Analysis of differential expression of hypoxia-inducible microRNA-210 gene targets in mild and severe preeclamptic patients.

Preeclampsia (PE) is a multi-system disorder that is specific to human pregnancy. Inadequate oxygenation of uterus and placenta is considered as one of the leading causes for the disease. MicroRNA-210(miR-210) is one of the prime molecules that has emerged in response to hypoxia. The objective of this study was to determine miR-210 expression patterns in plasma from severe PE and mild PE patients, and how that affects the expression of miR-210 target genes. The expression levels of miR-210 were validated using reverse transcription-quantitative PCR in plasma of severe PE (15) and mild PE (15) patients in comparison to controls subjects (15) with normal pregnancy. Then, the association between miR-210 and its downstream genes was validated by using human miR-210 targets RT2 profiler PCR Array. Both the categories (mild and severe) showed significantly high miR-210 expression levels. Also out of the 84 hypoxia miR-210 associated genes screened using mRNA, 18 genes were found to be differentially expressed in severe PE whereas 16 genes in mild PE cases with varying magnitude. All the genes in both the PE groups were found downregulated in comparison to controls. These downregulated genes expressed in both the cases were shown to be participating in immunosuppression, apoptosis, cell growth, signaling, angiogenesis, DNA repair. This study provides novel data on the genes that work downstream of miR-210 and how dysregulated expression of miR-210 can affect their expression and in turn functioning which can be associated with PE risk and severity. This study is the very first to determine the effect of miR-210 expression levels on associated genes in plasma samples.

Differentially expressed circulating microRNAs associated with idiopathic recurrent pregnancy loss.

Recurrent pregnancy loss (RPL) is a major pregnancy complication which reportedly affects 2-3% of all pregnancies. Currently, RPL lacks an effective therapy and a reliable diagnostic and prognostic biomarker. Circulating microRNAs were recently described as potential biomarkers of pregnancy-associated complications. The aim of this study was to determine microRNA expression patterns in the plasma of RPL patients as potential early biomarker of RPL. Study subjects comprised 20 women with early RPL (miscarriage at 8-12 weeks of gestation), and 20 age- and gestation-matched multiparous control women. Circulating microRNAs were extracted from maternal plasma, and the differential microRNA expression were determined using customized pathway-focused miRNA profiler kit. Of the 10 differentially-expressed microRNAs identified, Hsa-let-7e, Hsa-miR-221-3p, Hsa-miR-16, Hsa-miR-519d, Hsa-miR-184, Hsa-miR-410 were upregulated, while Hsa-miR-21, Hsa-miR-125, Hsa-let-7a, Hsa-let-7d were downregulated in RPL cases as compared to control women. Of these, 5 novel microRNAs were reported for the first time to be associated with RPL. These comprised Hsa-let-7e, Hsa-miR-519d, Hsa-miR-410 which were upregulated, and Hsa-let-7a, Hsa-let-7d which were downregulated in RPL. While its association with RPL was reported earlier, this study is also the first to report on the upregulation of Hsa-miR-184 in circulating fluids in association with RPL. The study provides for understanding circulating microRNAs expression pattern in RPL which may be involved in its pathogenesis and demonstrates their potential role as noninvasive diagnostic and prognostic biomarkers for RPL.

Characterization of Calcined Jade and its immunomodulatory effect on macrophage isolated from Swiss albino mice.

Calcined Jade (CJ) is a metasilicate frequently used in traditional system of medicine as tonic to vital organs with several other pharmacological activities. X-ray powder diffraction (XRPD), inductively coupled plasma mass spectrometry (ICP-MS), atomic absorption spectroscopy (AAS) and CHNS analyzer techniques were used to characterize CJ sample. CJ was administered orally to Swiss albino mice at a dose of 50, 75, 100 and 200 µg/kg body weight for 10 days and modulation of the macrophage mediated innate immune responses was studied. Flow cytometric analysis of TLR-2/4 on peritoneal macrophage revealed elevated expression of TLR-2 as compared to control. Significant increase in phagocytic activity was observed in peritoneal macrophage. The lymphoid organs weight and other toxicity parameters did not exhibit any harmful effect. To evaluate the presence of nanoparticles, CJ was dissolved in milli Q water, filtered and lyophilized. Transmission electron microscopic (TEM) analysis revealed the presence of spherical nanoparticles in CJ [14.7-142.0 nm dimension with average particle size of 64.6 nm]. In conclusion, we report stimulation of innate immune responses by CJ may partly be due to the formation of nanoparticles. Further experiments using isolated nanoparticles may further validate the role of nanoparticles.

Circulating microRNA expression as predictor of preeclampsia and its severity.

