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1.
J Immunol ; 212(1): 81-95, 2024 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-38038392

RESUMO

Antimicrobial peptides/proteins (AMPs) constitute a critical component of gut immunity in animals, protecting the gut from pathogenic bacteria. However, the interactions between AMPs and gut microbiota remain elusive. In this study, we show that leukocyte-derived chemotaxin-2 (LECT2)-b, a recently discovered AMP, helps maintain gut homeostasis in grass carp (Ctenopharyngodon idella), one of the major farmed fish species globally, by directly regulating the gut microbiota. Knockdown of LECT2-b resulted in dysregulation of the gut microbiota. Specifically, LECT2-b deficiency led to the dominance of Proteobacteria, consisting of proinflammatory bacterial species, over Firmicutes, which includes anti-inflammatory bacteria. In addition, the opportunistic pathogenic bacteria genus Aeromonas became the dominant genus replacing the probiotic bacteria Lactobacillus and Bacillus. Further analysis revealed that this effect was due to the direct and selective inhibition of certain pathogenic bacterial species by LECT2-b. Moreover, LECT2-b knockdown promoted biofilm formation by gut microbiota, resulting in tissue damage and inflammation. Importantly, LECT2-b treatment alleviated the negative effects induced by LECT2-b knockdown. These findings highlight the crucial role of LECT2-b in maintaining the gut microbiota homeostasis and mucosal health. Overall, our study provides important data for understanding the roles of AMPs in the regulation of gut homeostasis in animals.


Assuntos
Anti-Infecciosos , Microbioma Gastrointestinal , Probióticos , Animais , Bactérias , Homeostase
2.
Carbohydr Polym ; 320: 121188, 2023 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-37659786

RESUMO

Tuning 3D refractive indices of polymers is urgently needed for optical films, but it is quite challenging. Here, we proposed a simple constrained uniaxial stretch method that successfully tuned the 3D refractive indices in cellulose triacetate (TAC) films plasticized with triethyl citrate (TEC). Our results suggest that, under constrained uniaxial stretch, the main chains and side groups prefer to orientate in the stretch direction and the constrained direction, respectively. Such a unique chain arrangement differentiates the refractive indices in three directions of the film. The branched small molecule TEC is also crucial for tuning refractive indices, which promotes chain activity and enhances the chain orientation under stretching, leading to a considerable change in refractive indices before samples fracture. The polymer film we fabricated possesses a direction-dependent optical performance, where the refractive index in the film thickness direction is between that of the stretch direction and constrained direction. This work provides a fundamental understanding on the chain structure and optical performance of polymer films. The constrained uniaxial stretch method, in general, should also be applicable to tuning other 3D physical properties through tuning the direction-dependent orientation of polymers.

3.
Rev Sci Instrum ; 94(2): 023906, 2023 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-36859001

RESUMO

A biaxial stretching device is designed and developed for the real-time structural measurements of polymer films. This device adopts a vertical layout to perform real-time x-ray scattering measurements. It has a maximum stretching ratio of 8 × 8 in two perpendicular directions. Its maximum experimental temperature and stretching rate are 250 °C and 100 mm/s, respectively. The control accuracies of the experimental temperature and stretching rate are ±1 °C and 0.01 mm, respectively. All the parameters related to film biaxial processing, such as stretching speed, stretching ratio, and temperature, can be independently set. The device feasibility is demonstrated via a real-time experiment in a synchrotron radiation beamline. Wide-angle x-ray diffraction, small-angle x-ray scattering, and stress-strain data can be simultaneously obtained during various stretching modes. The proposed device fills the gap between the synchrotron radiation x-ray scattering technique and the biaxial stretching processing of polymer films. This device will play an important role in improving the understanding of the physics behind biaxial polymer processing.

4.
Carbohydr Polym ; 296: 119915, 2022 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-36087971

RESUMO

Cellulose acetate (CA) based films are widely used in liquid crystal displays due to their outstanding transparency, and a certain orientation birefringence of CA films is required when they are used as retardation films. The regulation of orientation birefringence is usually from the perspective of stretch-induced orientation, while the effects of crystallization behaviors of CA films remain obscure. In this study, the roles of crystallization and orientation on the orientation birefringence of CA films were elucidated. For cellulose diacetate films, the orientation birefringence is dominated by the orientation degree. In comparison, apart from the orientation degree, crystallinity is another key variable to regulate the orientation birefringence of cellulose triacetate and plasticized cellulose triacetate films, originating from the birefringence heterogeneity of the crystalline and amorphous phases. These results provide valuable guidelines for the production of CA-based optical films with excellent optical performance.


