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1.
Food Chem ; 441: 138310, 2024 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-38218143

RESUMO

This research investigates the dynamic alterations that occur in protein molecular structure during the fermentation process of feed. Fourier transform infrared spectroscopy (FTIR), coupled with deconvolution, second derivative and curve-fitting methodologies, was employed to comparatively analyse the protein molecular structures in fermented feed. At the 48-h fermentation mark, the α-helix and ß-sheet contents reached their peaks, while the random coil and ß-turn contents were at their lowest. Simultaneously, the ß-sheet/α-helix ratio was minimized. FTIR spectroscopy emerged as a comprehensive tool, revealing the nuanced changes in molecular structure throughout the fermentation process of corn-soybean meal feed. When integrated with spectral quantitative analysis, it provides a novel perspective for evaluating the nutritional value of fermented feed.


Assuntos
Farinha , Zea mays , Espectroscopia de Infravermelho com Transformada de Fourier/métodos , Estrutura Molecular , Estrutura Secundária de Proteína , Proteínas , Glycine max
2.
Biol Trace Elem Res ; 202(5): 2075-2084, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-37610602

RESUMO

This study was designed to assess whether selenium-chitosan (Se-CTS) can protect porcine endometrial epithelial cells (PEECs) against damage and apoptosis induced by zearalenone (ZEA) via modulating the JNK/SAPK signaling pathway. The cell cycle, mitochondrial membrane potential (MMP), reactive oxygen species (ROS), and apoptosis rates of porcine endometrial epithelial cells were determined, as well as the expression levels of genes related to the SAPK/JNK signaling pathway. The results showed that 3.0 µmol/L Se-CTS decreased the percentage of ZEA-induced G1 phase in PEECs (P < 0.01), whereas 1.5 and 3.0 µmol/L Se-CTS increased the percentage of ZEA-induced percentage of G2 phase of PEECs (P < 0.01). Further, Se-CTS at 1.5 and 3.0 µmol/L improved the ZEA-induced decrease in MMP (P < 0.01), whereas Se-CTS at 0.5, 1.5, and 3.0 µmol/L reduced the increase in ROS levels and apoptosis rate induced by ZEA in PEECs (P < 0.01 or P < 0.05). Furthermore, 3.0 µmol/L Se-CTS ameliorated the increase in the expression of c-Jun N-terminal kinase (JNK), apoptosis signal-regulated kinase (ASK1), and c-Jun induced by ZEA (P < 0.01) and the reduction in mitogen-activated protein kinase kinase 4 (MKK4) and protein 53 (p53) expression (P < 0.01), while 1.5 µmol/L Se-CTS improved the expression of ASK1 and c-Jun induced by ZEA (P < 0.05). The results proved that Se-CTS alleviates ZEA-induced cell cycle stagnation, cell mitochondrial damage, and cell apoptosis via decreasing ZEA-produced ROS and modulating the JNK/SAPK signaling pathway.


Assuntos
Quitosana , Selênio , Zearalenona , Animais , Suínos , Sistema de Sinalização das MAP Quinases , Selênio/farmacologia , Selênio/metabolismo , Zearalenona/toxicidade , Zearalenona/metabolismo , Quitosana/farmacologia , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Células Epiteliais/metabolismo , Apoptose
3.
Front Vet Sci ; 10: 1184969, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37261113

RESUMO

This paper assessed the positive effects of selenized-oligochitosan (SOC) on zearalenone(ZEN)-induced intestinal dysfunction in piglets. Sixty piglets were randomly divided into 4 groups. Group C was fed the basal diet as a control and Group Z was supplemented with 2 µg/g ZEN in the basal diet; Group ZS1 and ZS2 were supplemented with 0.3 or 0.5 µg/g SOC (calculated by selenium), in addition to 2 µg/g ZEN in the basal diet. After 42 days, ileal mucosal structure, digestive enzyme activities, tight junction protein mRNA expressions, plasma D-lactate and D-xylose contents, and plasma diamine oxidase activities were determined. Compare with Group C, ileal villus height, value of villus height/crypt depth, trypsin, lipase and α-amylase activities, occluding, claudin-1 and ZO-1 mRNA expressions, and plasma D-xylose levels were significantly decreased (p < 0.01) in piglets of group Z; while compare to Group C, ileal crypt depth, plasma D-lactate contents and diamine oxidase activities were significantly increased in piglets of group Z (p < 0.01 or p < 0.05). Compare with Group Z, ileal villus height, lipase and α-amylase activities, occluding, claudin-1 and ZO-1 mRNA expressions, and plasma D-xylose levels were significantly elevated in piglets of group ZS1 and ZS2 (p < 0.01); while compare to Group Z, plasma D-lactate and diamine oxidase contents were significantly reduced in piglets of group ZS1 and ZS2 (p < 0.01 or p < 0.05). Compare with Group Z, value of villus height/crypt depth and trypsin activity were significantly promoted in piglets of group ZS2 (p < 0.01); whereas ileal crypt depth was significantly reduced in piglets of group ZS2 (p <0.01).Thus, SOC can mitigate ZEN-induced intestinal dysfunction in piglets.