Preeclampsia (PE) is a pregnancy syndrome characterized by hypertension and proteinuria, and a leading cause of maternal and fetal morbidity and mortality, with poorly defined pathophysiological mechanisms remain. Circulating microRNAs (miRNAs) are small, noncoding RNA molecules, which negatively regulate gene expression, and considered as promising biomarkers for PE. The objective of the study was to evaluate circulating miRNA signatures in women with PE compared to healthy women, and in women sub-grouped per PE severity. This study assessed miRNA expression profile in the plasma of 15 women with PE (7 mild and 8 severe) compared to 7 women with uncomplicated pregnancies. Circulating miRNA was extracted from maternal plasma, and the differential expression of 84 miRNA species were determined using customized pathway-focused miRNA profiler kits. A set of 7 miRNAs that were differentially expressed in PE patients and in mild vs. severe PE cases subgroups. These included miR-215, miR-155, miR-650, miR-210, miR-21 which were upregulated, and miR-18a, miR-19b1 were downregulated in women with PE compared to control women, and between women with severe PE compared to mild PE. In addition, four novel miRNAs comprising miR-518b and miR-29a which were upregulated, and miR-144, miR-15b which were downregulated in severe PE compared to mild PE. This study for the first time presents the differential expression profile of circulating miRNAs according to the severity of the disease. The results confirm the contribution of miRNA to PE pathogenesis, as well as being predictors of the severity of PE.

MicroRNA expression pattern in pre-eclampsia (Review).

Pre-eclampsia (PE), a pregnancy complication, is a leading cause of maternal and fetal morbidity and mortality. Although its exact etiology and pathogenesis remain elusive, PE results from an interaction of inherited and non­inherited factors. The clinical symptoms of PE appear post­mid­stage of gestation and, at present, there are no early signs/markers for its onset and progression. MicroRNAs function as gene regulators, and are involved in development and pathology. A burgeoning number of studies have highlighted microRNAs as potential biomarkers for minimal invasive assessment. However, it remains a matter of debate as to which microRNA type is involved in PE onset and progression, as well as the clinical utility of testing for these species. In the present review, we have summarized the latest findings on the association of PE with the aberrant expression of placental microRNAs; in particular, those that are detectable in the blood. The current understanding of the mechanisms of microRNA­target gene interactions that underpin the involvement of microRNAs in the pathogenesis of PE is also discussed.

A procedure for the rapid screening of Maillard reaction inhibitors.

A procedure for the rapid screening of inhibitors of glycation reaction, based on their ability to protect RNase against sugar induced inactivation of the enzyme is described. Glycation is implicated in variety of disorders including diabetes, atherosclerosis various micropathies yet is a slow process both in vivo and in vitro. In order to speed up glycation, the reaction was carried out at 60 degrees C using a thermostable protein RNase and ribose, a sugar that is known to react rapidly than glucose in the glycation reaction. It was observed that incubation of RNase with ribose at 60 degrees C in rapid inactivation of the enzyme with a parallel decrease in tyrosine fluorescence, enhancement in new fluorescence and hyperchromicity in the UV-region. No such alterations in the enzyme activity were observed when the incubation was carried out in absence of the sugar. Compounds and drugs that are known to act as inhibitors of glycation reaction restricted the ribose-induced inactivation of RNase. RNase immobilized on CNBr-activated Sepharose was also sensitive to exposure to ribose and appeared a better system to screen inhibitors of glycation from natural sources that contain substances that interfere with the assay of enzyme as well as in the study of post Amadori inhibitors of glycation.

Immunoglobulin glycation with fructose: a comparative study.

BACKGROUND: Immunoglobulins undergo non-enzymatic glycation reaction with sugars both in vivo and in vitro. Effects of glycation on the ability of the antibodies to bind antigens are contradictory. Antibodies raised in various animals may also be exposed to high concentration of sugars that are added during freeze-drying/pasteurization for preservation. METHODS: IgG isolated from the sera of goat, human, rabbit, mouse, buffalo as well as IgY from hen egg yolk was subjected to in vitro glycation with fructose. The behavior of glycated IgG was investigated by SDS-PAGE, hyperchromicity at 280 nm, tryptophan fluorescence and new fluorescence. RESULTS: Marked variations were observed in the response of the immunoglobulins derived from various animals to incubation with fructose. Also, incubation of anti-glucoseoxidase (GOD) antibodies with fructose resulted in a rapid loss of their ability to bind the enzyme antigen as revealed by immunodiffusion and ELISA. DETAPAC and EDTA were quite protective but were unable to completely prevent the fructose-induced alterations. CONCLUSIONS: Immunoglobulins derived from goat, human, rabbit, mouse, buffalo and hen egg yolk undergo remarkable structural alterations on incubation with fructose. The susceptibility of the immunoglobulins to the modification however differed remarkably. The goat IgG was most recalcitrant while hen egg yolk IgY was most susceptible to the alterations. DETAPAC or EDTA restricted the fructose-induced alterations remarkably.