Expression In Day Research Articles

The Pattern of Gene Expression (Igf Family, Muscle Growth Regulatory Factors, and Osteogenesis-Related Genes) Involved in the Growth of Skeletal Muscle in Pikeperch (Sander lucioperca) During Ontogenesis

The pikeperch (Sander lucioperca) is an economically important freshwater fish and a valuable food with high market acceptance. It is undergoing important changes in growth and regulatory metabolism during the ontogeny. Hence, the current study aims to investigate the mRNA expression of the growth hormone (gh)/insulin-like growth factor (igf) axis (ghr, igfI, igfbp, igfr), muscle regulatory factors (pax7, myf5, myod, myogenin, mrf, mymk, mstn), and osteogenesis-related genes (colla1a, fib1a, on, op, ostn) from hatching through day 40th post-hatching (DPH). The average total length (TL) of larvae measured at hatching was 3.6 ± 0.4 mm (67 degree days), and at the end of the experiment (40 DPH, 777 degree days), it was 27.1 ± 1.1 mm. The results showed three phases of gene expression in day 0 (egg), larval, and juvenile stages of pikeperch, which can be a progression or transition from the initial state toward the juvenile state. The expression pattern of myf5, mymk, and fib1a genes showed the highest value at day 0. The growth hormone receptor gene (ghr) and igfbp5 were raised to 1 DPH, whereas increased expression of igfI, igfII, igf1bp4, igf1rb, myod2, and mrf4 was detected at 14 DPH. The myod1, pax7, op, ostc, on, igf1ra, and col1a1a genes were highly expressed at 21 DPH and juvenile stages. According to the PLS-DA model, the most relevant VIPs are myf5 and mymk as best markers of earlier stages and igf1ra, ostc, pax7, and ghr as markers of later stages of ontogeny. Results from this study suggest that basal metabolism, growth of body cells and muscles, and bone proliferation and development can be regulated by the dynamic changes in gene expression patterns in this species. The identified genes will help to understand the basic biological process of pikeperch larvae and development, which is very important in pikeperch farming

The disruptive effects of COPD exacerbation-associated factors on epithelial repair responses.

Exacerbations of chronic obstructive pulmonary disease (COPD) increase mortality risk and can lead to accelerated loss of lung function. The increased inflammatory response during exacerbations contributes to worsening of airflow limitation, but whether it also impacts epithelial repair is unclear. Therefore, we studied the effect of the soluble factor micro-environment during COPD exacerbations on epithelial repair using an exacerbation cocktail (EC), composed of four factors that are increased in COPD lungs during exacerbations (IL-1β, IL-6, IL-8, TNF-α). Mouse organoids (primary CD31-CD45-Epcam+ cells co-cultured with CCL206 fibroblasts) were used to study epithelial progenitor behavior. Mature epithelial cell responses were evaluated using mouse precision cut lung slices (PCLS). The expression of epithelial supportive factors was assessed in CCL206 fibroblasts and primary human lung fibroblasts. EC exposure increased the number and size of organoids formed, and upregulated Lamp3, Muc5ac and Muc5b expression in day 14 organoids. In PCLS, EC imparted no effect on epithelial marker expression. Pre-treatment of CCL206 fibroblasts with EC was sufficient to increase organoid formation. Additionally, the expression of Il33, Tgfa and Areg was increased in CCL206 fibroblasts from EC treated organoids, but these factors individually did not affect organoid formation or size. However, TGF-α downregulated Foxj1 expression and upregulated Aqp5 expression in day 14 organoids. EC exposure stimulates organoid formation and growth, but it alters epithelial differentiation. EC changes the epithelial progenitor support function of fibroblasts which contributes to observed effects on epithelial progenitors.

The Mask Diction/Sensitivity: An Analysis of Selected Poems from The Outcast

The mask is a covering for the face and/or making a replica of it. It is a pristine human phenomenon, the product of human’s spirit and artistry. It was at the core of the “early man’s” apprehension of life and its imponderable that is still used in the present. Its use is also in the mind and this has been subjected to writing especially in the diction of poetry, and is still intrinsic to modern humanity’s expression of day to day realities and impulses. This paper examines and explains Osaro et al’s sensitivity to the mask. It analyses the ways in which the poets exploit in a volcano of voices, the image of the mask to make social, cultural, political, artistic, oil exploration and exploitation statements in their entire poetic oeuvre. They aptly condemn and vehemently query the predators of the Niger Delta.

