scholarly journals Autism-associated synaptic vesicle transcripts are differentially expressed in maternal plasma exosomes of physiopathologic pregnancies

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Yangwu Fang ◽  
Chan Wan ◽  
Youlu Wen ◽  
Ze Wu ◽  
Jing Pan ◽  
...  
Keyword(s):  
Real Time ◽  
Fetal Brain ◽  
First Trimester ◽  
Maternal Plasma ◽  
Advanced Technology ◽  
Differentially Expressed ◽  
Autistic Children ◽  
Spontaneous Preterm Birth ◽  
Intrauterine Development ◽  
And Function

AbstractBackgroundDuring intrauterine development, the formation and function of synaptic vesicles (SVs) are thought to be fundamental conditions essential for normal development of the brain. Lacking advanced technology during the intrauterine period, such as longitudinal real-time monitoring of the SV-associated transcripts (SVATs), which include six pairs of lncRNA-mRNA, has limited acquisition of the dynamic gene expression profile (GEP) of SVATs. We previously reported the differential expression of SVATs in the peripheral blood of autistic children. The current study was designed to determine the dynamic profiles of differentially-expressed SVATs in circulating exosomes (EXs) derived from autistic children and pregnant women at different gestational ages.MethodsBlood samples were collected from autistic children and women with variant physiopathologic pregnancies. EXs were isolated with an ExoQuick Exosome Precipitation Kit and characterized by transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), and Western blotting. The expression of lncRNAs and lncRNA-targeted mRNAs were quantified using real-time PCR.ResultsSVAT-associated lncRNAs-mRNAs were detected in autistic children and differentially expressed from the first trimester of pregnancy to the term of delivery. Pathologic pregnancies, including spontaneous preterm birth (sPTB), preeclampsia (PE), and gestational diabetes mellitus (GDM), were compared to normal physiologic pregnancies, and shown to exhibit specific correlations between SVAT-lncRNA and SVAT-mRNA ofSTX8,SLC18A2, andSYPwith sPTB; SVAT-lncRNA and SVAT-mRNA ofSTX8with PE; and SVAT-lncRNA and SVAT-mRNA ofSV2Cas well as SVAT-mRNA ofSYPwith GDM.ConclusionVariant complications in pathologic pregnancies may alter the GEP of SVATs, which is likely to affect the intrauterine development of neural circuits and consequently influence fetal brain development.

scholarly journals Measurement of Allelic-Expression Ratios in Trisomy 21 Placentas by Quencher Extension of Heterozygous Samples Identified by Partially Denaturing HPLC

2008 ◽  
Vol 54 (2) ◽  
pp. 437-440 ◽  
Author(s):  
Attie T J I Go ◽  
Allerdien Visser ◽  
Ofir T Betsalel ◽  
John M G van Vugt ◽  
Marinus A Blankenstein ◽  
...  
Keyword(s):  
Real Time ◽  
Trisomy 21 ◽  
First Trimester ◽  
Maternal Plasma ◽  
Rapid Identification ◽  
Chromosome 21 ◽  
Wave System ◽  
Exon 2 ◽  
Reading Frame ◽  
Time Required

Abstract Background: Measuring the allelic ratios of placental transcripts in maternal plasma permits noninvasive prenatal detection of chromosomal aneuploidy. Current methods, however, require highly specialized equipment (MALDI-TOF), limiting the widespread implementation of this powerful RNA single-nucleotide polymorphism (SNP) strategy in routine diagnostic settings. We adapted and applied the Transgenomic WAVE System and quencher extension (QEXT) for this purpose. Methods: The expressed SNP (rs2187247) in exon 2 of the placentally expressed C21orf105 gene (chromosome 21 open reading frame 105) on chromosome 21 was tested in a trisomy 21 model system in which we obtained RNA selectively released from syncytiotrophoblasts of normal and trisomy 21 placentas during first trimester. Results: In identifying heterozygous samples, we observed an exact correspondence between sequencing results and results obtained with the WAVE System. With respect to the analysis time required, the WAVE System was superior. In addition, the real-time QEXT assay (as optimized and validated with calibration standards consisting of 262-bp C21orf105 cDNA amplicons) accurately measured allele ratios after we optimized fragment purification, concentrations of input DNA and quencher label, and calculations of reporter signals. Finally, the optimized and validated QEXT assay correctly distinguished normal placentas from trisomy 21 placentas in tests of the following clinically relevant combinations: diploid homozygous (CC), diploid heterozygous (AC), triploid homozygous (AAA), and triploid heterozygous (AAC or ACC). Conclusion: The QEXT method, which is directly adaptable to current real-time PCR equipment, along with rapid identification of informative samples with the WAVE System, may facilitate routine implementation of the RNA-SNP assay for noninvasive aneuploidy diagnostics.

scholarly journals Vasopressin: the missing link for preeclampsia?

