Screen Printed Particle-Based Microfluidics: Optimization and Exemplary Application for Heavy Metals Analysis.

In this work, a screen-printing method was developed to create porous particle-based materials as layers with specifically designed shape to produce microfluidics systems. Among several tested binding agents, xanthan gum was found to be an excellent choice for a printing mixture thickener as well as a durable binder for the resulting material. In addition to demonstrating control over the shape of the printed microfluidics chips, control over material thickness, wetting characteristics and general method accuracy were also investigated. The applicability of the introduced method was further demonstrated with a development of an exemplary microfluidics chip for quantitative detection of Fe (III), Ni (II), Cu (II), Cd (II), and Pb (II) from a mixed sample at millimolar levels. The novel approaches demonstrated in this article offer new perspective into creating multiplexed on-site chemical analysis tests.

A New Direction in Microfluidics: Printed Porous Materials.

In this work, the feasibility of a novel direction for microfluidics is studied by demonstrating a set of new methods to fabricate microfluidic systems. Similarly to microfluidic paper-based analytical devices, porous materials are being used. However, alternative porous materials and different printing methods are used here to give the material the necessary pattern to act as a microfluidic system. In this work, microfluidic systems were produced by the following three separate methods: (1) by curing a porous monolithic polymer sheet into a necessary pattern with photolithography, (2) by screen printing silica gel particles with gypsum, and (3) by dispensing silica gel particles with polyvinyl acetate binder using a modified 3D printer. Different parameters of the printed chips were determined (strength of the printed material, printing accuracy, printed material height, wetting characteristics, repeatability) to evaluate whether the printed chips were suitable for use in microfluidics. All three approaches were found to be suitable, and therefore the novel approach to microfluidics was successfully demonstrated.

Association between self-reported binge drinking and absenteeism in the Baltic countries.

Objective: Substantial loss of productivity due to absenteeism is associated with alcohol use. This study examined the associations between absenteeism in the workplace and in schools and binge drinking across various beverage types in the Baltic countries. Methods: We utilised a dataset of 3,778 individuals compiled from 2015 to 2016 and performed multiple negative binomial regression analysis with multiple imputations to deal with missing data. Self-reported measures were used for both absenteeism and binge drinking. Results: We found evidence to support the claim that absenteeism, in terms of self-reported absence days, is positively associated with self-reported binge drinking, specifically with beer bingeing. On average, beer bingers reported 49% (p < .05) more absences than people who drink alcohol but do not binge on beer. For wine and spirits variables, the estimates indicated positive but statistically insignificant associations. No group differences were identified across gender and education. Conclusions: A considerable proportion of days absent from work and from school can be associated with beer bingeing. Therefore, it should be acknowledged that beverage-specific alcohol policies that are more lenient toward beer than other types of alcohol can inadvertently increase absenteeism and decrease workplace productivity.

Quantifying the social costs and benefits of tobacco in Estonia.

BACKGROUND: The tobacco cost literature has predominantly focused on estimating direct healthcare costs and productivity losses from morbidity and mortality. This study places a greater focus on the effects that arise through the fiscal system, illicit trade and fire accidents to estimate the social costs and benefits of tobacco use in Estonia in 2018. METHODS: A prevalence-based cost-of-illness approach was used, relying on data from the 2017-2018 period. Age-specific tobacco-attributable fractions were employed to estimate the costs and benefits of tobacco-related morbidity and mortality. Data on mortality, morbidity and healthcare costs were received from the National Institute of Health Development and the Estonian Health Insurance Fund. We used data for offences and fires from the Estonian Rescue Board and the Estonian Tax and Customs Board. RESULTS: It was estimated that the total social cost attributable to tobacco in Estonia in 2018 was €174 million, representing 0.67% of the nation's GDP. If future benefits are also considered, the net social cost is €78 million. While the greatest cost components are productivity losses from mortality and morbidity, the level of tobacco-related law enforcement costs are comparable to direct healthcare costs, both of which exceed €10 million annually. CONCLUSIONS: Despite the substantial social benefits arising through the fiscal system, tobacco-related costs outweighed these benefits in Estonia in 2018. In addition, a considerable portion of the economic burden of tobacco is associated with illicit trade. Therefore, the healthcare and law enforcement sectors would both benefit from effective tobacco control policies.

Design, synthesis and application of carbazole macrocycles in anion sensors.

