On Genotyping Polymorphic HLA Genes — Ambiguities and Quality Measures Using NGS

The major histocompatibility complex (MHC) region of the human genome is the most polymorphic sequence part on chromosome 6; this roughly 4 Mbase long stretch contains many genes involved in immune response and disease association. The HLA genes have a crucial role in transplantation; patients receiving organs or bone marrow from matching donors have significantly higher chance for survival. NGS-based HLA typing brings the hope of accurate genomic consensus sequences by relatively cheap and simple laboratory workflow. Using either targeted or whole-genome sequencing data, there are a lot of possibilities to get ambiguous results (combinations of several alleles as a result instead of a single pair). These can be sample- or reference-related, or the results of artifacts generated during the targeting and amplifying step. NGS technology itself has additional artifacts leading to ambiguity listed in our paper. The final bioinformatics step will not be able to resolve all the ambiguities; we are also proposing quality control metrics to assess the final ambiguity and typing failure

Detection of Borrelia burgdorferi sensu lato in lizards and their ticks from Hungary

To investigate the involvement of lizard species in the natural cycle of Borrelia burgdorferi sensu lato (s.l.) in Hungary, a total of 186 reptiles belonging to three species—126 green lizards (Lacerta viridis), 40 Balkan wall lizards (Podarcis taurica), and 20 sand lizards (Lacerta agilis)—were captured in 2007 and 2008. All ticks removed from the lizards were Ixodes ricinus, either larvae (324=472; 68.6%) or nymphs (148=472; 31.4%). More than half (66=126; 52.4%) of L. viridis individuals were infested, and the prevalence of tick infestation on both the other two species was 35% each. All 472 I. ricinus ticks and tissue samples collected from 134 collar scales and 62 toe clips of lizards were further analyzed for the presence of B. burgdorferi s.l. with polymerase chain reaction. The amplification of B. burgdorferi s.l. DNA was successful in 8% (n¼92) of L. viridis, 9% (n¼32) of P. taurica, and 10% (n¼10) of L. agilis tissue samples. Restriction fragment length polymorphism genotyping identified the species Borrelia lusitaniae in all tested lizard samples. Prevalence of B. burgdorferi s.l. in ticks collected from L. viridis, P. taurica, and L. agilis was 8%, 2%, and 0%, respectively. Most of the infected ticks carried B. lusitaniae (74% of genotyped positives); however, Borrelia afzelii (5%) and B. burgdorferi sensu stricto (21%) were detected in ticks removed from green lizards and Balkan wall lizards, respectively. We conclude that lizards, particularly L. viridis, can be important hosts for I. ricinus larvae and nymphs; thus, they can be regarded as reservoirs of these important pathogen vectors. The role of green lizards has been confirmed, and the implication of Balkan wall lizards is suggested in the natural cycle of B. lusitaniae at our study site

Eco-epidemiology of Borrelia miyamotoi and Lyme borreliosis spirochetes in a popular hunting and recreational forest area in Hungary

