Biophysical and biochemical characterisation of DNA-based inhibitors of the cytosine-mutating APOBEC3 enzymes : a thesis presented in partial fulfilment of the requirements for the degree of Doctor of Philosophy in Biochemistry at Massey University, Palmerston North, New Zealand

Figures are re-used with permission or under a Creative Commons CC BY license.With the rise of antiviral and anticancer drug resistance, a new approach must be taken to overcome this burden. The APOBEC3 (A3) family of cytosine deaminases hypermutate cytosines to uracils in single-stranded DNA (ssDNA). These enzymes act as double-edged swords: on one side they protect humans against a range of retroviruses and other pathogens, but several A3s are exploited by viruses and cancer cells to increase their rate of evolution using the enzyme’s mutagenic actions. This latter mode permits escape of cancer cells from the adaptive immune response and leads to the development of drug resistance. In particular, APOBEC3B (A3B) is considered to be a main driving source of genomic mutations in cancer cells. Inhibition of A3B, while retaining the beneficial actions of the other A3 in the immune system, may be used to augment existing anticancer therapies. In this study, we showed for the first time that short ssDNAs containing cytosine analogue nucleosides, 2'-deoxyzebularine (dZ) or 5-fluoro-2'-deoxyzebularine (5FdZ) in place of the substrate 2'-deoxycytidine (dC) in the preferred 5'-TC motif, inhibit the catalytic activity of A3B. However, as most A3 enzymes (except A3G) prefer to deaminate ssDNA with a 5'-TC motif, selective A3B inhibition was uncertain. We noted that nucleotides adjacent to the 5'-CCC motif influence the dC deamination preference of A3A, A3B, and A3G’s. Replacement of the A3B’s preferred dC in the 5'-CCC motif with dZ (5'-dZCC) led to the first selective inhibitor of A3B, in preference to A3A and A3G. Furthermore, using small-angle X-ray scattering (SAXS) we obtained the first model of a full-length two-domain A3 in complex with a dZ-ssDNA inhibitor. Our model showed that the ssDNA was largely bound to the C-terminal domain (CTD) with limited contact to the N-terminal domain in solution, due to the high affinity of the dZ for the CTD active-site. Our work provides a new platform for use of ssDNA-based inhibitors in targeting the mutagenic action of the A3B. Further developments using more potent inhibitors will help to achieve inhibition in cellular studies, with the ultimate goal to complement anti-cancer and antiviral treatments

Features of reciprocal interaction as coordination of motor and cardiac functions

Vinnytsia National Pirogov Memorial Medical University, The 6th International Medical Congress for Students and Young DoctorsIntroduction: Reciprocation is the coordination of 2 activity centers. The majority of works were devoted to the reciprocal interactions of motor centers; but much less attention was given to the phenomenon of vegetative function regulation. Aim: To study the features of reciprocal interaction of sympathetic and parasympathetic divisions of autonomic nervous system. Materials and methodology: A group of male rats Wistar (n = 12) weighing 250-300 g was used for experiment. The first stage of the experiment was to check the heart rate during food procuring movements in normal rats (n = 8) with normal, stable motor skills. There was the phenomenon of shortterm motor bradycardia during the movement. The second stage of the experiment was to register the heart rate during food procuring movement in rats influenced by an intraperitoneal atropine (1.7 mg/kg) injection, in order to exclude vagal effect on heart function. Results: Pharmacological blockage of the vagus nerve by atropine causes a significant increase (p<0.05) in heart rate background 11% to the value of 487 ± 10.1 BPM, but not to “rule out” the phenomenon of short-term motor bradycardia during the food procuring movement. Conclusion: The reciprocal innervation of antagonistic muscles and heart have their similarities and differences. This can have 3 levels of regulation and 2 opposing processes (excitation and inhibition) in the reciprocal organization. The working body (or an interconnected group) (lower level) gets antagonistic innervation from two centers (average level). It can predict that the presence of a higher level which causes excitation of one center and simultaneous inhibition of the antagonistic center. Key words: Reciprocation, heart rate, rats, atropine

New school culture? The question on potential impact of distance education during the COVID-19 pandemy

