scholarly journals The Mediation of miR-34a/miR-449c for Immune Cytokines in Acute Cold/Heat-Stressed Broiler Chicken

Animals ◽  
2020 ◽  
Vol 10 (11) ◽  
pp. 2168
Author(s):  
Tao Li ◽  
Yiping Song ◽  
Xiuyu Bao ◽  
Jianqin Zhang
Keyword(s):  
Broiler Chicken ◽  
Target Genes ◽  
Growth Period ◽  
Study Groups ◽  
Interferon Γ ◽  
Luciferase Reporter ◽  
Receptor Interaction ◽  
Acute Heat Stress ◽  
Transcriptional Regulatory ◽  
Mirna Sequencing

An increasing amount of evidence has revealed that microRNAs (miRNAs) participated in immune regulation and reaction to acute cold and heat stresses. As a new type of post-transcriptional regulatory factor, miRNA has received widespread attention; However, the specific mechanism used for this regulation still needs to be determined. In this study, thirty broilers at the same growth period were divided into three groups and treated with different temperature and humidity of CS (10–15 °C and 90% Relative Humidity (RH)), HS (39 °C and 90% RH), and NS (26 °C and 50–60% RH) respectively. After 6 h, splenic tissues were collected from all study groups. miRNA sequencing was performed to identify the differentially expressed miRNAs (DEMs) between HS, CS, and NS. We found 33, 37, and 7 DEMs in the HS-NS, HS-CS, CS-NS group. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed that DEMs were significantly enriched in cytokine–cytokine receptor interaction and functioned as the cellular responders to stress. We chose two miRNA, miR-34a and miR-449c, from the same family and differential expressed in HS-CS and HS-NS group, as the research objects to predict and verify the target genes. The dual-luciferase reporter assay and quantitative real-time PCR (qRT-PCR) confirmed that two cytokines, IL-2 and IL-12α, were the direct target genes of miR-34a and miR-449c. To further understand the mediation mechanism of miRNAs in acute cold/heat-stressed broiler chicken, a splenic cytokines profile was constructed. The results showed that IL-1β was strongly related to acute heat stress in broiler chicken, and from this we predicted that the increased expression of IL-1β might promote the expression of miR-34a, inducing the upregulation of interferon-γ (INF-γ) and IL-17. Our finds have laid a theoretical foundation for the breeding of poultry resistance and alleviation of the adverse effects of stress.

scholarly journals Circular RNA circSDHC serves as a sponge for miR-127-3p to promote the proliferation and metastasis of renal cell carcinoma via the CDKN3/E2F1 axis

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Junjie Cen ◽  
Yanping Liang ◽  
Yong Huang ◽  
Yihui Pan ◽  
Guannan Shu ◽  
...  
Keyword(s):  
Renal Cell Carcinoma ◽  
Cell Carcinoma ◽  
Renal Cell ◽  
Target Genes ◽  
Expression Patterns ◽  
Luciferase Reporter ◽  
Circular Rnas ◽  
Proliferation And Metastasis

Abstract Background There is increasing evidence that circular RNAs (circRNAs) have significant regulatory roles in cancer development and progression; however, the expression patterns and biological functions of circRNAs in renal cell carcinoma (RCC) remain largely elusive. Method Bioinformatics methods were applied to screen for circRNAs differentially expressed in RCC. Analysis of online circRNAs microarray datasets and our own patient cohort indicated that circSDHC (hsa_circ_0015004) had a potential oncogenic role in RCC. Subsequently, circSDHC expression was measured in RCC tissues and cell lines by qPCR assay, and the prognostic value of circSDHC evaluated. Further, a series of functional in vitro and in vivo experiments were conducted to assess the effects of circSDHC on RCC proliferation and metastasis. RNA pull-down assay, luciferase reporter and fluorescent in situ hybridization assays were used to confirm the interactions between circSDHC, miR-127-3p and its target genes. Results Clinically, high circSDHC expression was correlated with advanced TNM stage and poor survival in patients with RCC. Further, circSDHC promoted tumor cell proliferation and invasion, both in vivo and in vitro. Analysis of the mechanism underlying the effects of circSDHC in RCC demonstrated that it binds competitively to miR-127-3p and prevents its suppression of a downstream gene, CDKN3, and the E2F1 pathway, thereby leading to RCC malignant progression. Furthermore, knockdown of circSDHC caused decreased CDKN3 expression and E2F1 pathway inhibition, which could be rescued by treatment with an miR-127-3p inhibitor. Conclusion Our data indicates, for the first time, an essential role for the circSDHC/miR-127-3p/CDKN3/E2F1 axis in RCC progression. Thus, circSDHC has potential to be a new therapeutic target in patients with RCC.

