Expression of placental glycans and its role in regulating peripheral blood NK cells during preeclampsia: a perspective.

Preeclampsia is a pregnancy-related multisystem disorder characterized by altered trophoblast invasion, oxidative stress, exacerbation of systemic inflammatory response, and endothelial damage. The pathogenesis includes hypertension and mild-to-severe microangiopathy in the kidney, liver, placenta, and brain. The main mechanisms involved in its pathogenesis have been proposed to limit trophoblast invasion and increase the release of extracellular vesicles from the syncytiotrophoblast into the maternal circulation, exacerbating the systemic inflammatory response. The placenta expresses glycans as part of its development and maternal immune tolerance during gestation. The expression profile of glycans at the maternal-fetal interface may play a fundamental role in physiological pregnancy changes and disorders such as preeclampsia. It is unclear whether glycans and their lectin-like receptors are involved in the mechanisms of maternal-fetal recognition by immune cells during pregnancy homeostasis. The expression profile of glycans appears to be altered in hypertensive disorders of pregnancy, which could lead to alterations in the placental microenvironment and vascular endothelium in pregnancy conditions such as preeclampsia. Glycans with immunomodulatory properties at the maternal-fetal interface are altered in early-onset severe preeclampsia, implying that innate immune system components, such as NK cells, exacerbate the systemic inflammatory response observed in preeclampsia. In this article, we discuss the evidence for the role of glycans in gestational physiology and the perspective of glycobiology on the pathophysiology of hypertensive disorders in gestation.

Preeclampsia association of placental nucleotide variations in eNOS, VEGFA, and FLT-1 genes in Latin American pregnant women.

INTRODUCTION: Preeclampsia is a leading cause of maternal and fetal morbidity in low- and middle-income countries, including those in Latin America. Placental vascular alterations are crucial in the pathophysiology of preeclampsia and few studies have evaluated nucleotide variations on genes associated with vascular regulation in the human placenta. This study aimed to evaluate whether placental nucleotide variations on eNOS, VEGFA, and FLT-1 genes are more frequently associated with preeclampsia in the Latin American population. METHODS: This case-control study included placental tissue from 88 controls and 82 cases that were genotyped through Taqman probes for eNOS, VEGFA, and FLT-1 genes. The intergroup comparisons were analyzed with the Mann-Whitney U test. Genotype and allele frequencies were compared by the X2 test. The association between the nucleotide variants with preeclampsia was evaluated through logistic regression analysis. RESULTS: A significant association was observed for VEGFA SNV rs2010963 (OR 1.95; CI 95% 1.13-3.37), after adjusting for population substructure. The allele combination T, G, G, C, C, C (rs2070744, rs1799983, rs2010963, rs3025039, rs699947 and rs4769613 respectively), showed a negative association with preeclampsia (OR 0.08; CI 95% 0.01-0.93). DISCUSSION: Placental SNV rs2010963 in the VEGFA gene was a risk factor for preeclampsia, while the allele combination T, G, G, C, C, C may represent potential protective factors for preeclampsia within Latin American women.

Obstetric Antiphospholipid Syndrome: An Approach from Glycans of the Immunoglobulin G.

This is a case report of women with pregnancy morbidity (PM), some of them associated with antiphospholipid syndrome (APS), in which the glycan patterns of immunoglobulin G (IgG) were investigated based on the theory of alteration of glycosylation in autoimmunity. We used lectin blot to determine changes in terminal glycosylation of polyclonal IgG from women with antiphospholipid (aPL) antibodies and PM plus vascular thrombosis (PM/VT) and seronegative-obstetric APS (SN-OAPS). In addition, we analyzed IgG from women with PM without aPL (PM/aPL-) and healthy women, as controls. Even though the SN-OAPS and PM/VT groups share the PM, only the SN-OAPS group showed a decreased expression of galactose compared to the healthy group. We also found the presence of mannosylated oligosaccharides in IgG from all patients being significantly higher in IgG from women of the PM/aPL- group. The differences in glycans presented here could relate to pathological mechanisms of PM associated with APS.

