High-Speed Generation of Microbubbles with Constant Cumulative Production in a Glass Capillary Microfluidic Bubble Generator.

This work reports a simple bubble generator for the high-speed generation of microbubbles with constant cumulative production. To achieve this, a gas-liquid co-flowing microfluidic device with a tiny capillary orifice as small as 5 µm is fabricated to produce monodisperse microbubbles. The diameter of the microbubbles can be adjusted precisely by tuning the input gas pressure and flow rate of the continuous liquid phase. The co-flowing structure ensures the uniformity of the generated microbubbles, and the surfactant in the liquid phase prevents coalescence of the collected microbubbles. The diameter coefficient of variation (CV) of the generated microbubbles can reach a minimum of 1.3%. Additionally, the relationship between microbubble diameter and the gas channel orifice is studied using the low Capillary number (Ca) and Weber number (We) of the liquid phase. Moreover, by maintaining a consistent gas input pressure, the CV of the cumulative microbubble volume can reach 3.6% regardless of the flow rate of the liquid phase. This method not only facilitates the generation of microbubbles with morphologic stability under variable flow conditions, but also ensures that the cumulative microbubble production over a certain period of time remains constant, which is important for the volume-dominated application of chromatographic analysis and the component analysis of natural gas.

Application of biological soil crusts for efficient cadmium removal from acidic mine wastewater.

Utilizing an acid-resistant biological soil crust (BSC) species that we discovered, we developed a device capable of efficiently removing cadmium (Cd) from mine wastewater with varying levels of acidity. Our research has demonstrated that this particular BSC species adapts to acidic environments by regulating the balance of fatty acids and acid-resistant enzymes. At a Cd concentration of 5 mg/L, the BSC grew well. When the initial Cd concentration was 2 mg/L, and the flow rate was set at 1 mL/min (at pH levels of 3, 4, and 5), BSC had a high removal rate of Cd, and the removal rate increased with the increase of pH (from 90% to 97%). Chemisorption is the primary removal mechanism in the initial stage, where the functional groups and minerals on the surface of the BSC play a significant role. In addition, BSC also adapts to Cd stress by changing bacterial community structure. It was discovered through infrared spectroscopy and two-dimensional correlation analysis that hydrophilic groups, specifically phosphate and carboxyl groups, exhibited the highest reactivity during the Cd binding process. Protein secondary structure analysis confirmed that as the pH increased, the adsorption capacity of the BSC increased; making biofilm formation easier. This study presents a novel approach for the treatment of acidic wastewater.

Fertilizer-induced manganese oxide formation enhances cadmium removal by paddy crusts from irrigation water.

Fertilization is a crucial agrological measure for agricultural production that can significantly impact the removal of Cd from irrigation water by paddy crusts (PC). In this study, laboratory and field experiments were conducted to investigate the impact of fertilization at low, medium, and high concentrations on the accumulation of Cadmium (Cd) in PC and the underlying mechanisms involved. The results showed that only low fertilizer concentration could promote the removal of Cd by PC, which reduced the Cd concentration in irrigation water from 19.52 µg/L to 5.35 µg/L. Conversely, medium and high fertilizer concentrations reduced the accumulation of Cd by PC. After fertilizer addition, the proportion of Fe-Mn oxidizable-Cd in PC reached 55 % (with low concentration of fertilizer treatment). The application of low concentration of fertilizer was found to stimulate the growth of filamentous green algae, leading to a significant increase in the relative abundance of sphingomonadaceae (by 1.39 %) and comamonadaceae (by 1.29 %). The XRD, SEM and correlation analysis show that a large amount of manganese oxide is formed on the surface of PC, which increases the fixation of Cd. These findings provide a new perspective for the remediation of heavy metal contamination in paddy fields.

CircARF3 Mitigates Allergic Rhinitis through Targeting microRNA-205-5p/Sirtuin 5 Axis.

