No / Evaluation of drug candidates in in-vitro assays of action potential duration (APD) is one component of preclinical safety assessment. Current assays are limited by technically-demanding, time-consuming electrophysiological methods. This study aimed to assess whether a voltage-sensitive dye-based assay could be used instead. Methods Optical APs were recorded using di-4-ANEPPS in electrically field stimulated Beagle left ventricular midmyocardial myocytes (LVMMs). Pharmacological properties of di-4-ANEPPS on the main cardiac ion channels that shape the ventricular AP were investigated using IonWorks™ and conventional electrophysiology. Effects of 9 reference drugs (dofetilide, E4031, d-sotalol, ATXII, cisapride, terfenadine, alfuzosin, diltiazem and pinacidil) with known APD-modulating effects were assessed on optically measured APD at 1 Hz. Results Under optimum conditions, 0.1 μM di-4-ANEPPS could be used to monitor APs paced at 1 Hz during nine, 5 s exposures without altering APD. di-4-ANEPPS had no effect on either hIERG, hINa, hIKs and hIto currents in transfected CHO cells (up to 10 µM) or ICa,L current in LVMMs (at 16 µM). di-4-ANEPPS had no effect on APs recorded with microelectrodes at 1 or 0.5 Hz over a period of 30 min di-4-ANEPPS displayed the sensitivity to record changes in optically measured APD in response to altered pacing frequencies and sequential vehicle additions did not affect the optically measured APD. APD data obtained with 9 reference drugs were as expected except (i) d-sotalol-induced increases in duration were smaller than those caused by other IKr blockers and (ii) increases in APD were not detected using low concentrations of terfenadine. Discussion Early in drug discovery, the di-4-ANEPPS-based method can reliably be used to assess drug effects on APD as part of a cardiac risk assessment strategy.

Methamphetamine (MA) is a well-known, easily accessible and powerful psychostimulant, and its abuse has become a global problem. MA abuse affects millions of people worldwide and places an enormous burden on public healthcare resources. Documented consequences of MA abuse include cardiotoxic, neurotoxic and teratogenic effects, as well as long-term consequences of chronic abuse including affective disorders such as schizophrenia and major depressive disorder (MDD). MDD is a highly prevalent mood disorder in both adults and children, documented to contribute to approximately 850 000 suicides annually. This disorder is projected to become the 2nd leading disease of global burden by 2020, preceded only by ischemic heart disease. Depressive-like behaviour is documented as a symptom of chronic MA abuse and particularly during extensive MA withdrawal. Also, MA abuse during pregnancy is documented to cause neurodevelopmental changes that persist into later life. However, current understanding thereof is limited and warrants further investigation of the effects of early-life exposure to MA on outcome in adulthood, particularly in terms of mood disorders. The aim of the current study was to determine the effect of chronic exposure to MA on the depressive-like behaviour later in life in stress-sensitive (Flinders Sensitive Line) and control (Flinders Resistant Line) rats. Rats were exposed during one of the following natal day (ND) age groups: prenatal (ND-13 to ND+02), postnatal (ND+03 to ND+18), prepuberty (ND+19 to ND+34) or puberty (ND+35 to ND+50). These age groups represent different stages in neurodevelopment, as also seen in humans. For prenatal exposure, pregnant dams received 5 mg/kg daily subcutaneously (s.c.), and pups from postnatal, prepuberty and puberty age groups received an escalating dose regimen to simulate “binge-dosing” commonly seen in humans abusing MA. After MA exposure, rats were housed normally until behavioural testing on postnatal day 60 (ND+60), which included the novel object recognition test (NOR), open field test (OFT) and forced swim test (FST), measuring cognitive function, locomotor activity and depressive-like behaviour respectively. The FST data showed increased immobility behaviour of saline-treated FSL rats relative to that of FRL rats, in line with previous data validating FSL rats as a genetic rodent model of depression. Practically significant MA-induced increases in immobility behaviour were observed in all FSL and FRL treatment groups in the FST, reaching statistical significance in prenatally treated FRL rats, and in postnatally, prepuberty and puberty treated FSL rats. The data suggest that early-life MA exposure may alter neurodevelopment to predispose the rats to display depressive-like behaviour in early adulthood, and suggests that this detrimental effect of MA may be more expressed in stress-sensitive rats. Furthermore, all FSL groups plus prenatally and puberty treated FRL rats revealed practically and statistically significant decreases in swimming behaviour in the FST, whereas decreases in swimming behaviour in