Otsides nõela heinakuhjast – endometrioosi biomarkerid

Endometrioosi diagnoosimiseks puuduvad siiani mitteinvasiivsed ja minimaalselt invasiivsed biomarkerid ning haiguse diagnoos põhineb peamiselt laparoskoopilisel operatsioonil ja endometrioosikollete histoloogilisel uuringul. Kuigi endometrioosi biomarkereid on vereplasmast ja -seerumist, uriinist, menstruaalverest, emakaõõne aspiraadist ja ka endomeetriumi koest aktiivselt otsitud, on leitud markerite usaldusväärsus endometrioosi diagnoosimisel jäänud siiski tagasihoidlikuks. Ülevaates on tehtud kokkuvõte endometrioosi biomarkerite otsingute hetkeseisust ja leitud markerite kasutatavusest kliinilises praktikas. Eesti Arst 2017; 96(6):335–34

Review on Autoimmune Reactions in Female Infertility: Antibodies to Follicle Stimulating Hormone

Female fertility can be affected by diseases or dysfunctions of reproductive tract, neuroendocrine system, and immune system. Reproductive autoimmune failure can be associated with overall activation of immune system or with immune system reactions specifically directed against ovarian antigens. Majority of the antiovarian autoantibodies are directed against β-subunit of follicle stimulating hormone (anti-FSH). This paper summarizes a current clinical classification of female infertility in the context of general activation of autoimmunity and antiovarian autoimmunity by describing serum anti-FSH. The presence of naturally occurring anti-FSH in healthy women will be discussed. In addition, the putative impairment of ovarian folliculogenesis in case of increased production of those antibodies in infertile women will be characterized

EGF-i, IL1B, IL4 ja IL6 geeni seosed skisofreenia esmase psühhootilise episoodi ja prodroomsete sümptomitega

Skisofreenia on multifaktoriaalne neuropsühhiaatriline häire, millel tekkel on olulised nii geneetilised eelsoodumused kui ka keskkonnast tingitud faktorid. Paljud uuringud on viidanud tsütokiinide olulisusele skisofreenia etioloogias ja patofüsioloogias. Käesoleva uurimustöö eesmärgiks oli uurida, kas EGF-i, IL1B, IL4 ja IL6 ühenukleotiidsed polümorfismid võiksid olla seotud skisofreenia esmase psühhootilise episoodi ja prodroomsete sümptomitega. Juht-kontroll uuringusse kaasati 239 isikut, kuhu kuulusid 103 esmase psühhootilise episoodiga patsienti ja 22 prodroomsete sümptomitega isikut ning 114 tervet kontrollisikut. Uuringu tulemusena leiti piiripealne seos IL4 geenis paikneva polümorfismi rs2070874-ga. Lisaks viidi läbi ka haplotüübianalüüs, mille tulemusena leiti statistiliselt oluline seos IL1B geeni rs1143627-rs16944 haplotüübiga. Käesolev uuring viitab, et Eesti populatsioonis võivad antud IL4 ja IL1B geenide polümorfismid olla seotud skisofreeniaga

Determination of biological activity of gonadotropins hCG and FSH by Forster resonance energy transfer based biosensors

Determination of biological activity of gonadotropin hormones is essential in reproductive medicine and pharmaceutical manufacturing of the hormonal preparations. The aim of the study was to adopt a G-protein coupled receptor (GPCR)-mediated signal transduction pathway based assay for quantification of biological activity of gonadotropins. We focussed on studying human chorionic gonadotropin (hCG) and follicle-stimulating hormone (FSH), as these hormones are widely used in clinical practice. Receptor-specific changes in cellular cyclic adenosine monophosphate (cAMP, second messenger in GPCR signalling) were monitored by a Forster resonance energy transfer (FRET) biosensor protein (T)Epac(VV) in living cells upon activation of the relevant gonadotropin receptor. The BacMam gene delivery system was used for biosensor protein expression in target cells. In the developed assay only biologically active hormones initiated GPCR-mediated cellular signalling. High assay sensitivities were achieved for detection of hCG (limit of detection, LOD: 5 pM) and FSH (LOD: 100 pM). Even the smallscale conformational changes caused by thermal inactivation and reducing the biological activity of the hormones were registered. In conclusion, the proposed assay is suitable for quantification of biological activity of gonadotropins and is a good alternative to antibody- and animal-testing-based assays used in pharmaceutical industry and clinical research.Peer reviewe

A novel hypothesis for histone-to-protamine transition in Bos taurus spermatozoa

DNA compaction with protamines in sperm is essential for successful fertilization. However, a portion of sperm chromatin remains less tightly packed with histones, which genomic location and function remain unclear. We extracted and sequenced histone-associated DNA from sperm of nine ejaculates from three bulls. We found that the fraction of retained histones varied between samples, but the variance was similar between samples from the same and different individuals. The most conserved regions showed similar abundance across all samples, whereas in other regions, their presence correlated with the size of histone fraction. This may refer to gradual histone-protamine transition, where easily accessible genomic regions, followed by the less accessible regions are first substituted by protamines. Our results confirm those from previous studies that histones remain in repetitive genome elements, such as centromeres, and added new findings of histones in rRNA and SRP RNA gene clusters and indicated histone enrichment in some spermatogenesis-associated genes, but not in genes of early embryonic development. Our functional analysis revealed significant overrepresentation of cGMP-dependent protein kinase G (cGMP-PKG) pathway genes among histone-enriched genes. This pathway is known for its importance in pre-fertilization sperm events. In summary, a novel hypothesis for gradual histone-toprotamine transition in sperm maturation was proposed. We believe that histones may contribute structural information into early embryo by epigenetically modifying centromeric chromatin and other types of repetitive DNA. We also suggest that sperm histones are retained in genes needed for sperm development, maturation and fertilization, as these genes are transcriptionally active shortly prior to histone-to-protamine transition.Peer reviewe