Glycan expression in chorionic villi from histocultures of women with early-onset preeclampsia: Immunomodulatory effects on peripheral natural killer cells.

New evidence suggests that glycan expression in placental cells of women with invasive disorders of pregnancy differs from that in normal pregnant women. Hypothesizing that modifications of glycan expression could account for the course of preeclampsia, we established placental villous histocultures and compared glycan expression in women with preeclampsia with that in normal pregnant women and also in syncytialized BeWo cells, and we tested the effect of glycan expression on the functional phenotypes of circulating natural killer (NK) cells. Histocultures of third-trimester placentae from women with preeclampsia and full-term placentae from healthy pregnant women and BeWo choriocarcinoma cells were assessed for the expression of terminal glycans by lectin-binding assays. Circulating NK cells from nonpregnant healthy donors were tested in vitro for their cytotoxic activity and intracellular cytokine content. Histocultures from women with preeclampsia expressed significantly more mannose than did those from healthy pregnant women. Both histocultures and BeWo cells expressed terminal fucose, mannose, sialic acid, and N -acetylgalactosamine, although mean fluorescence intensity (MFI) expression was lower in choriocarcinoma cells than in cells from histocultures. Cocultures of circulating NK cells with K562 target cells resulted in a dose-dependent cytotoxicity effect, but the use of BeWo cells as target reduced cytotoxic activity; this reduction was not affected by syncytialization. Histocultures of placental villous tissue of women with preeclampsia expressed high levels of terminal mannose. We proposethat placental glycans may modulate the functional activity of circulating NK cells in the context of systemic inflammatory response in preeclampsia.

Cistatina C vs. marcadores convencionales de función renal: una actualización / Cystatin C vs conventional markers of renal function: an update

RESUMEN Objetivo: Esta revisión busca proporcionar información actual de los avances de los últimos diez años reportados para mejorar el diagnóstico de enfermedad renal con marcadores como la cistatina C, en comparación con otros analitos convencionales utilizados en la práctica clínica. Materiales y métodos: Se llevó a cabo una búsqueda de artículos originales y de revisión en idioma inglés y español de los últimos diez años en las siguientes bases de datos: MedLine, Library Plus, ProQuest, NCBI y ScienceDirect. Resultados: Para la evaluación de la función renal se usan marcadores convencionales como la creatinina y el nitrógeno ureico en sangre. Sin embargo, estas pruebas están sometidas a diferentes fuentes de variabilidad biológica, razón por la cual se ve la necesidad de identificar otros marcadores que permitan detectar de forma precoz la lesión renal, y que no se vean influenciados por factores dependientes del paciente como la cistatina C, una proteína de bajo peso molecular propuesta como un marcador de función renal más exacto, específico y sensible que la creatinina en la estimación de la tasa de filtración glomerular. Conclusión: Se considera que la cistatina C cumple con características que la hacen un biomarcador más útil en la determinación de lesión renal, ya que muestra mayor utilidad diagnóstica que la creatinina sérica para detectar daño renal temprano, sin embargo, no se utiliza como prueba rutinaria en el diagnóstico de patologías renales por su alto costo al momento de realizar un montaje en el laboratorio clínico.

ABSTRACT Objective: To provide current information about the progress of the last ten years reported to improve the diagnosis of kidney disease with markers such as cystatin C, with other usual analytes used in clinical practice. Materials and methods: A search for original and review articles in english and spanish in the last ten years was carried out in the following databases: MedLine, Library Plus, Pro-Quest, NCBI and ScienceDirect. Results: For the evaluation of renal function markers are used such as creatinine and blood urea nitrogen. However, these tests are sometimes determined to different sources of biological variability, which is why the need to identify other markers that detect kidney damage early, and that is not influenced by pacific-dependent factors such as Cystatin C, a molecular protein of low molecular weight proposed as a marker of renal function more accurate, specific and sensitive than creatinine in modifying the glomerular filtration rate. Conclusion: Cystatin C meets with characteristics that make it a more useful biomarker in the determination of renal injury, since it shows greater diagnostic utility than serum creatinine to detect early renal damage, however, it is not used as a routine test in the diagnosis of renal pathologies due to its high cost when assembling in the clinical laboratory.