prepuberty treated FRL rats were practically significant but did not reach statistical significance. These data suggest that MA-induced depressive-like behaviour in FSL rats may be related to impaired serotonergic neurotransmission, and that this appears to be more robust in FSL rats. Climbing behaviour in the FST was generally not altered by early-life MA exposure, with a notable exception being a practically and statistically significant increase in puberty treated FRL rats. These data suggest that in general early-life MA exposure does not affect noradrenergic neurotransmission in early adulthood, except when normal rats were treated at puberty. The reason for the latter observation is not clear. The data from the NOR test revealed no discernible trends of MA-induced effects on memory and cognition, except for a small albeit practically significant increase in exploration time in prepuberty treated FRL rats and a practically and statistically significant decrease in exploration time in puberty-treated FRL rats. Lastly, locomotor activity in the OFT was mostly unaffected by MA treatments, except for practically significant decreases in locomotor activity in postnatally-and prepuberty-treated FRL rats and practically and statistically significant decreases in locomotor activity of prepuberty treated FSL rats. Altered locomotor activity is therefore not expected to explain any of the immobility results of the FST. In final conclusion, the study confirms that early-life MA exposure results in a depressogenic effect later in life in stress-sensitive (FSL) and control (FRL) rats, but appears to be more robust in stress-sensitive animals. Furthermore the data suggest that long-lasting MA-induced depressogenic effects may relate to impaired serotonergic neurotransmission. / MSc (Pharmacology), North-West University, Potchefstroom Campus, 2014

Methamphetamine (MA) is a well-known, easily accessible and powerful psychostimulant, and its abuse has become a global problem. MA abuse affects millions of people worldwide and places an enormous burden on public healthcare resources. Documented consequences of MA abuse include cardiotoxic, neurotoxic and teratogenic effects, as well as long-term consequences of chronic abuse including affective disorders such as schizophrenia and major depressive disorder (MDD). MDD is a highly prevalent mood disorder in both adults and children, documented to contribute to approximately 850 000 suicides annually. This disorder is projected to become the 2nd leading disease of global burden by 2020, preceded only by ischemic heart disease. Depressive-like behaviour is documented as a symptom of chronic MA abuse and particularly during extensive MA withdrawal. Also, MA abuse during pregnancy is documented to cause neurodevelopmental changes that persist into later life. However, current understanding thereof is limited and warrants further investigation of the effects of early-life exposure to MA on outcome in adulthood, particularly in terms of mood disorders. The aim of the current study was to determine the effect of chronic exposure to MA on the depressive-like behaviour later in life in stress-sensitive (Flinders Sensitive Line) and control (Flinders Resistant Line) rats. Rats were exposed during one of the following natal day (ND) age groups: prenatal (ND-13 to ND+02), postnatal (ND+03 to ND+18), prepuberty (ND+19 to ND+34) or puberty (ND+35 to ND+50). These age groups represent different stages in neurodevelopment, as also seen in humans. For prenatal exposure, pregnant dams received 5 mg/kg daily subcutaneously (s.c.), and pups from postnatal, prepuberty and puberty age groups received an escalating dose regimen to simulate “binge-dosing” commonly seen in humans abusing MA. After MA exposure, rats were housed normally until behavioural testing on postnatal day 60 (ND+60), which included the novel object recognition test (NOR), open field test (OFT) and forced swim test (FST), measuring cognitive function, locomotor activity and depressive-like behaviour respectively. The FST data showed increased immobility behaviour of saline-treated FSL rats relative to that of FRL rats, in line with previous data validating FSL rats as a genetic rodent model of depression. Practically significant MA-induced increases in immobility behaviour were observed in all FSL and FRL treatment groups in the FST, reaching statistical significance in prenatally treated FRL rats, and in postnatally, prepuberty and puberty treated FSL rats. The data suggest that early-life MA exposure may alter neurodevelopment to predispose the rats to display depressive-like behaviour in early adulthood, and suggests that this detrimental effect of MA may be more expressed in stress-sensitive rats. Furthermore, all FSL groups plus prenatally and puberty treated FRL rats revealed practically and statistically significant decreases in swimming behaviour in the FST, whereas decreases in swimming behaviour in