INTRODUCTION: Circular RNAs (circRNAs) are essential in the progression of allergic rhinitis (AR). The purpose of this research was to examine the role of circRNA ADP-ribosylation factor 3 (circARF3) in the pathogenesis of AR. METHODS: To generate an animal model of AR, mice were treated with house dust mite (HDM), and mice nasal epithelial cells (NEpCs) were treated with IL-4/IL-13 to imitate the inflammatory damage of AR in vitro. Sanger sequencing, qRT-PCR, and RNAse R digestion assays all validated the circularization structure of circARF3. The levels of circARF3, miR-205-5p, and sirtuin 5 (SIRT5) were determined by qRT-PCR or Western blotting. Luciferase reporter, RNA immunoprecipitation, and pull-down experiments were used to investigate the regulatory network. Flow cytometry was used to investigate the rate of cell apoptosis, and Western blotting was used to determine the levels of apoptotic-related proteins (cleaved caspase 3, cleaved polyadenosine-diphosphate-ribose polymerase) and HMGB1, TLR4, and MyD88. Enzyme-linked immunosorbent assay was used to assess the inflammatory response. Hematoxylin-eosin staining and TUNEL were used to detect the histology of injury and apoptosis of nasal mucosa tissues. RESULTS: CircARF3 and SIRT5 levels were reduced in HDM-treated animals and IL-4/IL-13-treated NEpCs, while miR-205-5p expression was increased. CircARF3 was generated by back-splicing exons 3-5 with a stable circular shape. CircARF3 overexpression mitigated IL-4/IL-13-induced apoptosis in NEpCs by inhibiting miR-205-5p. SIRT5 upregulation attenuated IL-4/IL-13-induced inflammatory injury in NEpCs, and SIRT5 knockdown induced opposite effects. miR-205-5p silencing reversed the effects of SIRT5 knockdown on IL-4/IL-13-induced inflammatory injury. Furthermore, circARF3 overexpression alleviated histological abnormalities, apoptosis, inflammatory response, and HMGB1/TLR4 signaling activation in HDM-treated animals. CONCLUSION: CircARF3 inhibited cell apoptosis and inflammation via the miR-205-5p/SIRT5 axis in IL-4/IL-13-treated NEpCs and HDM-treated mice.

Effects of Vitamin A on Immune Responses and Vitamin A Metabolism in Broiler Chickens Challenged with Necrotic Enteritis.

Necrotic enteritis (NE) is an important enteric inflammatory disease of poultry, and the effects of vitamin A (VitA) on NE birds are largely unknown. The present study was conducted to investigate the effects of VitA on the immune responses and VitA metabolism of NE broilers as well as the underlying mechanisms. Using a 2 × 2 factorial arrangement, 336 1-day-old Ross 308 broiler chicks were randomly assigned to 4 groups with 7 replicates. Broilers in the control (Ctrl) group were fed a basal diet without extra VitA supplementation. Broilers in the VitA group were fed a basal diet supplemented with 12,000 IU/kg of VitA. Birds in NE and VitA + NE groups were fed corresponding diets and, in addition, co-infected with Eimeria spp. and Clostridium perfringens on days 14 to 20. Samples of the blood, jejunum, spleen and liver were obtained on day 28 for analysis, and meanwhile, lesion scores were also recorded. The results showed that NE challenge increased lesion score in the jejunum and decreased serum glucose, total glyceride, calcium, phosphorus and uric acid levels (p < 0.05). VitA supplementation reduced the levels of serum phosphorus, uric acid and alkaline phosphatase in NE-challenged birds and increased serum low-density lipoprotein content and the activity of aspartate aminotransferase and creatine kinase (p < 0.05). Compared with the Ctrl group, the VitA and NE groups had higher mRNA expression of interferon-γ in the jejunum (p < 0.05). NE challenge up-regulated mRNA expression of interleukin (IL)-13, transforming growth factor-ß4, aldehyde dehydrogenase (RALDH)-2 and RALDH-3 in the jejunum, while VitA supplementation increased jejunal IL-13 mRNA expression and hepatic VitA content, but down-regulated splenic IL-13 mRNA expression (p < 0.05). The VitA + NE group had higher serum prostaglandin E2 levels and the Ctrl group had higher splenic RALDH-3 mRNA expression than that of the other three groups (p < 0.05). NE challenge up-regulated jejunal retinoic acid receptor (RAR)-ß and retinoid X receptor (RXR)-α as well as splenic RAR-α and RAR-ß mRNA expression (p < 0.05). VitA supplementation up-regulated jejunal RAR-ß expression but down-regulated mRNA expression of RXR-α, RXR-γ, signal transducers and activators of transcription (STAT) 5 and STAT6 in the spleen (p < 0.05). Moreover, compared with the Ctrl group, the VitA and NE groups had down-regulated mRNA expression of jejunal and splenic Janus kinase (JAK) 1 (p < 0.05). In conclusion, NE challenge induced jejunal injury and expression of Th2 and Treg cell-related cytokines and enhanced RALDH and RAR/RXR mRNA expression, mainly in the jejunum of broilers. VitA supplementation did not alleviate jejunal injury or Th2 cell-related cytokine expression; however, it improved hepatic VitA deposition and inhibited the expression of RALDH-3, RXR and the JAK/STAT signaling pathway in the spleen of broilers. In short, the present study suggested the modulatory effects of vitamin A on the immune responses and vitamin A metabolism in broiler chickens challenged with necrotic enteritis.