Glycosylation Profile of the Transferrin Receptor in Gestational Iron Deficiency and Early-Onset Severe Preeclampsia.

OBJECTIVE: To examine the expression of hypoxia-inducible factor-1α (HIF-1α), TfR1, and TfR1-attached terminal monosaccharides in placentas of women with IDAP and severe preeclampsia. METHODS: TfR1 and HIF-1α were detected by western blot. Immunoadsorption of TfR1 was performed to characterize the terminal monosaccharides by specific lectin binding. RESULTS: There was no difference in the expression of TfR1 and HIF-1α between groups. Lectin blot analysis pointed out an overexpression of galactose ß1-4 N-acetylglucosamine (Gal-GlcNAc) and mannose in severe preeclampsia. CONCLUSION: The increase in Gal-GlcNAc may be due to the increased presence of antennary structures and the mannose glycans of TfR1 may indicate the presence of misfolded or incomplete proteins. These findings may be associated with the low expression of placental TfR1 in women with preeclampsia.

Assessment of endocrine disruptor effects of levonorgestrel and its photoproducts: Environmental implications of released fractions after their photocatalytic removal.

The presence of levonorgestrel (LNG) in water bodies via direct discharge and human excretion has been reported worldwide, but its effects on the reproduction of aquatic species and humans are still unknown. Owing to its recalcitrant properties, LNG is not completely removed during wastewater treatment plants, and many species may be exposed to low traces of this compound from discharged effluents. Thus, in this study, a photocatalytic process for removing LNG along with screening of endocrine disruptor effects for risk assessment was applied. Although the removal rate of LNG by ultraviolet C (UV-C) radiation was >90%, reproductive toxicity testing using the BeWo cell line exposed to LNG and its degraded fraction showed the reduced production of basal human chorionic gonadotropin hormone (ß-hCG) by more than 73%, from 8.90 mIU mL-1 to <2.39 mIU mL-1, with both LNG and the degraded fraction. ß-hCG hormone has been implicated in the viability of trophoblastic cells during the first trimester of pregnancy; therefore, degraded fractions and waterborne LNG may affect reproduction in some aquatic species and humans with low level of exposure.

Characterization of the Venom of C. d. cumanesis of Colombia: Proteomic Analysis and Antivenomic Study.

The Colombian rattlesnake Crotalus durissus cumanensis is distributed in three geographic zones of the country: the Atlantic Coast, the upper valley of the Magdalena River, and the eastern plains of the Colombian Orinoquía. Its venom induces neurological symptoms, such as eyelid ptosis, myasthenic facies, and paralysis of the respiratory muscles, which can lead to death. Identification and analysis of C. d. cumanensis showed nine groups of proteins responsible for the neurotoxic effect, of which the crotoxin complex was the most abundant (64.71%). Immunorecognition tests of C. d. cumanensis showed that the use of a commercial antivenom manufactured in Mexico resulted in immunoreactivity.

Síndrome antifosfolipídico: una aproximación desde la glicobiología / Antiphospholipid syndrome: an approach from the glycobiology

Introducción: La glicosilación es una modificación postraduccional que tiene un papel protagónico en los procesos de comunicación célula-célula, célula-matrix extracelular, por lo tanto modula la interacción con antígenos y el sistema inmune. La inmunoglobulina G (IgG) tiene un sitio de N-glicosilación en la posición 297. Los sacáridos terminales de este glicano impactan la función efectora de la proteína, dirigiendo la respuesta hacia un efecto proinflamatorio o antiinflamatorio. Maverakis. et al (2015), propusieron que cada enfermedad autoinmune tiene una única marca de glicanos, especialmente en las diferentes clases de inmunoglobulinas (teoría de la alteración de los glicanos en la autoinmunidad).

