Ureaplasma spp., commonly identified in the vagina/cervix of pregnant women with spontaneous preterm delivery, are the most frequently detected microorganisms in amniotic fluid. To date, 14 U. spp. serotypes have been characterized; however, modern molecular biology methods can distinguish different U. spp genotypes. Considering these factors, a knowledge gap exists regarding the association between U. spp. genotypes and the risk of the ascension of U. spp. from the cervix to the amniotic cavity. To fill this gap, an expanded multilocus sequence-typing scheme of U. spp. was performed to assess the relationship between cervical and amniotic fluid U. spp. in pregnant women with spontaneous preterm delivery. This study included 109 and 69 pregnant women with spontaneous preterm labor (PTL) and preterm prelabor rupture of membranes (PPROM), respectively. U. spp. DNA in cervical fluid was identified in 49% and 55% of the women with PTL and PPROM, respectively. The concurrent presence of U. spp. DNA in amniotic fluid was observed in 17% and 59% of the pregnant women with PTL and PPROM, respectively. Among pregnant women with PTL and PPROM, 38 expanded sequence types of cervical U. spp. were identified. No associations were observed between specific genotypes, subgroups, or clusters of cervical U. spp. and the presence of amniotic fluid U. spp. in pregnant women with spontaneous preterm delivery.

• Klíčová slova
• Amniotic fluid, Genital mycoplasmas, Invasive sampling, Non-invasive sampling, Preterm birth,
• MeSH
• cervix uteri * mikrobiologie   MeSH
• DNA bakterií genetika   MeSH
• dospělí MeSH
• genotyp MeSH
• infekční komplikace v těhotenství mikrobiologie   MeSH
• lidé MeSH
• mladý dospělý MeSH
• multilokusová sekvenční typizace MeSH
• plodová voda mikrobiologie   MeSH
• předčasná porodní činnost mikrobiologie   MeSH
• předčasný odtok plodové vody mikrobiologie   MeSH
• předčasný porod * mikrobiologie   MeSH
• těhotenství MeSH
• Ureaplasma * genetika   izolace a purifikace   klasifikace   MeSH
• ureaplasmatické infekce * mikrobiologie   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mladý dospělý MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• DNA bakterií MeSH

The main aim of the study was to determine progranulin levels in amniotic and cervical fluid samples from pregnancies complicated by preterm prelabor rupture of membranes (PPROM) or preterm labor with intact membranes (PTL), with concomitant microbial invasion of the amniotic cavity and/or intra-amniotic inflammation. A total of 104 and 108 women with PPROM and PTL, respectively, were included. Paired amniotic and cervical fluid samples were obtained using transabdominal amniocentesis and Dacron polyester swabs, respectively. Progranulin levels were assessed with an enzyme-linked immunosorbent assay. Women with PPROM and PTL were divided into subgroups based on microbial invasion of the amniotic cavity and/or intra-amniotic inflammation. Differences in progranulin levels among the PPROM and PTL subgroups were found in amniotic fluid: (a) PPROM: intra-amniotic infection: 51.8 pg/mL, sterile intra-amniotic inflammation: 52.8 pg/mL, colonization: 36.4 pg/mL, and negative amniotic fluid: 35.0 pg/mL; p < 0.0001; (b) PTL: intra-amniotic infection: 75.3 pg/mL, sterile intra-amniotic inflammation: 54.0 pg/mL, and negative amniotic fluid: 39.1 pg/mL; p < 0.0001. The corresponding differences were not found in cervical fluid: (a) PPROM: p = 0.14; (b) PTL: p = 0.53. In conclusion, amniotic fluid progranulin levels increased in PPROM and PTL cases with concomitant intra-amniotic inflammation, regardless of whether microbial invasion of the amniotic cavity was present or absent.

