTX target

Laborpsychology in agricultural sector

This article concerns the application of psychology in agricultural sector as an example of farming in Ukraine. 

TRENDS IN THE REINSURANCE SECTOR IN THE CONTEXT OF FINANCIAL INTEGRATION

The article deals with the problems of insurance integration processes in the EAEU. The strategic directions of state regulation of the reinsurance market of the Republic of Belarus have been identified. It is determined that there is no a single legislative act regulating reinsurance activities in Belarusian legislation. State regulation of reinsurance relies on the current insurance legislation. The analysis of financial stability in EAEU is based on the Solvency I criteria. In the EU countries the risk-based approach to financial stability for the insurance sector is offered. However, neither of these approaches can be fully applied in the Republic of Belarus due to the methods of the analysis, which are based on hypothetical scenarios of market developments and may have a high degree of unreliability. It is determined that in reinsurance, it is necessary to assess the reliability of a reinsurance organization not only on the basis of an analysis of financial indicators, but to make a complex study of the financial stability of a reinsurer. It was concluded that the above methodology makes it possible to objectively analysis of a reinsurer reliability and can help to develop a set of measures aimed at raising it

CerS6 Is a Novel Transcriptional Target of p53 Protein Activated by Non-genotoxic Stress

Our previous study suggested that ceramide synthase 6 (CerS6), an enzyme in sphingolipid biosynthesis, is regulated by p53: CerS6 was elevated in several cell lines in response to transient expression of p53 or in response to folate stress, which is known to activate p53. It was not clear, however, whether CerS6 gene is a direct transcriptional target of p53 or whether this was an indirect effect through additional regulatory factors. In the present study, we have shown that the CerS6 promoter is activated by p53 in luciferase assays, whereas transcriptionally inactive R175H p53 mutant failed to induce the luciferase expression from this promoter. In vitro immunoprecipitation assays and gel shift analyses have further demonstrated that purified p53 binds within the CerS6 promoter sequence spanning 91 bp upstream and 60 bp downstream of the transcription start site. The Promo 3.0.2 online tool for the prediction of transcription factor binding sites indicated the presence of numerous putative non-canonical p53 binding motifs in the CerS6 promoter. Luciferase assays and gel shift analysis have identified a single motif upstream of the transcription start as a key p53 response element. Treatment of cells with Nutlin-3 or low concentrations of actinomycin D resulted in a strong elevation of CerS6 mRNA and protein, thus demonstrating that CerS6 is a component of the non-genotoxic p53-dependent cellular stress response. This study has shown that by direct transcriptional activation of CerS6, p53 can regulate specific ceramide biosynthesis, which contributes to the pro-apoptotic cellular response

INFLUENCE OF THE ECONOMIC SITUATION OF VILLAGERS OF UKRAINE ON THEIR MENTAL STATE

The author analyzes the results of the social survey of rural residents of Ukraine, on the basis of which the author tried to make a forecast of further mental state and behavior of the villagers

THEORETICAL AND CONCEPTUAL MODEL OF SOCIO-PSYCHOLOGICAL SUPPORT OF PROFESSIONAL FARMING

The author presents the theoretical and conceptual models of socio-psychological support of professional farming. The models are based on the research conducted during 2007-2012 in the territory of Ukraine

ANALYSIS OF DEPRESSIVENESS AND ANXIETY DEGREE OF UKRAINIAN VILLAGERS (2012 – 2013)

In the report the author analyzes the results of psychological diagnostics of Ukrainian villagers. On their basis the author tried to make a forecast concerning the future dynamics of the state and behavior of the villagers. This is the extension of the report previously printed last year