4.
Microbiol Spectr ; 11(3): e0299522, 2023 06 15.
Artigo em Inglês | MEDLINE | ID: mdl-37154706

RESUMO

Lactococcus garvieae is an emerging zoonotic pathogen, but there are few reports regarding bovine mastitis. The prevalence of L. garvieae represents an increasing disease threat and global public health risk. Thirty-nine L. garvieae isolates were obtained from 2,899 bovine clinical mastitis milk samples in 6 provinces of China from 2017 to 2021. Five clonal complexes were determined from 32 multilocus sequence types (MLSTs) of L. garvieae: sequence type 46 (ST46) was the predominant sequence type, and 13 novel MLSTs were identified. All isolates were resistant to chloramphenicol and clindamycin, but susceptible to penicillin, ampicillin, amoxicillin-clavulanic acid, imipenem, ceftiofur, enrofloxacin, and marbofloxacin. Based on genomic analyses, L. garvieae had 6,310 genes, including 1,015 core, 3,641 accessory, and 1,654 unique genes. All isolates had virulence genes coding for collagenase, fibronectin-binding protein, glyceraldehyde-3-phosphate dehydrogenase, superoxide dismutase, and NADH oxidase. Most isolates had lsaD and mdtA antimicrobial resistance (AMR) genes. Based on COG (Clusters of Orthologous Genes database) results, the functions of defense, transcription and replication, and recombination and repair were enhanced in unique genes, whereas functions of translation, ribosomal structure, and biogenesis were enhanced in core genes. The KEGG functional categories enriched in unique genes included human disease and membrane transport, whereas COG functional categories enriched in core genes included energy metabolism, nucleotide metabolism, and translation. No gene was significantly associated with host specificity. In addition, analysis of core genome single nucleotide polymorphisms (SNPs) implied potential host adaptation of some isolates in several sequence types. In conclusion, this study characterized L. garvieae isolated from mastitis and detected potential adaptations of L. garvieae to various hosts. IMPORTANCE This study provides important genomic insights into a bovine mastitis pathogen, Lactococcus garvieae. Comprehensive genomic analyses of L. garvieae from dairy farms have not been reported. This study is a detailed and comprehensive report of novel features of isolates of L. garvieae, an important but poorly characterized bacterium, recovered in the past 5 years in 6 Chinese provinces. We documented diverse genetic features, including predominant sequence type ST46 and 13 novel MLSTs. Lactococcus garvieae had 6,310 genes, including 1,015 core, 3,641 accessory, and 1,654 unique genes. All isolates had virulence genes coding for collagenase, fibronectin-binding protein, glyceraldehyde-3-phosphate dehydrogenase, superoxide dismutase, and NADH oxidase and resistance to chloramphenicol and clindamycin. Most isolates had lsaD and mdtA antimicrobial resistance genes. However, no gene was significantly associated with host specificity. This is the first report that characterized L. garvieae isolates from bovine mastitis and revealed potential host adaptations of L. garvieae to various hosts.


Assuntos
Anti-Infecciosos , Mastite Bovina , Feminino , Animais , Bovinos , Humanos , Fibronectinas , Clindamicina , Mastite Bovina/epidemiologia , Mastite Bovina/microbiologia , Cloranfenicol , Genômica , Antibacterianos/farmacologia
5.
Front Immunol ; 13: 1034727, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36341458

RESUMO

Abundant microbiota resides in the organs of the body, which utilize the nutrition and form a reciprocal relationship with the host. The composition of these microbiota changes under different pathological conditions, particularly in response to stress and digestive diseases, making the microbial composition and health of the hosts body interdependent. Probiotics are living microorganisms that have demonstrated beneficial effects on physical health and as such are used as supplements to ameliorate symptoms of various digestive diseases by optimizing microbial composition of the gut and restore digestive balance. However, the supplementary effect does not achieve the expected result. Therefore, a targeted screening strategy on probiotic bacteria is crucial, owing to the presence of several bacterial strains. Core bacteria work effectively in maintaining microbiological homeostasis and stabilization in the gastrointestinal tract. Some of the core bacteria can be inherited and acquired from maternal pregnancy and delivery; others can be acquired from contact with the mother, feces, and the environment. Knowing the genera and functions of the core bacteria could be vital in the isolation and selection of probiotic bacteria for supplementation. In addition, other supporting strains of probiotic bacteria are also needed. A comprehensive strategy for mining both core and supporting bacteria before its clinical use is needed. Using metagenomics or other methods of estimation to discern the typically differentiated strains of bacteria is another important strategy to treat dysbiosis. Hence, these two factors are significant to carry out targeted isolation and selection of the functional strains to compose the resulting probiotic preparation for application in both research and clinical use. In conclusion, precise probiotic supplementation, by screening abundant strains of bacteria and isolating specific probiotic strains, could rapidly establish the core microbiota needed to confer resilience, particularly in bacterial dysfunctional diseases. This approach can help identify distinct bacteria which can be used to improve supplementation therapies.


