Two patients with clinically distinct manifestation of pyruvate dehydrogenase deficiency due to mutations in PDHA1 gene.

The most common cause of pyruvate dehydrogenase complex (PDHc) deficiency is the deficit of the E1α-subunit. The aim of this study was to describe distinct course of the disease in two boys with mutations in PDHA1 gene and illustrate the possible obstacles in measurement of PDHc activity. Clinical data and metabolic profiles were collected and evaluated. PDHc and E1α-subunit activities were measured using radiometric assay. Subunits of PDHc were detected by Western blot. PDHA1 gene was analysed by direct sequencing. In patient 1, the initial hypotonia with psychomotor retardation was observed since early infancy. The child gradually showed symptoms of spasticity and arrest of psychomotor development. In patient 2, the disease manifested by seizures and hyporeflexia in the toddler age. The diagnosis was confirmed at the age of seven years after attacks of dystonia and clinical manifestation of myopathy with normal mental development. Brain MRI of both patients revealed lesions typical of Leigh syndrome. Enzymatic analyses revealed PDHc deficiency in isolated lymphocytes in the first but not in the second patient. The direct measurement of PDH E1-subunit revealed deficiency in this individual. In patient 1, a novel hemizigous mutation c.857C>T (Pro250Leu) was detected in the X-linked PDHA1 gene. Mutation c.367C>T (Arg88Cys) was found in patient 2. We present first two patients with PDHc deficit due to mutations in PDHA1 gene in the Czech Republic. We document the broad variability of clinical symptoms of this disease. We proved that normal PDHc activity may not exclude the disease.

Mitochondrial DNA content and expression of genes involved in mtDNA transcription, regulation and maintenance during human fetal development.

The mitochondrial biogenesis and adequate energy production are important for fetal growth and early postnatal adaptation. The aim of the study was to characterize mitochondrial DNA (mtDNA) content and expression patterns of POLG, TFAM, NRF1,NRF2 and PGC1 family of regulated coactivators (PGC1A, PGC1B and PRC) involved in the mtDNA transcription, regulation and maintenance in human fetal tissues during second trimester of gestation. Further the mRNA expression profiles of selected cytochrome c oxidase (COX) subunits were analysed. Moreover enzyme activities of COX and CS and protein levels of COX subunits were analysed. DNA, RNA and proteins were isolated from 26 pairs of fetal liver and muscle samples obtained at autopsy after termination of pregnancy for genetic indications unrelated to OXPHOS deficiency between 13th and 28th week of gestation. This work offers a broad view on the mtDNA content changes in two different tissues during the second trimester of gestation and in the corresponding tissues after birth. The important differences in expression of POLG, TFAM, NRF2 genes and family PGC1 coactivators were found between the fetal tissues. The significant tissue-specific changes in expression of selected COX subunits on mRNA level (COX4 and MTCO2) were observed. Further the considerable differences in enzyme activities of COX and CS are demonstrated between fetal and postnatal phase. In conclusion our study indicates that the fetal developing tissues might differ in the control of mitochondrial biogenesis depending on their energy demand and the age of gestation. Moreover the gene expression is changed mainly on transcriptional level through fetal period.

A pre-attachment aggregation pheromone in the adult metastriate tick Hyalomma dromedarii Koch (Acarina: Ixodidae).

In vitro investigation demonstrates the existence of pheromone-regulated movement in the metastriate tick H. dromedarii before host contact. Males exhibit significant assembly to material of females. The palps, not Haller's organ, are essential for pheromone detection. The material is soluble in water, not in ether, benzene, or acetone and it is relatively stable to heat and cold.

Arbovirus isolations from mosquitoes in South Slovakia.

In the years 1973 and 1975 mosquitoes and some other Diptera (Tabanidae, Simuliidae, Hippoboscidae) were tested for virus. 13,924 mosquitoes, 75 horseflies and 60 blackflies were processed in 1973. Five strains of Tahyna virus were isolated from mosquito species Aedes vexans. 3,378 mosquitoes and 12 sheep keds were tested for virus in 1975. Twelve strains of Calovo virus were isolated from Anopheles maculipennis and one strain of Tahyna virus was obtained from Aedes vexans mosquitoes.