Knockdown of Two Trehalase Genes by RNA Interference Is Lethal to the White-Backed Planthopper Sogatella furcifera (Horváth) (Hemiptera: Delphacidae).

Trehalase (Tre) is a crucial enzyme involved in trehalose metabolism, and it plays pivotal roles in insect development and metamorphosis. However, the biological function of Tre genes in Sogatella furcifera remains unclear. In the present study, two Tre genes-SfTre1 and SfTre2-were cloned and identified based on the S. furcifera transcriptome data. Bioinformatic analysis revealed that the full-length complementary DNA of SfTre1 and SfTre2 genes were 3700 and 2757 bp long, with 1728- and 1902-bp open reading frame encoding 575 and 633 amino acid residues, respectively. Expression analysis indicated that SfTre1 and SfTre2 were expressed at all developmental stages, with the highest expression in day two adults. Furthermore, the highest expression levels of SfTre1 and SfTre2 were observed in the ovary; enriched expression was also noted in head tissues. The knockdown of SfTre1 and SfTre2 via injecting double-stranded RNAs decreased the transcription levels of the corresponding mRNAs and led to various malformed phenotypes and high lethality rates. The results of our present study indicate that SfTre1 and SfTre2 play crucial roles in S. furcifera growth and development, which can provide referable information for Tre genes as a potential target for planthopper control.

Genome-Wide Identification and Characterization of Circular RNAs during Skeletal Muscle Development in Meat Rabbits.

Simple SummaryOur knowledge of circRNAs regulating skeletal muscle development remains largely unknown in meat rabbits. Therefore, we collected the leg muscle tissues of ZIKA rabbits at three key growth stages. A combination of circRNA assembly from a circRNA-seq library and the whole-transcriptome sequencing data identified credible circRNAs in our samples. We found these circRNAs were more conserved between rabbits and humans than between rabbits and mice. A prediction of circRNA–microRNA–mRNAs networks revealed that circRNAs might be the regulators that mainly functioned in rabbits’ muscle neuron development and metabolic processes. Our work provides a catalog of circRNAs regulating skeletal muscle development at key growth stages in rabbits and might give a new insight into rabbit breeding.Skeletal muscle development plays a vital role in muscle quality and yield in meat rabbits. Circular RNAs (circRNAs) are a new type of single-stranded endogenous non-coding RNAs involved in different biological processes. However, our knowledge of circRNAs regulating skeletal muscle development remains largely unknown in meat rabbits. In this study, we collected the leg muscle tissues of ZIKA rabbits at three key growth stages. By performing whole-transcriptome sequencing, we found the sequential expression of day 0- (D0-), D35-, and D70-selective mRNAs mainly functioned in muscle development, nervous development, and immune response during skeletal muscle development, respectively. Then, a combination of circRNA assembly from a circRNA-seq library and the whole-transcriptome sequencing data identified 6845 credible circRNAs in our samples. Most circRNAs were transcribed from exons of known genes, contained few exons, and showed short length, and these circRNAs were more conserved between rabbits and humans than between rabbits and mice. The upregulated circRNAs, which were synchronously changed with host genes, primarily played roles in MAPK signaling pathways and fatty acid biosynthesis. The prediction of circRNA–microRNA–mRNAs networks revealed that circRNAs might be the regulators that mainly functioned in rabbits’ muscle neuron development and metabolic processes. Our work provides a catalog of circRNAs regulating skeletal muscle development at key growth stages in rabbits and might give a new insight into rabbit breeding.

Exosomal gene fold expression estimation of Leishmania major by real time qPCR in vivo and in vitro

The aim of this research is to estimate the virulence factor L. major exosome by real time gene expression. The higher gene expression in footpad tissue was (29.34), the higher gene expression in day seven culture was (20.37), higher gene expression in day twelve culture was (12.25), highest gene expression in culture line control (8.33). The current work revealed that exosome gene expression was elevated in footpad in comparison to cultures media. The elevated levels were in passage three and lowest levels in passage one, in control culture line the level of gene expression was declined with progress of time.