Assuntos
Infecções Bacterianas , Enteropatias , Microbiota , Probióticos , Gravidez , Feminino , Humanos , Probióticos/uso terapêutico , Bactérias
6.
Front Vet Sci ; 9: 1036104, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36277059

RESUMO

This study assessed the protective effects of selenium-chitosan (SC) against antioxidant and immune function-related damage induced by zearalenone (ZEN) in mice. In total, 150 female mice were allotted to five groups for a 30-day study. Control mice were fed a basal diet. Mice in the ZEN, ZEN-Se1, ZEN-Se2 and ZEN-Se3 groups were fed the basal diet supplemented with same dose of ZEN (2 mg/kg) and different doses of SC, 0.0, 0.2, 0.4 and 0.6 mg/kg, respectively (calculated by selenium). After 30 days, the total antioxidant capacity (T-AOC) level, glutathione peroxidase (GSH-Px) activity, total superoxide dismutase (T-SOD) activity and malondialdehyde (MDA) content in plasma and liver, as well as Con A-induced splenocyte proliferation, plasma interleukins concentrations and liver interleukin mRNA expression levels were determined. The plasma and liver GSH-Px activities, liver T-AOC levels, Con A-induced splenocyte proliferation, interleukin (IL) contents and mRNA expression levels in the ZEN group were significantly lower than in the control group (P < 0.01 or P < 0.05), whereas plasma and liver MDA contents in the ZEN group were significantly higher than in the control group (P < 0.01 or P < 0.05). Additionally, plasma and liver GSH-Px activities, liver T-AOC levels, Con A-induced splenocyte proliferation, IL-1ß, IL-17A, IL-2 and IL-6 contents and mRNA expression levels in ZEN+Se2 and ZEN+Se3 groups were significantly higher than in the ZEN group (P < 0.01 or P < 0.05), whereas plasma and liver MDA contents in the ZEN+Se2 and ZEN+Se3 groups were significantly lower than in the ZEN group (P < 0.01 or P < 0.05). The plasma and liver GSH-Px activities, Con A-induced splenocyte proliferation, IL-1ß and IL-6 contents, IL-2 and IL-17A mRNA expression levels in the ZEN+Se1 group were also significantly higher than in the ZEN group (P < 0.01 or P < 0.05), whereas the plasma MDA content in the ZEN+Se1 group was also significantly lower than in the ZEN group (P < 0.01). Thus, SC may alleviate antioxidant function-related damage and immunosuppression induced by ZEN in mice.

7.
Food Funct ; 12(13): 5913-5926, 2021 Jul 05.
Artigo em Inglês | MEDLINE | ID: mdl-34028482

RESUMO

The aim of this study was to investigate the effects and potential signaling pathway of selenium-enriched Bacillus subtilis (SEBS) on beta defensin 1 (BD1) expression in chicken intestine. Chinese Huainan Partridge chickens (500 individuals) were randomly allocated into five groups, including control, inorganic Se, B. subtilis, SEBS, and a mixture of Se and B. subtilis (Se-BS). After 56 d of feeding, chicken ileal mucous membranes were harvested to detect differences in expression of BD1. The results indicated that BD1 was produced in intestinal crypt cells and secreted into the lumen through the villi brush border. BD1 was up-regulated in distal ileum segments colonized by SEBS and B. subtilis. Chicken primary intestinal crypt cells were cultured and grouped into control, inorganic Se, B. subtilis, SEBS, and Se-BS treatments to identify the receptor of B. subtilis. Results indicated that B. subtilis and SEBS were recognized by toll-like receptor 2 (TLR2), stimulating the NF-κB1 signaling pathway to increase expression of BD-1, which was further enhanced when combined with Se. Pro-inflammatory cytokines TNF-α, IL-1ß, and IL-6 were up-regulated with B. subtilis supplementation, and inhibited under the action of Se. In conclusion, B. subtilis and SEBS were recognized by the TLR2 receptor in the ileal mucous membrane, which activated the TLR2-MyD88-NF-κB1 signaling pathway to upregulate BD1 expression. In addition, Se enhanced recognition of B. subtilis and reduced levels of pro-inflammatory factors caused by estrogenic B. subtilis supplementation.


Assuntos
Bacillus subtilis/efeitos dos fármacos , Selênio/farmacologia , Transdução de Sinais/efeitos dos fármacos , Receptor 2 Toll-Like/metabolismo , beta-Defensinas/metabolismo , Animais , Galinhas , Citocinas/metabolismo , Duodeno/metabolismo , Íleo/patologia , Intestinos , NF-kappa B/metabolismo , Probióticos/farmacologia
8.
Oncol Lett ; 21(2): 115, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33376547

RESUMO

Melanoma is a common malignant skin tumor, which is the only fatal skin tumor at present. Melanoma has a high degree of malignancy and metastasis. The activity of modified Temporin-La (T-La) peptides from bullfrog skin were evaluated for antitumor activity and improved targeting in melanoma cells. The amino acid sequence of T-La was modified, resulting in the antitumor peptide, T-La (FS). T-La and T-La (FS) were coupled to the RGD small molecule polypeptide to form the chimeric peptides RGD-T-La and RGD-T-La (FS), respectively. The secondary structures for the peptides, evaluated using circular dichroism, were found to be α-helical. The structure of T-La was evaluated using bioinformatics. In addition, the antitumor effects of the modified peptide and the targeting of RGD chimeric peptide to the tumor in vivo and in vitro were analyzed. Antitumor activity was measured in vitro using the MTT assay. Tumor cells with high integrin αvß3 expression were detected using flow cytometry, and tumor cells were screened for sensitivity to RGD-T-La (FS) to establish a tumor model in nude mice. The effects of the peptides on tumor cells were measured using laser confocal microscopy in real-time. The mechanism of the peptide antitumor activity in tumor cells was evaluated with scanning electron microscopy. B16 melanoma cells were the most sensitive to the peptides, for which the cell survival rate was 24.65% for 10 µg/ml RGD-T-La (FS). RGD-La (FS) had a rapid effect on tumor cells. RGD chimeric polypeptides exhibited site-targeting cytotoxic effects in tumor cells. In the B16 melanoma mouse model, the peptides exhibited antitumor effects against early melanoma development and induced tumor apoptosis, possibly by inhibiting VEGF and promoting caspase-3 expression. Overall, the present study provides a scientific basis for the application of small molecule antimicrobial peptides as targeted antitumor agents and lays the foundation for the clinical application of these peptides as antitumor drugs.

