Whole Transcriptome Analysis (Rna-Seq) By Next-Generation Sequencing of hAF-EVs
Ontology highlight
ABSTRACT:
INSTRUMENT(S): NextSeq 550
ORGANISM(S): Homo sapiens
SUBMITTER: Fazio Grazia
PROVIDER: E-MTAB-14034 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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Project description:Amniotic fluid-derived products are a promising resource for cell therapy and tissue engineering due to their anti-inflammatory, angiogenic, and antifibrotic properties. Human amniotic fluid (hAF) has been used in medical applications such as wound healing, skin disorders, and ophthalmic conditions. Recently, we demonstrated that hAF is an effective treatment for myocardial ischemia-reperfusion injury in adult rats. However, the protein composition of full-term acellular hAF has remained poorly characterized. To uncover the biologically active components underlying hAF’s cardioprotective effects, we conducted a global proteomic analysis of hAF collected from six patients at full-term cesarean sections. Previously shown to improve cardiac function in ischemic rats, these samples were analyzed by using tandem mass spectrometry. We identified 657 proteins, including 148 unique to the deep learning platform Inferys. Bioinformatic analysis revealed that these proteins are involved in immunity, inflammatory responses, cell adhesion, and apoptotic signaling pathways. In addition, these proteins were highly modified, with methylation and deamidation being the most abundant modifications. This study represents the first mass-spectrometry-based characterization of full-term, acellular hAF, suggesting that hAF offers a wide array of immune-modulating proteins working together to provide robust cardioprotection and a valuable treatment for ischemia-reperfusion injury.
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Project description:To better undersand the effects of drought stress on wheat developing seeds, the transcription profile of early developing wheat seeds under control and drought stress conditions were comparatively analyzed by using the Affymetrix wheat geneChip. Drought stress is a major yield-limiting factor for wheat. Wheat yields are particularly sensitive to drought stress during reproductive development. Early seed development stage is an important determinant of seed size, one of the yield components. We specifically examined the impact of drought stress imposed during postzygotic early seed development in wheat. We imposed a short-term drought stress on plants with day-old seeds and observed that even a short-duration drought stress significantly reduced the size of developing seeds as well as mature seeds. Drought stress delayed the developmental transition from syncytial to cellularized stage of endosperm. Coincident with reduced seed size and delayed endosperm development, a subset of genes associated with cytoskeleton organization was misregulated in developing seeds under drought-stressed. Several genes linked to hormone pathways were also differentially regulated in response to drought stress in early seeds. Notably, drought stress strongly repressed the expression of wheat storage protein genes such as gliadins, glutenins and avenins as early as 3 days after pollination. Our results provide new insights on how some of the early seed developmental events are impacted by water stress, and the underlying molecular pathways that can possibly impact both grain size and quality in wheat. Winter wheat cultivar Redland, PI 502907 (Triticum aestivum L.) was used for this study. Seedlings were vernalized at 4°C for 6 weeks and then transplanted to a one gallon pot of soil-sand mixture (3:1, v/v) and grown in a growth chamber under the following conditions: relative humidity, 50–70%; 16-h light/8-h dark photoperiod; 21°C daytime temperature and 18°C nights. Plants were watered regularly twice daily at the rate of 100ml/ per pot. Because, wheat has an asynchronous fertilization pattern for ovlues in the inflorescence, each floret needs to be specifically marked for timing the fertilization and stress induction. After spikes developed, unfertilized ovules were monitored and observed for the fertilization process. Closed wheat spikes with anthers outside were marked as fertilized. Drought stress was imposed 24h after the fertilization (HAF). Drought stress treatment was initiated by discontinuing watering on the drought treatment plants while control plants were regularly watered twice daily. Stress treatment was applied at 48 HAF and relieved at 96 HAF. The microarray study focuses on 24 HAF to 72 HAF in control and drought stress conditions. We started to impose drought stress 24HAF.
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Project description:Whole Transcriptome Analysis (Rna-Seq) By Next-Generation Sequencing of hAF-EVs
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