A dominant variant in apoptosis-related gene XKR8 is relevant to hereditary auditory neuropathy.

BACKGROUND: Auditory neuropathy is an unusual type of hearing loss. At least 40% of patients with this disease have underlying genetic causes. However, in many hereditary auditory neuropathy cases, etiology remains undetermined. METHODS: We collected data and blood samples from a four-generation Chinese family. After excluding relevant variants in known deafness-related genes, exome sequencing was conducted. Candidate genes were verified by pedigree segregation, transcript/protein expression in the mouse cochlea, and plasmid expression studies in HEK 293T cells. Moreover, a mutant mouse model was generated and underwent hearing evaluations; protein localization in the inner ear was also assessed. RESULTS: The clinical features of the family were diagnosed as auditory neuropathy. A novel variant c.710G > A (p.W237X) in apoptosis-related gene XKR8 was identified. Genotyping of 16 family members confirmed the segregation of this variant with the deafness phenotype. Both XKR8 mRNA and XKR8 protein were expressed in the mouse inner ear, predominantly in regions of spiral ganglion neurons; Moreover, this nonsense variant impaired the surface localization of XKR8 in cells. Transgenic mutant mice exhibited late-onset auditory neuropathy, and their altered XKR8 protein localization in the inner ear confirmed the damaging effects of this variant. CONCLUSIONS: We identified a variant in the XKR8 gene that is relevant to auditory neuropathy. The essential role of XKR8 in inner ear development and neural homeostasis should be explored.

Recent Advances in Thallium Removal from Water Environment by Metal Oxide Material.

Thallium is widely used in industrial and agricultural development. However, there is still a lack of systematic understanding of its environmental hazards and related treatment methods or technologies. Here, we critically assess the environmental behavior of thallium in aqueous systems. In addition, we first discuss the benefits and limitations of the synthetic methods of metal oxide materials that may affect the practicality and scalability of TI removal from water. We then assess the feasibility of different metal oxide materials for TI removal from water by estimating the material properties and contaminant removal mechanisms of four metal oxides (Mn, Fe, Al, and Ti). Next, we discuss the environmental factors that may inhibit the practicality and scalability of Tl removal from water. We conclude by highlighting the materials and processes that could serve as more sustainable alternatives to TI removal with further research and development.

Effects of Dietary Supplementation with Vitamin A on Antioxidant and Intestinal Barrier Function of Broilers Co-Infected with Coccidia and Clostridium perfringens.