9.
Poult Sci ; 99(11): 6045-6054, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-33142523

RESUMO

The aim of this study was to investigate the effects of selenium (Se)-enriched Saccharomyces cerevisiae (SSC) on meat quality and to elucidate the underlying mechanisms in broilers. A total of 200 one-day-old Arbor Acres broiler chickens were randomly allocated to one of four treatments with 5 replications of 10 chickens each. Group 1 served as a control and was fed a basal diet without Se supplementation, while groups 2, 3, and 4 were fed the basal diet supplemented with S. cerevisiae (SC), sodium selenite (SS), and SSC, respectively. Breast muscle samples were collected to evaluate meat quality, selenium concentration, oxidative stability, and the mRNA levels of antioxidant enzyme genes on day 42. As compared with groups 1 and 2, SS and SSC supplementation increased Se concentration, glutathione peroxidase (GPx) and thioredoxin reductase (TR) activities, total antioxidant capacity, and the mRNA levels of GPx-1, GPx-4, TR-1, and TR-3 (P < 0.05) and decreased drip loss and malondialdehyde (MDA) content (P < 0.05). As compared with group 3, SSC supplementation increased pH, lightness, yellowness, Se concentration, GPx and superoxide dismutase activities, and the mRNA levels of GPx-1 and GPx-4 (P < 0.05) but decreased drip loss and MDA content (P < 0.05). Thus, SSC improved meat quality and oxidative stability by activating the glutathione and thioredoxin systems, which should be attributed to the combined roles of Se and SC.


Assuntos
Suplementos Nutricionais , Microbiologia de Alimentos , Glutationa , Carne , Saccharomyces cerevisiae , Selênio , Tiorredoxinas , Ração Animal/análise , Animais , Galinhas/microbiologia , Dieta/veterinária , Glutationa/genética , Glutationa Peroxidase/genética , Carne/microbiologia , Carne/normas , Distribuição Aleatória , Saccharomyces cerevisiae/metabolismo , Tiorredoxinas/genética
10.
Poult Sci ; 99(8): 3979-3986, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32731985

RESUMO

This study aimed to investigate the effects of selenide chitosan sulfate (Se-CTS-S) on glutathione (GSH) system in hepatocytes and chickens. Chitosan, sodium selenite (Na2SeO3), selenide chitosan, chitosan sulfate (CTS-S), and Se-CTS-S were added to the culture medium and the basal diets; glutathione peroxidase (GSH-Px) activity, GSH content, total antioxidant capacity (T-AOC), and mRNA levels of cellular GPx (GPx-1) and phospholipid hydroperoxide GPx (GPx-4) in vivo and in vitro were determined. The results showed that Se-CTS-S increased (P < 0.05) GPx-1 and GPx-4 mRNA levels in hepatocytes and livers, and GSH-Px activity, GSH content, and T-AOC in the medium, hepatocytes, plasma, and livers compared with the control and chitosan treatments. Compared with CTS-S, Se-CTS-S treatments increased (P < 0.05) GPx-1 and GPx-4 mRNA levels in hepatocytes and livers, and GSH-Px activity, GSH content, and T-AOC capacity in the medium, hepatocytes, and livers. Compared with Na2SeO3 and CTS-Se, Se-CTS-S increased (P < 0.05) GPx-1 mRNA levels in hepatocytes and livers, GPx-4 mRNA levels in hepatocytes and livers, GSH-Px activity in the medium, hepatocytes, and livers, GSH contents in plasma and livers, and T-AOC in the medium, plasma, and livers. Thus, Se-CTS-S showed better biological activity that mainly benefited from the synergistic effects of Se and sulfate on GSH system.


Assuntos
Galinhas , Quitosana , Hepatócitos , Selênio , Animais , Quitosana/farmacologia , Glutationa/metabolismo , Glutationa Peroxidase/genética , Hepatócitos/efeitos dos fármacos , Organismos Livres de Patógenos Específicos
11.
Biol Trace Elem Res ; 191(1): 183-188, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-30554384

