Cellular Modifications Research Articles

Transmission and regulation of biochemical stimulus via a nanoshell directly adsorbed on the cell membrane to enhance chondrogenic differentiation of mesenchymal stem cell.

A nanoscale artificial extracellular matrix (nanoshell) formed by layer-by-layer adsorption can enhance and modulate the function of stem cells by transferring biochemical stimulus to the cell directly. Here, the nanoshell composed of fibronectin (FN) and chondroitin sulfate (CS) is demonstrated to promote chondrogenic differentiation of mesenchymal stem cells (MSCs). The multilayer structure of nanoshell is formed by repeating self-assembly of FN and CS, and its thickness can be controlled through the number of layers. The expression of chondrogenic markers in MSCs coated with the FN/CS nanoshell was increased as the number of bilayers in the nanoshell increased until four, but when it exceeds five bilayers, the effect began to decrease. Finally, the MSCs coated with optimized four bilayers of FN/CS nanoshell have high chondrogenic differentiation efficiency and showed the potential to increase formation of cartilage tissue when it is transplanted into mouse kidney. So, the precise regulation of stem cell fate at single cell level can be possible through the cellular surface modification by self-assembled polymeric film.

Reproductive and Postmenopausal Histological Changes of Human Endometrial Cells: Electron Microscopic Study

Background: The ability of women to reproduce is well known to decline with age. Advanced maternal age is related to reduce fertility and adverse pregnancy outcomes. There is evident decline in human fertility with age due to several histological changes occur with progressive age. Objective: The research was intended to know that some cellular modifications happen during the reproductive and post-menopause phases in human endometrial cells. Materials and methods: 30 womb samples each from reproductive and postmenopausal women were collected from unembalmed human cadavers. Before being taken to electron microscopy, samples were resolved in glutaraldehyde solution and osmium tetroxide as fixative. Small sizes of endometrial specimens were drawn into a unique fixative (i.e., 5% glutaraldehyde in a buffer of 0.1μphosphate (PH 7.4), then washed three times in a buffer solution. The moved to prepare thin sections for electron microscopy. Samples were integrated at 60co for two days in Arledit –Rezen capsule. Thin parts for E.M. were then ready study Ultra-structural cell of endometrium Results: Many differences were clear and evident when comparison was made between reproductive and postmenopause stages of endometrial cell ultrastructurally. The structural changes involved shape, density, cell organelles aggregation in general, and were assigned to age, as well as degeneration, deposition, and irregularity. Conclusion: Present work results indicated that, increase the amount of cell organelles in uterine cells of young women, while the old ages characterize by less number of organelles and degenerated.

Type XVIII Collagen Modulates Keratohyalin Granule Formation and Keratinization in Oral Mucosa.

Epithelial keratinization involves complex cellular modifications that provide protection against pathogens and chemical and mechanical injuries. In the oral cavity, keratinized mucosa is also crucial to maintain healthy periodontal or peri-implant tissues. In this study, we investigated the roles of type XVIII collagen, a collagen-glycosaminoglycan featuring an extracellular matrix component present in the basement membrane, in oral mucosal keratinization. Histological analysis of keratinized and non-keratinized oral mucosa showed that type XVIII collagen was highly expressed in keratinized mucosa. Additionally, a 3D culture system using human squamous carcinoma cells (TR146) was used to evaluate and correlate the changes in the expression of type XVIII collagen gene, COL18A1, and epithelial keratinization-related markers, e.g., keratin 1 (KRT1) and 10 (KRT10). The results showed that the increase in COL18A1 expression followed the increase in KRT1 and KRT10 mRNA levels. Additionally, loss-of-function analyses using silencing RNA targeting COL18A1 mRNA and a Col18-knockout (KO) mouse revealed that the absence of type XVIII collagen induces a dramatic decrease in KRT10 expression as well as in the number and size of keratohyalin granules. Together, the results of this study demonstrate the importance of type XVIII collagen in oral mucosal keratinization.

