<p>Myasthenia gravis (MG) is an autoimmune neuromuscular disorder that causes fluctuating muscle weakness. MG may be divided into an ocular form and a generalized form based on the involved muscles. Treatment differs between these different MG forms. The majority (80%) of patients with generalized MG are seropositive for antibodies against the acetylcholine receptor (AChR-Ab). Recently a new antibody was detected against muscle specific tyrosine kinase (MuSK) in about 40% of patients who are AChR-Ab seronegative. A few patients with MuSK-Abs have muscular atrophies, as well as electrophysiological myopathy.</p><p>In this thesis we have characterized MuSK-Ab seropositive [MuSK(+)] patients using clinical parameters, including health-related quality of life (hrQoL), neurophysiology and muscle morphology, and compared them to patients with and without AChR-Abs. The question concerned which factors contribute to their muscle weakness. Additionally, we wanted to determine if single-fiber electromyography (SFEMG) in a limb muscle has any predictive value for generalization of ocular MG. </p><p>Our results suggest that MuSK(+) patients more often have a myopathic electromyography pattern, although this pattern is found also in other immunological subtypes of MG. The myopathic pattern may be associated with the frequently found mitochondrial abnormalities. However, disturbed neuromuscular transmission is the primary cause of muscle weakness in the majority of MuSK(+) patients, as well as in AChR-Ab seropositive patients. The disease-specific hrQoL MG questionnaire was successfully validated into Swedish and these scores correlated with disturbed neuromuscular transmission in a proximal arm muscle. Abnormal SFEMG findings occur also in muscles outside of the facial area in ocular MG, although this is not predictive of subsequent generalization. </p><p>MuSK (+) patients have little or no beneficial effect of acetylcholine esterase inhibitors (AChEI). On the contrary AChEI may produce profound adverse effects. We present the hypothesis that this effect of AChEI is due to abnormal receptor morphology in MuSK(+) patients.</p>

Myasthenia gravis (MG) is an autoimmune neuromuscular disorder that causes fluctuating muscle weakness. MG may be divided into an ocular form and a generalized form based on the involved muscles. Treatment differs between these different MG forms. The majority (80%) of patients with generalized MG are seropositive for antibodies against the acetylcholine receptor (AChR-Ab). Recently a new antibody was detected against muscle specific tyrosine kinase (MuSK) in about 40% of patients who are AChR-Ab seronegative. A few patients with MuSK-Abs have muscular atrophies, as well as electrophysiological myopathy. In this thesis we have characterized MuSK-Ab seropositive [MuSK(+)] patients using clinical parameters, including health-related quality of life (hrQoL), neurophysiology and muscle morphology, and compared them to patients with and without AChR-Abs. The question concerned which factors contribute to their muscle weakness. Additionally, we wanted to determine if single-fiber electromyography (SFEMG) in a limb muscle has any predictive value for generalization of ocular MG. Our results suggest that MuSK(+) patients more often have a myopathic electromyography pattern, although this pattern is found also in other immunological subtypes of MG. The myopathic pattern may be associated with the frequently found mitochondrial abnormalities. However, disturbed neuromuscular transmission is the primary cause of muscle weakness in the majority of MuSK(+) patients, as well as in AChR-Ab seropositive patients. The disease-specific hrQoL MG questionnaire was successfully validated into Swedish and these scores correlated with disturbed neuromuscular transmission in a proximal arm muscle. Abnormal SFEMG findings occur also in muscles outside of the facial area in ocular MG, although this is not predictive of subsequent generalization. MuSK (+) patients have little or no beneficial effect of acetylcholine esterase inhibitors (AChEI). On the contrary AChEI may produce profound adverse effects. We present the hypothesis that this effect of AChEI is due to abnormal receptor morphology in MuSK(+) patients.

Synthetic fragrances or MUSK compounds are artificialy synthetised organic substances, used as fragrant constituents in various homecare products, such as cosmetics, detergents, air fresheners etc. The aim of diploma thesis was the quantification of some selected MUSK compounds in the influent and efluent of the water treatment plants.The method of quantification of MUSK compounds in organic matrix was also optimised during the research. Two modifications of solid phase microextraction were tested and evaluated. The analytes were identified and quantified by gas chromatography - mass spectrometry (GC/MS). The results were used to evaluate the amount of MUSK compounds in sewage waters and the ability of various water treatment plants to eliminate this kind of pollution from the water during the treatment process.