RESUMO

Sixty Kunming mice were randomly assigned into three groups. Mice in a control group were fed a basal diet, while mice in AFB1 group and AFB1-Se group were fed the basal diet supplemented with 250 µg/kg AFB1 or the basal diet supplemented with 250 µg/kg AFB1 and 0.2 mg/kg selenium as selenium-enriched yeast, respectively. On day 30 of the experiment, growth performance, glutathione peroxidase (GSH-Px) activities, total antioxidant capacity (T-AOC) levels, and malondialdehyde (MDA) contents in liver, interleukin-2 (IL-2), and interferon-γ (IFN-γ) contents in serum, and cytochrome P3a11 (Cyp3a11), IL-2, IFN-γ, and GSH-Px1 mRNA levels in liver were determined. The results showed that final weights, weight gains, T-AOC levels, GSH-Px1, and IFN-γ mRNA levels in AFB1-Se group and control group were higher or significantly higher than those in AFB1 group (P < 0.05 or P < 0.01), respectively. Body length gains in AFB1 group were lower than those in the control group (P < 0.05), while there was no significant difference between the AFB1-Se and control groups (P > 0.05). IL-2 contents and liver IL-2 mRNA levels in AFB1-Se group were significantly higher than those in the AFB1 group and control group (P < 0.01), and IL-2 contents in the control group were also significantly higher than those in the AFB1 group (P < 0.01). IFN-γ contents in AFB1-Se group and AFB1 group were significantly higher than those in control group (P < 0.01), while IFN-γ contents in AFB1-Se group were significantly lower than those in AFB1 group (P < 0.01). Cyp3a11 mRNA levels in AFB1-Se group and AFB1 group were significantly higher than those in the control group (P < 0.01). The results indicated that selenium-enriched yeast could partly reduce the toxicity induced by AFB1 in mice, including improving growth performance, antioxidation capacity, IL-2 and IFN-γ contents, and enhancing IL-2, IFN-γ, and GSH-Px1 mRNA levels.


Assuntos
Aflatoxina B1/toxicidade , Regulação da Expressão Gênica/efeitos dos fármacos , Selenito de Sódio/farmacologia , Fermento Seco/farmacologia , Animais , Suplementos Nutricionais , Feminino , Masculino , Camundongos , Selênio/farmacologia
12.
Pak J Pharm Sci ; 31(3(Supplementary)): 1013-1020, 2018 May.
Artigo em Inglês | MEDLINE | ID: mdl-29731438

RESUMO

Lectin has attracted attention because of its ability to serve as a carrier for targeted drug delivery. Large lectins isolated from marine invertebrates and crustaceans have strong immunogenicity and adverse effects, which limit their usefulness. This study reports the identification of catesbeianalectin via screening a bullfrog skin cDNA library. The catesbeianalectin polypeptide has a molecular weight of 1.47 kD, making it the smallest known lectin in terms of molecular weight. Circular dichroism analysis showed a PPII helix secondary structure. Catesbeianalectin strongly induces agglutination of rabbit erythrocytes and a variety of pathogens include Staphylococcus aureus, Streptococcus suis type 2, Actinobacillus pleuropneumoniae, and piglet paratyphoid Salmonella. The mean serum titer in catesbeianalectin-immunized Balb/c mice was 1:25, which was significantly lower than that of positive controls immunized with wheat germ agglutinin. Surface plasmon resonance indicated an S-type lectin. 125I-labeled catesbeianalectin did not pass the blood-brain barrier. This study provides a basis for further research on the potential of catesbeianalectin as a carrier in targeted drug delivery.


Assuntos
Aglutinação/efeitos dos fármacos , Lectinas/metabolismo , Animais , Bactérias/imunologia , Eritrócitos/imunologia , Feminino , Radioisótopos do Iodo/química , Camundongos , Coelhos , Distribuição Tecidual
13.
Cancer Cell Int ; 15: 20, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25729329

RESUMO

BACKGROUND: The objective of this study was to determine the in vitro tumor-inhibitory effect of a recombinant adenovirus expressing a fusion protein of tumor necrosis factor (TNF) related apoptosis inducing ligand (TRAIL) and hemagglutinin-neuraminidase (HN) genes on the MSB-1 Marek's disease tumor cell line. METHODS: TRAIL and HN genes were amplified from lymphocytes in the peripheral blood of chickens and the LaSota strain of Newcastle disease virus (NDV), respectively, using RT-PCR. The two genes were connected with a 2A connecting peptide by site-directed mutagenesis and gene splicing by overlap extension (SOE). The target gene TRAIL-2A-HN was cloned into the shuttle vector pShuttle-CMV. Homologous recombination was carried out with the vector pAdeasy-1 in the bacterium BJ5183 to construct the recombinant adenovirus plasmid pAd-TRAIL-2A-HN. After linearization, the plasmid was transfected into AD293 cells and packaged. Real-time quantitative PCR (RT-PCR) and fluorescence microscopy confirmed the introduction of the recombinant adenovirus into AD293 cells. The TCID50 method (50% tissue culture infectious dose) was employed to determine viral titers for the exprimental and control viruses, which met criteria for use. The Marek's disease tumor cell line MSB-1 was transfected with the constructed recombinant adenovirus. The infectivity of the recombinant adenovirus and the expression levels of exogenous genes were detected with RT-PCR and western blotting. The effects of the recombinant adenovirus on the growth of MSB-1 cells and cellular apoptosis were determined using flow cytometry. RESULTS: The recombinant adenovirus infected the cultured cells in vitro, and replicated and expressed exogenous genes in the cells. The recombinant adenovirus Ad-TRAIL-2A-HN inhibited the growth of MSB-1 cells and induced apoptosis by expressing exogenous genes. The rate of induced MSB-1 cell apoptosis reached 11.61%, which indicated that TRAIL and HN produced synergistic tumor-inhibiting effects. CONCLUSION: The constructed TRAIL-2A-HN fusion gene combined the apoptosis-inducing function of TRAIL and the adsorptive capacity of HN from NDV for tumor cells, and the capacity of the recombinant adenovirus expressing this fusion gene to induce tumor cell apoptosis was reported. These results provide a basis for future in vivo tumor suppression studies using recombinant adenoviruses.