scholarly journals Screening Driving Transcription Factors in the Processing of Gastric Cancer

2016 ◽  
Vol 2016 ◽  
pp. 1-9 ◽  
Author(s):  
Guangzhong Xu ◽  
Kai Li ◽  
Nengwei Zhang ◽  
Bin Zhu ◽  
Guosheng Feng
Keyword(s):  
Gastric Cancer ◽  
Transcription Factors ◽  
Regulatory Network ◽  
Target Genes ◽  
Transcriptional Regulatory Network ◽  
Expression Profiles ◽  
Functional Enrichment ◽  
Regulatory Mechanisms ◽  
Integrated Analysis ◽  
Transcriptional Regulatory

Background. Construction of the transcriptional regulatory network can provide additional clues on the regulatory mechanisms and therapeutic applications in gastric cancer.Methods. Gene expression profiles of gastric cancer were downloaded from GEO database for integrated analysis. All of DEGs were analyzed by GO enrichment and KEGG pathway enrichment. Transcription factors were further identified and then a global transcriptional regulatory network was constructed.Results. By integrated analysis of the six eligible datasets (340 cases and 43 controls), a bunch of 2327 DEGs were identified, including 2100 upregulated and 227 downregulated DEGs. Functional enrichment analysis of DEGs showed that digestion was a significantly enriched GO term for biological process. Moreover, there were two important enriched KEGG pathways: cell cycle and homologous recombination. Furthermore, a total of 70 differentially expressed TFs were identified and the transcriptional regulatory network was constructed, which consisted of 566 TF-target interactions. The top ten TFs regulating most downstream target genes were BRCA1, ARID3A, EHF, SOX10, ZNF263, FOXL1, FEV, GATA3, FOXC1, and FOXD1. Most of them were involved in the carcinogenesis of gastric cancer.Conclusion. The transcriptional regulatory network can help researchers to further clarify the underlying regulatory mechanisms of gastric cancer tumorigenesis.

scholarly journals Rs56288038 (C/G) in 3'UTR of IRF-1 Regulated by MiR-502-5p Promotes Gastric Cancer Development

2016 ◽  
Vol 40 (1-2) ◽  
pp. 391-399 ◽  
Author(s):  
Bo Wang ◽  
Huan Yang ◽  
Liqin Shen ◽  
Ji Wang ◽  
Wangyang Pu ◽  
...  
Keyword(s):  
Gastric Cancer ◽  
Survival Rate ◽  
Target Genes ◽  
Diagnostic Value ◽  
Tumor Promoter ◽  
Luciferase Reporter ◽  
Nucleotide Polymorphisms ◽  
Susceptible Population ◽  
Poor Differentiation ◽  
Regulatory Capacity

Background/Aims: Interferon regulatory factor 1 (IRF-1) has been shown to function as a transcriptional activator or repressor of a variety of target genes. However, its upstream, non-coding RNA-related regulatory capacity remains unknown. In this study, we focus on the miRNA-associated single nucleotide polymorphisms (SNPs) in the 3′untranslated region (UTR) of IRF-1 to further investigate the functional relationship and potential diagnostic value of the SNPs and miRNAs among Chinese gastric cancer (GC) patients. Methods: We performed a case-control study with 819 GC patients and 756 cancer-free controls. Genotyping by realtime PCR assay, cell transfection, and the dual luciferase reporter assay were used in our study, and the 5-year overall survival rate and relapse-free survival rate in different groups were investigated. Results: We found that patients suffering from Helicobacter pylori (Hp) infection were the susceptible population compared to controls. SNP rs56288038 (C/G) in IRF-1 3′UTR was involved in the occurrence of GC by acting as a tumor promoter factor. SNP rs56288038 (C/G) could be up-regulated by miR-502-5p, which caused a down-regulation of IRF-1 in cell lines and decreased apoptosis induced by IFN-γ. Carrying the G genotype was related to significantly low expression of IRF-1 and Hp infection, poor differentiation, big tumor size, invasion depth, as well as the high probability of metastasis, and moreover, the C/G SNP was associated with shorter survival of GC patients with five years of follow-up study. Conclusions: our findings have shown that the SNP rs56288038 (C/G) in IRF-1 3′UTR acted as a promotion factor in GC development through enhancing the regulatory role of miR-502-5p in IRF-1 expression.