Cryopreservation of rooster semen: Evidence for the epigenetic modifications of thawed sperm

Although semen cryopreservation is an important, widely used technique for long-term sperm storage, it not only induces partially irreversible damages to sperm but might also deteriorate anatomical, biochemical, and structural organelles. These cellular and epigenetic modifications are the main reasons underlying the decline in sperm motility and fertility during the freeze-thaw process. Using the two Lake and Beltsville semen extenders, the present study aims to evaluate the epigenetic patterns (DNA methylation and histone modification), cellular parameters (e.g., membrane integrity, viability, DNA stability, mitochondria activity, and apoptosis status), and fertility potential of rooster semen collected from six mature roosters before and after cryopreservation according to a standard protocol. The results show that cryopreservation leads to significantly (P < 0.05) reduced values of the parameters examined when compared with those of fresh sperms. While the extenders used exhibit no difference with respect to DNA methylation (DNMT), the Lake extender leads to significant reductions (P < 0.05) in H3K9 acetylation (17.4 ± 1.8) and H3K4 methylation (42 ± 2.3) compared to the Beltsville (9.2 ± 1.8 and 23 ± 2.3, respectively). Compared to the Beltsville extender, the Lake one is also observed to yield a significantly (P < 0.05) superior sperm quality in terms of total motility (TM; 77.2 ± 1.6 vs. 68.3 ± 1.6), average path velocity (VAP; 71 ± 1.4 vs. 53 ± 1.4), and straight-line velocity (VSL; 52 ± 1.5 vs. 34 ± 1.5) as well as significantly (P < 0.05) higher viability (60 ± 1.69 vs. 51 ± 1.69) and membrane functionality (55 ± 3.2 vs. 46 ± 3.2). The Lake extender is also found to outperform the Beltsville one due to its significantly (P < 0.05) higher fertility rate (59.5% vs. 47.2%). The two extenders, however, exhibit no differences in DNA fragmentation, mitochondrial activity, or hatchability rate. The Beltsville extender showed to be superior to the Lake one due to its significantly greater reactive oxygen species percentage (ROS; 45.9 ±.3.2 vs. 28.5 ± 3.2) and apoptosis (29 ± 2.3 vs. 27 ± 2.3). It may be concluded that the Lake extender is capable of improving the cellular and epigenetic parameters of rooster sperms during cryopreservation due to the crucial roles it plays in the protection of sperms against cryo-damages.

The next generation of approaches to investigate the link between synaptic plasticity and learning.

Activity-dependent synaptic plasticity has since long been proposed to represent the subcellular substrate of learning and memory, one of the most important behavioral processes through which we adapt to our environment. Despite the undisputed importance of synaptic plasticity for brain function, its exact contribution to learning processes in the context of cellular and connectivity modifications remains obscure. Causally bridging synaptic and behavioral modifications indeed remains limited by the available tools to measure and control synaptic strength and plasticity in vivo under behaviorally relevant conditions. After a brief summary of the current state of knowledge of the links between synaptic plasticity and learning, we will review and discuss the available and desired tools to progress in this endeavor.

Computational Network Analysis Identifies Evolutionarily Conserved miRNA Gene Interactions Potentially Regulating Immune Response in Bovine Trypanosomosis.

Bovine trypanosomosis is a devastating disease that causes huge economic loss to the global cattle industry on a yearly basis. Selection of accurate biomarkers are important in early disease diagnosis and treatment. Of late, micro-RNAs (miRNAs) are becoming the most useful biomarkers for both infectious and non-infectious diseases in humans, but this is not the case in animals. miRNAs are non-coding RNAs that regulate gene expression through binding to the 3′-, 5′-untranslated regions (UTR) or coding sequence (CDS) region of one or more target genes. The molecular identification of miRNAs that regulates the expression of immune genes responding to bovine trypanosomosis is poorly defined, as is the possibility that these miRNAs could serve as potential biomarkers for disease diagnosis and treatment currently unknown. To this end, we utilized in silico tools to elucidate conserved miRNAs regulating immune response genes during infection, in addition to cataloging significant genes. Based on the p value of 1.77E-32, we selected 25 significantly expressed immune genes. Using prediction analysis, we identified a total of 4,251 bovine miRNAs targeting these selected genes across the 3′UTR, 5′UTR and CDS regions. Thereafter, we identified candidate miRNAs based on the number of gene targets and their abundance at the three regions. In all, we found the top 13 miRNAs that are significantly conserved targeting 7 innate immune response genes, including bta-mir-2460, bta-mir-193a, bta-mir-2316, and bta-mir-2456. Our gene ontology analysis suggests that these miRNAs are involved in gene silencing, cellular protein modification process, RNA-induced silencing complex, regulation of humoral immune response mediated by circulating immunoglobulin and negative regulation of chronic inflammatory response, among others. In conclusion, this study identifies specific miRNAs that may be involved in the regulation of gene expression during bovine trypanosomosis. These miRNAs have the potential to be used as biomarkers in the animal and veterinary research community to facilitate the development of tools for early disease diagnosis/detection, drug targeting, and the rational design of drugs to facilitate disease treatment.