14.
Biol Trace Elem Res ; 165(2): 145-52, 2015 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-25634140

RESUMO

One hundred and eighty Kunming mice were allotted to three groups in a randomized complete block design, including two treatments and one control. Mice in group 1 were fed a basal diet as control, while mice in groups 2 and 3 were fed the basal diet supplemented with 0.2 mg/kg selenium as sodium selenite (SS) or selenium-chitosan (SC), respectively. On day 28 of the experiment, blood selenium concentration, glutathione peroxidase (GPx) activity, plasma superoxide dismutase (SOD) activity, malondialdehyde (MDA) content, and Con A-induced splenocyte proliferation were determined, and plasma interleukin-2 (IL-2) and interferon-γ (IFN-γ) concentrations, splenic plaque-forming cell (PFC) responses, serum hemolysis level (HC50), and delayed-type hypersensitivity (DTH) responses were determined on day 15 of the experiment. The results showed that blood selenium concentration, GPx activity, splenic PFC response, and plasma IL-2 and IFN-γ concentrations in SC group were higher than those in the control and SS groups (P < 0.01 or P < 0.05), respectively. Plasma SOD activity, Serum hemolysis level, DTH responses, and Con A-induced splenocyte proliferation in SC group were higher than those in control (P < 0.01 or P < 0.05). Plasma SOD activity, serum hemolysis level, DTH responses, and Con A-induced splenocyte proliferation in SC group were also higher than those in SS group, while there was no significant difference between SC and SS groups (P > 0.05). Plasma MDA content in SC group was lower than those in the control and SS groups (P < 0.01 or P < 0.05). It is concluded that SC supplement can increase blood selenium concentration, antioxidation status, and cellular and humoral immunity, and SC has better biological activity than SS in mice.


Assuntos
Antioxidantes/metabolismo , Quitosana/química , Imunidade Celular/efeitos dos fármacos , Imunidade Humoral/efeitos dos fármacos , Compostos de Selênio/química , Selênio/sangue , Selenito de Sódio/química , Animais , Proliferação de Células , Eritrócitos/citologia , Feminino , Glutationa Peroxidase/metabolismo , Hemólise , Interferon gama/sangue , Interferon gama/metabolismo , Interleucina-2/sangue , Interleucina-2/metabolismo , Masculino , Camundongos , Distribuição Aleatória , Selênio/química , Ovinos , Baço/citologia , Superóxido Dismutase
15.
J Agric Food Chem ; 59(21): 11424-31, 2011 Nov 09.
Artigo em Inglês | MEDLINE | ID: mdl-21942342

RESUMO

A 35-day experiment was conducted to evaluate the effect of selenium-enriched probiotics (SP) on laying performance, egg quality, egg selenium (Se) content, and egg glutathione peroxidase (GPX) activity. Five hundred 58-week-old Rohman laying hens were randomly allotted to 5 dietary treatments of 100 each. Each treatment had 5 replicates, and each replicate had 5 cages with 4 hens per cage. The SP was supplemented to a corn-soybean-meal basal diet at 3 different levels that supplied total Se at 0.2, 0.5, and 1.0 mg/kg. The basal diet served as a blank control, while the basal diet with supplemental probiotics served as a probiotics control. The results showed that dietary SP supplementation not only increased (p < 0.05) the rate of egg laying, day egg weight, mean egg weight, egg Se content, and egg GPX activity but also decreased (p < 0.05) the feed:egg ratio and egg cholesterol content. The egg Se content was gradually increased (p < 0.05) along with the increasing level of dietary Se. The SP supplementation also slowed down (p < 0.05) the drop of Haugh units (HU) of eggs stored at room temperature. The egg GPX activity had a positive correlation (p < 0.01) with egg Se content and a negative correlation (p < 0.01) with egg HU drop. These results suggested that Se contents, GPX activity, and HU of eggs were affected by the dietary Se level, whereas the egg-laying performance and egg cholesterol content were affected by the dietary probiotics. It was concluded that this SP is an effective feed additive that combines the organic Se benefit for hen and human health with the probiotics benefit for laying hen production performance. It was also suggested that the eggs from hens fed this SP can serve as a nutraceutical food with high Se and low cholesterol contents for both healthy people and patients with hyperlipidemia, fatty liver, or cardiovascular disease.


Assuntos
Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal , Proteínas Aviárias/análise , Galinhas/fisiologia , Proteínas do Ovo/análise , Ovos/análise , Glutationa Peroxidase/análise , Oviparidade , Probióticos/administração & dosagem , Selênio/análise , Animais , Galinhas/crescimento & desenvolvimento , Suplementos Nutricionais/análise , Feminino , Probióticos/análise
16.
J Trace Elem Med Biol ; 23(1): 29-35, 2009.
Artigo em Inglês | MEDLINE | ID: mdl-19203714