Glicanos de la vellosidad trofoblástica en la anemia ferropénica y la preeclampsia grave / Glycans in villus trophoblast in iron deficiency anemia and early-onset severe preeclampsia

Glycoproteins attached to cell membrane of syncytiotrophoblast are in close contact with maternal blood, thus these molecules could participate in cell-to-cell communication and biological functions involving ligand-receptors in the maternal-fetal interphase. The attached glycans are involved in the stability, folding and exportation of the protein towards the cell membrane. The objective of this study was to characterize the glycan profile of third trimester placental villi obtained from pregnant women with early-onset severe preeclampsia and gestational anemia compared with normal pregnant women. Protein extracts from placental villi were used in lectin blot assays. -2,3 N- and O-linked sialic acid was over-expressed in villous of severe preeclamptic placentas measured by MAA lectin staining. High mannose glycans and Gal-GlcNAc patterns were also increased in severe preeclampsia compared with the other groups. These findings can explain changes in the cell membrane expression of glycoproteins.

Introducción: Las glicoproteínas de la membrana del sincitiotrofoblasto (STB) se encuentran en contacto con la sangre materna, por lo que pueden participar en la comunicación en la interface materno-fetal. Objetivo: caracterizar patrones de glicanos de la vellosidad trofoblástica de mujeres sanas, anémicas por deficiencia de hierro y preeclámpticas graves de inicio temprano. Materiales y métodos: se obtuvieron extractos proteínicos de vellosidad placentaria de tercer trimestre y se determinó la expresión de patrones de glicanos, usando lectinas. Para la comparación de los grupos se utilizó la prueba de Kruskal-Wallis. Resultados: Se encontró una sobreexpresión en los patrones de glicosilación Gal-GlcNAc, manosa y ácido siálico α2-3 en el grupo con preeclampsia. Conclusiones: El aumento en los patrones Gal-GlcNAc, alta manosilación y ácido siálico α2-3, en proteínas de vellosidad placentaria en los pesos moleculares encontrados, pudiera explicar cambios en la expresión de proteínas de membrana del STB.

Papel del receptor 1 de transferrina en la captación del hierro y su relación con la deficiencia gestacional de hierro y la preeclampsia / Function of Receptor 1 in uptaking transferrin and its relation to iron deficiency and iron gestational preeclampsia

La anemia ferropénica gestacional afecta al 48 porciento de las mujeres y se asocia con efectos deletéreos para la madre y el feto. Para la captación del hierro de la gestante es necesaria la expresión en el sincitiotrofoblasto de la glicoproteína receptor 1 de transferrina (TfR1). En ensayos celulares, en modelos animales y en humanos la deprivación de hierro se ha asociado a un aumento en la transcripción y expresión del TfR1, que se ha explicado como un mecanismo compensatorio para la captación del hierro a favor del feto. De otra parte, en alteraciones de la gestación como la preeclampsia se espera un aumento en la expresión del TfR1 placentario, sin embargo se ha evidenciado una reducción de este. Este evento se ha explicado como una regulación de tipo transcripcional relacionada con el factor de transcripción inducible por la hipoxia. El objetivo fue revisar evidencia que soporte que en la reducción de la expresión del TfR1 en preeclampsia, estén implicados cambios en la glicosilación como una modificación postraduccional relacionada con el adecuado plegamiento, maduración y exportación del receptor a la membrana celular. La base de datos Pubmed fue consultada para identificar los artículos más relevantes. Los descriptores usados fueron metabolismo de hierro, anemia, placenta, receptor de transferrrina, preeclampsia, glicosilación. Se propone una regulación postranscripcional relacionada con la glicosilación que explica como, pese al aumento en la expresión del RNA mensajero del TfR1 inducido por la hipoxia en la placenta preeclámptica, se genera una reducción en su expresión