The Effects of Heat Shock Protein 70 Addition in the Culture Medium on the Development and Quality of In Vitro Produced Heat Shocked Bovine Embryos.

Simple SummaryEuropean dairy cows are extremely sensitive to heat stress. During hot summers milk yield is suppressed, welfare status of the cows is aggravated, and their fertility is seriously compromised. The observed subfertility is attributed to endocrinological alterations and mainly to inability of the early embryo to develop uneventfully until the implantation stages. To protect the integrity and the functionality of their cells, all mammals have inherent defense mechanisms against external insults. A major representative of these molecules are the heat shock proteins (HSPs) that are secreted by almost all cells. In this study, we examined the effects of exogenous HSP70 on the developmental competence and quality of early embryos produced in vitro, after 24-h exposure to slightly elevated temperature. Our findings imply that the temperature rise is detrimental for the developmental potential of the early embryos, and that the HSP70 can partly mitigate the harmful effects of heat stress, by improving the embryo yield and by reducing the adverse effects of heat stress on some embryo quality characteristics.The aims of the present study were to examine the effects of HSP70 addition in the in vitro culture medium of day 3 embryos on their developmental competence and quality. Bovine oocytes (n = 1442) were in vitro matured, inseminated and cultured for the first two days according to standardized methods. The presumptive zygotes were randomly allocated in three experimental groups: Control, C (embryos cultured at 39 °C throughout the culture period), group C41 (temperature was raised to 41 °C from the 48th to 72nd h post insemination (p.i.) and then it returned at 39 °C for the remaining culture period), and group H41 (the temperature modification was the same as in C41 and during heat exposure, HSP70 was added in the culture medium). Cleavage and embryo yield were assessed 48 h p.i. and on days 7, 8, 9, respectively and gene expression in day 7 blastocysts was assessed by RT-PCR. Blastocyst yield was the highest in group C39; and higher in group H41 compared to group C41. From the gene expression analyses, altered expression of 11 genes was detected among groups. The analysis of the orchestrated patterns of gene expression differed between groups. The results of this study confirm the devastating effects of heat stress on embryo development and provide evidence that HSP70 addition at the critical stages can partly counterbalance, without neutralizing, the negative effects of the heat insult on embryos, acting mainly through mechanisms related to energy deployment.

Transcriptome Profiling of Atlantic Salmon Adherent Head Kidney Leukocytes Reveals That Macrophages Are Selectively Enriched During Culture

The Atlantic salmon (Salmo salar) is an economically important fish, both in aquaculture and in the wild. In vertebrates, macrophages are some of the first cell types to respond to pathogen infection and disease. While macrophage biology has been characterized in mammals, less is known in fish. Our previous work identified changes in the morphology, phagocytic ability, and miRNA profile of Atlantic salmon adherent head kidney leukocytes (HKLs) from predominantly “monocyte-like” at Day 1 of in vitro culture to predominantly “macrophage-like” at Day 5 of culture. Therefore, to further characterize these two cell populations, we examined the mRNA transcriptome profile in Day 1 and Day 5 HKLs using a 44K oligonucleotide microarray. Large changes in the transcriptome were revealed, including changes in the expression of macrophage and immune-related transcripts (e.g. csf1r, arg1, tnfa, mx2), lipid-related transcripts (e.g. fasn, dhcr7, fabp6), and transcription factors involved in macrophage differentiation and function (e.g. klf2, klf9, irf7, irf8, stat1). The in silico target prediction analysis of differentially expressed genes (DEGs) using miRNAs known to change expression in Day 5 HKLs, followed by gene pathway enrichment analysis, supported that these miRNAs may be involved in macrophage maturation by targeting specific DEGs. Elucidating how immune cells, such as macrophages, develop and function is a key step in understanding the Atlantic salmon immune system. Overall, the results indicate that, without the addition of exogenous factors, the adherent HKL cell population differentiates in vitro to become macrophage-like.