Diploma thesis summarizes available information about chiral analysis of synthetic musk compounds in the aquatic environment. In the theoretical part of the thesis chirality and importance of chiral analysis by means of examples of different environmental pollutants is documented. Further consideration is aimed on musk compounds, their classification, characteristics and their fate in the environment, especially in the aquatic environment. After that methods for chiral analysis of synthetic musk compounds are reviewed. The aim of the experimental part of this diploma thesis was to carry out the determination of thirteen representatives of musk compounds (2-cyclohexylethanol, allylcyklohexylpropionate, Arocet, Aroflorone, Citronellol, Fresco Menthe, Galaxolide, hexyl cinnamic aldehyde, HSA, isoamylsalicylate, isobornylacetate, Lilial and Linalool), out of which some compounds are chiral and some are achiral. For the analysis samples of wastewater from the wastewater treatment plant Brno - Modřice were taken. Target compounds were isolated by solid phase microextraction (SPME) and were subsequently analyzed by gas chromatography with flame ionization and mass spectrometric detection. On the basis of data evaluation the removal efficiency forgiven musk compounds in the wastewater treatment plant was assessed.

Classical biocontrol systems are sometimes treated as an exercise in community assembly. As such, they include multiple species interactions. This thesis explores multi-species aspects in classical weed biocontrol, using thistles as a case study. The abundance, phenology and impact of three biocontrol agents were followed on their target host, Carduus nutans L. and are described, for the first time in New Zealand for two of them (Urophora solstitialis L. and Trichosirocalus horridus sensu (Panzer)). Composition in New Zealand of the recently revised Trichosirocalus weevil species complex was surveyed nation-wide. One species only was found, albeit exhibiting a wider host range than anticipated from the published revision. Interspecific interactions and individual and combined effect of multiple biocontrol agents on C. nutans were tested in cage setups; the effect on the weed population was then estimated by manipulations of an existing matrix population model for this weed in New Zealand. The potentially better seed predator (U. solstitialis) was outcompeted by the worse seed predator (Rhinocyllus conicus (Froehlich)) which has similar niche preference. Urophora solstitialis was also adversely impacted by the crown-root feeder (T. horridus). Trichosirocalus horridus affected C. nutans survival, even at the medium density used, and significantly reduced potential seed production by 33%; in field densities, T. horridus is likely to affect C. nutans even more. Urophora solstitialis was estimated to destroy about 28% of the remaining seed in the absence of the other agents, and about 17% in the presence of T. horridus. The estimated combined effect of T. horridus and U. solstitalis on C. nutans population growth rate was greater than the effect of either agent alone. In the face of growing weed invasions, multiple thistle species were used to test ‘multi-targeting’ as a novel approach to target groups of ‘sleeper weeds’. Both in a field experiment and in a field survey, the seed predator R. conicus was found to attack and damage some ‘non-target’ thistle species more in the presence of the target species (C. nutans) than in its absence; however, levels of attack on non-target species were always modest. The ultimate goal of biocontrol is to reduce weed populations. A field survey revealed that current population densities of multiple thistle species in Canterbury are not obviously lower than in the mid 1980s, when only R. conicus was present. This may be because successful biocontrol has reduced the management input required to maintain the same thistle density.

ColQ est un collagène spécifique ancrant l'acétylcholinestérase (AChE) dans la fente synaptique de la jonction neuromusculaire (JNM). L'importance du complexe AChE-ColQ dans le fonctionnement de cette synapse a été révélée par l'identification de mutations dans le gène humain codant pour ColQ qui sont à l'origine du syndrome myasthénique congénital avec déficience en AChE (SMC-1c). Les défauts présents chez les patients et le phénotype des souris ColQ-/- modèles pour le SMC-1c sont complexes, la déficience en AChE ne rendant probablement pas compte de tous les symptômes. Nous avons donc fait l'hypothèse que ColQ pourrait réguler la formation de la JNM afin d'expliquer certains des défauts observés. J'ai montré que ColQ régule l'expression des sous-unités du récepteur de l'acétylcholine (RACh) via le récepteur tyrosine kinase MuSK, molécule clé dans la formation de la synapse et cela indépendamment de l'AChE. La conséquence de cette régulation est que ColQ contrôle la formation des agrégats de RACh impliqués dans la transmission synaptique. Par ailleurs, mes résultats ont révélé que l'absence de ColQ entraîne une réduction de l'expression de MuSK à la membrane. Cette observation prend tout son intérêt dans un contexte clinique montrant que certains symptômes sont communs aux patients atteints du SMC-1c et du SMC lié à des mutations de MuSK. De plus, des signes d'atrophie musculaire ont été découverts chez la souris ColQ-/- pouvant laisser penser à une dénervation au moins partielle du muscle en absence de ColQ. Mes recherches ont ainsi permis de mettre en évidence des modifications pouvant être à l'origine des défauts fonctionnels observés lorsque ColQ est muté ou absent.