Iron deficiency gestational anemia affects 48 per cent of women and it is associated with deleterious effects for the mother and her fetus. For mother iron uptake, the syncytiotrophoblast expression of glycoprotein transferrin receptor 1 (TfR1) is required. In cellular trails, in animal models and in humans, iron deprivation has been associated with an increase in TfR1 transcription and expression, which has been explained as a compensatory mechanism for the fetus iron uptake. On the other hand, pregnancy alterations such as preeclampsia are expected to increase in the expression of placental TfR1; however, a reduction of it has shown. This event has been explained as a type transcriptional regulatory factor related to the hypoxia-inducible transcription. Our objective was to review evidence to support that in the reduction of TfR1 preeclampsia expression, changes in glycosylation are involved as a posttranslational modification regarding the appropriate folding, maturation and receptor export to the cell membrane. PubMed database was consulted to identify the most relevant articles. The descriptors used were iron metabolism, anemia, placenta, transferrin receiver, preeclampsia, glycosylation. A posttranscriptional regulation related to glycosylation is proposed explaining how a reduction in expression is generated, despite of the increase in RNA expression of TfR1messenger induced by hypoxia in the preeclamptic placenta

Papel del receptor 1 de transferrina en la captación del hierro y su relación con la deficiencia gestacional de hierro y la preeclampsia / Function of Receptor 1 in uptaking transferrin and its relation to iron deficiency and iron gestational preeclampsia

La anemia ferropénica gestacional afecta al 48 porciento de las mujeres y se asocia con efectos deletéreos para la madre y el feto. Para la captación del hierro de la gestante es necesaria la expresión en el sincitiotrofoblasto de la glicoproteína receptor 1 de transferrina (TfR1). En ensayos celulares, en modelos animales y en humanos la deprivación de hierro se ha asociado a un aumento en la transcripción y expresión del TfR1, que se ha explicado como un mecanismo compensatorio para la captación del hierro a favor del feto. De otra parte, en alteraciones de la gestación como la preeclampsia se espera un aumento en la expresión del TfR1 placentario, sin embargo se ha evidenciado una reducción de este. Este evento se ha explicado como una regulación de tipo transcripcional relacionada con el factor de transcripción inducible por la hipoxia. El objetivo fue revisar evidencia que soporte que en la reducción de la expresión del TfR1 en preeclampsia, estén implicados cambios en la glicosilación como una modificación postraduccional relacionada con el adecuado plegamiento, maduración y exportación del receptor a la membrana celular. La base de datos Pubmed fue consultada para identificar los artículos más relevantes. Los descriptores usados fueron metabolismo de hierro, anemia, placenta, receptor de transferrrina, preeclampsia, glicosilación. Se propone una regulación postranscripcional relacionada con la glicosilación que explica como, pese al aumento en la expresión del RNA mensajero del TfR1 inducido por la hipoxia en la placenta preeclámptica, se genera una reducción en su expresión(AU)

Iron deficiency gestational anemia affects 48 per cent of women and it is associated with deleterious effects for the mother and her fetus. For mother iron uptake, the syncytiotrophoblast expression of glycoprotein transferrin receptor 1 (TfR1) is required. In cellular trails, in animal models and in humans, iron deprivation has been associated with an increase in TfR1 transcription and expression, which has been explained as a compensatory mechanism for the fetus iron uptake. On the other hand, pregnancy alterations such as preeclampsia are expected to increase in the expression of placental TfR1; however, a reduction of it has shown. This event has been explained as a type transcriptional regulatory factor related to the hypoxia-inducible transcription. Our objective was to review evidence to support that in the reduction of TfR1 preeclampsia expression, changes in glycosylation are involved as a posttranslational modification regarding the appropriate folding, maturation and receptor export to the cell membrane. PubMed database was consulted to identify the most relevant articles. The descriptors used were iron metabolism, anemia, placenta, transferrin receiver, preeclampsia, glycosylation. A posttranscriptional regulation related to glycosylation is proposed explaining how a reduction in expression is generated, despite of the increase in RNA expression of TfR1messenger induced by hypoxia in the preeclamptic placenta(AU)

Pregnancy outcome in women with antiphospholipid syndrome and alloimmunity: a case report.

