Elevated human placental heat shock protein 5 is associated with spontaneous preterm birth

Background: Specific heat shock proteins are associated with pregnancy complications, including spontaneous preterm birth (SPTB). Placental proteomics and whole exome sequencing recently suggested an association between heat shock protein HSPA5 and uncomplicated SPTB. In the present study, we investigated the localization of and possible roles for HSPA5 in SPTB. Methods: Western blot was performed to validate the result from the previously published proteomic analysis. We used qPCR to assess mRNA expression of genes and immunohistochemistry and immunoelectron microscopy to examine localization of HSPA5 in placental tissue. We silenced the HSPA5 gene in the HTR8/SVneo human trophoblast cell line to investigate possible functions of HSPA5. Results: HSPA5 was upregulated in placentas from SPTBs compared to spontaneous term births. We did not observe upregulation of HSPA5 mRNA in placental samples. The protein was localized in placental trophoblast in both spontaneous preterm and term placentas. Gene silencing of HSPA5 in human trophoblast cell culture affected the inflammatory response and decreased the expression of several proinflammatory genes. Conclusions: We suggest that upregulation of HSPA5 in the placenta is associated with spontaneous preterm labor. HSPA5 may promote the inflammatory response and alter the anti-inflammatory state of the placenta which could eventually lead to premature labor. Impact: We validated upregulation of HSPA5 in placentas from spontaneous preterm birth. HSPA5 was not upregulated at transcriptional level which suggests that it may be regulated post-translationally.Silencing HSPA5 in a human trophoblast–derived cell line suggested that HSPA5 promotes expression of proinflammatory cytokines. The emerging inflammation could lead to spontaneous preterm labor.Identifying inflammatory pathways and factors associated with spontaneous preterm birth increases knowledge of the molecular mechanisms of premature labor. This could provide cues to predict imminent premature labor and lead to information about how to safely maintain pregnancies.publishedVersionPeer reviewe

Хроматоденситометрическое изучение листьев брусники обыкновенной (Vaccinium vitis-idaea L.)

VACCINIUMБРУСНИКАРАСТЕНИЙ ЛИСТЬЯРАСТЕНИЯ СТРУКТУРНЫЕ ЧАСТИ НАДЗЕМНЫЕАРБУТИНХРОМАТОГРАФИЯ ЖИДКОСТНАЯ /ИСПХРОМАТОДЕНСИТОМЕТРИЧЕСКИЙ СПОСОБХРОМАТОГРАФИЧЕСКАЯ ЗОНАХРОМАТОГРАММАРАСТЕНИЙ ЭКСТРАКТЫРЕАКТИВ ГИББСАЛИНЕЙНАЯ ЗАВИСИМОСТЬПроанализированы листья брусники обыкновенной, заготовленные в местах естественного произрастания в окрестностях г. Витебска Республики Беларусь. С помощью хроматоденситометрического способа изучались условия проведения хроматоденситометрического определения арбутина в листьях брусники. Подобраны система растворителей, концентрация реагента-проявителя (реактив Гиббса) для хроматоденситометрического способа определения арбутина в листьях брусники обыкновенной. Изучен характер зависимости цветометрических характеристик пятен арбутина от его количества в хроматографической зоне. Выявлена линейная зависимость цветометрических характеристик зон арбутина от его количества в хроматографической зоне. Было проведено сравнение хроматоденситометрического способа определения арбутина в листьях брусники с методом ВЭЖХ. Установлено, что результаты ВЭЖХ сопоставимы с результатами хроматоденситометрического способа определения арбутина в листьях брусники обыкновенной, что дает основание сделать вывод о достаточно высокой точности хроматоденситометрического способа и делает возможным его применение для качественного и количественного определения арбутина в листьях брусники. Изучались различные условия экстракции арбутина из листьев брусники обыкновенной. Построены графики зависимости полноты экстракции арбутина из листьев брусники обыкновенной при различных условиях экстракции. Подобраны оптимальные условия экстракции арбутина (степень измельченности листьев брусники –2 мм, время экстракции – 40 минут, экстрагент – вода).The leaves of common lingonberry harvested in the places of its natural growth in the vicinity of the city of Vitebsk of the Republic of Belarus have been analyzed using the chromatodensitometric method. The conditions of the chromatodensitometric method use for determining arbutin in the common lingonberry leaves have been studied. The solvent system, the concentration of the reagent- developer (Gibbs reagent) for the chromatodensitometric method used to determine arbutin in the common lingonberry leaves have been selected. The character of the dependence of the colorimetric characteristics of arbutin spots on its quantity in the chromatographic zone has been studied. A linear dependence of the colorimetric characteristics of arbutin zones on its quantity in the chromatographic zone has been revealed. The chromatodensitometric method of the determination of arbutin in the common lingonberry leaves has been compared to that of HPLC. It has been established that the HPLC results are comparable with those obtained by means of the chromatodensitometric method of determining arbutin in the common lingonberry leaves, which gives grounds to conclude that the chromatodensitometric method is rather accurate and may be used for the quantitative and qualitative determination of arbutin in the common lingonberry leaves. Various conditions for the extraction of arbutin from the leaves of common lingonberry have been studied. The graphs of the dependence of the completeness of the common lingonberry leaves arbutin extraction under various extraction conditions have been constructed. Optimal conditions for the extraction of arbutin have been selected (the degree of crushing the lingonberry leaves – 2 mm, extraction time – 40 minutes, the extractant – water)