Necrotic enteritis (NE) impairs poultry production and causes great economic loss. The nutritional regulation of diets has the potential to alleviate NE. The present study was conducted to investigate the effects of dietary supplementation with vitamin A (VA) on the antioxidant and intestinal barrier function of broilers co-infected with coccidia and C. perfringens (CCP). In a 2 × 2 factorial arrangement, 336 one-day-old Ross 308 broilers were divided into four treatments with two levels of VA (0 or 12,000 IU/kg) and challenged with or without CCP. The animal trial lasted for 42 days. The results showed that dietary supplemental VA improved body weight gain (BWG) and the feed intake (FI), and the FI was negatively affected by CCP. Additionally, the levels of catalase (CAT) in the serum, total superoxide dismutase (T-SOD), and CAT in the jejunum and glutathione peroxidase (GSH-Px) in the liver decreased with the CCP challenge (p < 0.05). The mRNA levels of SOD, CAT, GSH-Px1, and GSH-Px3 in the liver and jejunum were upregulated by the CCP challenge (p < 0.05). In addition, the level of serum diamine oxidase (DAO), and the mRNA level of ZO-1 were also upregulated with the CCP challenge. Dietary supplementation with VA contributed to the intestinal villi height and the mRNA level of Mucin-2 in the jejunum (p < 0.05). Additionally, dietary VA had the ability to alleviate the upregulation of SOD in the liver and SOD, CAT, GSH-Px1, GSH-Px3, ZO-1, and claudin-1 in the jejunum with the CCP challenge (p < 0.05). However, the mRNA level of GSH-Px3 and the levels of SOD in the liver and jejunum were downregulated with the VA supplementation in the diet. In conclusion, dietary VA improved the growth performance and the intestinal barrier function; nonetheless, it failed to alleviate the negative effects of CCP on the antioxidant function in broilers.

Applications and influencing factors of the biochar-persulfate based advanced oxidation processes for the remediation of groundwater and soil contaminated with organic compounds.

Biochar (BC) is a low-cost material rich in carbon, which is being used increasingly as a catalyst in persulfate-based advanced oxidation processes (PS-AOPs) for the remediation of groundwater and soil contaminated with organic compounds. In this work, a general summary of preparation methods and applications of various BC (i.e., pristine BC, magnetic BC, and chemically modified BC) in PS-AOPs is presented. Different influence factors (e.g., pH, anions, natural organic matter) for the degradation of organic compounds are discussed. Meanwhile, the influence of external energy (e.g., solar irradiation, UV-Vis, ultrasonic) is also mentioned. Furthermore, the advantage of different BC in PS-AOPs are compared. Finally, potential problems, challenges, and prospects in the application of biochar-persulfate based advanced oxidation processes (BCPS-AOPs) are discussed in the conclusion and perspective.

Exosomal miR-106a-5p accelerates the progression of nasopharyngeal carcinoma through FBXW7-mediated TRIM24 degradation.

Nasopharyngeal carcinoma (NPC) is prevalent in East Asia and causes increased health burden. Elucidating the regulatory mechanism of NPC progression is important for understanding the pathogenesis of NPC and developing novel therapeutic strategies. Nasopharyngeal carcinoma and normal tissues were collected. Nasopharyngeal carcinoma cell proliferation, migration, and invasion were evaluated using CCK-8, colony formation, wound healing, and transwell assays, respectively. A xenograft mouse model of NPC was established to analyze NPC cell growth and metastasis in vivo. The expression of miR-106a-5p, FBXW7, TRIM24, and SRGN was determined with RT-qPCR and Western blot. MiR-106a-5p, TRIM24, and SRGN were upregulated, and FBXW7 was downregulated in NPC tissues and cells. Exosomal miR-106a-5p could enter NPC cells, and its overexpression promoted the proliferation, migration, invasion, and metastasis of NPC cells, which were suppressed by knockdown of exosomal miR-106a-5p. MiR-106a-5p targeted FBXW7 to regulate FBXW7-mediated degradation of TRIM24. Furthermore, TRIM24 regulated SRGN expression by binding to its promoter in NPC cells. Suppression of exosomal miR-106a-5p attenuated NPC growth and metastasis through the FBXW7-TRIM24-SRGN axis in vivo. Exosomal miR-106a-5p accelerated the progression of NPC through the FBXW7-TRIM24-SRGN axis. Our study elucidates novel regulatory mechanisms of NPC progression and provides potential exosome-based therapeutic strategies for NPC.

Ursolic acid alleviates heat stress-induced lung injury by regulating endoplasmic reticulum stress signaling in mice.

