CX3CL1/CX3CR1 Signaling Mediated Neuroglia Activation Is Implicated in the Retinal Degeneration: A Potential Therapeutic Target to Prevent Photoreceptor Death.

Purpose: Retinal degeneration (RD) is a large cluster of retinopathies that is characterized by the progressive photoreceptor death and visual impairments. CX3CL1/CX3CR1 signaling has been documented to mediate the microglia activation and gliosis reaction during neurodegeneration. We intend to verify whether the CX3CL1/CX3CR1 signaling is involved in the RD pathology. Methods: A pharmacologically induced RD mice model was established. AZD8797, a CX3CR1 antagonist, was injected into the vitreous cavity of an RD model to modulate the neuroglia activation. Then, the experimental animals were subjected to functional, morphological, and behavioral analysis. Results: The CX3CL1/CX3CR1 signaling mediated neuroglia activation was implicated in the photoreceptor demise of an RD model. Intravitreal injection of AZD8797 preserved the retinal structure and enhanced the photoreceptor survival through inhibiting the CX3CL1/CX3CR1 expressions. Fundus photography showed that the distribution of retinal vessel was clear, and the severity of lesions was alleviated by AZD8797. In particular, these morphological benefits could be translated into remarkable functional improvements, as evidenced by the behavioral test and electroretinogram (mf-ERG) examination. A mechanism study showed that AZD8797 mitigated the microglia activation and migration in the degenerative retinas. The Müller cell hyper-reaction and secondary gliosis response were also suppressed by AZD8797. Conclusions: The neuroinflammation is implicated in the photoreceptor loss of RD pathology. Targeting the CX3CL1/CX3CR1 signaling may serve as an effective therapeutic strategy. Future refinements of these findings may cast light into the discovery of new medications for RD.

The network regulation mechanism of the effects of heat stress on the production performance and egg quality of Jinding duck was analyzed by miRNA‒mRNA.

To explore the differential regulation mechanism of heat stress on the egg production performance and egg quality of Jinding ducks, 200 Jinding ducks (360-day-old) in good health and with similar body weights and a normal appetite were selected and randomly divided into a control (normal temperature [NT]) group (20°C-25°C) and a heat stress (HS) group (32°C-36°C), with 4 replicates in each group and 25 ducks in each replicate. The pretrial period was 1 wk, and the formal trial period was 4 wk. At the end of the 4th wk, 12 duck eggs were collected from each replicate to determine egg quality. Pituitary and ovarian tissues of Jinding ducks were collected, transcriptome sequencing was performed to screen differentially expressed miRNAs and mRNAs related to high temperature and heat stress, and a competitive endogenous RNA regulatory network was constructed. The sequencing data were verified by qRT‒PCR method. The following results were obtained: (1) Compared with the NT group, the HS group had a significantly lower laying rate, total egg weight, average egg weight, total feed intake, and feed intake per duck (P < 0.01), an extremely significantly higher feed-to-egg ratio (P < 0.01), and a higher mortality rate. (2) Compared with the NT group, the HS group had an extremely significantly lower egg weight, egg yolk weight, eggshell weight, and eggshell strength (P < 0.01) and an extremely significantly lower yolk ratio and eggshell thickness (P < 0.01, P < 0.05); however, there was no significant difference in the egg shape index, Haugh unit or protein height (P > 0.05). (3) A total of 1,974 and 1,202 genes were identified in the pituitary and ovary, respectively, and there were 5 significantly differentially expressed miRNAs. The differentially expressed genes were involved in the arginine and proline metabolism pathways, ether lipid metabolism pathway, and drug metabolism-cytochrome P450 pathway, which are speculated to be related to the egg production performance of Jingding ducks under high-temperature heat stress. (4) Novel_221 may target the PRPS1 gene to participate in egg production performance; novel_168 and novel_289 may target PIGW; novel_289 may target Q3MUY2; and novel_289 and novel_208 may target PIGN or genes that may be related to high-temperature heat stress. (5) In pituitary tissue, upregulated novel_141 (center of the network) formed a regulatory network with HSPB1 and HSP30A, and downregulated novel_366 (center of the network) formed a regulatory network with the JIP1 gene. In ovarian tissue, downregulated novel_289 (center of the network) formed a regulatory network with the ZSWM7, ABI3, and K1C23 genes, novel_221 formed a regulatory network with the IGF1, BCL7B, SMC6, APOA4, and FARP2 genes, and upregulated novel_40 formed a regulatory network with the HA1FF10 gene. In summary, heat stress affects the production performance and egg quality of Jinding ducks by regulating the secretion of endocrine-related hormones and the release of neurotransmitters as well as the expression of miRNAs and mRNAs in pituitary and ovarian tissues. The miRNA‒mRNA regulatory network provides a theoretical basis for the molecular mechanism that regulates the stress response in pituitary and ovarian tissues, egg quality, and production performance under heat stress.