• Klíčová slova
• Amniotic fluid, Intra-amniotic inflammation, Invasive sampling, Microbial invasion of the amniotic cavity, Non-invasive sampling, Preterm delivery,
• MeSH
• amniocentéza MeSH
• cervix uteri * metabolismus   MeSH
• chorioamnionitida metabolismus   MeSH
• dospělí MeSH
• lidé MeSH
• plodová voda * metabolismus   MeSH
• předčasná porodní činnost metabolismus   MeSH
• předčasný odtok plodové vody * metabolismus   MeSH
• předčasný porod * metabolismus   MeSH
• progranuliny * metabolismus   MeSH
• retrospektivní studie MeSH
• těhotenství MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• GRN protein, human MeSH Prohlížeč
• progranuliny * MeSH

The main aim of this study was to determine expanded sequence types (eSTs) of Ureaplasma species (U. spp.). DNA isolated from the amniotic fluid of pregnancies complicated by preterm prelabor rupture of membranes (PPROM) using an expanded multilocus sequence typing scheme. Additionally, the study sought to examine whether phylogenetic subgroups of U. spp. DNA differ with respect to maternal demographic and clinical parameters and selected aspects of short-term neonatal morbidity. This retrospective cohort study was focused on singleton pregnancies complicated by PPROM occurring between the gestational ages of 24+0 and 36+6 weeks, where amniocentesis was conducted to assess the intra-amniotic environment and the presence of U. spp. DNA in the amniotic fluid samples was confirmed. The stored aliquots of U. spp. DNA were used to assess differences in nucleotide sequences in six U. spp. genes (ftsH, rpL22, valS, thrS,ureG, and mba-np1) using the eMLST scheme. The expanded multilocus sequence typing scheme was performed in 73 samples of U. spp. DNA isolated from pregnancies complicated by PPROM. In total, 33 different U. spp. DNA eSTs were revealed, 21 (#20, 233-244, 248-251, 253, 255, 259, and 262) of which were novel. The most frequently identified eST was #41, identified in 18% (13/73) of the aliquots. Based on their genetic relationships, the U. spp. DNA was divided into two clusters and four subgroups [cluster I (U. parvum): A, 43% (n = 31); B, 15% (n = 11); and C, 26% (n = 19); cluster II (U. urealyticum): 1; 16% (n = 12)]. Cluster II had a higher rate of polymicrobial findings than cluster I (58% vs 16%; p = 0.005), while subgroup A had the highest rate of concomitant Mycoplasma hominis in the amniotic fluid samples (66%; p = 0.04). In conclusion, Ureaplasma spp. DNA obtained from PPROM consisted of 33 different eSTs of U. spp. DNA. No differences in maternal and neonatal characteristics were found among the phylogenetical subgroups of U. spp. DNA, except for a higher rate of polymicrobial amniotic fluid findings in those with U. urealyticumand the concomitant presence of M. hominis in the amniotic fluid in those with the presence of U. parvum.

• Klíčová slova
• Genital mycoplasma, Microbial invasion of the amniotic cavity, Molecular biology, Mollicutes, Morbidity, Neonates, Preterm delivery, Sequencing,
• MeSH
• DNA bakterií analýza   genetika   MeSH
• dospělí MeSH
• fylogeneze MeSH
• gestační stáří MeSH
• infekční komplikace v těhotenství mikrobiologie   MeSH
• lidé MeSH
• multilokusová sekvenční typizace * MeSH
• plodová voda * mikrobiologie   MeSH
• předčasný odtok plodové vody * mikrobiologie   MeSH
• retrospektivní studie MeSH
• těhotenství MeSH
• Ureaplasma * genetika   izolace a purifikace   MeSH
• ureaplasmatické infekce * mikrobiologie   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• DNA bakterií MeSH