A minor role of asparaginase in predisposing to cerebral venous thromboses in adult acute lymphoblastic leukemia patients

Cerebral venous thrombosis (CVT) covers up to a third of all venous thromboses (VTs) detected in patients with acute lymphoblastic leukemia (ALL). It usually hampers patients' lives and may also endanger efficient leukemia treatment. Although many factors have been suggested to account for an elevated risk of VTs in patients with ALL, there still is a lack of studies focusing on CVTs and especially in the setting of adult ALL patients. We studied in our retrospective population-based cohort the occurrence, characteristics, as well as risk factors for VTs in 186 consecutively diagnosed Finnish adult ALL patients treated with a national pediatric-inspired treatment protocol ALL2000. In the risk factor analyses for VTs we found a distinction of the characteristics of the patients acquiring CVT from those with other kinds of VTs or without thrombosis. In contrast to previous studies we were also able to compare the effects of asparaginase in relation to CVT occurrence. Notably, more than half of the CVTs were diagnosed prior the administration of asparaginase which accentuates the role of other risk factors on the pathophysiology of CVT compared to truncal or central venous line (CVL) VTs in adult ALL patients

Yhtenäiset vaatimukset lääketieteen ja hammaslääketieteen tohtorintutkinnolle Suomessa

Oulussa järjestettiin 13.6.2018 lääketieteen ja hammaslääketieteen tohtorintutkinnon konsensuskokous, jossa käytiin läpi vallitsevia käytäntöjä ja muutostarpeita. Ohjeellinen tutkinnon laajuus on neljä vuotta kokopäiväistä työtä, ja muodollisesti tutkinnon myöntää yhtä yliopistoa lukuun ottamatta aina vastaava tiedekunta. Ohjaajia on tyypillisesti kahdesta kolmeen, ja yksi on pääohjaaja. Seurantaryhmä on käytössä tai ollaan ottamassa käyttöön kaikissa yliopistoissa. Aktiivinen seurantaryhmä tukee merkittävästi ohjausprosessia. Väitöskirjojen asiantuntijoina toimivat esitarkastajat ja vastaväittäjät ovat yleensä muualta kuin suorituspaikan yliopistosta. Osittain tästä syystä suuria eroja eri yliopistojen tohtorintutkinnon vaatimusten välillä ei todettu. Haasteena väitöskirjatyössä on kliinisen työn paine ja nuorten kollegoiden ruuhkavuosien kuormittuminen, mitkä heikentävät mahdollisuuksia tutkimustyöhön. Myönteisiä muutoksia ovat olleet tohtoriopintojen systematisoituminen sekä ohjaamisen tason ja väitöskirjantekijöiden tuen paraneminen. Kaikki osallistuneet kannattivat käytäntöjen vakiinnuttamista ja yhtenäistämistä säännöllisellä konsensuskokoustyöskentelyllä.</p

Human placental proteomics and exon variant studies link AAT/SERPINA1 with spontaneous preterm birth

Background: Preterm birth is defined as live birth before 37 completed weeks of pregnancy, and it is a major problem worldwide. The molecular mechanisms that lead to onset of spontaneous preterm birth are incompletely understood. Prediction and evaluation of the risk of preterm birth is challenging as there is a lack of accurate biomarkers. In this study, our aim was to identify placental proteins that associate with spontaneous preterm birth. Methods: We analyzed the proteomes from placentas to identify proteins that associate with both gestational age and spontaneous labor. Next, rare and potentially damaging gene variants of the identified protein candidates were sought for from our whole exome sequencing data. Further experiments we performed on placental samples and placenta-associated cells to explore the location and function of the spontaneous preterm labor-associated proteins in placentas. Results: Exome sequencing data revealed rare damaging variants in SERPINA1 in families with recurrent spontaneous preterm deliveries. Protein and mRNA levels of alpha-1 antitrypsin/SERPINA1 from the maternal side of the placenta were downregulated in spontaneous preterm births. Alpha-1 antitrypsin was expressed by villous trophoblasts in the placenta, and immunoelectron microscopy showed localization in decidual fibrinoid deposits in association with specific extracellular proteins. siRNA knockdown in trophoblast-derived HTR8/SVneo cells revealed that SERPINA1 had a marked effect on regulation of the actin cytoskeleton pathway, Slit–Robo signaling, and extracellular matrix organization. Conclusions: Alpha-1 antitrypsin is a protease inhibitor. We propose that loss of the protease inhibition effects of alpha-1 antitrypsin renders structures critical to maintaining pregnancy susceptible to proteases and inflammatory activation. This may lead to spontaneous premature birth.publishedVersionPeer reviewe

