Differential expression of microRNAs in porcine placentas on Days 30 and 90 of gestation

2010 ◽  
Vol 22 (8) ◽  
pp. 1175 ◽  
Author(s):  
Lijie Su ◽  
Shuhong Zhao ◽  
Mengjin Zhu ◽  
Mei Yu

The porcine placenta is classified as a non-invasive epitheliochorial type. To meet the increasing demands for nutrients by the rapidly growing conceptus and/or fetus, the placental microscopic folds undergo significant morphological and biochemical changes during two periods critical for conceptus and/or fetus, namely Days 30–40 and after Day 90 of gestation. MicroRNAs (miRNAs) are a class of small non-coding RNAs that can modulate gene activity by inhibiting the translation or regulation of mRNA degradation. In the present study, we identified 17 differentially expressed miRNAs in porcine placenta on Days 30 and 90 of gestation using a locked nucleic acid (LNA) microRNA array. Stem–loop real-time reverse transcription–polymerase chain reaction confirmed the differential expression of eight selected miRNAs (miR-24, miR-125b, miR-92b, miR-106a, miR-17, let-7i, miR-27a and miR-20). Analysis of targets and the pathways in which these miRNAs are involved revealed that the differentially expressed miRNAs target many genes that are important in various processes, including cell growth, trophoblast differentiation, angiogenesis and formation and maintenance of adherens junctions. The results of the present study suggest potential roles for these differentially expressed miRNAs in porcine placental growth and function.

Author(s):  
Jaya Garg ◽  
Atul Garg ◽  
Anand Kumar

Introduction: Tuberculosis (TB) is global health problem threatening millions every year. There is urgent need of effective biomarkers for its diagnostics and circulating microRNAs (miRNAs) have recently emerged as novel and non-invasive molecular markers in blood. Aim: This study was done to evaluate serum miRNAs as a potential biomarker for the diagnosis of pulmonary mycobacterium tuberculosis infection. Materials and Methods: In this prospective cross-sectional study, acute pulmonary TB patients were recruited based on positive Mycobacterium tuberculosis Polymerase Chain Reaction (PCR) in sputum samples and Microarray expression profiling was performed on blood of these patients to study differentially expressed miRNAs. The results were validated by Real-Time Polymerase Chain Reaction (RT-PCR) on pulmonary TB cases and healthy controls. Student’s t-tests and analysis variance tests were used for statistical analysis. The p-value <0.05 was considered statistically significant. Results: Results of microarray expression profiling showed 75 differentially expressed miRNAs in cases of pulmonary TB; among these 5 upregulated and 2 downregulated miRNAs were evaluated by RT-PCR. miRNA 29a exhibited good distinguishing efficiency; followed by miRNA 384. ROC plot of expression data for miRNA in cases and control groups showed that AUC of miRNA-29a was 0.8268 (sensitivity=80%, specificity=82%) . Conclusion: This study suggests that altered levels of serum miRNAs have great potential to serve as non-invasive biomarkers for detection of Mycobacterium tuberculosisinfection. miRNA-29a can be used as an effective biomarker for diagnosis of pulmonary TB.


2021 ◽  
Vol 52 (1) ◽  
Author(s):  
Emilia Bagnicka ◽  
Ewelina Kawecka-Grochocka ◽  
Klaudia Pawlina-Tyszko ◽  
Magdalena Zalewska ◽  
Aleksandra Kapusta ◽  
...  

AbstractMicroRNAs (miRNAs) are short, non-coding RNAs, 21–23 nucleotides in length which are known to regulate biological processes that greatly impact immune system activity. The aim of the study was to compare the miRNA expression in non-infected (H) mammary gland parenchyma samples with that of glands infected with coagulase-positive staphylococci (CoPS) or coagulase-negative staphylococci (CoNS) using next-generation sequencing. The miRNA profile of the parenchyma was found to change during mastitis, with its profile depending on the type of pathogen. Comparing the CoPS and H groups, 256 known and 260 potentially new miRNAs were identified, including 32 that were differentially expressed (p ≤ 0.05), of which 27 were upregulated and 5 downregulated. Comparing the CoNS and H groups, 242 known and 171 new unique miRNAs were identified: 10 were upregulated (p ≤ 0.05), and 2 downregulated (p ≤ 0.05). In addition, comparing CoPS with H and CoNS with H, 5 Kyoto Encyclopedia of Genes and Genomes pathways were identified; in both comparisons, differentially-expressed miRNAs were associated with the bacterial invasion of epithelial cells and focal adhesion pathways. Four gene ontology terms were identified in each comparison, with 2 being common to both immune system processes and signal transduction. Our results indicate that miRNAs, especially miR-99 and miR-182, play an essential role in the epigenetic regulation of a range of cellular processes, including immunological systems bacterial growth in dendritic cells and disease pathogenesis (miR-99), DNA repair and tumor progression (miR-182).