prepuberty treated FRL rats were practically significant but did not reach statistical significance. These data suggest that MA-induced depressive-like behaviour in FSL rats may be related to impaired serotonergic neurotransmission, and that this appears to be more robust in FSL rats. Climbing behaviour in the FST was generally not altered by early-life MA exposure, with a notable exception being a practically and statistically significant increase in puberty treated FRL rats. These data suggest that in general early-life MA exposure does not affect noradrenergic neurotransmission in early adulthood, except when normal rats were treated at puberty. The reason for the latter observation is not clear. The data from the NOR test revealed no discernible trends of MA-induced effects on memory and cognition, except for a small albeit practically significant increase in exploration time in prepuberty treated FRL rats and a practically and statistically significant decrease in exploration time in puberty-treated FRL rats. Lastly, locomotor activity in the OFT was mostly unaffected by MA treatments, except for practically significant decreases in locomotor activity in postnatally-and prepuberty-treated FRL rats and practically and statistically significant decreases in locomotor activity of prepuberty treated FSL rats. Altered locomotor activity is therefore not expected to explain any of the immobility results of the FST. In final conclusion, the study confirms that early-life MA exposure results in a depressogenic effect later in life in stress-sensitive (FSL) and control (FRL) rats, but appears to be more robust in stress-sensitive animals. Furthermore the data suggest that long-lasting MA-induced depressogenic effects may relate to impaired serotonergic neurotransmission. / MSc (Pharmacology), North-West University, Potchefstroom Campus, 2014

As technology is increasingly used in mobile settings, energy and battery management is becoming a part of everyday life. Many have experienced how quickly a battery can be depleted in a smartphone, laptop or electric cars, sometimes causing much distress. An important question is how we can understand and work with energy as a factor in interaction design to enable better experiences for end-users. Through design-oriented research, I have worked with the specific case of electric cars, which is currently a domain where people struggle in terms of energy management. The main issue in this use case is that current driving range estimates cause distrust and anxiety among drivers. Through sketches, prototypes and studies, I investigated causes as well as possible remedies to this situation. My conclusion is that instead of providing black-boxed predictions, in-car interfaces should expose the logics of estimates so that drivers know how their own actions in e.g. driving style, climate control, and other equipment, affects energy use. Revealing such energy mechanisms will not only empower the driver, it will also acknowledge the impact of variables that cannot be predicted automatically. In this work, understanding the dynamic aspects of energy has emerged as central to interaction with systems. This points to a need to design energy sensitive interactions - focusing on supporting users to find the right balance between energy use and the experiential values sought for. To ease design of energy sensitive interactions, energy use is divided into three different categories with accompanying ideals. These are exergy (always needed to achieve the required interaction), intergy (controllable and changing over time and use, needs to be addressed in design), and anergy (always waste that needs to be reduced). This articulation highlights aspects of energy that are specific to interaction design, and possible aspects to expose to allow more energy-efficient interactions in use. / I takt med att vi använder alltmer teknik i mobila sammanhang blir energi- och batterihantering en allt större del av vår vardag. Många har erfarenheter av de besvär som ett plötsligt urladdat batteri i en mobiltelefon, laptop eller elbil kan orsaka. En central fråga för att uppnå bättre användarupplevelser är hur vi kan förstå och arbeta med energi som en faktor i design av interaktion med mobil teknik. Genom designdriven forskning har jag arbetat specifikt med interaktionen i elbilar, en situation där många brottas med just hantering och förståelse av begränsad energi. En specifik utmaning i denna kontext är den misstro som många upplever kring existerande system för räckviddsberäkning. Genom skisser, prototyper och användarstudier har jag undersökt orsaker och praktiska lösningar på detta problem. Min slutsats är att bilens gränssnitt bör exponera den inre logik som beräkningarna bygger på, så att föraren förstår hur egna handlingar, såsom körsätt och användning av t ex kupévärmare, påverkar energiförbrukning och räckvidd. Detta leder till ökad upplevelse av kontroll för föraren, och