An unusually high frequency of SCAD deficiency caused by two pathogenic variants in the ACADS gene and its relationship to the ethnic structure in Slovakia.

BACKGROUND: Short-chain acyl-CoA dehydrogenase deficiency (SCADD) represents a rare autosomal recessive inborn metabolic disorder of mitochondrial ß-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. METHODS: Biochemical 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. RESULTS: Our 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. CONCLUSIONS: Analogous 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.

Relation of blood pressure values in thirteen-years old adolescents to the mode of vitamin D prophylaxis during their infancy.

OBJECTIVE: The broad spectrum of extraosseal functions of vitamin D has recently been investigated. Although majority of recent studies have documented its hypotensive effect, some older studies warrant the risk of development the juvenile hypertension namely after pulse-fortified D vitamin supplementation during infancy. The aim of this study was to compare of the blood pressure in 13 year old groups of adolescents to the mode of D vitamin supplementation during their infancy. METHODS: Study group represent 1138 thirteen year old adolescents in complex health care of 21 primary care pediatricians (PCPs). This group was divided into three subgroups according their mode of D vitamin supplementation in infancy. 100 children were given the continuous daily supplementation of AD vitamin 200-400 IU /day - " regular subgroup 0", 933 children were given by bolus doses of D vitamin forte 450 000 IU every three months during first year of life - "fortified subgroup 2", and 105 children given by bolus doses of D vitamin during winter, and continuous daily AD drops during summer - " mixed subgroup 1". D vitamin supplementation was done approximately 13 years before blood pressure study. RESULTS: The mean systolic blood pressure was 110 mmHg, and mean diastolic one was 70 mmHg in all subgroups, regardless the mode of vitamin D supplementation. There were no statistic differences among subgroups, between girls/ boys, blood pressure levels were in normal limit. Although calculated daily doses of D2 vitamin during infancy exceeded RDA (200-400 IU) four to ten time, no difference in morbidity was found, concerning various chronic diseases (allergy, cardiovascular, renal, etc.). These results are in full accordance to recent knowledge of higher requirement and safety limit during vitamin D application. CONCLUSION: The mode of D vitamin prophylaxis during infancy (pulse or daily application) has no influence to blood pressure level in early adolescence. No adverse effects have been found despite that the calculated daily dose was exceeded from three to ten times the recommended daily allowance.

[Monitoring the prevalence of Streptococcus pneumoniae in various groups]. / Monitoring prevalencie Streptococcus pneumoniae v rôznych kolektívoch.

BACKGROUND: The aim of the present study was to monitor the nasopharyngeal presence of Streptococcus pneumoniae in different age groups (especially children) in Banská Bystrica, Slovakia. The purpose of this screening was to determine the prevalence of different serotypes and to follow up the presence of pneumococcus in these children after the vaccination with heptavalent protein-conjugate vaccine. A contribution of molecular biology techniques was the detection of S. pneumoniae DNA by PCR and also the typisation and comparison of pneumococcal strains by pulsed-field gel electrophoresis. METHODS: S. pneumoniae in nasopharyngeal swabs was detected by cultivation on blood agar plates. Serotypisation was performed by standard Quellung reaction. The commercial diagnostic kit was used for PCR detection of S. pneumoniae DNA. Pulsed-field electrophoresis was performed by modified scheme according to literature. RESULTS: The incidence of pneumococcus is decreasing and less significant with the increasing age. Among youngest children is relatively high prevalence of pneumococci and the relatedness of isolated strains is high as well. After the vaccination, the less invasive serotypes were detected, although the overall incidence of S. pneumoniae was similar. CONCLUSIONS: The monitoring of S. pneumoniae in population is important according to variability of this bacteria with respect to possible changes in pneumococcal types as a consequence of vaccination.

Polymerase chain reaction (PCR)--an efficient tool for diagnosis of influenza and acute respiratory illnesses during influenza season.

Many severe diseases of the respiratory tract lead to hospitalisation. These diseases are often caused by viral infections and may cause increased mortality. The most common viral pathogens involved in these cases, which are also associated with significant morbidity and mortality during the influenza seasons are influenza viruses. Rapid differential diagnosis of influenza viruses is therefore of great importance. Classical diagnosis of these viruses involves virus cultures. Of the rapid diagnostic methodologies which have been developed are RT-PCR, multiplex PCR, real-time PCR. In the present study we have monitored clinical samples from patients of different age groups from selected regions in Slovakia and compared the effectiveness of the classical and molecular biological diagnostic methods. The molecular biological methods proved to be rapid, accurate and effective. Application of these techniques in diagnosis of the respiratory illnesses should help in the prevention, therapy and disease control.

Congenital hypothyroidism in one of monozygotic twins: comparison of their long-term psychosomatic development.

Authors present the long-term development of two monozygotic twin sisters. One of them was detected by newborn screening as athyreotic (gemelus A) and healthy gemelus B sister. Therapy in athyreotic girl started at 4 weeks after birth and was monitored in order to maintain serum thyroxine and TSH in normal range. Both sisters spent their chilhood and adolescence together in the same family, school and work together as tailors in the same factory. Their development was very similar. In order to detect subclinical difference of their development, the school achievements were tested using their school certificates. Studies were divided to testing mechanical memory, logical thinking and skills. The evaluation of all classes of school attendance (13 years) reveals subtle differences - deficit of patient at approximately 6-15% level, mainly in studies demanding skills. However, dynamic evaluation after years reveals that the worse results of the patient (gemelus A) occurred during the first years of basic school whereas during adolescence having attended the special school tailor, she attained better results than her healthy sister. It seems that mental deficit in early treated hypothyroid patient is not severe and varies even during the life. Problem of the fetal hypothyroidism is probably more related to the iodine deficiency, which assures necessity of thyroid hormones for athyreotic child from mother's or twin's normally supplemented thyroid glands.