Spontaneous preterm delivery presents one of the most complex challenges in obstetrics and is a leading cause of perinatal morbidity and mortality. Although it is a common endpoint for multiple pathological processes, the mechanisms governing the etiological complexity of spontaneous preterm birth and the placental responses are poorly understood. This study examined placental tissues collected between May 2019 and May 2022 from a well-defined cohort of women who experienced spontaneous preterm birth (n = 72) and healthy full-term deliveries (n = 30). Placental metabolomic profiling of polar metabolites was performed using Ultra-High Performance Liquid Chromatography/Mass Spectrometry (UHPLC/MS) analysis. The resulting data were analyzed using multi- and univariate statistical methods followed by unsupervised clustering. A comprehensive metabolomic evaluation of the placenta revealed that spontaneous preterm birth was associated with significant changes in the levels of 34 polar metabolites involved in intracellular energy metabolism and biochemical activity, including amino acids, purine metabolites, and small organic acids. We found that neither the preterm delivery phenotype nor the inflammatory response explain the reported differential placental metabolome. However, unsupervised clustering revealed two molecular subtypes of placentas from spontaneous preterm pregnancies exhibiting differential enrichment of clinical parameters. We also identified differences between early and late preterm samples, suggesting distinct placental functions in early spontaneous preterm delivery. Altogether, we present evidence that spontaneous preterm birth is associated with significant changes in the level of placental polar metabolites. Dysregulation of the placental metabolome may underpin important (patho)physiological mechanisms involved in preterm birth etiology and long-term neonatal outcomes.

• Klíčová slova
• inflammation, metabolism, metabolomics, placenta, preterm birth,
• Publikační typ
• časopisecké články MeSH

OBJECTIVE: To assess the association between newborn birth weight and the presence of intra-amniotic infection, presence of sterile intra-amniotic inflammation, and absence of intra-amniotic inflammation in pregnancies with preterm labor with intact membranes. METHODS: A total of 69 pregnancies with preterm labor with intact membranes between gestational ages 22 + 0 and 34 + 6 weeks who delivered within seven days of admission were included in this retrospective cohort study. Transabdominal amniocentesis to determine the presence of microorganisms and/or their nucleic acids in amniotic fluid (through culturing and molecular biology methods) and intra-amniotic inflammation (according to amniotic fluid interleukin-6 concentrations) were performed as part of standard clinical management. The participants were further divided into three subgroups: intra-amniotic infection (presence of microorganisms and/or nucleic acids along with intra-amniotic inflammation), sterile intra-amniotic inflammation (intra-amniotic inflammation alone), and without intra-amniotic inflammation. Birth weights of newborns were expressed as percentiles derived from the INTERGROWTH-21st standards for (i) estimated fetal weight and (ii) newborn birth weight. RESULTS: No difference in birth weights, expressed as percentiles derived from the standard for estimated fetal weight, was found among the women with intra-amniotic infection, with sterile intra-amniotic inflammation, and without intra-amniotic inflammation (with infection, median 29; with sterile inflammation, median 54; without inflammation, median 53; p = 0.06). Differences among the subgroups were identified in the birth weight rates, expressed as percentiles derived from the standard for estimated fetal weight, which were less than the 10th percentile (with infection: 20%, with inflammation: 13%, without inflammation: 0%; p = 0.04) and 25th percentile (with infection: 47%, with inflammation: 31%, without inflammation: 9%; p = 0.01). No differences among the subgroups were observed when percentiles of birth weight were derived from the birth weight standard. CONCLUSIONS: The presence of intra-amniotic inflammatory complications in pregnancies with preterm labor with intact membranes prior to the gestational age of 35 weeks was associated with a higher rate of newborns with birth weight less than the 10th and 25th percentile, when percentiles of birth weight were derived from the standard for estimated fetal weight.

• Klíčová slova
• amniocentesis, amniotic fluid, estimated fetal weight, fetal growth, intergrowth, intra-amniotic inflammation, microbial invasion of the amniotic cavity, preterm birth,
• Publikační typ
• časopisecké články MeSH