Time matters when ultrasonicating beef: The best time for tenderness is not the best for reducing microbial counts

AbstractResearch on the effects of high‐intensity ultrasound (HIU) on meat quality properties shows contradictory results. The latter has been sometimes attributed to limited exposure time, not enough to cause cellular modifications. This study evaluated the effect of different exposure times (0, 10, 20, and 40 min) of HIU (37 kHz, 90 W cm−2) on physicochemical properties including; pH, color, tenderness, and microbial counts of two different portions of beef Longissimus lumborum (cranial–caudal). No significant effect of loin portion was observed (p &gt; .05). Ultrasonication time caused a 20.7% toughness reduction of beef (p &lt; .05). However, ultrasonication time also increased hue from 0.62 to 0.76 from red to orange values (p &lt; .05), but it did not affect important coordinates such as a* or C*. Ultrasound caused an increase in the pH from 5.46 to 5.6 (p &lt; .05), but pH was not affected by the time. The highest tenderness was achieved at 40 min of sonication. Significant reductions of mesophiles, psychrophiles, and coliforms were observed after ultrasonication when compared to control (p &lt; .05). The best ultrasonic condition for microbial reduction was at 10 min, when the lowest microorganism counts were achieved, with a subsequent growth when increasing ultrasonication time.Practical ApplicationsHigh‐intensity ultrasound application appears to be a promising method to increase tenderness in beef. However, optimum time, intensity, and methodology for its application have to be defined, because the variations may affect other quality characteristics such as color or microbial contamination. To find ideal parameter values may help the industry to take decisions on the design of specialized equipment for ultrasonication of meat.

Differential regulation of mycelial growth and aflatoxin biosynthesis by Aspergillus flavus under different temperatures as revealed by strand-specific RNA-Seq.

Although several regulatory pathways have been reported for Aspergillus flavus, the regulation of aflatoxin production and mycelial growth under different temperatures remains unclear. In this study, A. flavus differentially expressed genes (DEGs) and regulatory pathways were analyzed under three temperatures, by strand‐specific RNA‐Seq. Results show that a total of 2,428 and 1,474 DEGs were identified in fungal mycelia cultured at 20°C and 37°C, respectively, as compared with the control (28°C). Approximately ~ 79% of DEGs in the 37°C samples were up‐regulated genes, while ~ 63% of DEGs in the 20°C samples were down‐regulated genes. Most of the DEG pathways enriched by lower temperatures differed from those enriched by higher temperatures, while only a small portion of the pathways were shared by A. flavus grown under different temperatures. Aflatoxin biosynthesis, Butanoate metabolism, oxidation–reduction process, and benzene‐containing compound metabolic process were the shared down‐regulated pathways, while steroid biosynthesis, oxidoreductase activity, cellular protein modification process, DNA binding, protein complex were the shared up‐regulated pathways between lower and higher temperatures. The shared genes and pathways are the key regulatory candidates for aflatoxin biosynthesis with changes of temperature. In addition, the identification of both up‐regulated and down‐regulated genes provides a useful gene set for further investigation of the aflatoxin biosynthesis among Aspergillus.

Mass Spectrometry-Based Proteomics of Fungal Pathogenesis, Host-Fungal Interactions, and Antifungal Development.