RPD3 encodes a second factor required to achieve maximum positive and negative transcriptional states in Saccharomyces cerevisiae

1991 ◽  
Vol 11 (12) ◽  
pp. 6317-6327 ◽  
Author(s):  
M Vidal ◽  
R F Gaber
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Target Genes ◽  
Current Data ◽  
Regulatory Function ◽  
Fourfold Increase ◽  
Recessive Mutations ◽  
Activation Of Transcription ◽  
Transcriptional Regulatory ◽  
Low Levels ◽  
Full Activation

In Saccharomyces cerevisiae, TRK1 and TRK2 encode the high- and low-affinity K+ transporters, respectively. In cells containing a deletion of TRK1, transcription levels of TRK2 are extremely low and are limiting for growth in media containing low levels of K+ (Trk- phenotype). Recessive mutations in RPD1 and RPD3 suppress the TRK2, conferring an approximately fourfold increase in transcription. rpd3 mutations confer pleiotropic phenotypes, including (i) mating defects, (ii) hypersensitivity to cycloheximide, (iii) inability to sporulate as homozygous diploids, and (iv) constitutive derepression of acid phosphatase. RPD3 was cloned and is predicted to encode a 48-kDa protein with no extensive similarity to proteins contained in current data bases. Deletion of RPD3 is not lethal but confers phenotypes identical to those caused by spontaneous mutations. RPD3 is required for both full repression and full activation of transcription of target genes including PHO5, STE6, and TY2. RPD3 is the second gene required for this function, since RPD1 is also required. The effects of mutations in RPD1 and RPD3 are not additive, suggesting that these genes are involved in the same transcriptional regulatory function or pathway.

Screening candidate microR-15a- IRAK2 regulatory pairs for predicting the response to Staphylococcus aureus-induced mastitis in dairy cows

2019 ◽  
Vol 86 (4) ◽  
pp. 425-431 ◽  
Author(s):  
Zhi Chen ◽  
Jingpeng Zhou ◽  
Xiaolong Wang ◽  
Yang Zhang ◽  
Xubin Lu ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Differential Expression ◽  
High Throughput Sequencing ◽  
Target Genes ◽  
Interleukin 1 ◽  
Differentially Expressed ◽  
Luciferase Reporter ◽  
Expression Of Genes ◽  
Chinese Holstein Cows ◽  
Reporter Assays

AbstractWe established a mastitis model using exogenous infection of the mammary gland of Chinese Holstein cows with Staphylococcus aureus and extracted total RNA from S. aureus-infected and healthy mammary quarters. Differential expression of genes due to mastitis was evaluated using Affymetrix technology and results revealed a total of 1230 differentially expressed mRNAs. A subset of affected genes was verified via Q-PCR and pathway analysis. In addition, Solexa high-throughput sequencing technology was used to analyze profiles of miRNA in infected and healthy quarters. These analyses revealed a total of 52 differentially expressed miRNAs. A subset of those results was verified via Q-PCR. Bioinformatics techniques were used to predict and analyze the correlations among differentially expressed miRNA and mRNA. Results revealed a total of 329 pairs of negatively associated miRNA/mRNA, with 31 upregulated pairs of mRNA and 298 downregulated pairs of mRNA. Differential expression of miR-15a and interleukin-1 receptor-associated kinase-like 2 (IRAK2), were evaluated by western blot and luciferase reporter assays. We conclude that miR-15a and miR-15a target genes (IRAK2) constitute potential miRNA–mRNA regulatory pairs for use as biomarkers to predict a mastitis response.

scholarly journals Differential Expression Pattern of Goat Uterine Fluids Extracellular Vesicles miRNAs during Peri-Implantation

Cells ◽  
2021 ◽  
Vol 10 (9) ◽  
pp. 2308
Author(s):  
Yanshe Xie ◽  
Guangbin Liu ◽  
Xupeng Zang ◽  
Qun Hu ◽  
Chen Zhou ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Small Rnas ◽  
Target Genes ◽  
Embryo Implantation ◽  
Mature Embryo ◽  
Luciferase Reporter ◽  
Small Rna Sequencing ◽  
Transmission Electron ◽  
Differential Expression Pattern ◽  
Endometrial Epithelium

