Mikroszkópikus gombák (Aspergillus, Fusarium, Cryptococcus) mitokondriális genomszerveződésének összehasonlító elemzése = Study on the mitochondrial genome organisation of some microscopic fungi

Munkánk során meghatároztuk egy Aspergillus niger (1a típus, 31103 bp) és egy A. tübingensis (2b típus, 33656 bp) törzs teljes mitokondriális DNS szekvenciáját. A két genom géntartalma és a gének sorrendje megegyezik, különbséget mindössze a restrikciós enzimek hasítási mintázatában tapasztaltunk. Megállapítottuk, hogy a méretbeli eltérésekért a cox1, atp9 és a ndh4L gének intron-tartalma felelős. Elkészítettük a korábban már RFLP mintázat alapján elkülönített hat A. tübingensis mtDNS fizikai és funkcionális térképét. Eredményeink bizonyították, hogy a tapasztalt intraspecifikus polimorfizmus intron-szerzéssel illetve restrikciós hasítóhelyeket érintő pontmutációkkal magyarázható. Növény-patogén Fusarium törzsek biodiverzitását a mtDNS RFLP mintázata alapján tanulmányoztuk. Több haplotípust sikerült elkülönítenünk a vizsgált 3 fajban. Meghatároztuk e haplotípusok gazdanövény szerinti eloszlását. F. proliferatum lineáris, mitokondriumban lokalizált DNS plazmidjának teljes szekvenálását elvégeztük. A plazmid funkciójára jelenleg nincs bizonyítékunk. Cryptococcus neoformans két varietas-ának mtDNS szerveződését hasonlítottuk össze. Megállapítottuk, hogy a tapasztalt méretbeli különbséget a cox1, cob és LRNS gének eltérő intron-tartalma okozza. Egy másik vizsgált faj, a Trichosporon pullulans 18 kb méretű mtDNS-e a legkisebb, NADH géneket is tartalmazó mtDNS-nek bizonyult élesztőgombák között. | In the present work the complete mitochondrial DNA (mtDNA) of Aspergillus niger mtDNA type 1a (31103 bp) was sequenced and compared to the Aspergillus tubingensis type 2b (33656 bp) mtDNA. The patterns of restriction sites were similar, the gene content and order was identical. The size difference was principally attributed to the intron content of their cox1, atp9 and ndh4L genes. A. tubingensis isolates were earlier classified into six groups on the basis of the mtDNA RFLP pattern. The reason of the intraspecific mtDNA variability was investigated and proved that polymorphism due to intron acquisition and also sporadic point mutations affecting the recognition motifs of the restriction enzymes. Biodiversity of plant-pathogenic Fusarium isolates belonging to 3 different species were studied. On the basis of the RFLP pattern of their mtDNA several haplotypes were identified. The distribution of these haplotypes among host species was determined. The complete nucleotide sequence of a 10 kb linear DNA plasmid was identified, however, its function is still unknown. The functional map of the mtDNAs of two Cryptococcus neoformans varities was constructed. Characteristic sequences were cloned, sequenced and verified that the intron-content of coxI, cob and LRNA genes are responsible for the size differences of the two strains. The study of the organisation of Trichosporon pullulans mtDNA revealed that it is smallest known mtDNA among yeast carrying NADH dehidrogenase genes

Integrated Systems Biology Approach Identifies Novel Maternal and Placental Pathways of Preeclampsia

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Storage stability of five steroids and in dried blood spots for newborn screening and retrospective diagnosis of congenital adrenal hyperplasia

Congenital adrenal hyperplasia (CAH) is a severe inherited disorder of cortisol biosynthesis that is potentially lethal or can seriously affect quality of life. For the first time, we aimed to assess the stability of 21-deoxycortisol (21Deox), 11-deoxycortisol (11Deox), 4-androstenedione (4AD), 17-hydroxyprogesterone (17OHP) and cortisol (Cort), diagnostic for CAH, in dried blood spots (DBSs) during a 1 year storage at different temperatures. Spiked DBS samples were stored at room temperature, 4 °C, -20 °C or -70 °C, respectively and analyzed in triplicates using liquid chromatography-tandem mass spectrometry at Weeks 0, 1, 2, 3 and 4, Month 6 and Year 1. Analyte levels within ±15% vs the baseline were considered stable. Our observations show that 21Deox, 4AD and 17OHP were not significantly changed for 1 year even at room temperature at either analyte levels. In contrast, Cort required storage at 4 °C, -20 °C or -70 °C for long-term stability, being significantly decreased at room temperature from Month 6 (p<0.01) in both the 30(60) nM and the 90(180) nM samples. 11Deox was significantly decreased at room temperature at Year 1 (p<0.01) and only in the 30(60) nM samples. Thus, all biomarkers were stable for up to 1 year at 4 °C, -20 °C or -70 °C and at least for 4 weeks at room temperature. These findings have implications for analyses of stored DBS samples in 2nd-tier assays in newborn screening and for retrospective CAH studies