The prevalence of fungal diseases is increasing on a global scale, ranging from acute to systemic infections caused by commensal or pathogenic microorganisms, often associated with the immune status of the host. Morbidity and mortality rates remain high and our ability to treat fungal infections is challenged by a limited arsenal of antifungal agents and the emergence of drug resistant pathogens. There is a high demand for new approaches to elucidate the fungal mechanisms of pathogenesis and the interplay between host and pathogen to discover novel treatment options. Moreover, the need for improved drug efficacy and reduced host toxicity requires the identification and characterization of antifungal biological targets and molecular mechanisms of action. Mass spectrometry (MS)-based proteomics is a rapidly advancing field capable of addressing these priorities by providing comprehensive information on the dynamics of cellular processes, modifications, and interactions. In this Review, we focus on applications of MS-based proteomics in a diverse array of fungal pathogens and host systems to define and distinguish the molecular details of fungal pathogenesis and host–fungal interactions. We also explore the emerging role of MS-based proteomics in the discovery and development of novel antifungal therapies and provide insight into the future of MS-based proteomics in fungal biology.

Biochemical Changes in Human Cells Exposed to Low Concentrations of Gold Nanoparticles Detected by Raman Microspectroscopy.

The toxicological implications of nanoparticles deserve accurate scientific investigation for the protection of human health. Although toxic effects involve specific organs, the events that cause them have their origin from biochemical modifications of some cellular constituents. Therefore, a first analysis to evaluate the effects due to the action of nanoparticles is achieved by investigation of in vitro cells, which allows the identification of the cellular modifications caused by nanoparticles (NPs) even at much lower doses than the lethal ones. This work evaluated the Raman microspectroscopy capability to monitor biochemical changes occurring in human cells as a consequence of exposure to a suspension of gold nanoparticles with a non-cytotoxic concentration. Human keratinocyte cells were used as a model cell line, because they are mainly involved in environmental exposure. A trypan blue assay revealed that the investigated concentration, 650 ng/mL, is non-cytotoxic (about 5% of cells died after 48 h exposure). Specific Raman spectral markers to represent the cell response to nanoparticle exposure were found (at 1450 and 2865 cm−1) in the cytoplasm spectra, with the aid of ratiometric and principal component analysis.

Sub-cytotoxic doses of pharmaceutical silica nanoparticles show significant impact on the proteome of HepG2 cells

The ever-growing application of nanoparticles (NPs) in medical and pharmaceutical domains has brought issues related to their toxicity into focus. However, a profound analysis of non-acute, sub-lethal effects of engineered pharmaceutical NPs is often disregarded during such toxicological investigations. Here, two selected NPs were investigated in cultured HepG2 cells in terms of their intracellular localization and the associated impact on pharmacokinetically relevant CYP3A4 isoform, as well as the induced changes observed in the proteome of such cells. Using SILAC (Stable Isotope Labeling by Amino acids in Cell culture)-based mass spectrometry facilitated quantitative proteomics, significant proteomic changes in NP-treated hepatocytes were detected, which were subsequently analyzed via bioinformatic tools.Both, silica NPs (SiO2 NP) and cargo-free PEGylated stealth liposomes resulted in the induction of CYP3A4-activity up to 150% in a dose-dependent manner, with different time-dependent response-patterns as a function of NP-type after a single treatment. Proteomic analysis revealed that the observed metabolic alterations are only one aspect of the cellular response to NP-exposure. SiO2NPs (free in cytoplasm) caused extensive changes in the proteome, whereas liposomes (compartmentalized) seemed unproblematic as they accounted for minimal changes in the protein profile.Based on the obtained results, proteomic analyses were revealed to be highly important for the toxicological assessment of NPs. Although sub-toxic concentrations of many NPs are considered as uncritical based on standard toxicological assays, proteomic analysis indicated that drug-free NPs could cause fundamental cellular modifications, which were attributed to different causal networks and regulatory pathways.

MRNA transcription profile of potato (Solanum tuberosum L.) in response to explant cutting