samtidigt till mer tillförlitliga beräkningar då det tar hänsyn till variabler som inte kan förutsägas automatiskt. I arbetet har dynamiska aspekter av energi framträtt som centralt i användning av interaktiva system. Detta pekar på behovet av att designa energikänsliga interaktioner, som hjälper användaren att förstå balansen mellan energiåtgång och bruksvärde. För att stödja design av energikänsliga interaktioner artikuleras tre kategorier av energianvändning i interaktiva system. Dessa är exergi (behövs för att uppnå tänkt interaktion), intergi (kontrollerbar och föränderlig över tid och användning, måste adresseras med design), och anergi (är alltid ett slöseri som behöver reduceras). Denna artikulation belyser specifikt de aspekter av energiförbrukningen som varierar genom användning, och som skulle kunna exponeras för mer energieffektiv interaktion med ny teknik. / <p>QC 20160429</p>

Imaging membrane potential is a promising technique in the elucidation of the interactions of large networks of neurons. The membrane potential in a neuron varies as an action potential, the basic electrical signal of neuronal communication, travels along the length of the cell. Voltage sensitive dyes play a key role by providing an optical readout of the electric field generated across a neuron membrane by the action potential. However, none of the dyes reviewed in Chapter 1 generate sufficient signal change with changes in membrane potential; this sensitivity problem limits the ability of the imaging membrane potential technique to allow the high spatial and temporal resolution necessary for neuronal networks to be better understood. This thesis features two avenues of research that are expected to result in the necessary enhancements to voltage sensitive dyes to improve the signal change. The first avenue is based on the effect of an electric field upon the non-linear optical properties of a porphyrin macromolecule. The encouraging field sensitivity of a previous porphyrin monomer voltage sensor inspired an investigation which identified optimisations to enhance the voltage sensitivity (Chapter 2). The design, synthesis and initial characterisation of optimised porphyrin voltage sensors is detailed in Chapter 3. The second avenue is based on the effect of an electric field upon the rate of intermolecular electron transfer. In a suitably designed dye, the competition between electron transfer and fluorescence, following excitation by incoming light, allows the fluorescence intensity to act as an optical indicator of the electron transfer rate. New dyes were rationally designed and synthesised, as this effect had not been applied to voltage sensitive imaging before the research detailed in Chapter 4. The challenging purification of the new amphiphilic dyes synthesised also inspired research into a novel testing method which does not require amphiphilic dyes (Chapter 5).

The MCM helicase plays an important role in eukaryotic DNA replication, unwinding double stranded DNA ahead of the replication fork. MCM is a hetero-hexamer consisting of the six related proteins, Mcm2-Mcm7. The distantly related MCM-binding protein (MCM-BP) was first identified in a screen for proteins interacting with MCM2-7 in human cells and was found to specifically interact with Mcm3-7 but not Mcm2. It is conserved in most eukaryotes and seems to play an important role in DNA replication but its exact function is not clear yet. This study contributes to the understanding of the fission yeast homologue of MCM-BP, named Mcb1, but also of MCM-BP in general. Results presented in this thesis document the initial biochemical characterisation of the complex Mcb1 forms with Mcm proteins, the MCMᴹᶜᵇ¹ complex. Interactions of Mcb1 with Mcm proteins, potential interaction sites between the proteins and the size of the complex were analysed using a variety of methods, including tandem affinity purification, co-immunoprecipitation, sucrose gradients and in vitro pull-down assays. Sequence analysis and structure prediction were utilised to gain some insight into Mcb1 and MCM-BP ancestry and structure. Results presented here indicate that fission yeast Mcb1 shares homology with Mcm proteins and forms a complex with Mcm3-Mcm7 but not Mcm2 and thus replaces the latter in an alternative high molecular weight complex that is likely to have an MCM-like appearance. Deletion of mcb1⁺ showed that Mcb1 is essential in fission yeast. To examine the cellular function of the protein, temperature-sensitive mutants were generated. Inactivation of Mcb1 leads to an increase in DNA damage and cell cycle arrest in G2-phase depending on the activation of the Chk1 dependent DNA damage checkpoint. Similar observations were made when Mcb1 was overexpressed, indicating that certain levels of the protein are important for accurate DNA replication. Construction of truncated versions of Mcb1 suggested that almost the full-length protein is needed for proper function.

