Histones are an essential part of nucleosomes that regulate chromatin structure and function. Histone exchanges and modifications represent a scaffold for DNA transcription, repair, and replication. ...Studying histones and histone code is an important and fast-developing branch of epigenetic science. Here we propose a fast, efficient, and versatile assay for nucleosomal histone isolation from mammalian cells, without the use of acids or high salt solutions which are common for other histone isolation techniques. All components used in the protocol are common and inexpensive laboratory chemicals. The protocol has been evaluated on six commonly used cell lines and two animal tissue samples. The mild extraction conditions preserve delicate histone epigenetic changes, allowing its downstream analyses. We have demonstrated the assays' successful application during changes in the transcriptional activity of histone genes, cell cycle transitions, and DNA-damaging conditions. Histone fractions, obtained by the protocol, can be used for further applications, such as electrophoresis, immunoblot, and mass spectrometry. Therefore, the new proposed nucleosomal histone isolation method is sensitive, specific, and suitable for downstream applications of various kinds.
Nerolidol is a naturally occurring sesquiterpene alcohol with multiple properties, including antioxidant, antibacterial, and antiparasitic activities. A few studies investigating the antitumor ...properties of nerolidol have shown positive results in both cell culture and mouse models. In this study, we investigated the antitumor mechanism of
-nerolidol in bladder carcinoma cell lines. The results of our experiments on two bladder carcinoma cell lines revealed that nerolidol inhibited cell proliferation and induced two distinct cell death pathways. We confirmed that
-nerolidol induces DNA damage and ER stress. A mechanistic study identified a common cAMP, Ca
, and MAPK axis involved in signal propagation and amplification, leading to ER stress. Inhibition of any part of this signaling cascade prevented both cell death pathways. The two cell death mechanisms can be distinguished by the involvement of caspases. The early occurring cell death pathway is characterized by membrane blebbing and cell swelling followed by membrane rupture, which can be prevented by the inhibition of caspase activation. In the late cell death pathway, which was found to be caspase-independent, cytoplasmic vacuolization and changes in cell shape were observed.
-Nerolidol shows promising antitumor activity through an unorthodox mechanism of action that could help target resistant forms of malignancies, such as bladder cancer.
The purpose of this paper is to prove possibility of the application of the sociometric method in human resources management with the aim of appointing managers in one organization. Based on the some ...results, published in the scientific papers, authors established general and particular hypothesis about applying sociometry as a research method in human resources management. The concrete research of the applications of sociometry for appointing managers was conducted in one company in Serbia. Sociometric research shows real relations in a social group, as well as the projection of the position of individuals, mutual attraction and rejection within the group. Presented research is one of the "pioneering" attempts of broader application of the sociometric method in human resources management with the aim of appointing managers in one organization, especially in Serbia.
The paper presented a multidimensional model of correlation between human resource management practices and the quality of banks' operations as a support in financing agriculture in Serbia, tested ...twice which gives this analysis a longitudinal character - trend study. Considering that from the firstly surveyed research sample, Agrobank that had showed good results went bankrupt, this in some way brought into question the quality and validity of the tested model of correlation of human resource management practice and performance quality, so there was a need for its re-testing. The indisputable significance of this research is reflected in the fact that, based on the twice-tested multi-dimensional model of connection between these two entities, it was possible not only to examine how much but also the way in which these human resource management practices generate the quality of banks' operations, as well as the future trends of this practice.
Stari grad Novigrad na Dobri smješten je u Karlovačkoj županiji,
općini Netretić (Zagradci). Zaštićeno je kulturno dobro. Kaštel
pripada kasnom srednjem i ranom novome vijeku. Na prostoru
kaštela ...između 2009. i 2012. godine, pod vodstvom Sanjina Mihelića
iz Arheološkog muzeja u Zagrebu, provedena su zaštitna
arheološka istraživanja unutar kaštela koja su zahvatila prostor
sjevernog dijela dvorišta i cisternu. Cisterna pripada filter-tipu, a
prema radiokarbonskoj analizi, keramici i odnosu među stratigrafskim
jedinicama može se datirati u kraj 15. i početak 16. stoljeća.
Istraženi sustav vodoopskrbe na Starom gradu Novigradu
na Dobri sastoji se od cisterne kružnog oblika sa središnjim spremnikom
za vodu i filterom, kanala za dovod vode s prostora krovišta
kaštela i „kanala-tunela” za regulaciju viškova i dotok vode.
Cisterna je smještena u sjeveroistočnom dijelu dvorišta, a svojom
je izgradnjom presjekla ranije slojeve ukopavanjem u živu
stijenu. Bila je okružena galerijama, od kojih su ostale sačuvane
jedino baze kamenih stupaca koje su djelo kasnijih intervencija.
Ovaj tip cisterne pojavljuje se početkom razvijenog srednjega
vijeka na prostoru srednje Europe i Mediterana, a poradi svoje
praktičnosti, nastavlja se koristiti i u ranom novome vijeku na
fortifikacijama.