2015 ◽  
Vol 309 (9) ◽  
pp. R1062-R1064 ◽  
Author(s):  
Jeremy A. Sandgren ◽  
Sabrina M. Scroggins ◽  
Donna A. Santillan ◽  
Eric J. Devor ◽  
Katherine N. Gibson-Corley ◽  
...  
Keyword(s):  
Plasma Concentrations ◽  
Late Pregnancy ◽  
First Trimester ◽  
Cardiovascular Disorder ◽  
Maternal Plasma ◽  
Complex Interplay ◽  
Chronic Infusion ◽  
And Function ◽  
Tissue Rejection ◽  
Sustained Increase

Preeclampsia is a devastating cardiovascular disorder of late pregnancy, affecting 5–7% of all pregnancies and claiming the lives of 76,000 mothers and 500,000 children each year. Various lines of evidence support a “tissue rejection” type reaction toward the placenta as the primary initiating event in the development of preeclampsia, followed by a complex interplay among immune, vascular, renal, and angiogenic mechanisms that have been implicated in the pathogenesis of preeclampsia beginning around the end of the first trimester. Critically, it remains unclear what mechanism links the initiating event and these pathogenic mechanisms. We and others have now demonstrated an early and sustained increase in maternal plasma concentrations of copeptin, a protein by-product of arginine vasopressin (AVP) synthesis and release, during preeclampsia. Furthermore, chronic infusion of AVP during pregnancy is sufficient to phenocopy essentially all maternal and fetal symptoms of preeclampsia in mice. As various groups have demonstrated interactions between AVP and immune, renal, and vascular systems in the nonpregnant state, elevations of this hormone are therefore positioned both in time (early pregnancy) and function to contribute to preeclampsia. We therefore posit that AVP represents a missing mechanistic link between initiating events and established midpregnancy dysfunctions that cause preeclampsia.

First trimester HtrA1 maternal plasma level and spontaneous preterm birth

2020 ◽  
pp. 1-5
Author(s):  
Stefano Raffaele Giannubilo ◽  
Caterina Licini ◽  
Elena Picchiassi ◽  
Federica Tarquini ◽  
Giuliana Coata ◽  
...  
Keyword(s):  
Preterm Birth ◽  
Plasma Level ◽  
First Trimester ◽  
Maternal Plasma ◽  
Spontaneous Preterm Birth

scholarly journals Optimized Real-Time Quantitative PCR Measurement of Male Fetal DNA in Maternal Plasma

2005 ◽  
Vol 51 (9) ◽  
pp. 1598-1604 ◽  
Author(s):  
Bernhard Zimmermann ◽  
Ahmad El-Sheikhah ◽  
Kypros Nicolaides ◽  
Wolfgang Holzgreve ◽  
Sinuhe Hahn
Keyword(s):  
Real Time ◽  
Quantitative Pcr ◽  
Pcr Amplification ◽  
First Trimester ◽  
Single Copy ◽  
Maternal Plasma ◽  
Probit Regression ◽  
Real Time Quantitative Pcr ◽  
Fetal Dna ◽  
First Trimester Of Pregnancy

Abstract Background: Circulating fetal DNA (cfDNA) in maternal plasma has been measured to investigate its possible relationship with pregnancy-related disorders, including fetal trisomy 21 and preeclampsia. The circulating concentrations of single-copy fetal genes, however, are close to the detection limits of PCR methods. Methods: We optimized a protocol for the real-time quantitative PCR amplification of the multicopy sequence DYS14 on the Y-chromosome. This was compared with an established real-time PCR assay for the single-copy SRY gene. Results: By probit regression analysis, the measurements of male DNA by the DYS14 assay had a 10-fold lower detection limit (0.4 genome equivalents) than did measurements of SRY. For plasma samples from women in the first trimester of pregnancy, imprecision (CV) was 2%–22% when amplifying DYS14 compared with 26%–140% for SRY. Conclusions: The low copy numbers of fetal DNA in plasma of women in the first trimester of pregnancy cannot be measured precisely when targeting single-copy sequences. Better results are obtained by amplifying a sequence that is present in multiple copies per male genome.

Development of Neural Structure and Function in Autism Spectrum Disorder: Potential Implications for Learning Language

2020 ◽  
Vol 29 (4) ◽  
pp. 1783-1797
Author(s):  
Kelly L. Coburn ◽  
Diane L. Williams
Keyword(s):  
Autism Spectrum Disorder ◽  
Language Development ◽  
Diffusion Tensor ◽  
Autism Spectrum ◽  
Language Intervention ◽  
Spectrum Disorder ◽  
Autistic Children ◽  
Neural Structure ◽  
Complex Information ◽  
And Function