The explant is an important parameter in plant tissue culture. Most plant tissue culture studies in the literature have focused almost exclusively on the use of explants as a tool for organogenesis or (micro)propagation, and there is limited information that documents the changes that occur in explants in response to cutting during explant preparation just prior to or during culture. To better understand the molecular changes occurring during the wounding process that accompanies the preparation of an explant, this study examined the mRNA transcription profile of 4-week-old potato (Solanum tuberosum L. cv. Desirée) single-node stem segments. To achieve this, RNA-seq analysis was performed to identify significantly differentially expressed genes (DEGs) that existed between uncut in vitro plantlets and cut explants. DEGs corresponding to specific enzyme profiles were broadly connected to cellular processes in the extracellular region, nucleus, and plasma membrane, and were associated with biosynthesis, carbohydrate metabolism and catabolism, cellular protein modification, growth and development, and response to stress. The RNA-seq data of eight highly up- or down-regulated DEGs was validated by RT-qPCR, showing high positive Spearman and Pearson correlation coefficients (0.73 and 0.95, respectively) between SeqMonk LFC and RT-qPCR LFC. These results provide a novel perspective of the changes taking place within explants when they are cut and serve as a valuable basis for the study of explant-related stress in plant tissue culture in other species. In addition, our findings may contribute to an understanding of the success or failure of the tissue culture protocol and subsequent establishment of in vitro cultures by better appreciating the stress-related molecular changes taking place.

KEGG-expressed genes and pathways in intervertebral disc degeneration: Protocol for a systematic review and data mining.

miRNAs and genes play significant roles in the etiology and pathogenesis of intervertebral disc degeneration (IDD). This study aimed to identify aberrantly expressed miRNAs, genes, and pathways in IDD through a comprehensive bioinformatics analysis.Data of miRNAs expression microarrays (GSE63492) and genes microarrays (GSE23130) were obtained from GEO database. Similarly, aberrantly expressed miRNAs and genes were obtained using GEO2R. In addition, functional and enrichment analyses of selected miRNAs and genes were performed using the DAVID database. Meanwhile, protein–protein interaction (PPI) network was constructed using STRING, and then visualized in Cytoscape.A total of 98 upregulated miRNAs were identified. They were enriched in biological processes of response to organelle, ion binding, cellular nitrogen compound metabolic process, biosynthetic process, small molecule metabolic process, cellular protein modification process, catabolic process, molecular function, neurotrophin TRK receptor signaling pathway, and protein complex. In addition, 1405 high expression protein genes were detected. It indicated enrichment in biological processes, such as translational initiation, nonsense-mediated decay, viral transcription, cell–cell adhesion, rRNA processing, translation, RP-dependent cotranslational protein targeting to membrane, nuclear-transcribed mRNA catabolic process, regulation of mRNA stability, and mRNA splicing via spliceosome and extracellular matrix organization. In addition, pathway analysis exhibited the common enrichment in focal adhesion, Hippo signaling pathway, ECM-receptor interaction, Wnt signaling pathway, PI3K-Akt signaling pathway, endocytosis, proteoglycans in cancer, and so on. The top 10 central genes of PPI network were POTEE, PPP2CA, RPL17, HSP90AA1, POTEF, RPL13A, ACTB, RPL18, RPS24, and HSPA1A.In conclusion, our research proposed abnormally expressed miRNAs, genes, and pathways in IDD through bioinformatics methods, which may provide new insights into the pathogenesis of IDD. Thus, the Hub gene involving POTEE, PPP2CA, RPL17, HSP90AA1, POTEF, RPL13A, ACTB, RPL18, RPS24, and HSPA1A may be biomarkers for accurate diagnosis and treatment of IDD in the future.

Rapidly evolving PARP proteins act as both positive and negative regulators of the antiviral response

Abstract Viral manipulation of cellular post-translational modifications (PTMs) can be key to their survival and replication. One understudied PTM that has been implicated in the antiviral response is ADP-ribosylation. To manipulate this modification, several viral families encode for a macrodomain capable of binding and removing ADP-ribosylation. Importantly, disruption of this function can lead to reduction in pathogenesis or increases in sensitivity to the host interferon response. While the family of host proteins capable of catalyzing ADP-ribosylation, known as PARPs, is large, a subset show multiple signs of rapid evolution, including gene duplication and loss as well as recurrent sequence changes. Focusing in on these genes, we have found that the long isoform of PARP13 is acting directly to inhibit Sindbis virus, a member of the macrodomain-containing alphavirus family. The full antiviral activity requires the presence of the PARP domain in addition to proper localization. In contrast to this, the short isoform acts as a negative regulator of the interferon response by binding and degrading Interferon mRNAs. The balance between antiviral and proviral functions creates a novel dynamic that is potentially more resistant to viral antagonism. We are now exploring the role of ADP-ribosylation in modulating these two phenotypes.