Specialized Proresolving Mediators (SPMs) Help to Alleviate the Cold Platelet Storage Lesion

Introduction: Refrigeration of platelets in plasma is a promising alternative to conventional room temperature storage, but the cold temperature leads to increased levels of activation (i.e. P-Selectin and phosphatidylserine [PS]). Resolvins (Rv) are a class of specialized proresolving mediators (SPMs) that promote the resolution of inflammation and, more specifically, have been shown to regulate ADP activation of human platelets. In this study, we tested the hypothesis that the addition of Rv would help to reduce platelet activation during cold (4°C) storage and better preserve platelet function.Methods: Apheresis platelets were collected from healthy donors (n=3-4) using a Trima Accel collection system. On the day of collection (Day 0) platelets were aliquotted into 4 mini-bags (10 mL). Three of the bags received either RvE1 (100μM), RvD2 (100 μM), or Maresin 1 (MaR1; 100 μM), while the final bag received 0.1% EtOH (Vehicle control). Treated mini-bags were incubated for 15 minutes with compound or vehicle prior to long-term storage at 4°C. The following assays were performed on Day 0 and Day 7 of storage: platelet count, Multiplate aggregation (ADP, Collagen, TRAP), and flow cytometry (GPIb expression, P-Selectin, lactadherin [PS], and PAC-1 [activated GPIIb/IIIa]). Data were analyzed using paired t-tests. Significance was determined as P <0.05.Results: P-Selectin and PAC-1 expression in Day 7 Vehicle and compound-treated samples were significantly higher than Day 0 Vehicle levels (Fig. 1A, C). Compared to Vehicle at Day 7, RvD2- and MaR1-treated samples showed a significant reduction in PS expression (RvD2 vs Vehicle, P=0.031; MaR1 vs Vehicle, P=0.018) and PAC-I binding (RvD2 vs Vehicle, P=0.012; MaR1 vs Vehicle, P=0.036) (Fig. 1B, C). No differences in P-Selectin expression were observed for any of the compounds tested (Fig. 1A). TRAP response was higher in RvD2-treated samples compared to Day 7 Vehicle (P=0.047). ADP response was higher in MaR1-treated samples compared to Day 7 Vehicle (P=0.03). No differences in collagen aggregation were observed between samples. All SPM-treated bags resulted in higher platelet counts than Day 7 Vehicle.Conclusions: Cold storage of platelets at 4°C has gained increased attention as an alternative to room temperature storage due to the lower risk of bacterial contamination, better preservation of platelet function, and the potential for a prolonged shelf life; however, increased levels of platelet activation and platelet clumping in cold-stored platelets limit shelf life. Results from our study suggest that the addition of SPMs (RvE1, RvD2, and MaR1) to cold-stored platelets helps minimize platelet activation and even preserve platelet function (RvD2, MarR1) during long-term storage. A platelet product with an extended shelf life would improve platelet inventories and expand access to under-served patients. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Messenger RNA Expression of Albumin, Transferrin, Transthyretin, Asialoglycoprotein Receptor, Cytochrome P450 Isoform, Uptake Transporter, and Efflux Transporter Genes as a Function of Culture Duration in Prolonged Cultured Cryopreserved Human Hepatocytes as Collagen-Matrigel Sandwich Cultures: Evidence for Redifferentiation upon Prolonged Culturing.