A study of genes encoding cytokines (IL6, IL10, TNF), cytokine receptors (IL6R, IL6ST), and glucocorticoid receptor (NR3C1) and susceptibility to bronchopulmonary dysplasia

Peer reviewe

Differential Phagocytosis of White versus Opaque Candida albicans by Drosophila and Mouse Phagocytes

The human fungal pathogen Candida albicans resides asymptomatically in the gut of most healthy people but causes serious invasive diseases in immunocompromised patients. Many C. albicans strains have the ability to stochastically switch between distinct white and opaque cell types, but it is not known with certainty what role this switching plays in the physiology of the organism. Here, we report a previously undescribed difference between white and opaque cells, namely their interaction with host phagocytic cells. We show that both Drosophila hemocyte-derived S2 cells and mouse macrophage-derived RAW264.7 cells preferentially phagocytose white cells over opaque cells. This difference is seen both in the overall percentage of cultured cells that phagocytose white versus opaque C. albicans and in the average number of C. albicans taken up by each phagocytic cell. We conclude that susceptibility to phagocytosis by cells of the innate immune system is an important distinction between white and opaque C. albicans, and propose that one role of switching from the prevalent white form into the rarer opaque form may be to allow C. albicans to escape phagocytosis

Expression of CPPED1 in human trophoblasts is associated with timing of term birth

Understanding of timing of human parturition is incomplete. Therefore, we carried out proteomic analyses of full-term placentas from uncomplicated pregnancies to identify protein signatures associated with the onset of spontaneous delivery. We found quantitative associations of 10 proteins with spontaneous term birth, evident either in the basal or in the chorionic plates or in both. Additional 18 proteins were associated according to the location within placenta indicating local variations in protein amounts. Calcineurin-like phosphoesterase domain-containing 1 (CPPED1), a phosphatase previously suggested dephosphorylating AKT1/PKB, was one of the identified proteins. qRT-PCR revealed the mRNA level of CPPED1 was higher in elective caesarean deliveries than in spontaneous births, while immunohistochemistry showed CPPED1 in cytotrophoblasts, syncytiotrophoblasts and extravillous trophoblasts. Noteworthy, phosphorylation status of AKT1 did not differ between placentas from elective caesarean and spontaneous deliveries. Additionally, analyses of samples from infants indicated that single-nucleotide polymorphisms rs11643593 and rs8048866 of CPPED1 were associated with duration of term pregnancy. Finally, post-transcriptional silencing of CPPED1 in cultured HTR8/SVneo cells by siRNAs affected gene expression in pathways associated with inflammation and blood vessel development. We postulate that functions regulated by CPPED1 in trophoblasts at choriodecidual interphase have a role in the induction of term labour, but it may be independent of AKT1

Genotype and Gene Expression Associations with Immune Function in Drosophila

It is now well established that natural populations of Drosophila melanogaster harbor substantial genetic variation associated with physiological measures of immune function. In no case, however, have intermediate measures of immune function, such as transcriptional activity of immune-related genes, been tested as mediators of phenotypic variation in immunity. In this study, we measured bacterial load sustained after infection of D. melanogaster with Serratia marcescens, Providencia rettgeri, Enterococcus faecalis, and Lactococcus lactis in a panel of 94 third-chromosome substitution lines. We also measured transcriptional levels of 329 immune-related genes eight hours after infection with E. faecalis and S. marcescens in lines from the phenotypic tails of the test panel. We genotyped the substitution lines at 137 polymorphic markers distributed across 25 genes in order to test for statistical associations among genotype, bacterial load, and transcriptional dynamics. We find that genetic polymorphisms in the pathogen recognition genes (and particularly in PGRP-LC, GNBP1, and GNBP2) are most significantly associated with variation in bacterial load. We also find that overall transcriptional induction of effector proteins is a significant predictor of bacterial load after infection with E. faecalis, and that a marker upstream of the recognition gene PGRP-SD is statistically associated with variation in both bacterial load and transcriptional induction of effector proteins. These results show that polymorphism in genes near the top of the immune system signaling cascade can have a disproportionate effect on organismal phenotype due to the amplification of minor effects through the cascade