Genetic predisposition of suicidal behavior: variants in GRIN2B, GABRG2, and ODC1 genes in suicide attempt and completed suicide In two Balkan populations

Introduction: Suicidal behavior ranges between suicidal ideation and completed suicide. Completed suicide accounts for over 700,000 deaths worldwide, while attempted suicide is 20 times more frequent. Genetic background is an important factor contributing to suicidal behavior, and candidate genes linked to several neurotransmitter systems have been investigated. Alternations in glutamate, γ-aminobutyric acid (GABA) and polyamine systems have been detected in suicidal behavior. Our aim was to differentiate genetic predispositions underlying two different types of suicidal behavior, attempted and completed suicide, in two Balkan populations. Methods: The study sample included 173 suicide attempters with comorbid psychiatric disorders (major depressive disorder, bipolar affective disorder, or schizophrenia), 216 non-suicidal psychiatric patients and 172 healthy controls from Serbia, and 333 suicide completers and 356 non-suicidal autopsy controls from Slovenia. Variants in the genes GRIN2B (rs2268115 and rs220557), GABRG2 (rs424740), and ODC1 (rs1049500 and rs2302614) were genotyped by TaqMan assays and analyzed using PLINK. Results: The CA genotype of rs220557 in the GRIN2B gene increases the risk for completed suicide (OR=1.51, p=0.021), and particularly violent suicide (OR=1.49, p=0.037), compared to controls. In the ODC1 gene, the CA genotype of rs2302614 decreases the risk for completed suicide compared to suicide attempt (OR=0.32, p=0.012). Marginally, the AC haplotype for variants rs1049500-rs2302614 in the ODC1 gene decreases the risk for completed suicide compared to suicide attempt (OR=0.50, p=0.052). Conclusion: Specific genetic variants of the glutamate and the polyamine systems are differently distributed among diverse suicidal phenotypes, thus providing further information on the implication of these systems in suicidality

Isolation and characterization of recombinant protein erylysin B, produced in yeast Saccharomyces cerevisiae

Protein erilizin B je 59 kD velik protein iz užitne gobe kraljevi ostrigar (Pleurotus eryngii). Če ga rekombinantno izražamo v bakteriji Escherichia coli, se ne zvija pravilno in tvori inkluzijska telesca, posledično pa je doprinos topnega proteina zelo nizek. Erilizin B smo zato želeli izraziti v kvasovki Saccharomyces cerevisiae, saj smo pričakovali, da se bo protein izražal v topni obliki in da ga bomo izolirali v višjih koncentracijah. Izražanje rekombinantnega proteina erilizina B smo preverjali na več načinov. Spremljali smo hitrost rasti kvasovke S. cerevisiae in protein skušali zaznati s poliakrilamidno elektroforezo v prisotnosti natrijevega dodecil sulfata in s prenosom western. Ker je erilizin B v kombinaciji s proteinom ostreolizinom A6 hemolitičen, smo njegovo prisotnost preverili s hemolitičnim testom na govejih eritrocitih. V primeru, ko smo izražali erilizin B, označen z zelenim fluorescenčnim proteinom (EryB-eGFP), smo rast rekombinantnih kvasovk spremljali z merjenjem fluorescence in jih vzporedno opazovali s fluorescenčnim mikroskopom. Izražanja rekombinantnega proteina erilizina B nismo zaznali, zato sklepamo, da kvasovka S. cerevisiae v kombinaciji z ekspresijskim vektorjem pYES2 ni primeren ekspresijski sistem za pridobivanje rekombinantnega erilizina B.Erylysin B is a 59 kD protein from the king oyster mushroom (Pleurotus eryngii). When erylysin B is recombinantly expressed in Escherichia coli, it does not fold properly and forms inclusion bodies. The amount of soluble protein is therefore very low. The main goal of the MSc thesis was therefore to express erylysin B in the yeast Saccharomyces cerevisiae. We have expected the expression of recombinant protein in soluble form and in higher concentrations in contrast to the E. coli expression system. Therefore, the expression of the recombinant erylysin B was tested in several ways. We followed the growth curves of the transformed yeast cells and the protein expression was monitored using sodium dodecyl sulphate–polyacrylamide gel electrophoresis and Western blot analysis. Since erylysin B in the combination with ostreolysin A6 is haemolytic, its presence was also monitored by haemolytic assay on bovine erythrocytes. In the case when eGFP-labelled erylysin B was expressed, the fluorescence measurements were performed, as well as fluorescent microscopy in order to follow the expression of the protein in living cells. The expression of the recombinant erylysin B was not detected, so we concluded that the yeast S. cerevisiae in combination with the expression vector pYES2 is not a suitable expression system for obtaining recombinant erylysin B protein