MRNA transcription profile of potato (Solanum tuberosum L.) exposed to ultrasound during different stages of in vitro plantlet development

Key messageIn response to an ultrasound pulse, several hundred DEGs, including in response to stress, were up- or down-regulated in in vitro potato plantlets. Despite this abiotic stress, plantlets survived.Ultrasound (US) can influence plant growth and development. To better understand the genetic mechanism underlying the physiological response of potato to US, single-node segments of four-week-old in vitro plantlets were subjected to US at 35 kHz for 20 min. Following mRNA purification, 10 cDNA libraries were assessed by RNA-seq. Significantly differentially expressed genes (DEGs) were categorized by gene ontology or Kyoto Encyclopedia of Genes and Genomes identifiers. The expression intensity of 40,430 genes was studied. Several hundred DEGs associated with biosynthesis, carbohydrate metabolism and catabolism, cellular protein modification, and response to stress, and which were expressed mainly in the extracellular region, nucleus, and plasma membrane, were either up- or down-regulated in response to US. RT-qPCR was used to validate RNA-seq data of 10 highly up- or down-regulated DEGs, and both Spearman and Pearson correlations between SeqMonk LFC and RT-qPCR LFC were highly positive (0.97). This study examines how some processes evolved over time (0 h, 24 h, 48 h, 1 week and 4 weeks) after an abiotic stress (US) was imposed on in vitro potato explants, and provides clues to the temporal dynamics in DEG-based enzyme functions in response to this stress. Despite this abiotic stress, plantlets survived.

A Review in Research Progress Concerning m6A Methylation and Immunoregulation.

Over 100 types of cellular RNA modifications have been identified in both coding and a variety of non-coding RNAs. N6-methyladenosine (m6A) is the most prevalent and abundant post-transcriptional RNA modification on eukaryote mRNA, and its biological functions are mediated by special binding proteins (i.e., methyltransferases, demethylases, and effectors) that recognize this modification. The presence of m6A on transcripts contributes to diverse fundamental cellular functions, such as pre-mRNA splicing, nuclear transport, stability, translation, and microRNA biogenesis, implying an association with numerous human diseases. This review principally summarizes recent progress in the study of m6A methylation mechanisms and relevant roles they play in immunoregulation.

RNAi expression tuning, microfluidic screening, and genome recombineering for improved protein production in Saccharomyces cerevisiae.

The cellular machinery that supports protein synthesis and secretion lies at the foundation of cell factory-centered protein production. Due to the complexity of such cellular machinery, the challenge in generating a superior cell factory is to fully exploit the production potential by finding beneficial targets for optimized strains, which ideally could be used for improved secretion of other proteins. We focused on an approach in the yeast Saccharomyces cerevisiae that allows for attenuation of gene expression, using RNAi combined with high-throughput microfluidic single-cell screening for cells with improved protein secretion. Using direct experimental validation or enrichment analysis-assisted characterization of systematically introduced RNAi perturbations, we could identify targets that improve protein secretion. We found that genes with functions in cellular metabolism (YDC1, AAD4, ADE8, and SDH1), protein modification and degradation (VPS73, KTR2, CNL1, and SSA1), and cell cycle (CDC39), can all impact recombinant protein production when expressed at differentially down-regulated levels. By establishing a workflow that incorporates Cas9-mediated recombineering, we demonstrated how we could tune the expression of the identified gene targets for further improved protein production for specific proteins. Our findings offer a high throughput and semirational platform design, which will improve not only the production of a desired protein but even more importantly, shed additional light on connections between protein production and other cellular processes.

Transcriptome analysis to identify long non coding RNA (lncRNA) and characterize their functional role in back fat tissue of pig