Classical biocontrol systems are sometimes treated as an exercise in community assembly. As such, they include multiple species interactions. This thesis explores multi-species aspects in classical weed biocontrol, using thistles as a case study. The abundance, phenology and impact of three biocontrol agents were followed on their target host, Carduus nutans L. and are described, for the first time in New Zealand for two of them (Urophora solstitialis L. and Trichosirocalus horridus sensu (Panzer)). Composition in New Zealand of the recently revised Trichosirocalus weevil species complex was surveyed nation-wide. One species only was found, albeit exhibiting a wider host range than anticipated from the published revision. Interspecific interactions and individual and combined effect of multiple biocontrol agents on C. nutans were tested in cage setups; the effect on the weed population was then estimated by manipulations of an existing matrix population model for this weed in New Zealand. The potentially better seed predator (U. solstitialis) was outcompeted by the worse seed predator (Rhinocyllus conicus (Froehlich)) which has similar niche preference. Urophora solstitialis was also adversely impacted by the crown-root feeder (T. horridus). Trichosirocalus horridus affected C. nutans survival, even at the medium density used, and significantly reduced potential seed production by 33%; in field densities, T. horridus is likely to affect C. nutans even more. Urophora solstitialis was estimated to destroy about 28% of the remaining seed in the absence of the other agents, and about 17% in the presence of T. horridus. The estimated combined effect of T. horridus and U. solstitalis on C. nutans population growth rate was greater than the effect of either agent alone. In the face of growing weed invasions, multiple thistle species were used to test ‘multi-targeting’ as a novel approach to target groups of ‘sleeper weeds’. Both in a field experiment and in a field survey, the seed predator R. conicus was found to attack and damage some ‘non-target’ thistle species more in the presence of the target species (C. nutans) than in its absence; however, levels of attack on non-target species were always modest. The ultimate goal of biocontrol is to reduce weed populations. A field survey revealed that current population densities of multiple thistle species in Canterbury are not obviously lower than in the mid 1980s, when only R. conicus was present. This may be because successful biocontrol has reduced the management input required to maintain the same thistle density.

Synthetic fragrances (musk) are artificially produced organic compounds. It is used as fragrance ingredient in personal care products. Substances from this group have the ability to withstand the cleaning process in wastewater treatment plants. The physico-chemical properties of musk compounds cause their accumulation in the living and non-living components of the ecosystem, where they behave as persistent pollutants. The adsorption process is considered to be one of the most widely used separation and purification processes. The adsorption processes of a solution of sixteen synthetic fragrances took place with biochar and activated carbon. Solid phase microextraction (SPME) was used as a method for extracting analytes. Gas chromatography coupled with mass spectrometry (GC-MS) was applied for final analysis, time of flight analyzer was used as detector. The data were fitted with adsorption isotherms.

Congenital myasthenic syndromes (CMS) are a group of heterogeneous disorders caused by mutations in genes encoding for proteins that are essential for neuromuscular transmission. All CMS share the clinical feature of fatigable muscle weakness. The differential diagnosis of CMS is wide, with a range of diseases going from autoimmune myasthenia gravis to muscle disorders. In this thesis, it was shown that measuring antibodies to clustered acetylcholine receptors (AChRs) by cell-based assay is helpful in the differential diagnosis of CMS. The findings of the current investigations showed that mutations in COL13A1, encoding the Collagen Type XIII α1 chain, were responsible for the symptoms of several patients with previously undefined myasthenias. In addition, this work described the clinical and complementary features of a novel CMS subtype due to mutations in the glycosylation pathway gene GMPPB. Investigations on a novel MUSK missense mutation (p.Ala617Val) uncovered previously unrecognised mechanisms of how levels of MuSK phosphorylation are critical to maintain synaptic structure, and guided suitable treatment for the patient. The study on the clinical and molecular basis of stridor, a novel clinical feature recently identified in patients with DOK7-CMS, prompted the identification of a novel DOK7 isoform, which warrants further investigation to elucidate its role in AChR clustering. Finally, the therapy of patients with severe AChR-deficiency was optimised thanks to a case series study that showed a robust improvement following the addition of β2-adrenergic agonists to their long-term treatment regime that included pyridostigmine.