9 research outputs found

    In-vivo monitoring of catecholamines in rat brain microdialysates after the optimization of an HPLC-ECD and a CE-LIF method

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    -Measurement of the concentration of neurotransmitters such as dopamine (DA) and noradrenaline (NA) in the extracellular space of animals provides important information about their synthesis, release and metabolism in response to behavioral or pharmacological manipulations. Intracerebral microdialysis in conjunction with a highly sensitive analysis method such as high-performance liquid chromatography with electrochemical detection (HPLC-ECD) and capillary electrophoresis with laser-induced fluorescence (CE-LIF) detection was used to study the in-vivo release of catecholamines (DA and NA) and monoamine metabolites (3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA) and 5-hydroxyindole-3-acetic acid (5-HIAA)) within distinct brain regions of freely moving rats. Therefore, we optimized these methods and established a setup that enables separation and quantification of low DA and NA standard concentrations (1-200 nM). By using the CE-LIF system we were able to quantify DA and NA in low volume (<10 L) aqueous standards. In a next step we used HPLC-ECD for quantifying DA and NA and their metabolites in microdialysis samples from the rat nucleus accumbens (NAc). We were able to reliably measure basal extracellular levels of DA, NA and selected metabolites in microdialysates from this area. Moreover, the release of DA and NA was considerably enhanced in response to local depolarization by high potassium perfusion. Interestingly, we also found a significant increase of DA release in the NAc in response to swim stress indicating physiological functionality of released DA within this area under ethological relevant conditions. Thus, the present thesis has revealed the potential of combining microdialysis with highly sensitive analysis methods such as HPLC-ECD and probably also CE-LIF to detect and quantify extracellular levels of catecholamines (DA and NA) in microdialysates.submitted by Anita SillerUniversität Innsbruck, Diplomarbeit, 2016Innsbruck, Univ., Diplomarb., 2016(VLID)149189

    Seroprevalence, Waning and Correlates of Anti-SARS-CoV-2 IgG Antibodies in Tyrol, Austria: Large-Scale Study of 35,193 Blood Donors Conducted between June 2020 and September 2021.

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    There is uncertainty about the seroprevalence of anti-SARS-CoV-2 antibodies in the general population of Austria and about the waning of antibodies over time. We conducted a seroepidemiological study between June 2020 and September 2021, enrolling blood donors aged 18-70 years across Tyrol, Austria (participation rate: 84.0%). We analyzed serum samples for antibodies against the spike or the nucleocapsid proteins of SARS-CoV-2. We performed a total of 47,363 samples taken from 35,193 individuals (median age, 43.1 years (IQR: 29.3-53.7); 45.3% women; 10.0% with prior SARS-CoV-2 infection). Seroprevalence increased from 3.4% (95% CI: 2.8-4.2%) in June 2020 to 82.7% (95% CI: 81.4-83.8%) in September 2021, largely due to vaccination. Anti-spike IgG seroprevalence was 99.6% (95% CI: 99.4-99.7%) among fully vaccinated individuals, 90.4% (95% CI: 88.8-91.7%) among unvaccinated individuals with prior infection and 11.5% (95% CI: 10.8-12.3%) among unvaccinated individuals without known prior infection. Anti-spike IgG levels were reduced by 44.0% (95% CI: 34.9-51.7%) at 5-6 months compared with 0-3 months after infection. In fully vaccinated individuals, they decreased by 31.7% (95% CI: 29.4-33.9%) per month. In conclusion, seroprevalence in Tyrol increased to 82.7% in September 2021, with the bulk of seropositivity stemming from vaccination. Antibody levels substantially and gradually declined after vaccination or infection

    Seroprevalence of Anti-SARS-CoV-2 IgG Antibodies in Tyrol, Austria: Updated Analysis Involving 22,607 Blood Donors Covering the Period October 2021 to April 2022.

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    Because a large proportion of the Austrian population has been infected with SARS-CoV-2 during high incidence periods in winter 2021/2022, up-to-date estimates of seroprevalence of anti-SARS-CoV-2 antibodies are required to inform upcoming public health policies. We quantified anti-Spike IgG antibody levels in 22,607 individuals that donated blood between October 2021 and April 2022 across Tyrol, Austria (participation rate: 96.0%). Median age of participants was 45.3 years (IQR: 30.9−55.1); 41.9% were female. From October 2021 to April 2022, seropositivity increased from 84.9% (95% CI: 83.8−86.0%) to 95.8% (94.9−96.4%), and the geometric mean anti-Spike IgG levels among seropositive participants increased from 283 (95% CI: 271−296) to 1437 (1360−1518) BAU/mL. The percentages of participants in categories with undetectable levels and detectable levels at <500, 500−<1000, 1000−<2000, 2000−<3000, and ≥3000 BAU/mL were 15%, 54%, 15%, 10%, 3%, and 3% in October 2021 vs. 4%, 18%, 17%, 18%, 11%, and 32% in April 2022. Of 2711 participants that had repeat measurements taken a median 4.2 months apart, 61.8% moved to a higher, 13.9% to a lower, and 24.4% remained in the same category. Among seropositive participants, antibody levels were 16.8-fold in vaccinated individuals compared to unvaccinated individuals (95% CI: 14.2−19.9; p-value < 0.001). In conclusion, anti-SARS-CoV-2 seroprevalence in terms of seropositivity and average antibody levels has increased markedly during the winter 2021/2022 SARS-CoV-2 waves in Tyrol, Austria

    beta 2-subunit alternative splicing stabilizes Cav2.3 Ca2+ channel activity during continuous midbrain dopamine neuron-like activity

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    In dopaminergic (DA) Substantia nigra (SN) neurons Cav2.3 R-type Ca2+-currents contribute to somatodendritic Ca2+-oscillations. This activity may contribute to the selective degeneration of these neurons in Parkinson's disease (PD) since Cav2.3-knockout is neuroprotective in a PD mouse model. Here, we show that in tsA-201-cells the membrane-anchored beta 2-splice variants beta 2a and beta 2e are required to stabilize Cav2.3 gating properties allowing sustained Cav2.3 availability during simulated pacemaking and enhanced Ca2+-currents during bursts. We confirmed the expression of beta 2a- and beta 2e-subunit transcripts in the mouse SN and in identified SN DA neurons. Patch-clamp recordings of mouse DA midbrain neurons in culture and SN DA neurons in brain slices revealed SNX-482-sensitive R-type Ca2+-currents with voltage-dependent gating properties that suggest modulation by beta 2a- and/or beta 2e-subunits. Thus, beta-subunit alternative splicing may prevent a fraction of Cav2.3 channels from inactivation in continuously active, highly vulnerable SN DA neurons, thereby also supporting Ca2+ signals contributing to the (patho)physiological role of Cav2.3 channels in PD
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