Objectives: To develop a rat model of intra-amniotic inflammation, characterized by the concentration of interleukin-6 in the amniotic fluid, induced by an ultrasound-guided transabdominal administration of lipopolysaccharide into individual gestational sacs. Methods: An ultrasound-guided transabdominal intra-amniotic administration of lipopolysaccharide or phosphate-buffered saline (PBS) as control was performed in rats on embryonic day 18. Only accessible gestational sacs with precise recording of their positions were injected. Twenty-four hours later, individual amniotic fluid samples were collected from the gestational sacs of laparotomized animals. The gestational sacs were divided into four subgroups: (i) with lipopolysaccharide: injected gestational sacs from rats undergoing lipopolysaccharide administration; (ii) without lipopolysaccharide: non-injected gestational sacs from rats undergoing lipopolysaccharide administration; (iii) with PBS: injected gestational sacs from rats undergoing PBS administration; and (iv) without PBS: non-injected gestational sacs from rats undergoing PBS administration. The concentration of interleukin-6 in individual amniotic fluid samples was assessed using ELISA. Results: In the group of five animals receiving lipopolysaccharide, 24 (33%) and 48 (77%) gestational sacs were and were not injected, respectively. The amniotic fluid was obtained from 21 (88%) injected and 46 (95%) non-injected sacs. In the control group of five animals receiving phosphate-buffered saline, 28 (35%) and 52 (75%) gestational sacs were and were not injected, respectively. The amniotic fluid was obtained from 18 (64%) injected and 50 (96%) non-injected sacs. No labor occurred, and only one fetal death was observed in a gestational sac injected with lipopolysaccharide. Differences in concentrations of interleukin-6 in the amniotic fluid were found among the subgroups of the gestational sacs (with lipopolysaccharide: median 762 pg/ml; without lipopolysaccharide: median 35.6 pg/ml; with PBS: median 35.6 pg/ml; and without PBS: median 35.6 pg/ml; p < 0.0001). Concentrations of interleukin-6 in the amniotic fluid from the gestational sacs with lipopolysaccharide were significantly higher than those in the three remaining subgroups (p < 0.0001). No differences in concentrations of interleukin-6 in the amniotic fluid were identified between the three remaining subgroups. Conclusion: The ultrasound-guided transabdominal intra-amniotic administration of lipopolysaccharide with a subsequent collection and analysis of amniotic fluid samples is feasible in rats. The intra-amniotic administration of lipopolysaccharide led to the development of intra-amniotic inflammation without leading to fetal mortality or induction of labor.

• Klíčová slova
• amniocentesis, animal model, lipopolysaccharide, minimally invasive, preterm birth, preterm delivery,
• Publikační typ
• časopisecké články MeSH

Objectives: To determine the prevalence and load of Ureaplasma spp. DNA in the cervical fluid of women with singleton pregnancies complicated by preterm prelabor rupture of membranes (PPROM) with respect to intra-amniotic infection, sterile intra-amniotic inflammation, and colonization of the amniotic fluid. Methods: A total of 217 women with PPROM between gestational ages 24 + 0 and 33 + 6 weeks were included in this study. Paired amniotic and cervical fluid samples were collected at the time of admission via transabdominal amniocentesis and using a Dacron polyester swab, respectively. Microbial invasion of the amniotic cavity was diagnosed using a combination of culture and molecular biology methods. Intra-amniotic inflammation was determined based on the concentration of interleukin-6 in the amniotic fluid. Based on the presence or absence of these conditions, the women were stratified into the following subgroups: intra-amniotic infection (with both), sterile intra-amniotic inflammation (with inflammation only), colonization (with microorganisms only), and negative amniotic fluid (without either). The Ureaplasma spp. DNA load in the cervical fluid was assessed using PCR. Results: Ureaplasma spp. DNA in the cervical fluid was found in 61% (133/217) of the women. Women with negative amniotic had similar prevalence of Ureaplasma spp. DNA in cervical fluid (55%) to those with sterile intra-amniotic inflammation (54%) but lower than those with intra-amniotic infection (73%) and colonization (86%; p < 0.0001). Women with negative amniotic fluid had a lower load of Ureaplasma spp. DNA in their cervical fluid (median: 4.7 × 103 copies of DNA/ml) than those with intra-amniotic infection (median: 2.8 × 105 copies DNA/ml), sterile intra-amniotic inflammation (median: 5.3 × 104 copies DNA/ml), and colonization (median: 1.2 × 105 copies DNA/mL; p < 0.0001). Conclusion: In conclusion, in PPROM at <34 weeks, the presence of intra-amniotic infection, sterile intra-amniotic inflammation, or colonization of the amniotic fluid was associated with a higher prevalence and/or load of Ureaplasma spp. DNA in the cervical fluid than the absence of intra-amniotic complications.

