Magistro baigiamajame darbe atskleista, kad daugelis respondentų yra neapsisprendę dėl savo pozicijos GMO gamybos bei naudojimo atžvilgiu, tačiau beveik trečdalis asmenų GMO gamybai bei naudojimui nepritaria. Dauguma gyventojų mano, kad GMO ne tik kenkia žmogaus sveikatai, bet ir yra pavojingi, taip pat kenkia aplinkai, o GMO turinčių produktų naudojimas gali negrįžtamai pakeisti genetinį žmogaus kodą ir augalų bei gyvūnų populiacijose gali sukelti negrįžtamus mutacinius pokyčius. Vis dėlto GMO produkciją kai kurie gyventojai linkę naudoti dėl mažesnės jų kainos ir ilgesnio vartojimo galiojimo termino. Nors kai kurie respondentai ir buvo įsitikinę GMO teikiama nauda bei tolimesniu GMO gamybos bei naudojimo plitimu ateityje, tačiau beveik visi respondentai buvo įsitikinę ir tuo, jog GMO gamyba bei naudojimu labiausiai suinteresuotos yra kuo didesnio pelno siekiančios monopolinės kampanijos. Vienareikšmiškai galima teigti, jog nors gyventojai apie GMO informacijos daugiausiai ir gauna iš televizijos laidų, pokalbių su draugais pažįstamais ar giminaičiais bei iš spaudos leidinių arba naršydami internete, tačiau tos informacijos kiekis yra nepakankamas ir daugiau kaip pusė respondentų norėtų, jog informacijos apie GMO būtų pateikiama daugiau. / The Master’s final paper on GMO discovered that the majority of respondents have no strong opinion about production and use of GMO, but one third of responses on production and use of GMO were negative. Many people consider that GMO not only negatively affect human health, but also damage environment. Moreover, eating genetically-modified food can irreversibly change human’s genetic code as well as irreversible mutations in plants’ and animals’ populations. However, some people tend to use GM foods because they are cheaper and can be stored longer than other foods. Even though some of the respondents see benefits of GMO production and usage, almost all of the respondents claim that genetically engineered products interest mostly large companies that are seeking to profit from GMO. The most obvious finding to emerge from this study is that people learn about GMO from mass media and communicating with friends and relatives mostly, but they also admit the lack of credible information on GMO. More than a half of the respondents would like to receive more information about genetically modified organisms.

Legionella pneumophila est une bactérie aquatique qui prolifère en se répliquant à l’intérieur de cellules amibiennes. Elle peut persister dans ces environnements en vivant en communauté sous forme de biofilms. L’inhalation par l’Homme d’eau contaminée, vaporisée par les réseaux d’eau chaude ou les tours aéro‐réfrigérantes, peut mener à l’infection des macrophages pulmonaires qui se traduit par une grave pneumonie appelée légionellose. Le di‐GMP cyclique (diGMPc) est impliqué, chez diverses espèces bactériennes, dans la transition entre les modes de vie mobiles et sessiles, et chez certains pathogènes, dans la régulation de la virulence. Mon travail de thèse vise à démontrer l’implication des voies de signalisation à diGMPc dans le contrôle de la virulence et de la formation de biofilms par L. pneumophila. Cette implication a été étudiée grâce à l’inactivation systématique de chacun des gènes codant les protéines du métabolisme du diGMPc chez la souche L. pneumophila Lens. Notre étude a révélé que trois de ces protéines, Lpl0780, Lpl0922 et Lpl1118, sont spécifiquement requises pour le contrôle de la virulence et, plus particulièrement, pour la survie précoce lors de l’infection de cellules‐hôtes via l’orchestration de la sécrétion de facteurs de virulence dans la cellule‐hôte. Lpl1118 participerait également à la biogénèse de la vacuole de réplication. Cinq autres de ces protéines sont impliquées dans la régulation de la formation et de l’architecture des biofilms. L’une d’elles est, plus particulièrement, requise pour la formation de biofilms en présence d’oxyde nitrique. Ces résultats contribuent à une meilleure compréhension de l’organisation complexe et spécifique des voies de signalisation à diGMPc chez L. pneumophila et pourraient permettre d’envisager une lutte plus efficace contre ce pathogène / Legionella pneumophila is a bacterium that proliferates in fresh water environments through the replication within amoebas. These bacteria can persist in these environments through biofilm formation. The inhalation of aerosolized contaminated water through hot water systems or cooling towers can induce the infection of human lungs, leading to a severe pneumonia called legionellosis. Cyclic di‐GMP (c‐di‐GMP) in involved, in various bacterial species, in the motility‐to‐sessility transition, and in some pathogens, in virulence control. My work aims to demonstrate the involvement of signaling pathways that use c‐di‐GMP in virulence control and biofilm formation of L. pneumophila. This involvement was investigated by systematically inactivating each gene encoding a c‐di‐GMP‐metabolizing enzyme in L. pneumophila Lens strain. Our work revealed that 3 of these proteins, Lpl0780, Lpl0922 and Lpl1118 are specifically involved in virulence control and, particularly, in the early survival during host cell infection through the orchestration of virulence factors secretion within host cell. Lpl1118 is particularly required for replicative vacuole biogenesis. Five other proteins, participate in the formation and architecture of biofilms. One of them is more specifically involved in biofilm formation in the presence of nitric oxide. These results help to better understand the complexity and the specificity of c‐di‐GMP signaling pathways in L. pneumophila and should allow the exploration of more effective ways to fight this pathogen