Carboxylate sensing solid-contact ion-selective electrodes (ISEs) were created to provide a proof-of-concept ISE development process covering all aspects from in silico ionophore design to functional sensor characterization. The biscarbazolylurea moiety was used to synthesize methylene-bridged macrocycles of different ring size aiming to fine tune selectivity towards different carboxylates. Cyclization was achieved with two separate strategies, using either amide synthesis to access up to -[CH2]10- macrocycles or acyl halides to access up to -[CH2]14- macrocycles. Seventy-five receptor-anion complexes were modelled and studied with COSMO-RS, in addition to all free host molecules. In order to predict initial selectivity towards carboxylates, 1H NMR relative titrations were used to quantify binding in DMSO-d 6/H2O solvent systems of two proportions - 99.5%:0.5% m/m and 90.0%:10.0% m/m, suggesting initial selectivity towards acetate. Three ionophores were selected for successful sensor prototype development and characterization. The constructed ion-selective electrodes showed higher selectivity towards benzoate than acetate, i.e., the selectivity patterns of the final sensors deviated from that predicted by the classic titration experiments. While the binding constants obtained by NMR titration in DMSO-d 6/H2O solvent systems provided important guidance for sensor development, the results obtained in this work emphasize the importance of evaluating the binding behavior of receptors in real sensor membranes.

The effects of the lower ignition propensity cigarettes standard in Estonia: time-series analysis.

BACKGROUND: In 2011, the lower ignition propensity (LIP) standard for cigarettes was implemented in the European Union. Evidence about the impact of that safety measure is scarce. OBJECTIVE: The aim of this paper is to examine the effects of the LIP standard on fire safety in Estonia. METHODS: The absolute level of smoking-related fire incidents and related deaths was modelled using dynamic time-series regression analysis. The data about house fire incidents for the 2007-2013 period were obtained from the Estonian Rescue Board. RESULTS: Implementation of the LIP standard has reduced the monthly level of smoking-related fires by 6.2 (p<0.01, SE=1.95) incidents and by 26% (p<0.01, SE=9%) when estimated on the log scale. Slightly weaker evidence was found about the fatality reduction effects of the LIP regulation. All results were confirmed through counterfactual models for non-smoking-related fire incidents and deaths. CONCLUSIONS: This paper indicates that implementation of the LIP cigarettes standard has improved fire safety in Estonia.

Ala5-galanin (2-11) is a GAL2R specific galanin analogue.

It is over 30years since the regulatory peptide galanin was discovered by Professor Mutt and co-workers. Galanin exerts its effects by binding to three galanin G-protein coupled receptors, namely GAL1R, GAL2R and GAL3R. Each galanin receptor has a different distribution in the central nervous system and the peripheral nervous system as well as distinctive signaling pathways, which implicates that the receptors are involved in different biological- and pathological effects. The delineation of the galaninergic system is however difficult due to a lack of stable, specific galanin receptor ligands. Herein, a new short GAL2R specific ligand, Ala5-galanin (2-11), is presented. The galanin (2-11) modified analogue Ala5-galanin (2-11) was tested in 125I-galanin competitive binding studies for the three galanin receptors and the G-protein coupled receptor signaling properties was tested by the ability to influence second-messenger molecules like inositol phosphate and cyclic adenosine monophosphate. In addition, two different label-free real-time assays, namely EnSpire® based on an optical biosensor and xCELLigence® based on an electric biosensor, were used for evaluating the signaling properties using cell lines with different levels of receptor expression. Ala5-galanin (2-11) was subsequently found to be a full agonist for GAL2R with more than 375-fold preference for GAL2R compared to both GAL1R and GAL3R. The single amino acid substitution of serine to alanine at position 5 in the short ligand galanin (2-11) resulted in a ligand subsequently unable to bind neither GAL3R nor GAL1R, even at concentrations as high as 0.1mM.

Pharmacological stimulation of GAL1R but not GAL2R attenuates kainic acid-induced neuronal cell death in the rat hippocampus.

The neuropeptide galanin is widely distributed in the central and peripheral nervous systems and part of a bigger family of bioactive peptides. Galanin exerts its biological activity through three G-protein coupled receptor subtypes, GAL1-3R. Throughout the last 20years, data has accumulated that galanin can have a neuroprotective effect presumably mediated through the activation of GAL1R and GAL2R. In order to test the pharmaceutical potential of galanin receptor subtype selective ligands to inhibit excitotoxic cell death, the GAL1R selective ligand M617 and the GAL2R selective ligand M1145 were compared to the novel GAL1/2R ligand M1154, in their ability to reduce the excitotoxic effects of intracerebroventricular injected kainate acid in rats. The peptide ligands were evaluated in vitro for their binding preference in a competitive (125)I-galanin receptor subtype binding assay, and G-protein signaling was evaluated using both classical signaling and a label-free real-time technique. Even though there was no significant difference in the time course or severity of the kainic acid induced epileptic behavior in vivo, administration of either M617 or M1154 before kainic acid administration significantly attenuated the neuronal cell death in the hippocampus. Our results indicate the potential therapeutic value of agonists selective for GAL1R in the prevention of neuronal cell death.

Do alcohol excise taxes affect traffic accidents? Evidence from Estonia.