Ferroptosis in the ageing retina: A malevolent fire of diabetic retinopathy.

Ageing retina is prone to ferroptosis due to the iron accumulation and impaired efficiency of intracellular antioxidant defense system. Ferroptosis acts as a cell death modality that is characterized by the iron-dependent accumulation of lipid peroxidation. Ferroptosis is distinctively different from other types of regulated cell death (RCD) at the morphological, biochemical, and genetic levels. Diabetic retinopathy (DR) is a common microvascular complication of diabetes. Its prevalence and severity increase progressively with age. Recent reports have shown that ferroptosis is implicated in the pathophysiology of DR. Under hyperglycemia condition, the endothelial cell and retinal pigment epithelium (RPE) cell will undergo ferroptosis, which contributes to the increased vascular permeability and the disrupted blood retinal barrier (BRB). The underlying etiology of DR can be attributed to the impaired BRB integrity and subsequent damages of the neurovascular units. In the absence of timely intervention, the compromised BRB can ultimately cause profound visual impairments. In particular, the ageing retina is vulnerable to ferroptosis, and hyperglycemia will accelerate the progression of this pathological process. In this article, we discuss the contributory role of ferroptosis in DR pathogenesis, and summarize recent therapeutic trials that targeting the ferroptosis. Further study on the ferroptosis mediated damage would enrich our knowledge of DR pathology, and promote the development of clinical treatment for this degenerative retinopathy.

Intravitreal Injection of ZYAN1 Restored Autophagy and Alleviated Oxidative Stress in Degenerating Retina via the HIF-1α/BNIP3 Pathway.

Mitochondrial autophagy plays a contributary role in the pathogenesis of retina degeneration (RD). ZYAN1 is a novel proline hydroxylase domain (PHD) inhibitor that can enhance the expression of hypoxia-inducible factor 1-alpha (HIF-1α). This study investigated whether ZYAN1 could alleviate progressive photoreceptor loss and oxidative damage in a pharmacologically induced RD model via the modulation of mitophagy. ZYAN1 was injected into the vitreous body of the RD model, and the retinal autophagy level was analyzed. The therapeutic effects of ZYAN1 were evaluated via a function examination, a morphological assay, in situ reactive oxygen species (ROS) detection, and an immunofluorescence assay. It was shown that the thickness of the outer nuclear layer (ONL) increased significantly, and visual function was efficiently preserved via ZYAN1 treatment. The mitochondria structure of photoreceptors was more complete in the ZYAN1-treated mice, and the number of autophagosomes also increased significantly. Membrane disc shedding and ROS overproduction were alleviated after ZYAN1 treatment, and the axonal cilia were more structurally intact. A Western blot analysis showed that the expression levels of the autophagy-related proteins LC3-B, Beclin-1, and ATG5 increased significantly after ZYAN1 treatment, while the expression of P62 was down-regulated. Moreover, the expression levels of HIF-1α and BNIP3 were up-regulated after ZYAN1 treatment. Therefore, an intravitreal injection of ZYAN1 can act as part of the pharmacologic strategy to modulate mitophagy and alleviate oxidative stress in RD. These findings enrich our knowledge of RD pathology and provide insights for the discovery of a therapeutic molecule.

