Long noncoding RNAs and mRNAs profiling in ovary during laying and broodiness in Taihe Black-Bone Silky Fowls (Gallus gallus Domesticus Brisson).

BACKGROUND: Broodiness significantly impacts poultry egg production, particularly notable in specific breeds such as the black-boned Silky, characterized by pronounced broodiness. An understanding of the alterations in ovarian signaling is essential for elucidating the mechanisms that influence broodiness. However, comparative research on the characteristics of long non-coding RNAs (lncRNAs) in the ovaries of broody chickens (BC) and high egg-laying chickens (GC) remains scant. In this investigation, we employed RNA sequencing to assess the ovarian transcriptomes, which include both lncRNAs and mRNAs, in eight Taihe Black-Bone Silky Fowls (TBsf), categorized into broody and high egg-laying groups. This study aims to provide a clearer understanding of the genetic underpinnings associated with broodiness and egg production. RESULTS: We have identified a total of 16,444 mRNAs and 18,756 lncRNAs, of which 349 mRNAs and 651 lncRNAs exhibited significantly different expression (DE) between the BC and GC groups. Furthermore, we have identified the cis-regulated and trans-regulated target genes of differentially abundant lncRNA transcripts and have constructed an lncRNA-mRNA trans-regulated interaction network linked to ovarian follicle development. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) annotation analyses have revealed that DE mRNAs and the target genes of DE lncRNAs are associated with pathways including neuroactive ligand-receptor interaction, CCR6 chemokine receptor binding, G-protein coupled receptor binding, cytokine-cytokine receptor interaction, and ECM-receptor interaction. CONCLUSION: Our research presents a comprehensive compilation of lncRNAs and mRNAs linked to ovarian development. Additionally, it establishes a predictive interaction network involving differentially abundant lncRNAs and differentially expressed genes (DEGs) within TBsf. This significantly contributes to our understanding of the intricate interactions between lncRNAs and genes governing brooding behavior.

Identification and functional analysis of ovarian lncRNAs during different egg laying periods in Taihe Black-Bone Chickens.

Introduction: Long non-coding RNA (lncRNA) refers to a category of non-coding RNA molecules exceeding 200 nucleotides in length, which exerts a regulatory role in the context of ovarian development. There is a paucity of research examining the involvement of lncRNA in the regulation of ovary development in Taihe Black-Bone Chickens. In order to further investigate the egg laying regulation mechanisms of Taihe Black-Bone Chickens at different periods, transcriptome analysis was conducted on the ovarian tissues at different laying periods. Methods: This study randomly selected ovarian tissues from 12 chickens for RNA-seq. Four chickens were selected for each period, including the early laying period (102 days, Pre), the peak laying period (203 days, Peak), and the late laying period (394 days, Late). Based on our previous study of mRNA expression profiles in the same ovarian tissue, we identified three differentially expressed lncRNAs (DE lncRNAs) at different periods and searched for their cis- and trans-target genes to draw an lncRNA-mRNA network. Results and discussion: In three groups of ovarian tissues, we identified 136 DE lncRNAs, with 8 showing specific expression during the early laying period, 10 showing specific expression during the peak laying period, and 4 showing specific expression during the late laying period. The lncRNA-mRNA network revealed 16 pairs of lncRNA-target genes associated with 7 DE lncRNAs, and these 14 target genes were involved in the regulation of reproductive traits. Furthermore, these reproductive-related target genes were primarily associated with signaling pathways related to follicle and ovary development in Taihe Black-Bone Chickens, including cytokine-cytokine receptor interaction, TGF-beta signaling pathway, tyrosine metabolism, ECM-receptor interaction, focal adhesion, neuroactive ligand-receptor interaction, and cell adhesion molecules (CAMs). This study offers valuable insights for a comprehensive understanding of the influence of lncRNAs on poultry reproductive traits.

Transcriptomic and metabolomic analyses of the ovaries of Taihe black-bone silky fowls at the peak egg-laying and nesting period.

The poor reproductive performance of most local Chinese chickens limits the economic benefits and output of related enterprises. As an excellent local breed in China, Taihe black-bone silky fowl is in urgent need of our development and utilization. In this study, we performed transcriptomic and metabolomic analyses of the ovaries of Taihe black-bone silky fowls at the peak egg-laying period (PP) and nesting period (NP) to reveal the molecular mechanisms affecting reproductive performance. In the transcriptome, we identified five key differentially expressed genes (DEGs) that may affect the reproductive performance of Taihe black-bone silky fowl: BCHE, CCL5, SMOC1, CYTL1, and SCIN, as well as three important pathways: the extracellular region, Neuroactive ligand-receptor interaction and Cytokine-cytokine receptor interaction. In the metabolome, we predicted three important ovarian significantly differential metabolites (SDMs): LPC 20:4, Bisphenol A, and Cortisol. By integration analysis of transcriptome and metabolome, we identified three important metabolite-gene pairs: "LPC 20:4-BCHE", "Bisphenol A-SMOC1", and "Cortisol- SCIN". In summary, this study contributes to a deeper understanding of the regulatory mechanism of egg production in Taihe black-bone silky fowl and provides a scientific basis for improving the reproductive performance of Chinese local chickens.