Purpose Neurodevelopmental processes that begin during gestation and continue throughout childhood typically support language development. Understanding these processes can help us to understand the disruptions to language that occur in neurodevelopmental conditions, such as autism spectrum disorder (ASD). Method For this tutorial, we conducted a focused literature review on typical postnatal brain development and structural and functional magnetic resonance imaging, diffusion tensor imaging, magnetoencephalography, and electroencephalography studies of the neurodevelopmental differences that occur in ASD. We then integrated this knowledge with the literature on evidence-based speech-language intervention practices for autistic children. Results In ASD, structural differences include altered patterns of cortical growth and myelination. Functional differences occur at all brain levels, from lateralization of cortical functions to the rhythmic activations of single neurons. Neuronal oscillations, in particular, could help explain disrupted language development by elucidating the timing differences that contribute to altered functional connectivity, complex information processing, and speech parsing. Findings related to implicit statistical learning, explicit task learning, multisensory integration, and reinforcement in ASD are also discussed. Conclusions Consideration of the neural differences in autistic children provides additional scientific support for current recommended language intervention practices. Recommendations consistent with these neurological findings include the use of short, simple utterances; repetition of syntactic structures using varied vocabulary; pause time; visual supports; and individualized sensory modifications.

Enhancing Clinical Supervision Using High-Definition Real-Time Digital Recording, Annotation, and Bluetooth Technology

2019 ◽  
Vol 4 (2) ◽  
pp. 356-362
Author(s):  
Jennifer W. Means ◽  
Casey McCaffrey
Keyword(s):  
Graduate Students ◽  
Data Collection ◽  
Real Time ◽  
Clinical Supervision ◽  
Video Recording ◽  
Advanced Technology ◽  
High Definition ◽  
Recording Technology ◽  
Self Confidence ◽  
Additional Data Collection

Purpose The use of real-time recording technology for clinical instruction allows student clinicians to more easily collect data, self-reflect, and move toward independence as supervisors continue to provide continuation of supportive methods. This article discusses how the use of high-definition real-time recording, Bluetooth technology, and embedded annotation may enhance the supervisory process. It also reports results of graduate students' perception of the benefits and satisfaction with the types of technology used. Method Survey data were collected from graduate students about their use and perceived benefits of advanced technology to support supervision during their 1st clinical experience. Results Survey results indicate that students found the use of their video recordings useful for self-evaluation, data collection, and therapy preparation. The students also perceived an increase in self-confidence through the use of the Bluetooth headsets as their supervisors could provide guidance and encouragement without interrupting the flow of their therapy sessions by entering the room to redirect them. Conclusions The use of video recording technology can provide opportunities for students to review: videos of prospective clients they will be treating, their treatment videos for self-assessment purposes, and for additional data collection. Bluetooth technology provides immediate communication between the clinical educator and the student. Students reported that the result of that communication can improve their self-confidence, perceived performance, and subsequent shift toward independence.

Online Measurement of Glucose Consumption from HepG2 Cells Using an Integrated Bioreactor and Enzymatic Assay

2018 ◽  
Author(s):  
Anna Adams ◽  
Radha Krishna Murthy Bulusu ◽  
Nikita Mukhitov ◽  
Jose Mendoza-Cortes ◽  
Michael Roper
Keyword(s):  
Real Time ◽  
Glucose Concentration ◽  
Hepg2 Cells ◽  
Glucose Consumption ◽  
Microfluidic System ◽  
Structure And Function ◽  
Online Measurement ◽  
Fluorescent Assay ◽  
And Function ◽  
Integrated Bioreactor

In this work, we developed a microfluidic bioreactor for optimizing growth and maintaining structure and function of HepG2, and when desired, the device could be removed and the extracellular output from the bioreactor combined with enzymatic glucose reagents into a droplet-based microfluidic system. The intensity of the resulting fluorescent assay product in the droplets was measured, and was directly correlated to glucose concentration, allowing the effect of insulin on glucose consumption in the HepG2 cells to be observed and quantified online and in near real-time.

Online Measurement of Glucose Consumption from HepG2 Cells Using an Integrated Bioreactor and Enzymatic Assay

2018 ◽  
Author(s):  
Anna Adams ◽  
Radha Krishna Murthy Bulusu ◽  
Nikita Mukhitov ◽  
Jose Mendoza-Cortes ◽  
Michael Roper
Keyword(s):  
Real Time ◽  
Glucose Concentration ◽  
Hepg2 Cells ◽  
Glucose Consumption ◽  
Microfluidic System ◽  
Structure And Function ◽  
Online Measurement ◽  
Fluorescent Assay ◽  
And Function ◽  
Integrated Bioreactor

In this work, we developed a microfluidic bioreactor for optimizing growth and maintaining structure and function of HepG2, and when desired, the device could be removed and the extracellular output from the bioreactor combined with enzymatic glucose reagents into a droplet-based microfluidic system. The intensity of the resulting fluorescent assay product in the droplets was measured, and was directly correlated to glucose concentration, allowing the effect of insulin on glucose consumption in the HepG2 cells to be observed and quantified online and in near real-time.

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