Although the origins of remote education date back to the 18th century (Kentnor, 2015) however, only recently, in the second decade of the 21st century, due to a natural experiment that the COVID-19 pandemic has become, remote learning began to play a key role in mass global education. In these unprecedented times of reorganisation of time and space in order to ensure the continuity of social life, the school, subjected to a major organisational transformation, changed its form from the traditional setting (characterised by a clear division of time and space, i.e. lessons in the school classroom) to the virtual setting, escaping this division. The aim of the conducted research study is to describe to what extent the functioning of the institution of the school has changed (if at all) due to its virtual environment, with particular reference to the school culture understood as “the beliefs, views, attitudes and relationships, and the written and unwritten rules that shape every aspect of the school as an institution (...) and community” (Ward, Burke, 2004:1). In other words, the question considered is how and what has changed in the functioning of the school, with particular regards to school rituals, teaching strategies, and the interactions of school actors, as a result of remote education? Has online education changed the way teachers and students behave and perceive school? The results of the study reveal that the functioning of the "virtual school" as compared to the "traditional school" in its essence remains unchanged due to the strong, long-standing cultural codes instilled by and within the institution of the school. In the virtual classroom, as in the material classroom, assessment and grading are teachers' strategies for recalling the definition of the school situation; moreover, they often become a survival strategy to maintain control over the group, in order to sustain ontological security (Giddens, 1984) in the performance of the school roles of teacher and student

The impact of other family member's well being on expatriate's success - A Literature review The impact of other family members' well-being on expatriates' well-being, adjustment, performance and success.

The literature review shows existing trends in international assignments and focuses on the potential impact of the family members' well-being on the expatriates' success. The research is done in the quest for reliable publications which are further analyzed in two parts, based on the general impact of the family on the expatriates and based on the well-being factor of the family influencing the expatriates. The thesis shows an issue of the family members being often a significant failure factor based on their problems and issues related to the functioning in the host country. However, the family's state of well-being, consistent with the cross-over theory, will influence the expatriates and the international assignment success positively.The literature review shows existing trends in international assignments and focuses on the potential impact of the family members' well-being on the expatriates' success. The research is done in the quest for reliable publications which are further analyzed in two parts, based on the general impact of the family on the expatriates and based on the well-being factor of the family influencing the expatriates. The thesis shows an issue of the family members being often a significant failure factor based on their problems and issues related to the functioning in the host country. However, the family's state of well-being, consistent with the cross-over theory, will influence the expatriates and the international assignment success positively.

The expiration of an administrative decision

Niniejsza praca została poświęcona tematyce wygaśnięcia decyzji administracyjnej. Autorka w pierwszej kolejności podjęła analizę w zakresie istoty pojęcia decyzji administracyjnej i jej poszczególnych rodzajów. Następnie rozważania przeniesiono na grunt zasady trwałości decyzji administracyjnej. Główna część pracy została poświęcona analizie przyczyn wygaśnięcia decyzji administracyjnej na podstawie przypadków zachodzących na gruncie materialnego prawa administracyjnego. Najpierw skupiono się na aspekcie podmiotowym wygaśnięcia decyzji administracyjnej, w dalszej kolejności analizie został poddany aspekt przedmiotowy. Ostatni rozdział stanowi omówienie instytucji stwierdzenia wygaśnięcia decyzji administracyjnej z art. 162 ust.1 KPA, ze szczególnym zwróceniem uwagi na charakter decyzji stwierdzającej wygaśnięcie.This paper was devoted to the issue of the expiration of an administrative decision. Firstly, the author of this paper analizes the essence of an administrative decision’s term and its particular type. Secondly, the considerations concern the principle of the durability of administrative decision. In the main part of the paper were discussed causes of the decision’s expiration which are based on cases from the administrative substantive law. The author starts with the subjective aspects of decision’s expiration, afterwards analysis is connected with objective aspects of decision’s expiration. The institution of revocation of decision’s expiration under Rule 162 (1) the Code of Administrative Procedure was discussed in the last chapter

How does the interaction between the Filamin A repeat 10 domain and F-actin lead to severe OPD skeletal disorders? : a thesis presented in partial fulfilment of the requirements for the degree of Master of Science in Biochemistry at Massey University, Palmerston North, New Zealand