• Klíčová slova
• genital mycoplasma, intra-amniotic inflammation, microbial invasion of the amniotic cavity, non-invasive sample, preterm delivery,
• Publikační typ
• časopisecké články MeSH

To determine the main clinical characteristics of preterm prelabor rupture of membranes (PPROM) complicated by colonization of the amniotic cavity (microbial invasion of the amniotic cavity without intra-amniotic inflammation). A total of 302 women with PPROM were included. Transabdominal amniocentesis was performed and amniotic fluid was assessed. Based of microbial invasion of the amniotic cavity and intra-amniotic inflammation (interleukin-6 ≥ 3000 pg/mL), the women were divided into following groups: intra-amniotic infection, sterile intra-amniotic inflammation, colonization of the amniotic cavity, and negative amniotic fluid. Colonization was found in 11% (32/302) of the women. The most common bacteria identified in the amniotic fluid were Ureaplasma spp. with a lower burden than those with intra-amniotic infection (p = 0.03). The intensity of intra-amniotic inflammatory response measured by interleukin-6 was higher in women with colonization than in those with negative amniotic fluid (medians: 961 pg/mL vs. 616 pg/mL; p = 0.04). Women with colonization had higher rates of acute inflammatory placental lesions than those with negative amniotic fluid. In PPROM, colonization, caused mainly by microorganisms from the lower genital tract, might represent an early stage of microbial invasion of the amniotic cavity with a weak intra-amniotic inflammatory response.

• MeSH
• chorioamnionitida * mikrobiologie   MeSH
• interleukin-6 MeSH
• lidé MeSH
• novorozenec MeSH
• placenta MeSH
• plodová voda mikrobiologie   MeSH
• předčasný odtok plodové vody MeSH
• retrospektivní studie MeSH
• těhotenství MeSH
• zánět komplikace   MeSH
• Check Tag
• lidé MeSH
• novorozenec MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• interleukin-6 MeSH

Spontaneous preterm birth is a serious medical condition responsible for substantial perinatal morbidity and mortality. Its phenotypic characteristics, preterm labor with intact membranes (PTL) and preterm premature rupture of the membranes (PPROM), are associated with significantly increased risks of neurological and behavioral alterations in childhood and later life. Recognizing the inflammatory milieu associated with PTL and PPROM, here, we examined expression signatures of placental tryptophan metabolism, an important pathway in prenatal brain development and immunotolerance. The study was performed in a well-characterized clinical cohort of healthy term pregnancies (n = 39) and 167 preterm deliveries (PTL, n = 38 and PPROM, n = 129). Within the preterm group, we then investigated potential mechanistic links between differential placental tryptophan pathway expression, preterm birth and both intra-amniotic markers (such as amniotic fluid interleukin-6) and maternal inflammatory markers (such as maternal serum C-reactive protein and white blood cell count). We show that preterm birth is associated with significant changes in placental tryptophan metabolism. Multifactorial analysis revealed similarities in expression patterns associated with multiple phenotypes of preterm delivery. Subsequent correlation computations and mediation analyses identified links between intra-amniotic and maternal inflammatory markers and placental serotonin and kynurenine pathways of tryptophan catabolism. Collectively, the findings suggest that a hostile inflammatory environment associated with preterm delivery underlies the mechanisms affecting placental endocrine/transport functions and may contribute to disruption of developmental programming of the fetal brain.

• MeSH
• biologické markery MeSH
• lidé MeSH
• metabolické sítě a dráhy MeSH
• náchylnost k nemoci MeSH
• placenta metabolismus   MeSH
• předčasný porod diagnóza   etiologie   metabolismus   MeSH
• regulace genové exprese MeSH
• rizikové faktory MeSH
• stanovení celkové genové exprese MeSH
• těhotenství MeSH
• transkriptom * MeSH
• tryptofan metabolismus   MeSH
• výpočetní biologie metody   MeSH
• výsledek těhotenství MeSH
• zánět komplikace   etiologie   MeSH
• Check Tag
• lidé MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• biologické markery MeSH
• tryptofan MeSH