CONTEXT: Patients with antiphospholipid syndrome and alloimmunity have poor pregnancy outcomes. Several diagnostic and therapeutic options exist for these disorders, although there is no consensus as to the best treatment. CASE REPORT: We present here the clinical course and treatment of a woman with a history of two miscarriages who joined our program 10 years ago and has been followed up ever since. After antiphospholipid syndrome and alloimmune failure were diagnosed, she was given preconceptional treatment using unfractionated heparin, aspirin, prednisone and lymphocyte immunizations. She delivered two premature babies in the following two pregnancies. At present both children are healthy and are attending school. The fifth pregnancy was unsuccessful, in spite of having undergone a similar but postconceptional therapeutic scheme. We discuss this case focusing on the pathogenic mechanisms and the therapeutic aspects of these disorders.

Pregnancy outcome in women with antiphospholipid syndrome and alloimmunity: a case report

CONTEXTO: Mulheres com síndrome do anticorpo antifosfolípide e aloimunidade têm um mau prognóstico de gravidez. Existem muitas opções para o diagnóstico e o tratamento destas doenças, embora o tratamento mais adequado não esteja estabelecido. RELATO DE CASO: Apresentamos a evolução clínica e o tratamento de uma mulher com antecedentes de dois abortos e que vem sendo acompanhada, há 10 anos, em nosso serviço. Após o diagnóstico da síndrome do anticorpo antifosfolípide e de aloimunidade, a paciente recebeu tratamento com heparina, aspirina e prednisona, além de imunizações com linfócitos. Nas duas gravidezes subseqüentes, teve dois bebês prematuros, que evoluíram com crescimento e desenvolvimento normais. A quinta gravidez não foi bem-sucedida, apesar de a paciente ter recebido um tratamento semelhante ao utilizado previamente embora após o início da gravidez, não planejada. Na discussão deste caso, enfatizamos os mecanismos patogênicos e o tratamento destas doenças.

Partenogénesis: Un modelo para el estudio de los eventos tempranos del desarrollo mamífero / Parthenogenesis: a model for the study of early events of mammalian development

Las nuevas tecnologías en reproducción animal han abierto líneas de investigación y con ello se han planteado conceptos que revolucionan el campo de la biología reproductiva y de la genética. Una de éstas es la partenogénesis, la cual ha permitido revelar algunos mecanismos moleculares del desarrollo embrionario. Se la puede definir como la generación de un individuo capaz de reproducirse sin la participación del genoma paterno: su éxito depende de una adecuada activación del oocito y de la normal embriogénesis. Se presenta una revisión de la literatura de los fenómenos asociados a la inducción de la partenogénesis y las potencialidades de uso en la investigación de los eventos tempranos de la biología reproductiva de humanos y animales. La partenogénesis en asocio de las nuevas tecnologías de manipulación de embriones in vitro permite aclarar y entender los mecanismos de repartición de los cromosomas, del desarrollo embrionario temprano, el estudio de nuevas formas de herencia y, gracias a la clonación, la interacción del citoplasma y el núcleo en modelos embrionarios.

The Development Of New Reproductive technologies has opened new research lines and allowed to propose concepts in the field of reproductive biology and genetics; one of them is parthenogenesis, defined as the birth of a reproduction-capable individual without the participation of the paternal genome. The successful results depend on the normal activation of the oocyte and the embryonic development. The aim of this paper is to review the molecular events related to the induction of parthenogenesis and their potential use in studying the early events of development in humans and animals. The induction of parthenogenesis associated with new embryo micromanipulation technologies and clonation allow to study chromosome separation, early development, centrosomes and new forms of inheritance and nucleus-cytoplasm interactions