Die fünf CSS-Domänen Phosphodiesterasen aus Escherichia coli K12 (E. coli) gehören zu den weit verbreiteten c-di-GMP-PDEs. Ein Vertreter, PdeC, wurde bereits von Herbst et al. (2018) charakterisiert. Durch DsbA/DsbB geförderte Disulfidbrückenbindung (DSB) in der CSS-Domäne von PdeC wird die PDE-Aktivität des Proteins gehemmt. Gegenteilig ist die freie Thiolform, in Abhängigkeit von der TM2 als Dimerisierungs-Domäne, enzymatisch aktiver. Diese Form wird von den periplasmatischen Proteasen DegP und DegQ prozessiert. Ein irreversibel aktiviertes TM2+EAL-Fragment wird generiert, dass durch weitere Proteolyse langsam entfernt wird. Die Reduktion der CSS-Domäne von PdeC zur der freien Thiolform stimuliert die PDE-Aktivität der EAL-Domäne in vitro. Zusammen mit den Daten von Herbst et al. (2018) wird die CSS-Domäne in dieser Arbeit als eine neue sensorische Domäne charakterisiert, dessen Aktivität durch einen DSB/Thiol-Schaltmechanismus reguliert wird. Alle fünf CSS-Domänen-PDEs von E. coli K12 weisen eine ähnliche Domänenarchitektur auf, jedoch unterscheiden sich Redox-Biochemie, Proteolyse und PDE-Aktivität innerhalb dieser Proteinfamilie. Auf Basis der PDE-Aktivität von Nicht-DSB-Varianten wurden PdeB, PdeC und PdeG als aktivierbare (Reduktion steigert die PDE-Aktivität) und PdeD und PdeN als nicht aktivierbare (Reduktion inaktiviert PDEs) charakterisiert. Ein weiterer Vertreter de CSS-Domänen PDEs, PdeN, scheint nicht über die Ausbildung einer DSB in der CSS-Domäne reguliert und aktiviert zu werden. Nach erfolgter Proteinbiosynthese wird die Proteinkonzentration vielmehr über den N-Terminus reguliert, wobei saure Wachstumsbedingungen das Protein maßgeblich induzieren und die Aktivität erhöhen. Wird das Protein erfolgreich in die Membran eingelagert, kann es bedingt durch die strukturelle DSB seine PDE-Aktivität entfalten und die Biofilmmatrixproduktion maßgeblich beeinflussen. / The five CSS domain phosphodiesterases from Escherichia coli K12 (E. coli) belong to the widespread group of c-di-GMP PDEs. One representative, PdeC, has already been characterized by Herbst et al. (2018). DsbA/DsbB promoted disulfide bond (DSB) formation in the CSS domain of PdeC inhibits the PDE activity of the protein. On the contrary, the free thiol form is more enzymatically active, depending on the TM2 as the dimerization domain. This form is processed by the periplasmic proteases DegP and DegQ. An irreversibly activated TM2 + EAL fragment is generated that is slowly removed by further proteolysis. The reduction of the CSS domain of PdeC to the free thiol form stimulates the PDE activity of the EAL domain in vitro. Together with the data from Herbst et al. (2018) the CSS domain is characterized as a new sensory domain whose activity is regulated by a DSB / thiol switch mechanism. All five E. coli K12 CSS domain PDEs share a similar domain architecture, but redox biochemistry, proteolysis, and PDE activity differ within the protein family. On the basis of the PDE activity of non-DSB variants, PdeB, PdeC and PdeG were characterized as activatable (reduction increases PDE activity) and PdeD and PdeN as non-activatable (reduction inactivated PDE activity). Another representative of the CSS domain PDEs, PdeN, does not seem to be regulated and activated by forming a DSB in the CSS domain. After protein biosynthesis the protein concentration is rather regulated via the N-terminus, with acidic growth conditions significantly inducing the protein and increasing its activity. If the protein is successfully inserted in the membrane, it can develop its PDE activity due to the structural DSB and influence the biofilm matrix production significantly.