Hepatic gene expression as a function of culture duration was evaluated in prolonged cultured human hepatocytes. Human hepatocytes from seven donors were maintained as near-confluent collagen-Matrigelsandwich cultures, with messenger RNA expression for genes responsible for key hepatic functions quantified by real-time polymerase chain reaction at culture durations of 0 (day of plating), 2, 7, 9, 16, 23, 26, 29, 36, and 43 days. Key hepatocyte genes were evaluated, including the differentiation markers albumin, transferrin, and transthyretin; the hepatocyte-specific asialoglycoprotein receptor 1 cytochrome P450 isoforms CYP1A2, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP3A4, and CYP3A7; uptake transporter isoforms SLC10A1, SLC22A1, SLC22A7, SLCO1B1, SLCO1B3, and SLCO2B1; efflux transporter isoforms ATP binding cassette (ABC)B1, ABCB11, ABCC2, ABCC3, ABCC4, and ABCG2; and the nonspecific housekeeping gene hypoxanthine ribosyl transferase 1 (HPRT1). The well established dedifferentiation phenomenon was observed on day 2, with substantial (>80%) decreases in gene expression in day 2 cultures observed for all genes evaluated except HPRT1 and efflux transporters ABCB1, ABCC2, ABCC3 (<50% decrease in expression), ABCC4 (>400% increase in expression), and ABCG2 (no decrease in expression). All genes with a >80% decrease in expression were found to have increased levels of expression on day 7, with peak expression observed on either day 7 or day 9, followed by a gradual decrease in expression up to the longest duration evaluated of 43 days. Our results provide evidence that cultured human hepatocytes undergo redifferentiation upon prolonged culturing. SIGNIFICANCE STATEMENT: This study reports that although human hepatocytes underwent dedifferentiation upon 2 days of culture, prolonged culturing resulted in redifferentiation based on gene expression of differentiation markers, uptake and efflux transporters, and cytochrome P450 isoforms. The observed redifferentiation suggests that prolonged (>7 days) culturing of human hepatocyte cultures may represent an experimental approach to overcome the initial dedifferentiation process, resulting in "stabilized" hepatocytes that can be applied toward the evaluation of drug properties requiring an extended period of treatment and evaluation.

Combining ability and gene action in relation to alloplasmic isonuclear lines in pearl millet [Pennisetum glaucum (L.) R. Br.

The Line × Tester analysis involving two CMS lines with diverse (A1, A4 and A5) cytoplasmic sources and six testers (males) of pearl millet was carried out at JAU, Jamnagar (Gujarat) during Kharif 2018 to identify crosses and good combiners for developing new hybrids to achieve higher yield. The variance due to GCA and SCA showed that the non-additive components were pre-dominant for the expression of days to 50% flowering, plant height (cm), number of effective tillers per plant (no.), ear head diameter (cm), grain yield per plant (g) and Fe and Zn content (ppm) Whereas, additive components were predominant for the expression of days to maturity and ear head diameter (cm). Among the female parents ICMB 96222 were identified as good general combiner for grain yield per plant and some other component traits. Among the male parents J-2597, J-2598 and J-2603 was good general combiner for most of the characters. Among the 36 hybrids, five crosses (ICMA5 95222 x J-2597, ICMA4 95222 x J-2597 ICMA5 96222 x J-2596, ICMA4 96222 x J-2584 and ICMA1 96222 x J-2597) were identified as good specific combiners based on significant and positive sca effect for grain yield per plant.

Effect of hyperoxia exposure on expression of microRNA-125b and tumor necrosis factor alpha, interleukin-6 in lung tissues of premature newborn rats

Objective To observe the expression of microRNA-125b (miR-125), tumor necrosis factor alpha (TNF-α) and interleukin-6(IL-6) in premature rats exposed to hyperoxia. Methods Eighty 1-day old Sprague Dawley (SD) rats were randomly divided into an air group and a hyperoxia group.The rats in the hyperoxia group were continuously exposed to oxygen chamber for 1-3 L/min, oxygen volume fraction was maintained at (800±50) mL/L, and the rats in air group were placed in the same room with the oxygen volume fraction at 210 mL/L.The feeding conditions were same in 2 groups.Lung tissues of premature rats were extracted at different time (1, 4, 7, 10, 14 days). The pathologic changes in the lung tissues were observed by hematoxylin-eosin (HE) staining.The levels of miR-125b and TNF-α, IL-6 in lung tissues were detected by reverse transcription polymerase chain reaction (qRT-PCT) and enzyme-linked immunosorbent assay (ELISA). Results Compared with the air group, miR-125b in the hyperoxia group increased slowly after day 1, reached the highest in day 10 (2.554±0.323), and the relative expression in day 14 decreased slightly(2.329±0.263), and there were significant differences between 2 groups at di-fferent time (all P 0.05), but there was significant correlation between miR-125b and IL-6 in hyperoxia groups(r=0.439, P<0.05). Conclusions The levels of miR-125b, TNF-α and IL-6 are involved in the pathological process of bronchopulmonary dysplasia induced by hyperoxia, and IL-6 may be the key factor for miR-125b. Key words: Hyperoxia exposure; Rat, premature; Lung injury; MicroRNA-125b; Tumor necrosis factor alpha; Interleukin-6

Genetic architecture of yield and quality traits in wheat (Triticum aestivum L.)