2009 ◽  
Vol 19 (9) ◽  
pp. 1545-1549 ◽  
Author(s):  
Yi Miao ◽  
Bin Cai ◽  
Ling Liu ◽  
Yixia Yang ◽  
Xiaoping Wan

Objective:To investigate the genes that were differentially expressed between clear cell carcinoma (CCC) and serous carcinoma (SAC) of the ovary with complementary DNA microarray.Methods:Complementary DNA microarray was carried out in 8 CCCs and 8 SACs of the ovary. Differentially expressed genes were identified and verified by real-time polymerase chain reaction. The expression of the protein was also verified with immunohistochemistry and Western blot in cells and tissues of ovarian CCC.Results:Comparison of the gene expression profiling identified 21 genes with more than 2-fold different expression between CCC and SAC of the ovary. The up-regulated and down-regulated genes were 9 and 12, respectively. The verification of Annexin IV in the cell line and tissues was in accordance with the result of the microarray.Conclusions:The complementary DNA microarray technique is a feasible way to explore the difference of the gene expression profiling between the 2 types of ovarian carcinoma. The overexpression of Annexin IV may be an ovarian CCC-specific molecular marker.Abbreviations:CCC- clear cell carcinoma, SAC- serous carcinoma, PCR- polymerase chain reaction, RT-PCR- reverse transcriptase PCR, ABCF2- ATP-binding cassette, sub-family F- member 2, HNF-1b- hepatocyte nuclear factor 1β


2012 ◽  
Vol 7 (12) ◽  
pp. 1934578X1200701 ◽  
Author(s):  
Lili Huang ◽  
Hong Wang ◽  
Hechun Ye ◽  
Zhigao Du ◽  
Yansheng Zhang ◽  
...  

cDNAs encoding Hypericum sampsonii benzophenone synthase (HsBPS) and chalcone synthase (HsCHS) were isolated and functionally characterized. Differential expressions of HsBPS and HsCHS were monitored using quantitative polymerase chain reaction (PCR). In the vegetative stage, HsBPS was highly expressed in the roots; its transcript level was approx. 100 times higher than that of HsCHS. Relatively high transcript amounts of HsBPS were also detected in older leaves, whereas the youngest leaves contained higher transcript amounts of HsCHS. In the reproductive stage, maximum HsCHS expression was detected in flowers, the transcript level being approx. 5 times higher than that of HsBPS. The inversed situation with a 10-fold difference in the expression levels was observed with fruits. High transcript amounts for both proteins were found in roots.


2020 ◽  
Vol 13 (1) ◽  
Author(s):  
Kameshwar P. Singh ◽  
Krishna P. Maremanda ◽  
Dongmei Li ◽  
Irfan Rahman

Abstract Background Electronic cigarettes (e-cigs) vaping, cigarette smoke, and waterpipe tobacco smoking are associated with various cardiopulmonary diseases. microRNAs are present in higher concentration in exosomes that play an important role in various physiological and pathological functions. We hypothesized that the non-coding RNAs transcript may serve as susceptibility to disease biomarkers by smoking and vaping. Methods Plasma exosomes/EVs from cigarette smokers, waterpipe smokers and dual smokers (cigarette and waterpipe) were characterized for their size, morphology and TEM, Nanosight and immunoblot analysis. Exosomal RNA was used for small RNA library preparation and the library was quantified using the High Sensitivity DNA Analysis on the Agilent 2100 Bioanalyzer system and sequenced using the Illumina NextSeq 500 and were converted to fastq format for mapping genes. Results Enrichment of various non-coding RNAs that include microRNAs, tRNAs, piRNAs, snoRNAs, snRNAs, Mt-tRNAs, and other biotypes are shown in exosomes. A comprehensive differential expression analysis of miRNAs, tRNAs and piRNAs showed significant changes across different pairwise comparisons. The seven microRNAs that were common and differentially expressed of when all the smoking and vaping groups were compared with non-smokers (NS) are hsa-let-7a-5p, hsa-miR-21-5p, hsa-miR-29b-3p, hsa-let-7f-5p, hsa-miR-143-3p, hsa-miR-30a-5p and hsa-let-7i-5p. The e-cig vs. NS group has differentially expressed 5 microRNAs (hsa-miR-224-5p, hsa-miR-193b-3p, hsa-miR-30e-5p, hsa-miR-423-3p, hsa-miR-365a-3p, and hsa-miR-365b-3p), which are not expressed in other three groups. Gene set enrichment analysis of microRNAs showed significant changes in the top six enriched functions that consisted of biological pathway, biological process, molecular function, cellular component, site of expression and transcription factor in all the groups. Further, the pairwise comparison of tRNAs and piRNA in all these groups revealed significant changes in their expressions. Conclusions Plasma exosomes of cigarette smokers, waterpipe smokers, e-cig users and dual smokers have common differential expression of microRNAs which may serve to distinguish smoking and vaping subjects from NS. Among them has-let-7a-5p has high sensitivity and specificity to distinguish NS with the rest of the users, using ROC curve analysis. These findings will pave the way for the utilizing the potential of exosomes/miRNAs as a novel theranostic agents in lung injury and disease caused by tobacco smoking and vaping.