BACKGROUND: Borrelia miyamotoi, the newly discovered human pathogenic relapsing fever spirochete, and Borrelia burgdorferi sensu lato are maintained in natural rodent populations. The aim of this study was to investigate the natural cycle of B. miyamotoi and B. burgdorferi s.l. in a forest habitat with intensive hunting, forestry work and recreational activity in Southern Hungary. METHODS: We collected rodents with modified Sherman-traps during 2010–2013 and questing ticks with flagging in 2012. Small mammals were euthanized, tissue samples were collected and all ectoparasites were removed and stored. Samples were screened for pathogens with multiplex quantitative real-time polymerase chain reaction (qPCR) targeting a part of flagellin gene, then analysed with conventional PCRs and sequencing. RESULTS: 177 spleen and 348 skin samples of six rodent species were individually analysed. Prevalence in rodent tissue samples was 0.2 % (skin) and 0.5 % (spleen) for B. miyamotoi and 6.6 % (skin) and 2.2 % (spleen) for B. burgdorferi s.l. Relapsing fever spirochetes were detected in Apodemus flavicollis males, B. burgdorferi s.l. in Apodemus spp. and Myodes glareolus samples. Borrelia miyamotoi was detected in one questing Ixodes ricinus nymph and B. burgdorferi s.l in nymphs and adults. In the ticks removed from rodents DNA amplification of both pathogens was successful from I. ricinus larvae (B. miyamotoi 5.6 %, B. burgdorferi s.l. 11.1 %) and one out of five nymphs while from Ixodes acuminatus larvae, and nymph only B. burgdorferi s.l. DNA was amplified. Sequencing revealed B. lusitaniae in a questing I. ricinus nymph and altogether 17 B. afzelii were identified in other samples. Two Dermacentor marginatus engorged larva pools originating from uninfected hosts were also infected with B. afzelii. CONCLUSIONS: This is the first report of B. miyamotoi occurrence in a natural population of A. flavicollis as well as in Hungary. We provide new data about circulation of B. burgdorferi s.l. in rodent and tick communities including the role of I. acuminatus ticks in the endophilic pathogen cycle. Our results highlight the possible risk of infection with relapsing fever and Lyme borreliosis spirochetes in forest habitats especially in the high-risk groups of hunters, forestry workers and hikers

Crosstalk between the Arabidopsis Glutathione Peroxidase-Like 5 Isoenzyme (AtGPXL5) and Ethylene

Glutathione peroxidases (GPXs) are important antioxidant enzymes in animals. Plants contain GPX-like (GPXL) enzymes, which-in contrast to GPXs-contain cysteine in their active site instead of selenocysteine. Although several studies proved their importance in development and stress responses, their interaction with ethylene (ET) signalling is not known. Our aim was to investigate the involvement of AtGPXL5 in ET biosynthesis and/or signalling using Atgpxl5 mutant and AtGPXL5 cDNA-overexpressing (OX-AtGPXL5) lines. Four-day-old dark-grown Atgpxl5 seedlings had shorter hypocotyls and primary roots, while OX-AtGPXL5 seedlings exhibited a similar phenotype as wild type under normal conditions. Six-week-old OX-AtGPXL5 plants contained less H2O2 and malondialdehyde, but higher polyamine and similar ascorbate- and glutathione contents and redox potential (E-GSH) than the Col-0. One-day treatment with the ET-precursor 1-aminocyclopropane-1-carboxylic acid (ACC) induced the activity of glutathione- and thioredoxin peroxidases and some other ROS-processing enzymes. In the Atgpxl5 mutants, the E-GSH became more oxidised; parallelly, it produced more ethylene after the ACC treatment than other genotypes. Although the enhanced ET evolution measured in the Atgpxl5 mutant can be the result of the increased ROS level, the altered expression pattern of ET-related genes both in the Atgpxl5 and OX-AtGPXL5 plants suggests the interplay between AtGPXL5 and ethylene signalling

Műtét közben:hit, flow és sebészet

Munkámban a családi rítusok, a szülői bánásmód és a serdülők közösségbe történő beilleszkedésének összefüggéseit, egymással való kapcsolatát vizsgálom. A családi rituálék fontos részei a család életének, hiszen azon túl, hogy az együtt töltött idő a családtagok számára kellemes élményt nyújt, számos pszichés tényezővel is összefüggésben van. A családtagok számára fontos védőfaktort jelentenek a külvilággal szemben. A családi rituálék ápolásának szerepe –a rendszeresség, a szerepvállalás- a biztonság és a stabil háttér nyújtásában nyilvánul meg, ami egy gyermek életében is elsődleges fontosságú. Az érzelmileg érett, az érzéseket felvállaló és kimutató közegben, biztonságos környezetben, megfelelő szülői bánásmód esetén, válaszkész szülők jelenlétében szocializálódó gyermekek maguk is érzelmileg kompetensek lesznek. A kutatásom során a családi szokásokat, a szülői bánásmódot és a serdülők iskolai beilleszkedésének egymással való kapcsolatát vizsgáltam.BSc/BAPszichológi