Acute lung injury has been reported to be associated with heat stress in various animals. Ursolic acid is a natural pentacyclic triterpenoid compound with multiple bioactivities. However, it remains unknown whether ursolic acid supplementation alleviates heat stress-induced lung injury. In the present study, male Institute of Cancer Research mice were left untreated under a normal temperature condition (23±1°C), receiving orally administrated with vehicle (phosphate buffered saline) or ursolic acid (40 mg/kg BW-1·d-1 for 2 d), and then were subjected to high temperature (41±1°C) for 2 h. Histological alterations, activities of antioxidative enzymes, apoptosis, generation of reactive oxygen species, abundance of inflammatory cytokines, and endoplasmic reticulum stress-related proteins were analyzed. Compared with the controls, heat stress treatment led to enhanced apoptosis, increased H2O2 production, and upregulated protein levels of inflammatory cytokines in the serum, including tumor necrosis factor alpha, interleukin-6, and interleukin-1 beta. Activities of malondialdehyde, lactate dehydrogenase, and myeloperoxidase were increased, while the activities for superoxide dismutase and catalase were reduced in lung tissues of mice. All these alterations were significantly prevented by ursolic acid administration. Further study showed that heat stress led to activation of protein kinase-like ER kinase eukaryotic initiation factor 2 alpha -the transcription factor CCAAT-enhancer-binding protein homologous protein (CHOP) signaling, which was attenuated by ursolic acid supplementation. These findings indicated that ursolic acid pretreatment protected lung tissues against heat stress-induced injury by regulating inflammatory cytokines and unfolded protein response in mice. Ursolic acid supplementation might be a therapeutic strategy to alleviate high temperature-induced lung injury in humans and animals.

Electrophysiologic Characteristics of Various Degrees of Selective Outer Hair Cell Loss in Rats.

HYPOTHESIS: The characteristics of auditory brainstem response (ABR), electrocochleogram (ECochG), and distortion product otoacoustic emissions (DPOAE) of different degrees of selective outer hair cells (OHCs) loss may be helpful for clinicians to evaluate the pathogeny, diagnosis, and rehabilitation of individuals' hearing loss. BACKGROUND: How many OHCs are necessary to maintain cochlear amplifier function remains unknown. The electrophysiologic characteristics may indicate different degrees of OHCs loss. METHODS: Electrophysiological characteristics were tested using 8-kHz pure-tone stimulus and OHCs counted specifically in the region of the cochlea corresponding to 8-kHz. Rat models of selective OHCs loss were established by injecting kanamycin (KM) at various dosages, and the region of 8-kHz was obtained by 8-kHz pure-tone exposure. RESULTS: The ABR thresholds were affected slightly with OHCs loss < 30%, and were increased dramatically with OHCs loss ranging from 30 to 70%, but the thresholds did not increase further when OHCs loss exceeded 70%. As OHCs loss increased, the compound action potential (CAP) amplitude decreased. The CAP amplitude and OHCs loss were negatively correlated. Moreover, the summating potential (SP)/action potential (AP) increased as OHCs loss increased. DPOAE and cochlear microphonics (CM) exhibited reduced amplitudes when OHCs loss < 30%. CONCLUSIONS: Electrophysiologic characteristics may indicate different degrees of OHCs loss. While OHCs loss > 70%, the cochlear amplification may lose completely, but it is difficult to detect OHCs loss < 30%, because the ABR or DPOAE may reveal "normal" at this level. Moreover, the decreased CAP amplitude or increased SP/AP may be indicators for OHCs loss.

Enhancement of repeated applications of chelates on phytoremediation of uranium contaminated soil by Macleaya cordata.

A greenhouse pot experiment was performed to investigate the enhancement of repeated applications of citric acid (CA), ethylenediamine disuccinic acid (EDDS), and Oxalic acid (OA) on phytoremediation of uranium (U) contaminated soil by Macleaya Cordata. The chelates followed the order CA > EDDS > OA in terms of the enhancement on uranium uptake by M. cordata. The repeated applications of the chelates were found to be more effective than the one time application at the equal dose as the U concentration of soil solution increased significantly from the 8th to 14th day. The repeated applications of 10 mmol kg-1 CA promoted the solubilization of U in the U-contaminated soil by significantly decreasing the pH of soil solution, achieved the maximum U concentration of soil solution (1463.6 µg L-1), bioconcentration factors (BCFs, 11.4), bioaccumulation factors (BAFs, 21.4) and transfer factors (TFs, 1.9), which were 215.2, 5.7, 30.6 and 16.3 times as compared with the control group, respectively. The three applied chelates significantly affected the activities of the antioxidant enzymes in the leaves. Repeated applications of CA further enhanced the activities of the antioxidant enzymes in the leaves of M. cordata as compared with the control, EDDS and OA, mitigated the oxidative stress induced by uranium and chelates, and maximized the enhancement on the uranium uptake, which will be beneficial for the enhancement on the phytoremediation of uranium contaminated soil by U hyperaccumulating plants. These results indicated that the phytoavailability of uranium in soil solution as well as the accumulation of U by M. cordata were both significantly increased after repeated applications of CA, and that the repeated applications of 10 mmol kg-1 CA increased the activities of the antioxidant enzymes and promoted U accumulation by M. cordata. The study provided an environmentally friendly alternative for the enhancement on the phytoremediation of uranium contaminated soil using M. cordata.

