BackgroundShort-chain acyl-CoA dehydrogenase deficiency (SCADD) represents a rare autosomal recessive inborn metabolic disorder of mitochondrial β-oxidation of monocarboxylic acids. Clinical symptoms can vary from a severe life-threatening condition to an asymptomatic state, reported in the majority of cases. Since the expansion of newborn screenings, more than three hundred probands were admitted for molecular-genetic analysis, most selected because of elevated values of C4-acylcarnitine detected in newborn screenings in Slovakia. Searching for the principal genomic changes led us to the selection of sixty-two patients in whom the presence of sequence variants in the ACADS gene was analysed and correlated with the available biochemical and clinical data.MethodsBiochemical and molecular genetic tests were performed. Acylcarnitine profiles focused on an elevated level of C4-acylcarnitine, which was analysed via tandem mass spectrometry. Urinary organic acids, specifically a quantity of ethylmalonic acid, were determined by gas chromatography/mass spectrometry. The entire coding region of the ACADS gene was sequenced. A low-cost restriction fragment length polymorphism of PCR amplified fragments analysis (PCR-RFLP) of pathogenic variants was introduced and implemented for the molecular-genetic algorithm appropriate for the Slovak population.ResultsOur molecular genetic study was performed on sixty-two patients with a pathological biochemical pattern related to short-chain acyl-CoA dehydrogenase deficiency. In this cohort, we discovered a high occurrence of two rare pathogenic variants—the deletion c.310_312delGAG and the substitution c.1138C>T, with allelic frequencies of 64% and 31%, respectively. Up to 86% of investigated individuals belong to the Roma ethnic group.ConclusionsAnalogous to other countries, SCADD is not included in the newborn screening programme. Based on the exceeded levels of the specific biomarker C4-acylcarnitine as well as ethylmalonic acid, we revealed a high prevalence of short-chain acyl-CoA dehydrogenase deficiency cases, confirmed by the findings of two rare pathogenic variants. A deletion c.310_312delGAG and c.1138C > T substitution in the ACADS gene appear with a high frequency in the Roma ethnic group of Slovakia. Due to the uncertainty of the pathogenicity and clinical consequences, it is important to follow up the morbidity and mortality in these patients over time and evaluate SCADD in relation to clinical outcomes and preventive healthcare recommendations.

The proposed work deals with the utilization of three carbon-based electrode substrates such as boron-doped diamond, glassy carbon and carbon paste for the preparation of in situ bismuth film modified electrodes. Such modified electrodes were subsequently used for the differential pulse anodic stripping voltammetric determination of heavy metal cations (Zn2+, Cd2+ and Pb2+) individually and simultaneously using similar experimental conditions (0.1 mol l−1 acetic buffer solution of pH 4.5 as supporting electrolyte with the addition of 0.1 mmol l−1 Bi3+, deposition potential of −1.4 V and deposition time of 120 s). The results showed that the modification step mostly enhanced the deposition and stripping process of studied cations when compared to the bare electrode substrates. A boron-doped diamond electrode was selected as the substrate for modification and the procedure was applied to the real sample analysis including water sample (certified reference material) and wastewater sample. Using the standard addition method the concentrations of particular heavy metals were quantified and the determined values were in a good agreement with those obtained by the reference method — high resolution atomic absorption spectroscopy with electrothermal atomisation and continual radiation source. This fact highlights that the developed in situ bismuth film modified boron-doped diamond electrode is a suitable electrochemical sensor to be applied to routine analysis of water samples containing heavy metals.

Electrochemical methods have many inherent advantages such as simplicity, low cost and amenability to miniaturization. A new, sensitive and selective electrochemical method for the caffeine determination using boron doped diamond electrode was developed. It was found by cyclic voltammetry that caffeine provided highly reproducible and well-defi ned irreversible oxidation peak, at very positive potential of +1.55 V vs. Ag/AgCl electrode. The effect of pH and scan rate on the voltammetric response of caffeine oxidation were studied to select the optimum experimental conditions. Linear response of peak current on the concentration in the range from 4 × 10 -7 to 2.5 × 10 -5 mol L -1, good repeatability (RSD of 2.1 %) and the detection limit of 1.5 × 10 -7 mol L -1 without any chemical modifi cations and electrochemical surface pretreatment were observed by differential pulse voltammetry in 0.4 mol L -1 perchloric acid. The effect of possible interfering compounds appeared to be negligible which evidently proved very good selectivity. The proposed method was successfully applied for the caffeine determination in commercially available beverage samples, with results in a close statistical agreement to these declared by manufacturer.