Long non coding RNAs (lncRNA) have been previously found to be involved in important cellular activities like epigenetics, implantation, cell growth etc. in pigs. However, comprehensive analysis of lncRNA in back fat tissues at different developmental stages in pigs is still lacking. In this study we conducted transcriptome analysis in the back fat tissue of a F1 crossbred Korean Native Pig (KNP) × Yorkshire Pig to identify lncRNA. We investigated their role in 16 pigs at two different growth stages; stage 1 (10 weeks, n = 8) and stage 2 (26 weeks, n = 8). After quality assessment of sequencing reads, we got a total of 1,641,165 assembled transcripts out of eight paired end read from each stage. Among them, 6808 lncRNA transcripts were identified by filtering on the basis of multiple parameters like read length ≥ 200 nucleotides, exon numbers ≥2, FPKM ≥0.5, coding potential score < 0 etc. PFAM and RFAM were used to filter out all possible protein coding genes and housekeeping RNAs respectively. A total of 103 lncRNAs and 1057 mRNAs were found to be differentially expressed (DE) between the two stages (|log2FC| > 2, q < 0.05). We also identified 306 genes located around 100 kb upstream and 234 genes downstream around these DE lncRNA transcripts. The expression of top eleven DE lncRNAs (COL4A6, LY7S, MYH2, OXCT1, SMPDL3A, TMEM182, TTC36, RFOOOO4, RFOOO15, RFOOO45, CADM2) had been validating by qRT-PCR. Pathway and GO terms analysis showed that, positive regulation of biosynthetic process, Wnt signaling pathway, cellular protein modification process, and positive regulation of nitrogen compound were differentially enriched. Our results suggested that, KEGG pathways such as protein digestion and absorption, Arrhythmogenic right ventricular cardiomyopathy (ARVC) to be significantly enriched in both DE lncRNAs as well as DE mRNAs and involved in back fat tissues development. It also suggests that, identified lncRNAs are involved in regulation of important adipose tissues development pathways.

Cellular interactions during implantation in domestic ruminants

Implantation is a critical step in the progress of pregnancy, during which the conceptus acquires a fixed position within the uterine lumen, and leads to the establishment of the placental structures. This process implies some cellular modifications of both the uterine epithelium and the trophoblast to ensure cell adhesion between the two tissues. In ruminants, the implantation process is characterized by three main steps: a long pre-attachment period lasting 2-3 weeks during which the conceptus elongates considerably, an apposition stage when cellular contacts are established between the trophoblast and the uterine epithelium, and an adhesion stage which ends the process and gives rise to the cellular structure of an epithelio-chorial placenta. Trophoblast apposition begins in the vicinity of the embryo by day 15, 18 and 19 in sheep, goats and cows, respectively. The trophoblast cells surrounding the embryo show morphological and functional changes. These modifications are local within the conceptus since non-implanted areas of trophoblast still display the morphological and functional features that characterized this tissue during the pre-attachment period. As the implantation process spreads towards the extremity of the elongated conceptus, these cellular transformations progressively affect the whole trophoblast. Apposition is completed by a close adhesion between the interdigitating uterine microvilli and the trophoblastic plasma membrane. By this stage, trophoblastic binucleate cells migrate through the trophoblast monolayer and fuse with individual uterine cells to form a syncytial tissue. During this process placental lactogen hormones and pregnancy serum proteins (PSP) produced by the binucleate cells are transported to the endometrial tissues and then to the maternal blood circulation.

Cardiomyopathy in obesity, insulin resistance and diabetes.

The prevalence of obesity, insulin resistance and diabetes is increasing rapidly. Most patients with these disorders have hypertriglyceridaemia and increased plasma levels of fatty acids, which are taken up and stored in lipid droplets in the heart. Intramyocardial lipids that exceed the capacity for storage and oxidation can be lipotoxic and induce non-ischaemic and non-hypertensive cardiomyopathy, termed diabetic or lipotoxic cardiomyopathy. The clinical features of diabetic cardiomyopathy are cardiac hypertrophy and diastolic dysfunction, which lead to heart failure, especially heart failure with preserved ejection fraction. Although the pathogenesis of the cardiomyopathy is multifactorial, diabetic dyslipidaemia and intramyocardial lipid accumulation are the key pathological features, triggering cellular signalling and modifications of proteins and lipids via generation of toxic metabolic intermediates. Most clinical studies have shown no beneficial effect of anti-diabetic agents and statins on outcomes in heart failure patients without atherosclerotic diseases, indicating the importance of identifying underlying mechanisms and early interventions for diabetic cardiomyopathy. Here, we summarize the molecular mechanisms of diabetic cardiomyopathy, with a special emphasis on cardiac lipotoxicity, and discuss the role of peroxisome proliferator-activated receptor α and dysregulated fatty acid metabolism as potential therapeutic targets.