Transcriptome Analysis of the Ovaries of Taihe Black-Bone Silky Fowls at Different Egg-Laying Stages.

The poor egg-laying performance and short peak egg-laying period restrict the economic benefits of enterprises relating to the Taihe black-bone silky fowl. Ovaries are the main organ for egg production in poultry. Unlike that of mammals, the spawning mechanism of poultry has rarely been reported. As a prominent local breed in China, the reproductive performance of Taihe black-bone silky fowls is in urgent need of development and exploitation. To further explore the egg-laying regulation mechanism in the different periods of Taihe black-bone silky fowls, the ovarian tissues from 12 chickens were randomly selected for transcriptome analysis, and 4 chickens in each of the three periods (i.e., the pre-laying period (102 days old, Pre), peak laying period (203 days old, Peak), and late laying period (394 days old, Late)). A total of 12 gene libraries were constructed, and a total of 9897 differential expression genes (DEGs) were identified from three comparisons; the late vs. peak stage had 509 DEGs, the pre vs. late stage had 5467 DEGs, and the pre vs. peak stage had 3921 DEGs (pre-stage: pre-egg-laying period (102 days old), peak-stage: peak egg-laying period (203 days old), and late-stage: late egg-laying period (394 days old)). In each of the two comparisons, 174, 84, and 2752 differentially co-expressed genes were obtained, respectively, and 43 differentially co-expressed genes were obtained in the three comparisons. Through the analysis of the differential genes, we identified some important genes and pathways that would affect reproductive performance and ovarian development. The differential genes were LPAR3, AvBD1, SMOC1, IGFBP1, ADCY8, GDF9, PTK2B, PGR, and CD44, and the important signaling pathways included proteolysis, extracellular matrices, vascular smooth muscle contraction, the NOD-like receptor signaling pathway and the phagosome. Through the analysis of the FPKM (Fragments Per Kilobase of exon model per Million mapped fragments) values of the genes, we screened three peak egg-laying period-specific expressed genes: IHH, INHA, and CYP19A1. The twelve genes and five signaling pathways mentioned above have rarely been reported in poultry ovary studies, and our study provides a scientific basis for the improvement of the reproductive performance in Taihe black-bone silky fowls.

Electrochemical removal of nitrate from a Donnan dialysis waste stream.

The objective of this work was to investigate electrochemical removal of nitrate from a high salinity waste stream generated by Donnan dialysis. Donnan dialysis for nitrate removal is a promising technique. It produces a distinctive composition of a high salinity waste stream of NaCl or Na2SO4 that requires a viable disposal method. The waste stream has the full anionic composition of contaminated groundwater, but the only cation is sodium. Experiments were conducted in a batch system setup. A copper cathode was chosen over brass, aluminum and graphite cathodes. A dimensionally stable anode (DSA), Ti/PbO2, was selected over a Ti/Pt anode. Electrochemical denitrification of high salinity Donnan dialysis nitrate wastes was successfully achieved, with different behavior exhibited in high salinity NaCl solution than in high salinity Na2SO4 solution. NaCl inhibited nitrate removal at high salinities while Na2SO4 did not. The maximum removals after 4 h operation in the high salinity wastes were 69 and 87% for the NaCl and Na2SO4 solutions respectively.

A fungal ABC transporter FgAtm1 regulates iron homeostasis via the transcription factor cascade FgAreA-HapX.

Iron homeostasis is important for growth, reproduction and other metabolic processes in all eukaryotes. However, the functions of ATP-binding cassette (ABC) transporters in iron homeostasis are largely unknown. Here, we found that one ABC transporter (named FgAtm1) is involved in regulating iron homeostasis, by screening sensitivity to iron stress for 60 ABC transporter mutants of Fusarium graminearum, a devastating fungal pathogen of small grain cereal crops worldwide. The lack of FgAtm1 reduces the activity of cytosolic Fe-S proteins nitrite reductase and xanthine dehydrogenase, which causes high expression of FgHapX via activating transcription factor FgAreA. FgHapX represses transcription of genes for iron-consuming proteins directly but activates genes for iron acquisition proteins by suppressing another iron regulator FgSreA. In addition, the transcriptional activity of FgHapX is regulated by the monothiol glutaredoxin FgGrx4. Furthermore, the phosphorylation of FgHapX, mediated by the Ser/Thr kinase FgYak1, is required for its functions in iron homeostasis. Taken together, this study uncovers a novel regulatory mechanism of iron homeostasis mediated by an ABC transporter in an important pathogenic fungus.

Improving the secretion of cadaverine in Corynebacterium glutamicum by cadaverine-lysine antiporter.