2013 ◽  
Vol 2013 ◽  
pp. 1-6 ◽  
Author(s):  
Wan Himratul Aznita Wan Harun ◽  
Nur Alyaa Jamil ◽  
Nor Hazwani Jamaludin ◽  
Mohd-Al-Faisal Nordin

The study aimed to identify theHWP1gene in non-Candida albicans Candidaspecies and the differential expression ofHWP1following treatment withPiper betleandBrucea javanicaaqueous extracts. All candidal suspensions were standardized to1×106 cells/mL. The suspension was incubated overnight at 37 °C (C. parapsilosis, 35°C). Candidal cells were treated with each respective extract at 1, 3, and 6 mg/mL for 24 h. The total RNA was extracted and reverse transcription-polymerase chain reaction was carried out with a specific primer ofHWP1.HWP1mRNAs were only detected inC. albicans,C. parapsilosis, andC. tropicalis. Exposing the cells to the aqueous extracts has affected the expression ofHWP1transcripts.C. albicans,C. parapsilosis, andC. tropicalishave demonstrated different intensity of mRNA. Compared toP. betle,B. javanicademonstrated a higher suppression on the transcript levels ofHWP1in all samples.HWP1was not detected inC. albicansfollowing the treatment ofB. javanicaat 1 mg/mL. In contrast,C. parapsilosisandC. tropicaliswere shown to haveHWP1regulation. However, the expression levels were reduced upon the addition of higher concentration ofB. javanicaextract.P. betleandB. javanicahave potential to be developed as oral health product.


2020 ◽  
Vol 21 (3) ◽  
pp. 785
Author(s):  
Yanfeng Jia ◽  
Chunrong Li ◽  
Quanlin Li ◽  
Pengcheng Liu ◽  
Dongfeng Liu ◽  
...  

MicroRNAs (miRNAs) are crucial player in plant-pathogen interaction. While the evidence has demonstrated that rice miRNAs mediate immune response to pathogens invasion, the roles of miRNAs on Xanthomonas oryzae pv. oryzae (Xoo) attack remain be in place. Herein, we monitored the responsive changes of rice miRNAs at 0, 8, 24 h across Xoo strain PXO86 infection in its compatible rice variety IR24 and incompatible variety IRBB5 by small RNA sequencing, and the genes targeted by miRNAs were also detected via degradome technology. The faithfulness of sequencing data was validated through quantitative real-time stem-loop reverse transcription-polymerase chain reaction assay. Bioinformatic analysis showed that the differentially expressed miRNAs could be divided into three immunity-related clusters, and 80 regulatory units were emerged in infection process, which comprises 29 differentially expressed known miRNAs and 38 cleaved targets. Furthermore, the miRNA presumptive function of separate immunity cluster in rice-Xoo interplay was confirmed through overexpressing osa-miR164a, osa-miR167d and osa-miR159b, and the disruption of regulatory units, osa-miR164a/OsNAC60, osa-miR167d-5p/OsWD40-174 and osa-miR159b/OsMYBGA, OsLRR-RLK2, OsMPK20-4, may reset rice defense response to Xoo infestation in a controllable manner. These findings provide new insights into the complex roles of characteristic miRNAs and their targets in rice-Xoo interactions.


2019 ◽  
Vol 32 (11) ◽  
pp. 1207-1215
Author(s):  
Babak Emamalizadeh ◽  
Yousef Daneshmandpour ◽  
Abbas Tafakhori ◽  
Sakineh Ranji-Burachaloo ◽  
Sajad Shafiee ◽  
...  