Cyclic 3',5'-guanosine monophosphate (cGMP) is a key signaling molecule involved in a myriad of physiological processes, including vascular smooth muscle (VSM) tone, water- and electrolyte homeostasis, platelet aggregation, airway smooth muscle tone, smooth muscle proliferation and bone formation. Increased occurrence of vascular disorders including erectile dysfunction, hypertension, stroke and coronary artery disease, have made it increasingly important to study the dynamic interplay between cGMP synthesis and hydrolysis in VSM cells. This dissertation examines the spatial distribution of intracellular cGMP, [cGMP]i, in response to NO and atrial natriuretic peptide (ANP) in VSM cells. To investigate the spatial patterning of [cGMP]i, we have developed a new generation of non-FRET (fluorescence resonance energy transfer) cGMP biosensors that are suitable to monitor [cGMP]i in response to physiological (low-nanomolar) NO and ANP concentrations and that qualify for real-time, confocal imaging techniques. We have termed these indicators FlincGs, for green fluorescent indicators of cGMP. For the development of FlincGs, we made use of the specific cGMP binding characteristics of PKG. We utilized site-specific mutagenesis, kinetic cGMP binding, dissociation and kinase assays, as well as crystallography, in order to investigate PKG activation and cGMP binding dynamics in greater detail. Based on these studies, our novel, non-FRET cGMP biosensors were designed by attaching cGMP binding fragments of PKG to the N-terminus of circular permutated green fluorescent protein. We applied FlincGs in cultured VSM cells as well as in intact tissue to determine whether two spatially distinct populations of guanlylyl cyclase (cytosolic versus membrane bound) underlie the generation of spatiotemporally-specific patterns of [cGMP]i formation.

Thesis (Ph. D.)--University of Washington, 2001. / Vita. Includes bibliographical references (leaves 113-137).

Thesis (Ph. D.)--University of Washington, 2001. / Vita. Includes bibliographical references (leaves 118-135).