RESUMO

The experiment was conducted to compare the effect of different selenium sources on the expression of glutathione peroxidase 1 (GPx1) and iodothyronine deiodinase 1 (Dio1) mRNA in mice by quantitative real-time PCR. A total of 60 male Kunming mice at average body weight of 20 g were allotted to three groups in a randomized complete block design, namely two treatments and one control. Mice in Group 1 were fed a basal diet as control, while mice in Groups 2 and 3 were fed the basal diet supplemented with 0.1mg/kg selenium as sodium selenite or selenized yeast, respectively. Whole feeding experiment lasted for 30 d. At the end of the feeding trial, liver mRNA levels of GPx1 and Dio1 were determined by quantitative real-time PCR, as well as growth performance, body composition, blood and GPx activity were determined. The results showed that no significant differences in overall growth performance and body composition, including body weight, body length, heart weight, kidney weight and liver weight, were found between the experimental groups (P>0.05). Blood GPx activity increased in all of the selenium supplemented groups compared with control group (P<0.01). However, blood GPx activity in selenized yeast group was higher than that in sodium selenite group (P<0.05). Liver mRNA levels of GPx1 and Dio1 also increased in the two selenium supplemented groups compared with the control group (P<0.05), while there was no significant difference between the sodium selenite and selenized yeast groups (P>0.05). In conclusion, selenium increased the mRNA expression of GPx1 and Dio1 genes in murine liver, and there was no significant difference between the organic or inorganic form of selenium used.


Assuntos
Antioxidantes/administração & dosagem , Glutationa Peroxidase/genética , Iodeto Peroxidase/genética , Fígado/enzimologia , Selenito de Sódio/administração & dosagem , Fermento Seco/administração & dosagem , Animais , Glutationa Peroxidase/metabolismo , Iodeto Peroxidase/metabolismo , Fígado/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos , RNA Mensageiro/metabolismo , Glutationa Peroxidase GPX1
17.
Dig Dis Sci ; 54(2): 246-54, 2009 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-18612820

RESUMO

The purpose of this study was to evaluate the inhibitory activity of selenium-enriched probiotics against pathogenic Escherichia coli (E. coli) in vitro and in vivo. Escherichia coli was co-cultured in vitro with each probiotic strain individually, and a mixture of the four strains and its population was counted at various time points. We also collected a cell-free culture supernatant (CFCS) of each probiotic strain and the four-strain mix to examine their antibacterial activity, using the cylinder plate method. Results demonstrated that co-culture with probiotics significantly reduced the number of E. coli. The different sizes of the inhibition zones made by each CFCS proved that E. coli was inhibited by the metabolites of the probiotics. In vivo, Kunming mice were allocated to different groups supplemented with selenium-enriched and other probiotics. After 28 days, the mice were inoculated with pathogenic E. coli so that we could compare mortality rates and inspect other indexes of each treatment. The mortality of the group with selenium-enriched probiotics was the lowest. In addition, the organic antioxidant status improved, immunity was fortified, and the internal environment of the intestinal tract was enhanced with selenium-enriched probiotic supplementation. In conclusion, selenium-enriched probiotics can strongly antagonize pathogenic E. coli in vitro and in vivo.


Assuntos
Escherichia coli/efeitos dos fármacos , Bactérias Gram-Positivas/metabolismo , Probióticos/farmacologia , Compostos de Selênio/farmacologia , Selênio/metabolismo , Animais , Antioxidantes/metabolismo , Candida/metabolismo , Técnicas de Cocultura , Meios de Cultivo Condicionados/farmacologia , Diarreia/microbiologia , Diarreia/prevenção & controle , Sinergismo Farmacológico , Infecções por Escherichia coli/patologia , Infecções por Escherichia coli/prevenção & controle , Feminino , Trato Gastrointestinal/química , Trato Gastrointestinal/microbiologia , Bactérias Gram-Positivas/crescimento & desenvolvimento , Concentração de Íons de Hidrogênio , Lactobacillus acidophilus/crescimento & desenvolvimento , Lactobacillus acidophilus/metabolismo , Lacticaseibacillus rhamnosus/crescimento & desenvolvimento , Lacticaseibacillus rhamnosus/metabolismo , Dose Letal Mediana , Masculino , Malondialdeído/sangue , Camundongos , Probióticos/uso terapêutico , Selênio/uso terapêutico , Compostos de Selênio/metabolismo , Compostos de Selênio/uso terapêutico , Baço/patologia , Streptococcus thermophilus/crescimento & desenvolvimento , Streptococcus thermophilus/metabolismo , Timo/patologia
18.
Vet Parasitol ; 150(1-2): 13-7, 2007 Nov 30.
Artigo em Inglês | MEDLINE | ID: mdl-17954011

RESUMO

The present study was undertaken to compare the viability and infectivity of Cryptosporidium parvum oocysts that had been stored for 1, 4, 7, 10, 13, 16, 20, 25 and 30 months at 4 degrees C in 2.5% potassium dichromate (Cr) or chlorinated tap water, respectively. An excystation protocol was performed in vitro to evaluate viability. One hundred and eighty female BABL/c mice were used to evaluate the infectivity of oocysts by investigating the prepatent period of C. parvum infection, the quantity of oocysts excreted, and the number of parasites that colonized the villi of the ileum. The results showed that C. parvum oocysts preserved in Cr for 1-16 months or in water for 1-13 months were capable of excystation in vitro and infection of mice. The excystation rates of oocysts and the prepatent periods in mice infected by oocysts stored in Cr and water were not significantly different (p>0.05), and there was a strong correlation between prepatent period and duration of oocyst storage (Cr: R2=0.92; water: R2=0.98). There were no significant differences in oocyst shedding from feces or parasitism of the terminal ilea of mice by Cryptosporidia between the two storage media (p>0.05). In conclusion, C. parvum oocysts may be stored at 4 degrees C in water instead of Cr for the purposes of laboratory research. However, the presence of viable C. parvum oocysts in water is a severe challenge to the drinking water treatment industry.