Crosstalk Between Microglia and Müller Glia in the Age-Related Macular Degeneration: Role and Therapeutic Value of Neuroinflammation.

Age-related macular degeneration (AMD) is a progressive neurodegeneration disease that causes photoreceptor demise and vision impairments. In AMD pathogenesis, the primary death of retinal neurons always leads to the activation of resident microglia. The migration of activated microglia to the ongoing retinal lesion and their morphological transformation from branching to ameboid-like are recognized as hallmarks of AMD pathogenesis. Activated microglia send signals to Müller cells and promote them to react correspondingly to damaging stimulus. Müller cells are a type of neuroglia cells that maintain the normal function of retinal neurons, modulating innate inflammatory responses, and stabilize retinal structure. Activated Müller cells can accelerate the progression of AMD by damaging neurons and blood vessels. Therefore, the crosstalk between microglia and Müller cells plays a homeostatic role in maintaining the retinal environment, and this interaction is complicatedly modulated. In particular, the mechanism of mutual regulation between the two glia populations is complex under pathological conditions. This paper reviews recent findings on the crosstalk between microglia and Müller glia during AMD pathology process, with special emphasis on its therapeutic potentials.

Interaction of exopolysaccharide produced by Lactobacillus plantarum YW11 with whey proteins and functionalities of the polymer complex.

Exopolysaccharide (EPS)-producing lactic acid bacteria have been widely used in fermented milk, but interaction between the EPS and milk proteins has not been well studied. In this study, interaction between the EPS from Lactobacillus plantarum YW11 (EPS-YW11) and whey proteins (WP), and functional properties of the EPS-YW11/WP were investigated. The results showed that EPS-YW11 tended to encase WP by ζ-potential analysis with a decrease in the surface charge of the protein fraction (from -26.00 mV to 15.30 mV), and an increase in the melting temperature of the protein fraction (from 76.31 °C to 84.48 °C) as shown by differential scanning calorimetry. Circular dichroism spectrometry showed that the EPS could induce structural change of WP, that is, increment in the content of α-helixes and random coils, There was stronger interaction between EPS-YW11 and WP at higher temperatures (60 °C, 90 °C) due to formation of intermolecular H-bonds and OH stretching vibration as indicated by infrared spectral analysis. A significant improvement in the texture (hardness, springiness, gumminess, resilience, cohesiveness, and chewiness) of the EPS-YW11/WP complex was also observed when compared to that of the EPS or WP alone. This was confirmed by microstructural observation of the EPS-YW11/WP complex that formed branched and porous structures, and it became more complex and stable with increased temperature treatment. Due to the strong interaction the EPS-YW11/WP exhibited improved functionality. This study identifies the potential of the EPS-YW11 to serve as a functional agent in the processing of fermented dairy products with enhanced textural stability and bioactivities such as cholesterol-lowering, antioxidant, and antibiofilm.

Application of luteinizing hormone-releasing hormone-ferrosoferric oxide nanoparticles in targeted imaging of breast tumors.

BACKGROUND: Ferrosoferric oxide (Fe3O4) nanoparticles are a commonly used magnetic resonance imaging (MRI) reagent. Luteinizing hormone-releasing hormone (LHRH) is highly expressed on the surfaces of tumors, but its expression is low or absent in the corresponding normal tissues, allowing it to be used for targeted imaging and treatment. METHODS: We prepared Fe3O4 nanoparticles using a chemical co-precipitation method, performed coupling with chitosan to prepare LHRH-Fe3O4 nanoparticles, and explored the application value of LHRH-Fe3O4 nanoparticles in targeted imaging and treatment of breast tumors through in vitro and in vivo experiments. RESULTS: The particle size of the LHRH-Fe3O4 nanoparticles was 10 nm, and they could be taken in by human MCF-7 breast cancer cells. The nanomaterial had low cytotoxicity. In vivo MRI experiments showed that LHRH-Fe3O4 could effectively concentrate on the tumor under the action of a magnetic field. It also had a good negative enhancement effect that significantly reduced the signal intensity of the T2 field, allowing it to be used as a contrast agent of the T2 field. CONCLUSION: LHRH-Fe3O4 nanoparticles serve the purpose of targeting contrast agents to target sites and are expected to be used for targeted imaging and treatment of cancers with high LHRH expression.