An artificially constructed Syngonium podophyllum-Aspergillus niger combinate system for removal of uranium from wastewater.

Aspergillus niger was inoculated to the roots of five plants, and the Syngonium podophyllum-A. niger combinate system (SPANCS) was found to be the most effective in removing uranium from hydroponic liquid with initial uranium concentration of 5 mg L(-1). Furthermore, the hydroponic experiments on the removal of uranium from the hydroponic liquids with initial uranium concentrations of 0.5, 1.0, and 3.0 mg L(-1) by the SPANCS were conducted, the inhibitory effect of A. niger on the growth of S. podophyllum in the SPANCS was studied, the accumulation characteristics of uranium by S. podophyllum in the SPANCS were analyzed, and the Fourier transform infrared (FT-IR) and extended X-ray absorption fine structure (EXAFS) spectra were measured. The results show that the removal of uranium by the SPANCS from the hydroponic liquids with initial uranium concentrations of 0.5, 1.0, and 3.0 mg L(-1) reached 98.20, 97.90, and 98.50%, respectively, after 37 days of accumulation of uranium; that the uranium concentrations in the hydroponic liquids decreased to 0.009, 0.021, and 0.045 mg L(-1), respectively, which are lower than the stipulated concentration for discharge of 0.050 mg L(-1) by the People's Republic of China; that A. niger helped to generate more groups in the root of S. podophyllum which can improve the complexing capability of S. podophyllum for uranium; and that the uranium accumulated in the root of S. podophyllum was in the form of phosphate uranyl and carboxylic uranyl.

DNA methylation and histone modification patterns during the late embryonic and early postnatal development of chickens.

Early mammalian embryonic cells have been proven to be essential for embryonic development and the health of neonates. A series of epigenetic reprogramming events, including DNA methylation and histone modifications, occur during early embryonic development. However, epigenetic marks in late embryos and neonates are not well understood, especially in avian species. To investigate the epigenetic patterns of developing embryos and posthatched chicks, embryos at embryonic day 5 (E5), E8, E11, E14, E17, and E20 and newly hatched chicks on day of life 1 (D1), D7, D14, D21 were collected. The levels of global DNA methylation and histone H3 at lysine 9 residue (H3K9) modifications were measured in samples of liver, jejunum, and breast skeletal muscles by Western blotting and immunofluorescence staining. According to our data, decreased levels of proliferating cell nuclear antigen expression were found in the liver and a V-shaped pattern of proliferating cell nuclear antigen expression was found in the jejunum. The level of proliferating cell nuclear antigen in muscle was relatively stable. Caspase 3 expression gradually decreased over time in liver, was stable in the jejunum, and increased in muscle. Levels of DNA methylation and H3K9 acetylation decreased in liver over time, while the pattern was N-shaped in jejunal tissue and W-shaped in pectoral muscles, and these changes were accompanied by dynamic changes of DNA methyltransferases, histone acetyltransferases 1, and histone deacetylase 2. Moreover, dimethylation, trimethylation, and acetylation of H3K9 were expressed in a time- and tissue-dependent manner. After birth, epigenetic marks were relatively stable and found at lower levels. These results indicate that spatiotemporal specific epigenetic alterations could be critical for the late development of chick embryos and neonates.

Inflammatory responses to a Clostridium perfringens type A strain and α-toxin in primary intestinal epithelial cells of chicken embryos.