The cytoskeleton network allows cells to differentiate, divide, and move in response to the external environment creating a mechanoprotection system against cell stress. The actin cytoskeleton is stabilised and tightly regulated by various actin-binding proteins, one of which are the family of Filamin (FLN) proteins that crosslink F-actin into three-dimensional networks. Filamins also link the actin cytoskeleton to the cellular membrane through interactions with transmembrane proteins and function as a molecular scaffold for signalling molecules. In addition to an actin binding domain, each monomer contains a rod region of 24 immunoglobulin-like repeat domains with dimerisation of the monomers occurring at repeat 24. The human filamin family contains three FLN isoforms; FLNA, FLNB, and FLNC which are differentially expressed where FLNA is identified as the dominant isoform located on the X-chromosome essential for mammalian development. Mutations in Filamin A (FLNA) have been identified to cause distinctly different human diseases affecting the central nervous system, vascular system, or skeletal muscles; however, the molecular mechanisms of FLNA leading to these diseases remain unclear. Mutations cluster in distinct FLNA domains, suggesting their functional importance for mediating correct functions. Mutations in the FLNA repeat 10 domain are correlated with severe forms of the skeletal disorders Otopalatodigital syndrome spectrum disorders (OPD) thought to be due to an altered or gain-of-function phenotype. The aim of this study was to provide an insight into the biochemical properties of FLNA repeat 10 domain by better understanding how mutations in this domain lead to OPD. Initially, recombinant wildtype (Wt) and mutant (V1249A and A1188T) FLNA repeat 10 domain proteins (FLNAR10) were purified then compared by in vitro biochemical studies to investigate secondary structure, stability, and affinity towards F-actin. The FLNAR10 protein was revealed to have relatively weak binding affinity towards F-actin, consistent with being an additional contributor in the filamin protein to bind F-actin. Mutations in the FLNAR10 protein exhibited a slight increase in affinity towards F-actin, accompanied by a slight reduction of thermostability in comparison to the Wt protein, but no significant changes in the secondary structure were observed. This slight increase in the affinity of the mutant FLNA repeat 10 proteins towards F-actin is consistent with a gain-of function mechanism for the disease phenotype. Overall, these results contribute towards a better understanding of the FLNA function, providing further evidence towards a gain-of function mechanism for OPD

Wonders of the Invisible World

https://stars.library.ucf.edu/diversefamilies/2289/thumbnail.jp

The verification of Michael Leiter’s Model: the relationship between burn out syndrome and inadequacy to work by hypermarket’s workers.

Celem pracy magisterskiej była weryfikacja modelu Michaela Leitera, zależności występujących między wypaleniem zawodowym a niedostosowaniem do pracy w grupie zawodowej pracowników marketów. Badania zostały przeprowadzone w marketach na terenie województwa małopolskiego powiatu suskiego na grupie 80 osób. Pomiaru dokonano posługując się Kwestionariuszem AWS autorstwa Leitera, który sprawdza dopasowanie pracownika do sześciu obszarów życia zawodowego (obciążenie pracą, poczucie kontroli, wynagrodzenie, relacje interpersonalne, sprawiedliwość i wartość) oraz Kwestionariuszem MBI-GS autorstwa Maslach, który bada trzy wymiary wypalenia zawodowego (wyczerpanie, cynizm i poczucie efektywności zawodowej. Do analizy statystycznej zastosowano metodę analizy korelacji i regresji. Wyniki badań zostały oparte na analizie głównych obszarów, w których pojawiają się oznaki wypaleniaThe aim of this master thesis is the verification of Michael’s Leiter Model: the relationship between burn out syndrome and inadequacy to work by hypermarket’s workers. The research was done in hypermarkets located in Małopolska voivodeship, Suski county with a group of eighty person. Surveys were made using AWS (Areas of Worklife Survey) questionnaire developed by Michael Leiter, which tests the worker’s matching to six areas of professional life (workload, feeling of beeing controled, salary, interpersonal relations, fairness and value) and using MBI-GS questionnaire developed by Maslach, which tests three aspects of burn out syndrome (exhaustion, cynicism and feeling of professional efficiency). The analyses of correlation and regression were applied. Results of the research were based on the analysis of the main areas, where symptoms of burn out syndrome appear

A Pharmacy Business Management Simulation Exercise as a Practical Application of Business Management Material and Principles

Objective. To implement a pharmacy business management simulation exercise as a practical application of business management material and principles and assess students’ perceived value. Design. As part of a pharmacy management and administration course, students made various calculations and management decisions in the global categories of hours of operation, inventory, pricing, and personnel. The students entered the data into simulation software and a realistic community pharmacy marketplace was modeled. Course topics included accounting, economics, finance, human resources, management, marketing, and leadership. Assessment. An 18-item posttest survey was administered. Students’ slightly to moderately agreed the pharmacy simulation program enhanced their knowledge and understanding, particularly of inventory management, cash flow statements, balance sheets, and income statements. Overall attitudes toward the pharmacy simulation program were also slightly positive and students also slightly agreed the pharmacy simulation program enhanced their learning of pharmacy business management. Inventory management was the only area in which students felt they had at least “some” exposure to the assessed business management topics during IPPEs/internship, while all other areas of experience ranged from “not at all” to “a little.” Conclusion. The pharmacy simulation program is an effective active-learning exercise and enhanced students’ knowledge and understanding of the business management topics covered