Characterization and Antioxidant Activity of the Exopolysaccharide Produced by Bacillus amyloliquefaciens GSBa-1.

The exopolysaccharide (EPS) produced by Bacillus amyloliquefaciens GSBa-1 was isolated and purified by ethanol precipitation, and DEAE-cellulose and Sepharose CL-6B chromatographies. The molecular mass of the purified EPS was determined to be 54 kDa. Monosaccharide analysis showed that the EPS was composed of predominantly glucose, and it was further confirmed by NMR spectroscopy to be α-glucan that consisted of a trisaccharide repeating unit with possible presence of two α-(1→3) and one α-(1→6) glucosidic linkages. Microstructural analysis showed that the EPS appeared as ellipsoid or globose with a smooth surface. The EPS had a degradation temperature at 240°C. Furthermore, the EPS had strong DPPH and hydroxyl radical scavenging activities, and moderate superoxidant anion scavenging and metal ion-chelating activities. This is the first characterization of a glucan produced by B. amyloliquefaciens with strong antioxidant activity. The results of this study suggest the potential of the EPS from B. amyloliquefaciens GSBa-1 to serve as a natural antioxidant for application in functional products.

Antioxidant status and gut microbiota change in an aging mouse model as influenced by exopolysaccharide produced by Lactobacillus plantarum YW11 isolated from Tibetan kefir.

This study investigated the effect of exopolysaccharide (EPS) produced by Lactobacillus plantarum YW11 on the oxidative status and gut microbiota in an aging mouse model induced with d-galactose. The in vitro assay of the antioxidant activity of the EPS showed concentration-dependent (0.25-3.0 mg/mL) activities. At 3.0 mg/mL, the EPS reached the highest scavenging activities with half maximal inhibitory concentration values against hydroxyl radicals at 75.10% and 1.22 mg/mL, superoxide anion at 62.71% and 1.54 mg/mL, 2, 2-diphenyl-1-picrylhydrazyl at 35.11% and 0.63 mg/mL, and the maximal chelating rate on ferrous ion and the half-maximal chelating concentration of the EPS at 41.09% and 1.07 mg/mL, respectively. High doses of EPS (50 mg/kg per day) effectively relieved the oxidative stress in the aging mice with increased levels of glutathione peroxidase, superoxide dismutase, catalase, and total antioxidant capacity in mice serum by 21.55, 33.14, 61.09, and 38.18%, respectively, and decreased malondialdehyde level from 11.69 to 5.89 mmol/mL compared with those in the untreated aging mice model. The analysis of pyrosequencing sequence data from the gut microbiota revealed that the EPS could recover the microbiota diversity and phylotypes decreased or eliminated by the d-galactose treatment. The EPS could selectively decrease the abundance of Flexispira (37.5 fold), and increase the abundance of Blautia (36.5 fold) and Butyricicoccus (9.5 fold), which correspondingly decreased the content of nitrogen oxides to 9.87% and increased the content of short-chain fatty acids by 2.23 fold, thereby improving the oxidative and health conditions of the host intestinal tract. Further correlation analysis of core-microbiota variation induced by different treatments showed a strong correlation with oxidative phenotypes [catalase, goodness of prediction (Q2) = 0.49; total antioxidant capacity, Q2 = 0.45; nitrogen oxides, Q2 = 0.67; short-chain fatty acids, Q2 = 0.55]. The fermented milk with L. plantarum YW11 containing EPS also showed favorable antioxidant and gut microbiota regulating activities. The present finding provided new insights into the functional mechanism of probiotics bioactivity.