Role of a preschool teacher in a hospital ward

Vzgojitelji predšolskih otrok lahko delo opravljajo tudi v bolnišničnih oddelkih, kjer se v sklopu javnih vrtcev prav tako izvaja predšolska vzgoja. Pri svojem delu si poleg Kurikuluma za vrtce pomagajo s Konceptom dela v bolnišničnih oddelkih. Narava dela od vzgojiteljev zahteva veliko prilagodljivosti, saj v primerjavi z rednimi oddelki tu nimajo stalne skupine, otroci pa imajo lahko različna bolezenska stanja. V diplomskem delu sem se osredinila na organizacijo dela v bolnišničnih oddelkih vrtca, skrbi vzgojiteljev za otrokove potrebe, na vlogo vzgojiteljev pri otrocih brez staršev, koliko lahko bolnišnica nadomesti vrtec za otroke, ki v bolnišnici preživijo več časa, kakšna je otroška igra v bolnišnicah in kako je epidemija koronavirusa vplivala na delo v bolnišničnih vrtcih. Pri raziskovanju sem uporabila deskriptivno in kazalno-neeksperimentalno metodo raziskovanja. S pomočjo kvalitativne tehnike zbiranja podatkov (nestandardiziran intervju) sem v raziskavo vključila šest vzgojiteljic, ki delajo v različnih bolnišničnih oddelkih v Sloveniji. Rezultati so pokazali, da imajo vzgojitelji v bolnišnicah oteženo načrtovanje, večji poudarek je na sprotni prilagodljivosti in kasnejši evalvaciji. V bolnišnicah se trudijo, da se otroci v najkrajšem možnem času vrnejo domov, zato ležalne dobe niso dolge, otroci pa imajo v večini primerov ob sebi starša oz. skrbnika, ki sobiva z njim. Otroška igra v bolnišnicah se v največji meri prilagaja zdravstvenemu stanju otroka, omogočena pa mu je vedno, ko nima zdravstvenih pregledov ali preiskav. Na delo v bolnišnicah pa je močno vplivala epidemija koronavirusaskozi čas so se ukrepi zelo spreminjali, nekaj vzgojiteljev je ostalo brez igralnice, nošenje mask je postala obvezna stalnica, igrače in materiali pa se še strožje razkužujejo.Preschool teachers can also work in hospital wards, where preschool education is provided as part of public preschool education. In addition to the Kindergarten Curriculum, they base their work on the Concept of Work in Hospital Wards. Preschool teachers must be very adaptable, as, compared to regular groups, they do not teach the same group of children, and children can have different medical conditions. In this paper, I focused on work organisation in preschool hospital wards, preschool teachers’ care for children’s needs, role of preschool teachers in working with children in the absence of their parents, the extent to which the hospital can replace the preschool from the perspective of children who spend a lot of time in the hospital, children’s play in the hospital, and the way in which the coronavirus epidemic affected the work in hospital wards. The research was based on a descriptive as well as a causal and non-experimental research method. By using a qualitative method of collecting data (non-standardised interview), I included in the research six preschool teachers who work in different hospital wards across Slovenia. The results showed that preschool teachers find it hard to plan their work, and they emphasise the need for constant adaptation and later evaluation. In hospitals, they try to make sure that children return home as soon as possible, so inpatient stay is not long, and children are normally accompanied by their parents or guardian. Children’s play in hospitals adapts to their medical conditions to a large extent, and children can play whenever they do not have examinations or checks. Work in the hospital was severely affected by the coronavirus epidemic. The measures changed through the epidemic, some playrooms were closed, mask-wearing became obligatory, and toys and materials needed to be disinfected more meticulously

Role of a preschool teacher in a hospital ward

Preschool teachers can also work in hospital wards, where preschool education is provided as part of public preschool education. In addition to the Kindergarten Curriculum, they base their work on the Concept of Work in Hospital Wards. Preschool teachers must be very adaptable, as, compared to regular groups, they do not teach the same group of children, and children can have different medical conditions. In this paper, I focused on work organisation in preschool hospital wards, preschool teachers’ care for children’s needs, role of preschool teachers in working with children in the absence of their parents, the extent to which the hospital can replace the preschool from the perspective of children who spend a lot of time in the hospital, children’s play in the hospital, and the way in which the coronavirus epidemic affected the work in hospital wards. The research was based on a descriptive as well as a causal and non-experimental research method. By using a qualitative method of collecting data (non-standardised interview), I included in the research six preschool teachers who work in different hospital wards across Slovenia. The results showed that preschool teachers find it hard to plan their work, and they emphasise the need for constant adaptation and later evaluation. In hospitals, they try to make sure that children return home as soon as possible, so inpatient stay is not long, and children are normally accompanied by their parents or guardian. Children’s play in hospitals adapts to their medical conditions to a large extent, and children can play whenever they do not have examinations or checks. Work in the hospital was severely affected by the coronavirus epidemic. The measures changed through the epidemic, some playrooms were closed, mask-wearing became obligatory, and toys and materials needed to be disinfected more meticulously

Anti-Vimentin Nanobody Decreases Glioblastoma Cell Invasion In Vitro and In Vivo

Purpose: Glioblastoma (GBM) is the most common primary brain tumour and one of the deadliest cancers. In addition to late diagnosis and inadequate treatment, the extremely low survival rate is also due to the lack of appropriate therapeutic biomarkers and corresponding therapeutic agents. One of the potential therapeutic biomarkers is the intermediate filament vimentin, which is associated with epithelial-mesenchymal transition (EMT). The purpose of this study was to analyse the effect of the anti-vimentin nanobody Nb79 on cell invasion in vitro and in vivo. To further our understanding of the mechanism of action, we investigated the association between Nb79 and EMT in GBM and GBM stem cells by analysing the expression levels of key EMT-related proteins. Methods: The expression of vimentin in glioma tissues and cells was determined by RT-qPCR. An invasion assay was performed on differentiated glioblastoma cell line U-87 MG and stem cell line NCH421k in vitro as well as in vivo in zebrafish embryos. The effect of Nb79 on expression of EMT biomarkers beta-catenin, vimentin, ZEB-1 and ZO1 was determined by Western blot and immunocytochemistry. Results: Our study shows that vimentin is upregulated in glioblastoma tissue compared to lower grade glioma and non-tumour brain tissue. We demonstrated that treatment with Nb79 reduced glioblastoma cell invasion by up to 64% in vitro and up to 21% in vivo. In addition, we found that the tight junction protein ZO-1 had higher expression on the cell membrane, when treated with inhibitory anti-vimentin Nb79 compared to control. Conclusion: In conclusion, our results suggest that anti-vimentin nanobody Nb79 is a promising tool to target glioblastoma cell invasion