Posttraumatic stress disorder (PTSD) is a severe anxiety disorder characterised by hypothalamic-pituitary-adrenal (HPA)-axis abnormalities, hyperarousal, anxiety, flashbacks of trauma memories and avoidance. Increasing evidence is now accumulating that the disorder is also associated with shrinkage of the hippocampus and cognitive dysfunction that may have its origin in stress-induced excitotoxicity. Animal studies have indeed highlighted a potential role of the excitotoxic glutamatenitric oxide (NO) pathway in the stress response. Since PTSD appears to be an illness that progresses and worsens over time after an initial severe traumatic event, this study has used an animal model that emphasises repeated trauma to investigate the effect of stress on hippocampal NO synthase (NOS) activity, the release of the nitrogen oxide metabolites of NO (NOx), and also the evoked release of cGMP. Furthermore, the modulation and dependency of these responses on glutamate, NO and cGMP activity using drugs selective for these targets, will also be investigated. Rats (n=10/group) were exposed to repeated stress together with saline or drug administration immediately after the stress procedure and continuing for one week post-stress. The animals were then sacrificed for assay of hippocampal NOS activity, NO, and cGMP accumulation. Animals received either the glutamate-NMDA receptor antagonist, memantine (MEM;5mg/kg ip/d), the neuronal NOS selective inhibitor, 7- nitroindazole monosodium salt (7-NINA;20mg/kg ip/d), the cGMP-specific PDE inhibitor, sildenafil (SIL;10mg/kg ip/d) or the NFkb antagonist, pyrollidine dithiocarbamate (PDTC;70mg/kg ip/d). The latter inhibits the nuclear transcription factor, NFkb, responsible for inducing the expression of iNOS, while it also appears to mediate the glutamatergic actions on NOS expression, Stress significantly increased hippocampal NOS activity, as well as significantly increased hippocampal cGMP and NO, levels. These increases were blocked by pretreatment with either PDTC or 7-NINA, while memantine was without effect. Sildenafil significantly augmented stress induced NO, accumulation, as well as cGMP. although the latter failed to reach significance. 7-NINA and memantine significantly blocked the increase in cGMP evoked by time-dependent sensitisation (TDS)-stress, with PDTC attenuating this response, but not significantly. Additionally, administration of each drug separately for seven days without exposure to stress, did not evoke significant changes in NOx levels, compared to the control group. However, significant increases in cGMP levels, compared to the control group, were found with all four drugs. Repeated trauma therefore activates the NO-cGMP pathway, possibly involving actions on both nNOS and iNOS. The NMDA receptor appears less involved after chronic repeated stress, and may have limited therapeutic implications. Sub-cellular NO-modulation, however, may represent an important therapeutic strategy in preventing the effects of severe stress and in treating PTSD. / Thesis (M.Sc. (Pharmacology))--North-West University, Potchefstroom Campus, 2005.

The use of mercury in artisanal mines is posing serious environmental and human health risks. Marcello Veiga is aiming to introduce global standards to regulate its use.

Posttraumatic stress disorder (PTSD) is a severe anxiety disorder characterised by hypothalamic-pituitary-adrenal (HPA)-axis abnormalities, hyperarousal, anxiety, flashbacks of trauma memories and avoidance. Increasing evidence is now accumulating that the disorder is also associated with shrinkage of the hippocampus and cognitive dysfunction that may have its origin in stress-induced excitotoxicity. Animal studies have indeed highlighted a potential role of the excitotoxic glutamatenitric oxide (NO) pathway in the stress response. Since PTSD appears to be an illness that progresses and worsens over time after an initial severe traumatic event, this study has used an animal model that emphasises repeated trauma to investigate the effect of stress on hippocampal NO synthase (NOS) activity, the release of the nitrogen oxide metabolites of NO (NOx), and also the evoked release of cGMP. Furthermore, the modulation and dependency of these responses on glutamate, NO and cGMP activity using drugs selective for these targets, will also be investigated. Rats (n=10/group) were exposed to repeated stress together with saline or drug administration immediately after the stress procedure and continuing for one week post-stress. The animals were then sacrificed for assay of hippocampal NOS activity, NO, and cGMP accumulation. Animals received either the glutamate-NMDA receptor antagonist, memantine (MEM;5mg/kg ip/d), the neuronal NOS selective inhibitor, 7- nitroindazole monosodium salt (7-NINA;20mg/kg ip/d), the cGMP-specific PDE inhibitor, sildenafil (SIL;10mg/kg ip/d) or the NFkb antagonist, pyrollidine dithiocarbamate (PDTC;70mg/kg ip/d). The latter inhibits the nuclear transcription factor, NFkb, responsible for inducing the expression of iNOS, while it also appears to mediate the glutamatergic actions on NOS expression, Stress significantly increased hippocampal NOS activity, as well as significantly increased hippocampal cGMP and NO, levels. These increases were blocked by pretreatment with either PDTC or 7-NINA, while memantine was without effect. Sildenafil significantly augmented stress induced NO, accumulation, as well as cGMP. although the latter failed to reach significance. 7-NINA and memantine significantly blocked the increase in cGMP evoked by time-dependent sensitisation (TDS)-stress, with PDTC attenuating this response, but not significantly. Additionally, administration of each drug separately for seven days without exposure to stress, did not evoke significant changes in NOx levels, compared to the control group. However, significant increases in cGMP levels, compared to the control group, were found with all four drugs. Repeated trauma therefore activates the NO-cGMP pathway, possibly involving actions on both nNOS and iNOS. The NMDA receptor appears less involved after chronic repeated stress, and may have limited therapeutic implications. Sub-cellular NO-modulation, however, may represent an important therapeutic strategy in preventing the effects of severe stress and in treating PTSD. / Thesis (M.Sc. (Pharmacology))--North-West University, Potchefstroom Campus, 2005.