The aim of this study was to investigate gene effects and genetic variability by generation mean analysis for some yield and quality traits in three diverse crosses of wheat (JW 3288 × HD 8864, MP 3269 x GW 173 andJW 3020 × GW 366). The parents P1 and P1 and F1 , F2 , BC1 and BC2 populations were grown in a randomized complete block design with three replications during 2017-2018 crop seasons. Both additive and non-additive gene effects were involved in the expression of days to ear emergence, plant height, days to maturity, number of grains per spike, grain yield, flour recovery, husk content, protein content, lysine content and amylose content. The epistatic interaction was observed for all the ten traits in said crosses. In general, magnitude of dominance effect (h) has a greater value than additive effect (d) in all the traits. Digenic interaction indicated complex nature of inheritance means non-additive gene action. The improvement by selection in early generation could be advisable.

Cryoprotectant role of exopolysaccharide of Pseudomonas sp. ID1 in the vitrification of IVM cow oocytes.

Biological molecules isolated from organisms that live under subzero conditions could be used to protect oocytes from cryoinjuries suffered during cryopreservation. This study examined the cryoprotectant role of exopolysaccharides of Pseudomonas sp. ID1 (EPS ID1) in the vitrification of prepubertal and adult cow oocytes. IVM oocytes were vitrified and warmed in media supplemented with 0, 1, 10, 100 or 1000µgmL-1 EPS ID1. After warming, oocytes were fertilised and embryo development, spindle morphology and the expression of several genes in Day 8 blastocysts were assessed. Vitrification led to significantly lower proportion of prepubertal oocytes exhibiting a normal spindle configuration. In fresh control oocytes and most groups of vitrified adult oocytes, similar percentages of oocytes with a normal spindle configuration were observed. Percentages of Day 8 blastocysts were similar for prepubertal oocytes vitrified in the absence or presence of 1 or 10µgmL-1 EPS ID1 and for adult oocytes vitrified in the presence of 10µgmL-1 EPS ID1 compared with non-vitrified oocytes. EPS ID1 supplementation had no effect on solute carrier family 2 member 3 (SLC2A3), ubiquitin-conjugating enzyme E2A (UBE2A) and histone deacetylase 1 (HDAC1) expression in Day 8 blastocysts form adult oocytes. However, supplementation with 10 and 100µgmL-1 EPS ID1 led to increased expression of genes involved in epigenetic modifications (DNA methyltransferase 3 alpha (DNMT3A) and K (lysine) acetyltransferase 2A (KAT2A)) and apoptosis (BCL2 associated X apoptosis regulator (BAX) and BCL2-like 1 (BCL2L1)). The lowest BAX:BCL2L1 ratio was found in the 10µgmL-1 EPS ID1-supplemented group. The results suggest that 10µgmL-1 EPS ID1 added to vitrification and warming media may help protect bovine oocytes against cryodamage.

PSVII-20 Differential gene expression of day 15 endometrium of pregnant Bos indicus beef cows according to expression of estrus at timed-AI.

PSVII-20 Differential gene expression of day 15 endometrium of pregnant Bos indicus beef cows according to expression of estrus at timed-AI.

Transcriptome analysis of immune genes in peripheral blood mononuclear cells of young foals and adult horses.