The causative pathogen of necrotic enteritis is the Gram-positive bacterium Clostridium perfringens. Its main cell wall component, peptidoglycan (PGN), can be recognized by Toll-like receptor 2 and nucleotide-binding oligomerization domain (NOD). Consequently, the immune response is initiated via activation of nuclear factor kappa B (NF-κB) signalling pathway. An in vitro study was conducted to investigate chicken intestinal inflammatory responses to C. perfringens type A and one of its virulence factors, α-toxin. In primary intestinal epithelial cells, C. perfringens as well as commercially available PGN and α-toxin challenge upregulated mRNA expression of interleukin (IL)-6, IL-8 and inducible nitric oxide synthase (iNOS) with a dosage-dependent manner at 3 h post infection (p.i.; P ≤ 0.001). Time-course effects of three stimulators at high concentration were further examined. C. perfringens infection elevated IL-6, IL-8 and iNOS levels from 1 h to 9 h p.i., while PGN treatment increased IL-6 and IL-8 expression at 1 h and 3 h p.i. (P < 0.05). Bacterial and PGN treatments induced NOD1 expression at 6 h p.i. and only bacterial infection boosted NF-κB p65 expression at 6 h and 9 h p.i. (P < 0.05). α-Toxin treatment upregulated IL-6 and IL-8 expression throughout infection, as well as iNOS, TNF-α and NF-κB p65 expression at later hours p.i. (P < 0.05). In conclusion, both C. perfringens and α-toxin challenge induced intense cytokine expression associated with NF-κB activation in chicken intestinal epithelial cells. The receptors for the recognition of PGN component of C. perfringens need further investigation.

Phytoextraction of uranium from contaminated soil by Macleaya cordata before and after application of EDDS and CA.

This is the first report on using Macleaya cordata for phytoextraction of uranium from the uranium contaminated soil in the greenhouse. Macleaya M. cordata was found to increase uranium concentration in the soil solution by increasing the dissolved organic carbon (DOC). The amendment experiments with citric acid (CA) and [S,S]-ethylenediamine disuccinic acid (EDDS) at the rates of 1.0, 2.5, 5.0, and 10.0 mmol kg(-1) dry weight (DW) soil showed that EDDS was more efficient to increase uranium concentration in the shoot than CA when they were applied at the same rate. The applications of 5.0 mmol kg(-1) EDDS and 10.0 mmol kg(-1) CA were most appropriate for increasing uranium concentrations in the shoot of M. cordata. CA was more efficient to increase the solubility of uranium at the same application rates except for 2.5 mmol kg(-1) application rate. There was a linear correlation between the uranium concentration in the shoot and the average uranium concentration of one planted pot during 14 days in soil solution after the application of different rates of EDDS and CA, respectively (r(2) = 0.972, P < 0.01; r (2) = 0.948, P < 0.01), indicating that uranium uptake was dependent on the soluble uranium concentration. The Fe-U-DOC and Mn-U-DOC complexes were probably formed after the application of CA. Soil solution pH and Fe, Mn, Ca, and DOC concentrations in soil solution were found to be changed by the chelates.

Maternal high-zinc diet attenuates intestinal inflammation by reducing DNA methylation and elevating H3K9 acetylation in the A20 promoter of offspring chicks.

A20 is an anti-inflammatory protein that suppresses ubiquitin-dependent nuclear factor κB (NF-κB) signaling, which can be regulated by the microelement zinc (Zn). In mammals, Zn deficiency contributes to a decrease in A20 abundance, which impairs the gut mucosa barrier. However, it is unclear whether the epigenetic reprogramming of the A20 promoter is involved in enhanced Zn-induced intestinal immunity, especially in avian species. Herein, we show that maternal organic Zn exposure resulted in significantly improved intestinal morphological characteristics, increased mucin 2 (MUC2) abundance and secretory IgA (sIgA) production in progeny jejunums. Maternal and offspring Zn supplementation partially alleviated Zn-deficiency-induced inflammatory response, accompanied by repression of NF-κB signaling. Additionally, we observed DNA hypomethylation and histone H3 at lysine 9 (H3K9) hyperacetylation at the A20 promoter region and subsequent activated A20 expression in Zn-supplemented hens compared with control. Notably, maternal dietary organic Zn exposure exhibited greater attenuation of gut impairment, along with increased MUC2 expression and sIgA level, and decreased the abundance of TNF-α and A20 relative to the inorganic-Zn group. Furthermore, enhanced acetylated H3K9 and A20 transcription at day 14 was found in the offspring adequate dietary Zn group. Thus, A20 may be a novel inflammatory-suppressed factor of chick gut that is persistently promoted by dietary Zn supplementation via epigenetic modifications at A20 promoter.