Endoplasmic reticulum (ER) stress, prompted by the accumulation of misfolded or unfolded proteins, triggers the activation of the unfolded protein response (UPR) pathway to restore ER homeostasis. ...This stress response is implicated in the development of hepatocellular carcinoma (HCC). A biallelic mutation in SPRTN is currently the only known single-gene mutation implicated in the early onset of HCC. However, the exact mechanism linking SPRTN mutations to HCC remains unclear. In our study, we analyzed SPRTN and UPR in 21 human HCC tissue samples using RT-qPCR, immunoblot, and immunohistochemistry. We found alterations in the expression levels of SPRTN and UPR-related genes and proteins in HCC samples. The impact of SPRTN on the ER stress response was assessed in SPRTN-depleted HepG2 cells through RNA sequencing, pull-down assay, comet assay, and mitotic index calculation. We demonstrated that SPRTN interacts with the UPR sensor GRP78. Furthermore, we observed a decrease in SPRTN levels during ER stress, and increased sensitivity to ER stress in SPRTN-depleted cells. These findings suggest an essential role for SPRTN in the ER stress response and provide new insights into HCC pathogenesis. This newly discovered function of SPRTN could significantly enhance our understanding and treatment of HCC.
Abstract
DNA-protein crosslinks (DPCs), formed by the covalent conjugation of proteins to DNA, are toxic lesions that interfere with DNA metabolic processing and transcription. The development of an ...accurate biochemical assay for DPC isolation is a priority for the mechanistic understanding of their repair. Here, we propose the STAR assay for the direct quantification of DPCs, sensitive to physiologically relevant treatment conditions. Implementing the STAR assay revealed the formation of small cross-linked peptides on DNA, created by the proteolytic degradation of DPCs by SPRTN. The initial proteolytic degradation of DPCs is required for the downstream activation of DNA repair, which is mediated through the phosphorylation of H2Ax. This leads to the accumulation of DNA repair factors on chromatin and the subsequent complete removal of the cross-linked peptides. These results confirmed that the repair of DPCs is a two-step process, starting with proteolytic resection by SPRTN, followed by the repair of the underlying damage to the DNA.
Graphical Abstract
Graphical Abstract
The first step in DPC repair is SPRTN-mediated proteolysis. The remaining cross-linked peptides are finally resolved by the activation of DNA damage repair.
The aim of this paper is to investigate the differences in managers’ motivation with short-term incentives in the banking sector of the Republic of Serbia. The research study examined whether ...short-term incentives differently affect managers’ motivation, depending on hierarchy and age. For this purpose the survey was conducted, using a questionnaire method. The analysis of various literature sources was performed, emphasizing the importance of short-term incentives, as compensation component, for managers’ motivation. Based on insights into existing studies, research hypotheses were established. The empirical results were analyzed quantitatively, using standard statistical methods. The results indicated that there are no significant differences in managers’ motivation with short-term incentives, in relation to their current hierarchical position. Further, the survey results revealed significant relationship between motivation with short-term incentives and managers’ age, that is, the findings show that short-term incentives are more effective in motivating older managers. Therefore, the results are important for compensation strategies in contemporary business organizations. From the aspect of designing executive compensation packages, organizations need to adapt them to different subgroups of managers, considering their motivational preferences.
Background
The potentially inappropriate medications (PIMs) and drug–drug interactions (DDIs) can significantly affect patient safety in the elderly, especially at transition of health care.
...Objective
The aim of this study is to evaluate PIMs involved in potentially clinically significant DDIs in prescribed pharmacotherapy of elderly patients at hospital discharge.
Setting
Internal Medicine Clinic of University Hospital Dubrava, Zagreb, Croatia.
Method
During a 16-month period, the pharmacotherapy data were assessed using Lexicomp Online screening software to identify category C (monitor drug therapy), D (consider therapy modification) and X (avoid combination) DDIs. The European Union (EU)(7)-PIM criteria were applied to detect inappropriately prescribed medications involved in DDIs. Clinical pharmacists obtained data from patients’ medical records and patient/caregiver interviews.
Main outcome measure
The incidence of PIMs involved in potentially clinically significant DDIs.
Results
A total of 364 consecutive elderly patients were enrolled in the study. The mean number of prescription medications at discharge was 9.3. Overall, 2833 potentially clinically significant DDIs were identified: 2445 (86.3%) of them were category C, 347 (12.3%) category D and 41 (1.4%) were category X interactions. A total of 1164 PIMs were involved in 31.2% of category C interactions, 60.2% of category D interactions and 43.9% of category X interactions. The most frequent PIMs involved in potentially clinically significant DDIs were tramadol, benzodiazepines, moxonidine, vildagliptin and metoclopramide.
Conclusion
A very high incidence of DDIs in elderly patients and a high incidence of PIMs involved in DDIs was determined at hospital discharge.
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