Is alumina suitable for solid phase extraction of catecholamines from brain tissue?

Occupational and environmental toxicology specialists find catecholamine fluctuations in brain tissue relevant for research of neurotoxicity, such as that induced by manganese or zinc, pesticides, industrial solvents, plastic, air pollution, or irradiation. Considering that catecholamine tissue concentrations are generally very low, their extraction requires a reliable and optimal method that will achieve maximum recovery and minimise other interferences. This study aimed to evaluate whether the aluminium (III) oxide (Al2O3, alumina) based cartridges designed for catecholamine isolation from plasma could be used for solid-phase extraction (SPE) of catecholamine from the brain tissue. To do that, we homogenised Wistar rat brain tissue with perchloric acid and compared three extraction techniques: SPE, the routine filtration through a 0.22 µm membrane filter, and their combination. In the extracts, we compared relative chromatographic catecholamine mobility measured with high performance liquid chromatography with electrochemical detection. Chromatographic patterns for norepinephrine and epinephrine were similar regardless of the extraction technique, which indicates that the alumina cartridge is good enough to isolate them from brain tissue. However, the dopamine pattern was unsatisfactory, and further experiments are needed to identify the issue and optimise the protocol.

Vitamin D Serum Levels and Vitamin D Receptor Genotype in Patients with Parkinson's Disease.

BACKGROUND: Vitamin D is a steroid hormone, known to be involved in the pathogenesis of various neurodegenerative disorders, including Parkinson's disease (PD). We aimed to clarify the relationship between hypovitaminosis D and the predisposition for PD and its clinical presentation. An additional aim was to examine the specific gene polymorphisms associated with vitamin D level. MATERIAL AND METHODS: Total level of 25(OH)-vitamin D (25(OH)D) was measured in the serum of parkinsonian patients (n = 113) and controls (n = 82) using a commercial immunoassay. Genetic analyses were performed using Taqman assays on Real Time PCR amplification system. RESULTS: Higher frequency of vitamin D deficiency (<50 nmol/L) was observed in PD patients, compared to controls (40.7% and 23.2%, respectively, P = 0.010). It was also a positive predictive marker of PD (OR, 2.27; 95% CI, 1.206-4.298; P < 0.011). Significantly higher UPDRS (35.85 ± 1.35 and 32.09 ± 0.99, respectively, P = 0.023) and HY scores (2(1.5-2.5) and 1.5(1.0-2.0), respectively, P = 0.005) were present in patients with 25(OH)D level < 50 nmol/L compared to patients with 25(OH)D level ≥ 50 nmol/L. Despite some trends observed, differences in allelic and genotypic distribution between controls and patients, as well as between subgroups, did not reach the level of significance (P > 0.05). CONCLUSIONS: Findings of this study confirm the hypothesis of a significant relationship between hypovitaminosis D and PD. We demonstrated higher prevalence of vitamin D deficiency in PD patients, as well as its predictive potential for the onset and progression of PD.

Antiphospholipid antibodies in patients with Graves' orbitopathy: preliminary data.

PURPOSE: Graves' orbitopathy (GO) is an inflammatory autoimmune disorder of the orbit and while the antiphospholipid antibodies (aPL) Abs were associated with the markers of inflammation in the antiphospholipid syndrome (APS), there is no literature that investigate the presence of aPL Abs in GO. We analyzed the prevalence of aPL Abs and the differences between aPL (+) and aPL (-) subgroups of GO patients. METHODS: Study included consecutive patients with GO (66 with Graves' (GD), 10 with Hashimoto (HD), and 8 were euthyroid). Anticardiolipin (aCL) and anti-beta 2glycoprotein I (aß2gpI) Abs were measured by ELISA. RESULTS: aPL Abs were present in 9/84 (10.71%) patients. The IgM aß2gpI Abs were present in 8/66 and in 1/10 patients with GD and HD. The IgG aCL Abs were present in one GD patient, and IgM aCL were present in 3/66 GD and in 1/10 patients with HD. In GD group, anti-Tg Abs were in positive correlation with aß2gpI IgG (p = 0.000) and with anti-TPO Abs (p = 0.016). In HD group, anti-Tg Abs were in positive correlation with IgM aCL (p = 0.042), while anti-TPO Abs were in positive correlation with aß2gpI IgM (p = 0.014). CONCLUSION: This study is the first report of the aPL Abs presence in GO patients. The anti-thyroid Abs were linked to aPL suggesting that their presence is not the sole consequence of hyperstimulation of autoreactive B-lymphocytes. Larger studies are necessary to confirm potential cause-effect relations.

Alpha-melanocyte-stimulating hormone during exercise recovery has prognostic value for coronary artery disease.

PURPOSE: Alpha-melanocyte-stimulating hormone (alpha-MSH) has proven cardiovascular effects and plays a significant role as an endogenous countermeasure to ischemia-reperfusion injury. The aim of the current study was to examine the response of alpha-MSH during exercise in patients diagnosed with coronary artery disease (CAD) and evaluate its value in the assessment of severity and prognosis. METHODS: Forty subjects with documented CAD (i.e., lesions on coronary angiography ≥ 50%) were included. Cardiopulmonary exercise testing (CPET) on a treadmill (TM) and recumbent ergometer (RE) were performed on two visits, 2-4 days apart, during 2 months of coronary angiography; subsequently, the subjects were followed up for 32 ± 10 months. At rest, at peak CPET, and after 3 min of recovery, plasma levels of alpha-MSH were measured by enzyme-linked immunosorbent assay technique. RESULTS: Mean ejection fraction was 56.7 ± 9.6%. Alpha-MSH similarly increased from rest to peak CPET on both modalities. There were no significant differences in alpha-MSH values during testing in patients with 1,2- and 3-vesel CAD, nor in patients with a SYNTAX score 0.05). Among CPET and hormonal parameters, ∆alpha-MSH recovery/peak during RE CPET was the best predictor of cardiac event occurrence (chi-square 6.67, HR = 0.51, CI = 0.25-1.02, p = 0.010). CONCLUSION: ∆alpha-MSH recovery/peak during RE CPET has predictive value for CAD prognosis, demonstrating involvement of alpha-MSH in CAD and a link between stress hormones and cardiac events.