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Viso pasaulio ir atskirų valstybių ekonomika pasižymi tuo, jog pagaminamos produkcijos kiekis kasmet kinta ir ilgu laikotarpiu būna nepastovus. Vienais metais produkcijos pagaminama daugiau, kitais mažiau. Pradėjusi mažėti gamybos apimtis sukelia daugybę šalutinių efektų: ima mažėti įmonių pajamos ir pelnai, mažiau pinigų lieka investicijoms, mažėja produktyvumas, didėja prekių atsargos sandėliuose, galiausiai tenka atleisti dalį darbuotojų ar sumažinti darbo užmokestį. Žmonės, gaunantys mažesnes pajamas, išleidžia mažiau pinigų ir taip priverčia įmones dar labiau riboti savo išlaidas. Tokiu būdu patenkama į užburtą ratą, kai priežastys tampa pasekmėmis ir jos tarpusavy kartojasi, tačiau ne visos ūkio šakos vienodai stipriai reaguoja į ekonomikos svyravimus. Darbo objektas – skirtingų ūkio sektorių jautrumas verslo ciklams. Darbo tikslas – nustatyti individualų skirtingų ūkio sektorių jautrumą verslo ciklams. Darbo uždaviniai: • Išanalizuoti verslo ciklo sąvoką, siekiant suvokti jo įtaką visai ekonomikai ir įmonėms • Nustatyti ciklinių svyravimų priežastis bei pasekmes, kurios atsiranda ekonomikai esant žemiausioje verslo ciklo fazėje • Remiantis moksliniais darbais bei specializuotų agentūrų informacija, išnagrinėti verslo ciklų indikatorius, nustatant jų reikšmę verslo ciklų analizėje • Išanalizuoti įmonėje dėl ekonomikos cikliškumo susidarantį užburtą ratą, kuris leistų suprasti procesus, vykstančius ciklų metu • Remiantis mokslininkų tyrimais, nustatyti daugiausiai ir... [toliau žr. visą tekstą] / The economy of all the world and different counties is characterized by the annual changes of production volume, that in the long-term is erratic. One year the amount of production is higher, others lower. The decrease in volume of production causes many side effects such as declining corporate earnings and profits, less money for investment, declining productivity, increasing inventory of goods in warehouses, then it leads firing workers or reducing wages. People with lower levels of income, spend less money and that makes companies even more to limit their costs. In this way, we get into the vicious circle, where causes become results and they inter-repeat, but not all economic sectors equally respond to economic fluctuations. Object – different economic sectors sensitivity to business cycles. The aim – to identify the individual sensitivity of different economic sectors to business cycles. Objectives: • To explore the concept of the business cycle, in order to understand it‘s impact on the economy and companies. • To find out the causes and the consequences of the cyclical fluctuations, that appears then the economy is in the lowest stage of the business cycle. • On the basis of scientific works and information of the specialized agencies, to analyze the business cycle indicators determining their value in business cycle analysis. • To analyze a vicious circle in the company, forming because of cycles of economy, which would allow to understand the processes during the... [to full text]

Timeliness in delivering packets for delay-sensitive applications is an important QoS (Quality of Service) measure in many systems, notably those that need to provide real-time performance. In such systems, if delay-sensitive traffic is delivered to the destination beyond the deadline, then the packets will be rendered useless and dropped after received at the destination. Bandwidth that is already scarce and shared between network nodes is wasted in relaying these expired packets. This thesis proposes that a deterministic per-hop delay can be achieved by using a dynamic queue threshold concept to bound delay of each node. A deterministic per-hop delay is a key component in guaranteeing a deterministic end-to-end delay. The research aims to develop a generic approach that can constrain network delay of delay-sensitive traffic in a dynamic network. Two adaptive queue management schemes, namely, DTH (Dynamic THreshold) and ADTH (Adaptive DTH) are proposed to realize the claim. Both DTH and ADTH use the dynamic threshold concept to constrain queuing delay so that bounded average queuing delay can be achieved for the former and bounded maximum nodal delay can be achieved for the latter. DTH is an analytical approach, which uses queuing theory with superposition of N MMBP-2 (Markov Modulated Bernoulli Process) arrival processes to obtain a mapping relationship between average queuing delay and an appropriate queuing threshold, for queue management. While ADTH is an measurement-based algorithmic approach that can respond to the time-varying link quality and network dynamics in wireless ad hoc networks to constrain network delay. It manages a queue based on system performance measurements and feedback of error measured against a target delay requirement. Numerical analysis and Matlab simulation have been carried out for DTH for the purposes of validation and performance analysis. While ADTH has been evaluated in NS-2 simulation and implemented in a multi-hop wireless ad hoc network testbed for performance analysis. Results show that DTH and ADTH can constrain network delay based on the specified delay requirements, with higher packet loss as a trade-off.