Cadaverine (1,5-pentanediamine, diaminopentane), the desired raw material of bio-polyamides, is an important industrial chemical with a wide range of applications. Biosynthesis of cadaverine in Corynebacterium glutamicum has been a competitive way in place of petroleum-based chemical synthesis method. To date, the cadaverine exporter has not been found in C. glutamicum. In order to improve cadaverine secretion, the cadaverine-lysine antiporter CadB from Escherichia coli was studied in C. glutamicum. Fusion expression of cadB and green fluorescent protein (GFP) gene confirmed that CadB could express in the cell membrane of C. glutamicum. Co-expression of cadB and ldc from Hafnia alvei in C. glutamicum showed that the cadaverine secretion rate increased by 22 % and the yield of total cadaverine and extracellular cadaverine increased by 30 and 73 %, respectively. Moreover, the recombinant strain cultured at acid and neutral pH separately hardly had any difference in cadaverine concentrations. These results suggested that CadB could be expressed in the cell membrane of C. glutamicum and that recombinant CadB could improve cadaverine secretion and the yield of cadaverine. Moreover, the pH value did not affect the function of recombinant CadB. These results may be a promising metabolic engineering strategy for improving the yield of the desired product by enhancing its export out of the cell.

Construction of an engineering strain producing high yields of α-transglucosidase via Agrobacterium tumefaciens-mediated transformation of Asperillus niger.

In this study, Agrobacterium tumefaciens-mediated transformation (ATMT) was used in breeding industrial strains for the purpose of improving α-transglucosidase production. Firstly, an efficient ATMT system for Asperillus niger was established by optimization of several influencing factors, in which transformation efficiency was improved up to 14-fold compared with the initial conditions. Furthermore, binary vector pBI-Glu containing an α-transglucosidase expression cassette was constructed and transferred into Agrobacterium tumefaciens LBA4404 in order to infect A. niger. By the efficient ATMT method, the gene for α-transglucosidase, driven by strong promoter PglaA (the glucoamylase gene promoter), had a high expression level in A. niger A-8 (25.02 U/mL). The optimized ATMT system was found to be effective and suitable for A. niger, and should be a useful tool for studying the function of A. niger genes and for industrial breeding of this strain.

Highly sensitive protein detection using enzyme-labeled gold nanoparticle probes.

A highly sensitive protein detection method based on a novel enzyme-labeled gold nanoparticle (AuNP) probe has been developed. In this method, we firstly prepared the enzyme-labeled AuNP probe by coating AuNP with antibody, single-stranded DNA (ssDNA), and horseradish peroxidase (HRP). Magnetic microparticle (MMP) functionalized with another antibody was used as capture probe. Then, target protein was sandwiched by the enzyme-labeled AuNP probe and the capture probe through immunoreaction. The target immunoreaction event could be sensitively transduced via the enzymatically amplified optical signal. By using this strategy, carcinoembryonic antigen (CEA), as a model protein, was detected with high sensitivity and good specificity. The detection limit for this approach was 12 ng L(-1), which was approximately 130-fold more sensitive than the conventional enzyme-linked immunosorbent assay (ELISA). The practical application of the proposed immunoassay was carried out for determination of CEA in serum samples. The demonstrated capability of the proposed method shows potentially applications for early diagnoses of diseases.

[Effect of (-)-epigallocatechin-3-gallate and 5-AZA-CdR on expression of X-linked inhibitor of apoptosis protein-associated factor 1 in human leukemia HL-60 and K562 cells].

OBJECTIVE: To study the expression of X-linked inhibitor of apoptosis protein (XIAP)-associated factor 1 (Xaf1) in human leukemia HL-60 and K562 cells treated with interferon alpha (INFalpha) and the demethylating agent 5-AZA-CdR, and observe the synergetic antitumor effect of Xaf1 inducer and (-)-epigallocatechin-3-gallate (EGCG). METHODS: Human leukemia HL-60 and K562 cells were treated for 48 h with 1000 U/ml INFalpha and different doses of 5-AZA-CdR, and the mRNA expressions of both Xaf1 and XIAP were measured by RT-PCR. The leukemia cells were also treated with the optimal Xaf1 inducer in combination with EGCG, after which flow cytometry was employed to examine the changes in the members of the Bcl-2 family, mitochondrial transmembrane potential and apoptosis. RESULTS: As the dose increased, 5-AZA-CdR dose-dependently up-regulated the mRNA expression of Xaf1 in HL-60 and K562 cells; INFalpha treatment also resulted in increased Xaf1 expression, but 5 micromol/L 5-AZA-CdR showed the most potent effect. Neither INFalpha nor 5-AZA-CdR caused significant changes in XIAP expression. Combined treatment with 5-AZA-CdR and EGCG altered the expressions of Bcl-2 (Bcl-xl) and Bax in HL-60 and K562 cells, decreased the mitochondrial transmembrane potential and induced cell apoposis, and the two agents exhibited obvious synergistic effect. CONCLUSION: INFalpha and 5-AZA-CdR can induce Xaf1 mRNA expressions in HL-60 and K562 cells, and the effect of 5-AZA-CdR was dose-dependent. 5-AZA-CdR and EGCG induces apoptosis of leukemia cells in vitro, and they exhibits obvious synergetic effects.