During the neonatal period, the ability to generate immune effector and memory responses to vaccines or pathogens is often questioned. This study was undertaken to obtain a global view of the natural differences in the expression of immune genes early in life. Our hypothesis was that transcriptome analyses of peripheral blood mononuclear cells (PBMCs) of foals (on day 1 and day 42 after birth) and adult horses would show differential gene expression profiles that characterize natural immune processes. Gene ontology enrichment analysis provided assessment of biological processes affected by age, and a list of 897 genes with ≥2 fold higher (p<0.01) expression in day 42 when compared to day 1 foal samples. Up-regulated genes included B cell and T cell receptor diversity genes; DNA replication enzymes; natural killer cell receptors; granzyme B and perforin; complement receptors; immunomodulatory receptors; cell adhesion molecules; and cytokines/chemokines and their receptors. The list of 1,383 genes that had higher (p<0.01) expression on day 1 when compared to day 42 foal samples was populated by genes with roles in innate immunity such as antimicrobial proteins; pathogen recognition receptors; cytokines/chemokines and their receptors; cell adhesion molecules; co-stimulatory molecules; and T cell receptor delta chain. Within the 742 genes with increased expression between day 42 foal and adult samples, B cell immunity was the main biological process (p = 2.4E-04). Novel data on markedly low (p<0.0001) TLR3 gene expression, and high (p≤0.01) expression of IL27, IL13RA1, IREM-1, SIRL-1, and SIRPα on day 1 compared to day 42 foal samples point out potential mechanisms of increased susceptibility to pathogens in early life. The results portray a progression from innate immune gene expression predominance early in life to adaptive immune gene expression increasing with age with a putative overlay of immune suppressing genes in the neonatal phase. These results provide insight to the unique attributes of the equine neonatal and young immune system, and offer many avenues of future investigation.

Characterization of human pre-implantation embryonic exosomes

To isolate and characterize exosomes secreted from human embryos. Prospective experimental study Spent IVF culture media was collected prospectively and pooled from 50 embryos at two different stages of early embryonic development on day 3 and at blastocyst. The 4 groups assessed were: control media without protein supplement (plasmanate), control media with plasmanate, day 3 embryo media, and blastocyst media. Exosome size and number were measured using nanosight tracking analysis (NTA). Characterization of embryonic exosome cargo was performed through mass spectrometry proteomic analysis. Exosomal protein cargo unique to embryos was submitted to bioinformatic analysis for molecular pathways and biological processes. NTA identified exosomes in control media with plasmanate, pooled day 3 embryo media, and pooled blastocyst media, which had significantly higher concentrations compared to control media without plasmanate. The particle size distribution for all 3 groups showed isolated peaks in the known exosome size range of 50-150 nm. Moreover, day 5 embryo media had significantly higher protein amount per mL media and per exosome compared to day 3 media and control media with plasmanate. Mass spectrometry proteomic analysis identified 273 distinct proteins in the control with plasmanate, day 3, and day 5 embryonic exosomes, of which 193 were shared by all groups. We identified 30 unique proteins with increased expression in day 3 and/or day 5 exosomes. These proteins were enriched in components of the WNT and p53 pathways, which have been shown in mouse and human models to play vital roles in embryonic development and implantation. Importantly, 11 of these genes have been shown previously via knockout models to play roles in blastocyst development, implantation and early embryogenesis. Furthermore, 10 of these unique embryonic proteins are involved in biological processes of immune cell activation and vesicle mediated exocytosis, which further supports their potential immune function and packaging of these proteins into exosomes. By analyzing spent IVF media, we have shown that pre-implantation embryos secrete exosomes with unique cargo that may serve potential roles in implantation, early embryogenesis and immune regulation. Future directions include molecular functioning studies with isolated embryonic exosomes as well as correlating exosome proteomic profile to embryonic outcomes such as blastocyst morphology and aneuploidy.

Genomic profiling of bovine corpus luteum maturation.