Early pregnancy failure occurs when a mature embryo attaches to an unreceptive endometrium. During the formation of a receptive endometrium, extracellular vesicles (EVs) of the uterine fluids (UFs) deliver regulatory molecules such as small RNAs to mediate intrauterine communication between the embryo and the endometrium. However, profiling of small RNAs in goat UFs’ EVs during pregnancy recognition (day 16) has not been carried out. In this study, EVs were isolated from UFs on day 16 of the estrous cycle or gestation. They were isolated by Optiprep™ Density G radient (ODG) and verified by transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), and Western blotting. Immunostaining demonstrated that CD63 was present both in the endometrial epithelium and glandular epithelium, and stain intensity was greater in the pregnant endometrium compared to the non-pregnant endometrium. Small RNA sequencing revealed that UFs’ EVs contained numerous sRNA families and a total of 106 differentially expressed miRNAs (DEMs). Additionally, 1867 target genes of the DEMs were obtained, and miRNA–mRNA interaction networks were constructed. GO and KEGG analysis showed that miRNAs were significantly associated with the formation of a receptive endometrium and embryo implantation. In addition, the fluorescence in situ hybridization assay (FISH) showed that chi-miR-451-5p was mainly expressed in stromal cells of the endometrium and a higher level was detected in the endometrial luminal epithelium in pregnant states. Moreover, the dual-luciferase reporter assay showed that chi-miR-451-5p directly binds to PSMB8 and may play an important role in the formation of a receptive endometrium and embryo implantation. In conclusion, these results reveal that UFs’ EVs contain various small RNAs that may be vital in the formation of a receptive endometrium and embryo implantation.

scholarly journals The Role of BCL6 in Glioblastoma

2021 ◽  
Author(s):  
◽  
Nicole Jones
Keyword(s):  
Dna Damage ◽  
Cell Death ◽  
Target Genes ◽  
Therapy Resistance ◽  
Response To Therapy ◽  
Transcriptional Analysis ◽  
Luciferase Reporter ◽  
Mobility Shift ◽  
Response To Chemotherapy ◽  
Targeted Inhibition

<p>Glioblastoma (GBM) is the most common and most deadly brain tumour to occur in adults. Initially patients respond to radiation and chemotherapy, which primarily work by causing large amounts of DNA damage, leading to cell death. However, this process does not happen effectively in GBM and understanding how these cells resist cell death in response to therapy is key to improving the efficacy of treatment. BCL6 is a transcription factor that stops cell death in response to DNA damage, primarily through repressing transcription of DNA damage response genes. Recent work in our lab has shown BCL6 to be present in untreated GBM tumours and up-regulated in GBM cells treated with chemotherapy or radiation, and inhibition of BCL6 leads to a profound loss in proliferative activity. These results indicate that BCL6 may be used as a mechanism of therapy resistance in GBM cells. The objective of this study was to establish a role for BCL6 in GBM cells using luciferase reporter assays, electrophoretic mobility shift assays (EMSA), quantitative chromatin immunoprecipitation (qChIP) and targeted inhibition of BCL6 with subsequent transcriptional analysis by RNA sequencing. We observed that BCL6 appeared to be a transcriptional activator in GBM, as shown by increased luciferase activity in GBM cells treated with radiation. EMSA experiments revealed that overexpressed BCL6 formed complexes with co-repressors, but endogenous BCL6 did not. qChIP experiments revealed that BCL6 was not bound to tradtional BCL6 target genes. Analysis of transcriptional profiles has identified a unique subset of genes which are downregulated when BCL6 is inhibited and upregulated in response to chemotherapy, and these genes were related to cell survival. These changes indicate that these genes may be regulated by BCL6 in chemotherapy treated cells. Together, these results illustrate that BCL6 appears to have an active and unique function in GBM cells, and reinforces this transcription factors position as an attractive therapeutic target in GBM.</p>

scholarly journals Study on Genetic Variance of miR-541 in Type 1 Diabetes

2012 ◽  
Vol 2012 ◽  
pp. 1-6 ◽  
Author(s):  
Bei Han ◽  
Xing Shi ◽  
Quan Peng ◽  
Wentao Gao
Keyword(s):  
Type 1 Diabetes ◽  
Healthy Volunteers ◽  
Target Genes ◽  
Direct Sequencing ◽  
Clinical Analysis ◽  
Genetic Variations ◽  
Luciferase Reporter ◽  
Putative Promoter ◽  
Putative Promoter Region

Genetic susceptibility plays a key role in type 1 diabetes development. Because miR-541 gene was located within the associated chromosome loci and its target genes include the diabetes-associated gene neurogenin3, this study aimed to investigate whether miR-541 had type 1 diabetes-associated genetic variations. Type 1 diabetes children and healthy volunteers were recruited; direct sequencing was performed in initial 69 patients and 46 volunteers. We identified 1 reported SNP (rs12893725) and 3 novel genetic variations, for the candidate -404 G→T variation, restriction fragment length polymorphism (RFLP) was performed in total 247 diabetes children and 212 healthy volunteers, a different distribution trait of allele frequencies was found between the two groups, and further clinical analysis found no significant correlation between clinical parameter and genotypes among patients. In addition, by luciferase reporter assay, -404 was found to be within putative promoter region of pre-miR-541; although mutation of G→T has no effect on promoter activity, a significant secondary structure alteration may possibly influence its processing and transcription. In conclusion, we identified 3 novel genetic variations in putative promoter of miR-541 in type 1 diabetes patients; -404 G→T of miR-541 is a potential T1D-associated genetic variation.