Care for patient\u27s intimacy - influence of nurse\u27s gender

Uvod: Moški so se že v zgodovini pojavljali kot negovalci. To dokazujejo zapisi iz Indije, Rimskega cesarstva in srednjega veka. Dandanes število moških diplomantov v zdravstveni negi narašča. Številke iz obdobja 2006 – 2016 kažejo na povečanje deleža moških z 9% na 17%. Pri izvajanju zdravstvenih storitev prihajamo v stik s pacientom (fizično in psihično), ta stik pa je v literaturi opisan z izrazi, kot so pacientov intimni prostor, dostojanstvo in bližina. Namen: Namen diplomskega dela je proučiti odnose med pacienti in zdravstvenimi delavci s poudarkom na pacientovem doživljanjem posega, s katerim posegamo v intimnost pacienta in dostojanstvo ter raziskati vpliv spola zdravstvenega delavca na ta odnos. Metode dela: Uporabljeni sta dve metodi dela: vsebinski pregled literature ter empirična metoda dela. Anketa je sestavljena iz 6 vprašanj o neprijetnih dogodkih pri pregledih, o zaupanju raznim zdravstvenim delavcem ter o vplivu spola zdravstvenega delavca pri izvajanju zdravstvene nege pacienta. Anketiranci so razdeljeni glede na demografske podatke (spol, starost in izobrazbo). Pri starosti so razvrščeni v starostne skupine po 10 let od vključno 18. leta dalje. Rezultati: K anketi je bilo povabljenih 165, sodelovalo je 161 anketirancev (98 žensk in 63 moških). Največ anketirancev je bilo v starostni skupini 18-29 let, večina je imela končano srednjo strokovno šolo ali gimnazijo. Najbolj pogost odgovor pri vprašanju o zadnji neprijetni izkušnji je fizikalni pregled (45%). V anketi je izpostavljen neprimeren odnos zdravstvenih delavcev (20%), nekateri anketiranci še niso imeli neprijetnih izkušenj (31%). Najbolj neprijetne izkušnje anketirancev so porazdeljene: primarna raven (41%), sekundarna raven (19%), terciarna raven zdravstvene dejavnosti (13%). Anketiranci najbolj zaupajo svoje težave zdravnikom in medicinskim sestram. 52% anketirancev meni, da je fizični pregled najbolj neprijeten. Pri vprašanju o vplivu spola zdravstvenega delavca je 42% anketirancev odgovorilo, da naj jih neguje oseba istega spola, 51% anketirancem se je odločilo za odgovor: Mi je vseeno. Razprava in zaključek: Število zaposlenih moških v zdravstveni negi se povečuje. Pri obravnavi se moramo zavedati vstopa v pacientov intimni prostor. Ta vstop je velikokrat spregledan. Raziskava je pokazatelj pacientovega doživljanja intimnega prostora, prav tako pa je tudi pokazatelj odnosa zdravstvenega delavca do pacienta.Introduction: Many sources in history that exist, which indicate that men have already been caregivers in the past. Nowadays the number of graduated males in nursing is increasing. Such are the facts from the year 2006 – 2016, where the percentage of males increased to 17%. As health workers we get in touch with patients (physically and psychologically). This contact is often described with words, such as intimate space, dignity and closeness. Purpose: The purpose is to study the relationship between patients and health workers with the emphasis on patient\u27s point of view on the interventions into his/her intimate space and dignity. Methods: Two methods were used in diploma work. First was short literature review, the second was the empirical research. The survey consisted of 6 questions about uncomfortable experiences at medical examinations and the influence of the health worker\u27s gender while providing intimate care. Respondents were organized into groups while taking the demographic data into account (gender, age and education). There were age groups made from the age of 18 onwards. Results: There were 161 respondents in total (98 women and 63 men). The majority was located in the age group 18-29, the most frequent level of education was finished secondary school. The most frequent answer about the last uncomfortable medical examination respondents chose is physical examination (45%). Answers about inappropriate health worker\u27s behaviour must not be overlooked (20%), 31% of respondents had no such experience respectively. The answers about the most uncomfortable experience were organised into three groups: primary (41%), secondary (19%) and tertiary health system (13%). Respondents trust doctors and nurses the most when discussing personal (intimate) issues. 52% of respondents thought that physical examination is the most uncomfortable. 42% of all respondents answered that they should receive help from the caregiver of the same gender. 51% of all respondents chose the answer I don\u27t care. Discussion and conclusion: The number of male nurses is increasing. While taking care of patients we need to remember how to approach them, because it is the significant indicator of the good nurse-patient relationship

Metabolic Reprogramming by Folate Restriction Leads to a Less Aggressive Cancer Phenotype

Folate coenzymes are involved in biochemical reactions of one-carbon transfer, and deficiency of this vitamin impairs cellular proliferation, migration and survival in many cell types. Here the effect of folate restriction on mammary cancer was evaluated using three distinct breast cancer subtypes differing in their aggressiveness and metastatic potential: non-invasive basal-like (E-Wnt), invasive but minimally metastatic claudin-low (M-Wnt), and highly metastatic claudin-low (metM-Wntliver) cell lines, each derived from the same pool of MMTV-Wnt-1 transgenic mouse mammary tumors. NMR-based metabolomics was used to quantitate 41 major metabolites in cells grown in folate-free medium versus standard medium. Each cell line demonstrated metabolic reprogramming when grown in folate-free medium. In E-Wnt, M-Wnt and metM-Wntliver cells 12, 29, and 25 metabolites, respectively, were significantly different (p<0.05 and at least 1.5-fold change). The levels of eight metabolites (aspartate, ATP, creatine, creatine phosphate, formate, serine, taurine and β-alanine) were changed in each folate-restricted cell line. Increased glucose, decreased lactate, and inhibition of glycolysis, cellular proliferation, migration and invasion occurred in M-Wnt and metM-Wntliver cells (but not E-Wnt cells) grown in folate-free versus standard medium. These effects were accompanied by altered levels of several folate-metabolizing enzymes, indicating that the observed metabolic reprogramming may result from both decreased folate availability and altered folate metabolism. These findings reveal that folate restriction results in metabolic and bioenergetic changes and a less aggressive cancer cell phenotype