OBJECTIVE: This article examines the association between alcohol excise tax rates and alcohol-related traffic accidents in Estonia. METHODS: Monthly time series of traffic accidents involving drunken motor vehicle drivers from 1998 through 2013 were regressed on real average alcohol excise tax rates while controlling for changes in economic conditions and the traffic environment. Specifically, regression models with autoregressive integrated moving average (ARIMA) errors were estimated in order to deal with serial correlation in residuals. Counterfactual models were also estimated in order to check the robustness of the results, using the level of non-alcohol-related traffic accidents as a dependent variable. RESULTS: A statistically significant (P <.01) strong negative relationship between the real average alcohol excise tax rate and alcohol-related traffic accidents was disclosed under alternative model specifications. For instance, the regression model with ARIMA (0, 1, 1)(0, 1, 1) errors revealed that a 1-unit increase in the tax rate is associated with a 1.6% decrease in the level of accidents per 100,000 population involving drunk motor vehicle drivers. No similar association was found in the cases of counterfactual models for non-alcohol-related traffic accidents. CONCLUSIONS: This article indicates that the level of alcohol-related traffic accidents in Estonia has been affected by changes in real average alcohol excise taxes during the period 1998-2013. Therefore, in addition to other measures, the use of alcohol taxation is warranted as a policy instrument in tackling alcohol-related traffic accidents.

Novel systemically active galanin receptor 2 ligands in depression-like behavior.

Neuropeptide galanin and its three G-protein coupled receptors, galanin receptor type 1-galanin receptor type 3 (GalR1-GalR3), are involved in the regulation of numerous physiological and disease processes, and thus represent tremendous potential in neuroscience research and novel drug lead development. One of the areas where galanin is involved is depression. Previous studies have suggested that activation of GalR2 leads to attenuation of depression-like behavior. Unfortunately, lack of in vivo usable subtype specific ligands hinders testing the role of galanin in depression mechanisms. In this article, we utilize an approach of increasing in vivo usability of peptide-based ligands, acting upon CNS. Thus, we have synthesized a series of novel systemically active galanin analogs, with modest preferential binding toward GalR2. We have shown that specific chemical modifications to the galanin backbone increase brain levels upon i.v. injection of the peptides. Several of the new peptides, similar to a common clinically used antidepressant medication imipramine, exerted antidepressant-like effect in forced swim test, a mouse model of depression, at a surprisingly low dose range (< 0.5 mg/kg). We chose one of the peptides, J18, for more thorough study, and showed its efficacy also in another mouse depression model (tail suspension test), and demonstrated that its antidepressant-like effect upon i.v. administration can be blocked by i.c.v. galanin receptor antagonist M35. The effect of the J18 was also abolished in GalR2KO animals. All this suggests that systemically administered peptide analog J18 exerts its biological effect through activation of GalR2 in the brain. The novel galanin analogs represent potential drug leads and a novel pharmaceutical intervention for depression.

Galanin, through GalR1 but not GalR2 receptors, decreases motivation at times of high appetitive behavior.

Galanin is a 29/30-amino acid long neuropeptide that has been implicated in many physiological and behavioral functions. Previous research has shown that i.c.v. administration of galanin strongly stimulates food intake in sated rats when food is freely available, but fails to stimulate this consumption when an operant response requirement is present. Using fixed ratio (FR) schedules, we sought to further clarify galanin's role in motivated behavior by administering galanin i.c.v. to rats working on fixed ratio schedules requiring either a low work condition (FR1) or higher work conditions (FR>1) to obtain a 0.2% saccharin reward. Rats in the FR>1 group were assigned to either an FR3, FR5 or FR7 schedule of reinforcement. The rate of reinforcement decreased for only the FR>1 group as compared to saline controls. Furthermore, injections of GalR1 receptor agonist M617 led to a similar, marginally significant decrease in the number of reinforcers received in the FR>1 condition, but a decrease was not seen after injections of GalR2 receptor agonist M1153. Taken together, these results show that galanin may be playing a role in decreasing motivation at times of high appetitive behavior, and that this effect is likely mediated by the GalR1 receptor.

Novel galanin receptor subtype specific ligand in depression like behavior.

Neuropeptide galanin and its three receptors, galanin receptor type 1-galanin receptor type 3, are known to be involved in the regulation of numerous psychological processes, including depression. Studies have suggested that stimulation of galanin receptor type 2 (GalR2) leads to attenuation of the depression-like behavior in animals. However, due to the lack of highly selective galanin subtype specific ligands the involvement of different receptors in depression-like behavior is yet not fully known. In the present study we introduce a novel GalR2 selective agonist and demonstrate its ability to produce actions consistent with theorized GalR2 functions and analogous to that of the anti-depressant, imipramine.

Cutting edge: IL-13Rα1 expression in dopaminergic neurons contributes to their oxidative stress-mediated loss following chronic peripheral treatment with lipopolysaccharide.