Darbo tikslas. Įvertinti greitosios medicinos pagalbos pasirengimą ekstremalių situacijų atvejais. Darbo uždaviniai: 1. Įvertinti VšĮ Kėdainių PSPC GMP skyriaus pasirengimą veikti ekstremalių situacijų atvejais kitų panašias paslaugas teikiančių įstaigų kontekste. 2. Įvertinti VšĮ Kėdainių PSPC GMP skyriaus pasirengimo veiklai ES atveju pokyčius 2013-2014m., įdiegus pasirengimo veiklai ir planavimo ES metodiką. 3. Ištirti įstaigos operatyvumo rodiklių pokyčius įdiegus naują GMP kvietimo valdymo sistemą. 4. Įstaigos patirties veikti ekstremaliose situacijose analizė. Darbo metodika: GMP įstaigų vadovų struktūrizuota apklausa. Tiesioginis Kėdainių GMP pasirengimo veikti ekstremaliose situacijose vertinimas. Įstaigos operatyvumo rodiklių pokyčių analizė. Kėdainių GMP skyriaus darbuotojų, dalyvavusių teikiant pagalbą masinių nelaimių atvejais struktūrizuotas interviu. Rezultatai: Tyrime dalyvavusių įstaigų bendrasis saugumo indeksas skirtingų dydžių GMP tarpe pasiskirstė tolygiai. Daugumos vadovų nuomone, jų įstaigų bendrasis saugumo indeksas yra pakankamas (72 proc. – 91proc.). VšĮ Kėdainių PSPC, GMP skyriuje, vertinant 2013 – 2014m. pasirengimo veiklai ES atveju, įvyko teigiami pokyčiai. Bendras saugomo indeksas nuo 0,78 išaugo iki 0,93. Įdiegus GMP iškvietimų ir pajėgų valdymo sistemą, operatyvumo rodikliai tapo tikslesni. Lyginant 2012m. ir 2013m. iškvietimų, neatitinkančių minimalių laiko kriterijų, skaičius mieste padidėjo nuo 3,9 iki 5,3 proc. (p=0,002). 2013m. pirmajame... [toliau žr. visą tekstą] / Aim of the study: to assess disaster preparedness of health care institutions providing emergency medical services (EMS). Objectives: 1. To assess disaster preparedness of Kėdainiai EMS in context of other similar institutions. 2. To evaluate changes of disaster preparedness of Kedainiai EMS after implementation of guidelines for disaster preparedness and planning in 2013-2014. 3. To evaluate changes of operative performance indicators after implementation of the new dispatch and ambulances’ management system in Kedainiai EMS. 4. To analyze experience of Kėdainiai EMS in recent disasters. Methods:Structured interview of EMS institutions managers. Direct measurement of Kėdainiai EMS preparedness to respond in emergency situations. Analysis of changes of operative performance indicators. Structured interview of Kėdainiai EMS staff who had experience in recent disasters. Results: In opinion of the majority of managers, their institutions safety index is sufficient (72% - 91%). There were positive changes in disaster preparedness in Kedainiai EMS in 2013-2014: Overall safety index increased from 0,78 to 0,93. Operative performance indicators became more accurate after installation of dispatch and ambulance management system. Proportion of calls, when ambulance response time that does not meet minimum arrival time requirements, increased from 3.9% to 5.3% in the year 2012 – 2013 (p=0,002). In the first quarter of 2013 right after installation of the system, this performance... [to full text]