scholarly journals Broiler surface temperature distribution of 42 day old chickens

2010 ◽  
Vol 67 (5) ◽  
pp. 497-502 ◽  
Author(s):  
Irenilza de Alencar Nääs ◽  
Carlos Eduardo Bites Romanini ◽  
Diego Pereira Neves ◽  
Guilherme Rodrigues do Nascimento ◽  
Rimena do Amaral Vercellino
Keyword(s):  
Temperature Distribution ◽  
Surface Temperature ◽  
Air Temperature ◽  
Broiler Chickens ◽  
Broiler Chicken ◽  
Growth Period ◽  
The Body ◽  
Body Parts ◽  
Surface Temperature Distribution ◽  
Thermographic Image

Broiler chickens in Brazil are generally reared from 1 to 42 days when they are exposed to procedures such as fasting, harvesting, crating and transport to slaughter. Maintaining homeostasis is of great importance for broiler survival under harsh environment especially prior to slaughter. Heat loss varies in the distinct parts of the body during the growth period, and it is related to the air temperature of the environment and to the amount of feather covering. This research aimed to study the surface temperature distribution using infrared thermographic image processing to characterize 42 day old broiler chicken surface temperature prior to slaughter. Broilers were reared for 42 days and prior to harvest and transport to slaughter the infrared surface temperature was recorded along the day. Data from the thermograms taken in feather and featherless regions were compared during the 42nd day of growth. High correlation between featherless regions and air temperature was found showing that these areas respond fast to changes in the rearing environment. Two functions were developed for predicting both surface temperature for featherless and feather covered areas of the broiler body parts.

PHYSIOLOGICAL AND BIOCHEMICAL CHANGES IN ADOLESCENT JUDO ATHLETES CAUSED BY TRAINING DURING START PERIOD

2017 ◽  
Vol 27 (78) ◽  
pp. 57-64
Author(s):  
Jan Jaszczanin ◽  
Wojciech Przybylski ◽  
Waldemar Moska ◽  
Egle Kemeryte-Riaubiene ◽  
Grzegorz Chruściński
Keyword(s):  
Age Groups ◽  
Anaerobic Capacity ◽  
Growth Period ◽  
Study Groups ◽  
Functional Adaptation ◽  
Anaerobic Performance ◽  
Oxygen Intake ◽  
Training Experience ◽  
Maximum Oxygen Intake ◽  
Judo Athletes

Study aim: The goal of the present study was to estimate and compare dynamics of physical fitness indices of judo athletes and non-sporting persons aged 11-17 years during this stage ontogeny and their importance of the body’s functional adaptation. Study material: The studies involved 47 judo athletes, 11–17 years old, who were divided into three age groups GP 11-13 years n=16; GP 14-15 years n=16; GP 16-17 years n=15, and 48 schoolchildren not involved in sports GK 11-13 years n=16; GK 14-15 years n=15; GK 16-17 years n=17. Aerobic and anaerobic capacity was studied in all groups. The initial studies were carried out in January and follow-up studies were conducted six months later. Results: Power indicators increased in all groups, but judo athletes’ anaerobic capacity was significantly higher comparing to other groups. Judo athletes’ simulation fights resulted in increased La concentrations, pH changes, and heart rate alterations, whereas the level of changes depended on athletes’ age, training, and training experience. Comparison of maximum oxygen intake parameters of judo players and untrained children of the same age did not reveal significant differences between these groups. Wrestlers aged 12 and 16 years presented significantly higher anaerobic prevalence in comparison with untrained children. The differences indicate that anaerobic performance potential in older judo athletes (16 years old and above) is increased, which is reflected by a higher intensity and elevated exercise loads as well as training experience. Conclusions: No significant differences were reported in terms of VO2 max between the study groups. The indicators of anaerobic performance of children training in judo (W/kg, W average /kg, Time achieved max power, Time to maintain max power) were significantly better in comparison with untrained peers. The maximum loads (Wingate test, especially in simulative judo fights) caused a significant increase in La levels accompanied by a decrease in pH in the chosen period of growth period.

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