Inflammation and its mediators, including cytokines and reactive oxygen species, are thought to contribute to neurodegeneration. In the mouse brain, we found that IL-13Rα1 was expressed in the dopaminergic (DA) neurons of the substantia nigra pars compacta, which are preferentially lost in human Parkinson's disease. Mice deficient for Il13ra1 exhibited resistance to loss of DA neurons in a model of chronic peripheral inflammation using bacterial LPS. IL-13, as well as IL-4, potentiated the cytotoxic effects of t-butyl hydroperoxide and hydrogen peroxide on mouse DA MN9D cells. Collectively, our data indicate that expression of IL-13Rα1 on DA neurons can increase their susceptibility to oxidative stress-mediated damage, thereby contributing to their preferential loss. In humans, Il13ra1 lies on the X chromosome within the PARK12 locus of susceptibility to Parkinson's disease, suggesting that IL-13Rα1 may have a role in the pathogenesis of this neurodegenerative disease.

Novel galanin receptor subtype specific ligands in feeding regulation.

Galanin a 29/30-residue neuropeptide has been implicated in several functions in the central nervous system, including the regulation of food consumption. Galanin and its analogues administered intraventricularly or into the hypothalamic region of brain have been shown to reliably and robustly stimulate the consumption of food in sated rodents. Three galanin receptor subtypes have been isolated, all present in the hypothalamus, but little is known about their specific role in mediating this acute feeding response. Presently, we introduce several novel GalR2 selective agonists and then compare the most selective of these novel GalR2 subtype selective agonists to known GalR1 selective agonist M617 for their ability to stimulate acute consumption of several foods shown to be stimulated by central administration of galanin. GalR1 selective agonist M617 markedly stimulated acute consumption of high-fat milk, but neither GalR2 selective agonist affected either high-fat milk or cookie mash intake. The present results are consistent with the involvement of GalR1 in mediating the acute feeding consumption by galanin and suggest an approach applicable to exploring galanin receptor specificity in normal and abnormal behavior and physiology.

A novel GalR2-specific peptide agonist.

The galanin peptide family and its three receptors have with compelling evidence been implicated in several high-order physiological disorders. The co-localization with other neuromodulators and the distinct up-regulation during and after pathological disturbances has drawn attention to this neuropeptide family. In the current study we present data on receptor binding and functional response for a novel galanin receptor type 2 (GalR2) selective chimeric peptide, M1145 [(RG)(2)-N-galanin(2-13)-VL-(P)(3)-(AL)(2)-A-amide]. The M1145 peptide shows more than 90-fold higher affinity for GalR2 over GalR1 and a 76-fold higher affinity over GalR3. Furthermore, the peptide yields an agonistic effect in vitro, seen as an increase in inositol phosphate (IP) accumulation, both in the absence or the presence of galanin. The peptide design with a N-terminal extension of galanin(2-13), prevails new insights in the assembly of novel subtype specific ligands for the galanin receptor family and opens new possibilities to apply the galanin system as a putative drug target.

The social costs of alcohol misuse in Estonia.

The aim of this study was to estimate the social costs of alcohol misuse in Estonia in 2006. Using a prevalence-based cost-of-illness approach, both direct and indirect costs were considered, including tangible costs associated with health care, criminal justice, rescue services, damage to property, premature mortality, incarceration, incapability of working due to illnesses, and lower labor productivity. The results show that alcohol misuse cost Estonia more than EUR 200 million in 2006. The costs involved are estimated to represent 1.6% of the gross domestic product (GDP), which is relatively high in comparison with many other countries. In addition, the state receives less receipts from the alcohol excise tax than the costs that it incurs as a consequence of alcohol misuse, which points to the existence of economic inefficiency with respect to the alcohol market. The results of this study suggest that there is definitely a need for further cost-benefit analysis to reach a conclusion regarding the possible utility of government intervention.

Road traffic mortality in Estonia: alcohol as the main contributing factor.

Traffic fatalities are the leading cause of death among the young and middle-aged population in Estonia. The objective of this study was to reveal the pattern of traffic fatalities among the population aged 15 - 64 years and to determine the role of alcohol in their fatalities. The data were collected from post-mortem reports at the Estonian Bureau of Forensic Medicine from 2000 to 2002. Alcohol-related deaths were those with a blood alcohol concentration (BAC) equal or above 0.05 g/100 ml. Out of 512 victims, 401 were males and 111 were females. The greatest group were car occupants (58%) followed by pedestrians (31%). The portion of alcohol-related deaths was 70% among men and 44% among women. The mean BAC and percentage of alcohol-related deaths was significantly higher in pedestrian than in driver fatalities. Alcohol intoxication was identified as the most powerful contributing factor to traffic fatalities. The results provide more evidence for politicians to tackle alcohol abuse and unsafe traffic environments.