Assuntos
Sobrevivência Celular/efeitos dos fármacos , Cloro/farmacologia , Cryptosporidium parvum/citologia , Oocistos/efeitos dos fármacos , Oocistos/fisiologia , Dicromato de Potássio/farmacologia , Água/química , Animais , Cloro/química , Cryptosporidium parvum/efeitos dos fármacos , Cryptosporidium parvum/fisiologia , Fezes/parasitologia , Camundongos , Camundongos Endogâmicos BALB C , Preservação Biológica , Soluções , Manejo de Espécimes , Fatores de Tempo
19.
Biol Trace Elem Res ; 116(1): 91-102, 2007 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-17634631

RESUMO

Thirty-two wether lambs of Tan sheep were randomly assigned into four dietary treatment groups (eight per group) for an 8-wk study and then fed a basal diet deficient in Se (0.06 mg/kg) or diets supplemented to provide 0.10 mg/kg Se from sodium selenite, selenized yeast, and seleniumenriched probiotics, respectively. Blood samples were collected at d 0, 28, and 56 of the experiment and tissue samples were collected at experiment termination. Tissue and blood Se concentrations, blood glutathione peroxidase (GSH-Px) activities, and plasma interleukin levels were analyzed. The results showed that the concentrations of Se in the kidney, liver, and muscle increased in all of the supplemented groups (p < 0.01) compared with the control group. However, the Se concentrations in the kidney, liver, and muscle in the groups supplemented with Se yeast and Se-enriched probiotics were higher than those in the group supplemented with sodium selenite (p < 0.01). The activities of GSH-Px and the concentrations of Se in blood also increased in all of the supplemented groups during the period of supplementation (p < 0.01) compared with the control group. The activities of GSH-Px and the concentrations of Se in the whole blood of the lambs fed with selenized yeast and Se-enriched probiotics were higher than those of lambs fed with sodium selenite (p<0.01 or p<0.05). The concentrations of interleukin-1 and interleukin-2 in plasma significantly increased in all of the supplemented groups during the entire period of experiment (p<0.01) compared with the control group, but had no significant differences among all of the supplemented groups. In conclusion, a diet supplemented with Se for finishing lambs was able to increase the concentrations of Se in tissue and blood, activities of GSH-Px in blood, and levels of interleukins in plasma. Organic Se sources (selenized yeast and Se-enriched probiotics) were more effective than the inorganic Se source (sodium selenite) in increasing tissue and blood Se concentrations and blood GSH-Px activities of lambs. However, there were no significant differences in plasma interleukin levels of lambs between organic and inorganic Se sources.


Assuntos
Glutationa Peroxidase/sangue , Interleucinas/sangue , Selênio/sangue , Selênio/farmacologia , Selenito de Sódio/sangue , Selenito de Sódio/farmacologia , Ração Animal , Animais , Suplementos Nutricionais , Glutationa Peroxidase/metabolismo , Probióticos/química , Distribuição Aleatória , Selênio/metabolismo , Compostos de Selênio , Ovinos , Distribuição Tecidual , Oligoelementos/química
20.
J Agric Food Chem ; 55(3): 1027-32, 2007 Feb 07.
Artigo em Inglês | MEDLINE | ID: mdl-17263508

RESUMO

The present study was conducted in a 2 x 4 factorial arrangement in a randomized complete block (RCB) design to compare the effects of a commercial inorganic Se source (sodium selenite, SS) with a commercial organic Se source (Se-enriched yeast, SY) on tissue Se distribution and blood and whole-egg Se concentrations in laying hens. Both Se sources were added into the basal diet at 0, 0.2, 0.5, and 1.0 mg/kg of Se. Seven hundred 68 week old Rohman laying hens were fed with a basal diet containing 0.15 mg/kg DM (dry matter) of Se for 2 weeks, and then, they were allocated randomly into seven groups and were investigated for 28 days. Each group was replicated five times with five cages of four hens per cage in each replicate. During the experiment, two eggs per replicate from each treatment were collected every 7 days and blood was sampled on days 0, 14, and 28 for whole-egg and whole-blood Se analyses. At the end of the experiment, two hens per replicate from each treatment were slaughtered, and muscle (cardiac and breast muscles), liver, spleen, and kidney were sampled for the determination of Se concentrations. The results showed that the addition of Se from either source caused a significant increase in whole-egg and whole-blood Se concentrations (p < 0.01) and Se concentrations in liver, kidney, spleen, and cardiac and breast muscles (p < 0.05) of hens in comparison to the control. Both Se sources and Se levels significantly influenced (p < 0.01) Se concentrations in egg, blood, and the above-mentioned tissues. There was a more significant increase in the Se concentrations in egg (p < 0.01), spleen (p < 0.05), and breast muscle (p < 0.01) and a decrease (p < 0.01) in whole-blood and kidney from hens fed SY than those from hens fed SS. The order of Se distribution was liver > kidney > spleen > cardiac muscle > egg > blood > breast muscle, irrespective of the addition level or source. It was concluded that meat and eggs from hens fed commercial SY are a potential source of Se for humans.


Assuntos
Galinhas/metabolismo , Dieta , Ovos/análise , Selênio/administração & dosagem , Selênio/farmacocinética , Animais , Feminino , Selênio/análise , Selenito de Sódio/administração & dosagem , Distribuição Tecidual
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