Abstract Background X-linked adrenoleukodystrophy (X-ALD), the most common peroxisomal disorder, is caused by mutations in the ABCD1 gene located on Xq28. X-ALD is characterized by a spectrum of different manifestations varying in patients and families. Methods Four pedigrees with X-ALD consisting of patients and healthy members were selected for investigation of ABCD1 gene mutations. The mutation analysis was performed by polymerase chain reaction (PCR) followed by direct sequencing of all exons. The identified mutations were investigated using bioinformatics tools to predict their effects on the protein product and also to compare the mutated sequence with close species. Results One previously known missense mutation (c.1978 C > T) and three novel mutations (c.1797dupT, c.879delC, c.1218 C > G) were identified in the ABCD1 gene, each in one family. Predicting the effects of the mutations on protein structure and function indicated the probable damaging effect for them with significant alterations in the protein structure. We found three novel mutations in the ABCD1 gene with damaging effects on its protein product and responsible for X-ALD.


2016 ◽  
Vol 22 (4) ◽  
pp. 201-209 ◽  
Author(s):  
Tzu-Yun Wang ◽  
Sheng-Yu Lee ◽  
Yi-Lun Chung ◽  
Shiou-Lan Chen ◽  
Chia-Ling Li ◽  
...  

Background: Different drug dependencies may have unique genetic vulnerabilities. Changes in serotonin availability and function have been linked to addiction. We investigated whether 2 serotonergic polymorphisms, TPH1 A218C (rs1800532) and 5-HTT-linked promoter region (5-HTTLPR) (rs25531), are differently associated with alcohol or opiate dependence. Methods: Alcohol-dependent patients (n = 292), opiate-dependent patients (n = 309), and healthy controls (n = 301) were recruited from the Han Chinese population in Taiwan. Genotypes of TPH1 A218C and 5-HTTLPR polymorphisms were analyzed using a polymerase chain reaction with restriction fragment length polymorphism. Results: The genotype frequencies of the TPH1 A218C polymorphisms were not significantly different in the 3 groups. The genotype frequencies of the 5-HTTLPR S+ (S/S, S/LG, LG/LG) polymorphisms were significantly higher in opiate-dependent patients (χ2 = 8.77, p = 0.01), but not after controlling for the covariates of age, gender, and interaction effect in logistic regression analysis. Moreover, there was a significant interaction between the TPH1 A218C A/C and 5-HTTLPR S+ gene polymorphisms in opiate-dependent (OR 2.72, p = 0.01), but not in alcohol-dependent patients. Conclusions: Our data suggested that there may be a differential genetic vulnerability in serotonergic genes for alcohol and opiate addiction. However, replications of our findings are still needed.


2020 ◽  
Vol 7 (2) ◽  
pp. 35 ◽  
Author(s):  
Ahmed M. El-Sebaey ◽  
Pavel N. Abramov ◽  
Fatma M. Abdelhamid

Computed tomography angiography (CTA) and biochemical parameters cannot specify liver pathologies in dogs with congenital portosystemic shunts (CPSS) that are easily determined by invasive histopathology. This study aims to assess the possibility of using circulating serum canine familiaris (cfa) microRNAs (miRNAs) as novel non-invasive serum-based fingerprints for liver injuries associated with various morphologies of extrahepatic and intrahepatic portosystemic shunts (EHPSS and IHPSS). Data were obtained from 12 healthy dogs and 84 dogs confirmed to have EHPSS (splenocaval, splenophrenic, splenoazygos, right gastrocaval (RGC), right gastrocaval with caudal loop (RGC–CL)) and IHPSS (right divisional and left divisional) using CTA. Hepatic pathologies were determined by histopathology. Serum expression of miRNAs was assessed by real-time polymerase chain reaction. Based on the nature of liver injuries in each shunt type, cfa-miR-122 was significantly upregulated in all CPSS groups. Meanwhile, serums cfa-miR-34a and 21 were not significantly expressed in splenophrenic or splenoazygos groups, but they were extensively upregulated in splenocaval, RGC, RGC–CL groups and less frequently in right or left divisional groups. Also, serum cfa-miR126 was significantly upregulated in both IHPSS groups but less significantly expressed in RGC, RGC–CL, and splenocaval groups. Overall, estimated cfa-miRNAs could serve as novel biomarkers to mirror the histopathological and molecular events within the liver in each shunt type.


Sign in / Sign up

Export Citation Format

Share Document