To unveil novel global changes associated with corpus luteum (CL) maturation, we analyzed transcriptome data for the bovine CL on days 4 and 11, representing the developing vs. mature gland. Our analyses revealed 681 differentially expressed genes (363 and 318 on day 4 and 11, respectively), with ≥2 fold change and FDR of <5%. Different gene ontology (GO) categories were represented prominently in transcriptome data at these stages (e.g. days 4: cell cycle, chromosome, DNA metabolic process and replication and on day 11: immune response; lipid metabolic process and complement activation). Based on bioinformatic analyses, select genes expression in day 4 and 11 CL was validated with quantitative real-time PCR. Cell specific expression was also determined in enriched luteal endothelial and steroidogenic cells. Genes related to the angiogenic process such as NOS3, which maintains dilated vessels and MMP9, matrix degrading enzyme, were higher on day 4. Importantly, our data suggests day 11 CL acquire mechanisms to prevent blood vessel sprouting and promote their maturation by expressing NOTCH4 and JAG1, greatly enriched in luteal endothelial cells. Another endothelial specific gene, CD300LG, was identified here in the CL for the first time. CD300LG is an adhesion molecule enabling lymphocyte migration, its higher levels at mid cycle are expected to support the transmigration of immune cells into the CL at this stage. Together with steroidogenic genes, most of the genes regulating de-novo cholesterol biosynthetic pathway (e.g HMGCS, HMGCR) and cholesterol uptake from plasma (LDLR, APOD and APOE) were upregulated in the mature CL. These findings provide new insight of the processes involved in CL maturation including blood vessel growth and stabilization, leucocyte transmigration as well as progesterone synthesis as the CL matures.

Combining Ability and Genetic Control of Pod Yield and Component Traits in Dolichos Bean

ABSTRACTThe major limitation of dolichos bean (Dolichos lablab L.) improvement is the narrow genetic base for useful traits present in available germplasm that may be overcome through interspecific hybridization. Five diverse genotypes involving two each of indeterminate and semi-indeterminate, and one determinate type, were crossed in a 5 × 5 half diallel mating design to estimate combining ability, mode of gene action, and extent of heterosis for 12 quantitative traits. The preponderance of additive genetic effects was evident for pod weight and number of pods/inflorescence; additive and non-additive gene effects for number of pods/plant, and non-additive gene action in expression of days to 50% flowering, raceme length, number nodes/inflorescence, number inflorescences/plant, number flower buds/inflorescence, pod length, pod diameter, pod thickness, and pod yield/plant. The parents “BCDB 10” and “BCDB-2” were identified as potential donors for breeding. The crosses “BCDB-2×BCDB-15” and “BCDB-10×BCDB-5” exhibited high heterobeltiosis for pod yield/plant to the extent of 114.26 and 86.63%, respectively. They could not be exploited at the commercial level. These crosses involved one parent having high general combining ability effects for pod yield and other important horticultural traits that could produce desirable transgressive segregants in subsequent generations.

Global assessment of imprinted gene expression in the bovine conceptus by next generation sequencing

ABSTRACTGenomic imprinting is an epigenetic mechanism that leads to parental-allele-specific gene expression. Approximately 150 imprinted genes have been identified in humans and mice but less than 30 have been described as imprinted in cattle. For the purpose of de novo identification of imprinted genes in bovine, we determined global monoallelic gene expression in brain, skeletal muscle, liver, kidney and placenta of day ∼105 Bos taurus indicus × Bos taurus taurus F1 conceptuses using RNA sequencing. To accomplish this, we developed a bioinformatics pipeline to identify parent-specific single nucleotide polymorphism alleles after filtering adenosine to inosine (A-to-I) RNA editing sites. We identified 53 genes subject to monoallelic expression. Twenty three are genes known to be imprinted in the cow and an additional 7 have previously been characterized as imprinted in human and/or mouse that have not been reported as imprinted in cattle. Of the remaining 23 genes, we found that 10 are uncharacterized or unannotated transcripts located in known imprinted clusters, whereas the other 13 genes are distributed throughout the bovine genome and are not close to any known imprinted clusters. To exclude potential cis-eQTL effects on allele expression, we corroborated the parental specificity of monoallelic expression in day 86 Bos taurus taurus × Bos taurus taurus conceptuses and identified 8 novel bovine imprinted genes. Further, we identified 671 candidate A-to-I RNA editing sites and describe random X-inactivation in day 15 bovine extraembryonic membranes. Our results expand the imprinted gene list in bovine and demonstrate that monoallelic gene expression can be the result of cis-eQTL effects.