Maternal zinc supplementation enhanced skeletal muscle development through increasing protein synthesis and inhibiting protein degradation of their offspring.

Previous studies proved that maternal zinc supplementation had no significant effect on body weight (BW) of the offspring, but the effects of maternal zinc supplementation on skeletal muscle development of the offspring are poorly defined. Here, broiler breeders at 46 weeks old were allocated into three treatments with six replicates of 40 hens each and fed with diets supplemented with zinc from ZnSO4 at 0 (group Zn/C), 50 mg/kg (group Zn/L), and 300 mg/kg (group Zn/H) respectively for 6 weeks. The male offspring from each dietary treatment were divided into seven cages of ten birds each and fed with a commercial diet with supplemental zinc from ZnSO4 at 20 mg/kg. Results indicated that with the increase of zinc supplementation in hen's diet, the zinc levels were significantly elevated (P < 0.05) in the egg yolk. Compared with the control group, the breast muscle yield and muscle fiber width were significantly (P < 0.05) higher and larger in the broilers from group Zn/H at 2 and 5 weeks post-hatch, the phosphorylation of AKT at serine 473 residue (Ser 473), mammalian target of rapamycin (mTOR) at serine 2448 residue (Ser 2448), and FOXO at serine 256 residue (Ser 256) in skeletal muscles of the birds from various dietary treatments at two different age post-hatch were significantly (P < 0.05) increased. The phosphorylation of mTOR and FOXO was usually related to protein synthesis and degradation. In conclusion, supplemental zinc into the breeders' diet could increase protein synthesis and decrease protein degradation, which, in turn, enhance breast muscle development of the offspring.

Xylanase supplementation of a wheat-based diet improved nutrient digestion and mRNA expression of intestinal nutrient transporters in broiler chickens infected with Clostridium perfringens.

Necrotic enteritis caused by Clostridium perfringens has become prevalent in the European Union due to the withdrawal of antibiotics in poultry feed. In an experiment with a 2 × 2 factorial arrangement, 336 one-day-old male broiler chicks (Ross 308) were assigned to 4 groups with or without C. perfringens challenge and fed wheat-based diets supplemented with or without xylanase at 5,500 U/kg of diet. The study aimed to investigate effects of xylanase addition on growth performance as well as nutrient digestion and absorption of C. perfringens-infected broilers. Before challenge (d 0-14), xylanase-supplemented birds had greater ADG and lower feed conversion ratio (FCR; P < 0.05). During infection (d 14-21), challenge tended to decrease ADG (P = 0.063) and significantly increased FCR (P < 0.05), whereas xylanase addition greatly reduced FCR (P < 0.05). Clostridium perfringens infection decreased AME values and apparent ileal digestibility of DM of diets (P < 0.05). Xylanase supplementation increased AME values regardless of infection and apparent ileal digestibility of CP in challenged birds (P < 0.05). Activities of duodenal α-amylase and chymotrypsin and pancreatic trypsin were decreased by C. perfringens infection (P < 0.05). Xylanase supplementation elevated pancreatic chymotrypsin activity and reduced duodenal α-amylase and trypsin activities (P < 0.05). It also decreased jejunal α-amylase activity and increased pancreatic α-amylase as well as jejunal sucrase activities in uninfected birds (P < 0.05). The duodenal mRNA expression of sodium glucose cotransporter 1 (SGLT1), H(+)-dependent peptide transporter 1 (PepT1), and liver fatty acid-binding protein (L-FABP) were downregulated (P < 0.05), but ileal SGLT1 gene expression was increased by infection (P < 0.05). Xylanase addition upregulated expression of jejunal SGLT1, PepT1, and L-FABP genes as well as ileal PepT1 and L-FABP genes in challenged broilers (P < 0.05). In conclusion, xylanase supplementation of wheat-based diets improved FCR and AME in birds irrespective of C. perfringens infection and elevated apparent ileal digestibility of CP and mRNA expression of nutrient transporters in challenged birds.