Assuntos
Cristalização , Birrefringência , Celulose/análogos & derivados
5.
Front Microbiol ; 13: 792532, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35464985

RESUMO

A recent study showed that patients with coronavirus disease 2019 (COVID-19) have gastrointestinal symptoms and intestinal flora dysbiosis. Yeast probiotics shape the gut microbiome and improve immune homeostasis. In this study, an oral candidate of yeast-derived spike protein receptor-binding domain (RBD) and fusion peptide displayed on the surface of the yeast cell wall was generated. The toxicity and immune efficacy of oral administration were further performed in Institute of Cancer Research (ICR) mice. No significant difference in body weights, viscera index, and other side effects were detected in the oral-treated group. The detectable RBD-specific immunoglobulin G (IgG) and immunoglobulin A (IgA) of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and more complex microbiota were detected from oral administration mice compared with those of the control group. Interestingly, the recombinant yeast was identified in female fetal of the high-dose group. These results revealed that the displaying yeast could fulfill the agent-driven immunoregulation and gut microbiome reconstitution. The findings will shed light on new dimensions against SARS-CoV-2 infection with the synergistic oral agents as promising non-invasive immunization and restoring gut flora.

6.
Biol Trace Elem Res ; 200(3): 1127-1139, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33895963

RESUMO

Renal fibrosis is the final result of the progression of chronic kidney disease (CKD) to end-stage renal disease (ESRD). Earlier studies confirmed that selenium (Se) displays a close association with kidney diseases. However, the correlation between Se and fibrosis has rarely been explored. Thus, this article mainly aimed to investigate the effect of Se deficiency on renal fibrosis and the Wnt/ß-catenin signaling pathway. Twenty BALB/c mice were fed a diet containing 0.02-mg/kg Se (Se-deficient diet) or 0.18-mg/kg Se (standard diet) for 20 weeks. A human glomerular mesangial cell (HMC) cell line was transfected with lentiviral TRNAU1AP-shRNA vector to establish a stable Se deficiency model in vitro. As indicated in this study, the glutathione (GSH) content in the Se-deficient group displayed an obvious decline compared with that in the control group, whereas the content of malondialdehyde (MDA) was obviously elevated. The results of Masson staining showed fibrosis around the renal tubules, and the results of immunohistochemistry showed that the area of positive fibronectin expression increased. In the Se-deficient group, the levels of collagen I, collagen III, matrix metalloproteinase 9 (MMP9), and other fibrosis-related proteins changed significantly in vivo and in vitro. Compared with the control group, the TRNAU1AP-shRNA group showed markedly reduced cell proliferation and migration abilities. Our data indicate that Se deficiency can cause kidney damage and renal fibrosis. Furthermore, the Wnt pathway is critical for the development of tissue and organ fibrosis. The data of this study demonstrated that the expression of Wnt5a, ß-catenin, and dishevelled 1 (Dvl-1) was significantly upregulated in the Se-deficient group. Therefore, the Wnt/ß-catenin pathway may play an important role in renal fibrosis caused by Se deficiency.


Assuntos
Insuficiência Renal Crônica , Selênio , Via de Sinalização Wnt , Animais , Fibrose , Túbulos Renais/patologia , Camundongos , Insuficiência Renal Crônica/patologia , beta Catenina/metabolismo
7.
Front Oncol ; 11: 791943, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34926308

RESUMO

BACKGROUND: Breast cancer (BRCA) is a malignant tumor with a high mortality rate and poor prognosis in patients. However, understanding the molecular mechanism of breast cancer is still a challenge. MATERIALS AND METHODS: In this study, we constructed co-expression networks by weighted gene co-expression network analysis (WGCNA). Gene-expression profiles and clinical data were integrated to detect breast cancer survival modules and the leading genes related to prognostic risk. Finally, we introduced machine learning algorithms to build a predictive model aiming to discover potential key biomarkers. RESULTS: A total of 42 prognostic modules for breast cancer were identified. The nomogram analysis showed that 42 modules had good risk assessment performance. Compared to clinical characteristics, the risk values carried by genes in these modules could be used to classify the high-risk and low-risk groups of patients. Further, we found that 16 genes with significant differential expressions and obvious bridging effects might be considered biological markers related to breast cancer. Single-nucleotide polymorphisms on the CYP24A1 transcript induced RNA structural heterogeneity, which affects the molecular regulation of BRCA. In addition, we found for the first time that ABHD11-AS1 was significantly highly expressed in breast cancer. CONCLUSION: We integrated clinical prognosis information, RNA sequencing data, and drug targets to construct a breast cancer-related risk module. Through bridging effect measurement and machine learning modeling, we evaluated the risk values of the genes in the modules and identified potential biomarkers for breast cancer. The protocol provides new insight into deciphering the molecular mechanism and theoretical basis of BRCA.

8.
Front Vet Sci ; 8: 727166, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34660764

RESUMO

Tick-borne diseases (TBDs) can cause serious economic losses and are very important to animal and public health. To date, research on TBDs has been limited in Qinghai-Tibet Plateau, China. This epidemiological investigation was conducted to evaluate the distribution and risk factors of Anaplasma spp. and Ehrlichia chaffeensis in livestock in Qinghai. A total of 566 blood samples, including 330 yaks (Bos grunniens) and 236 Tibetan sheep (Ovis aries) were screened. Results showed that A. bovis (33.3%, 110/330) and A. phagocytophilum (29.4%, 97/330) were most prevalent in yaks, followed by A. ovis (1.2%, 4/330), A. capra (0.6%, 2/330), and E. chaffeensis (0.6%, 2/330). While A. ovis (80.9%, 191/236) and A. bovis (5.1%, 12/236) infection was identified in Tibetan sheep. To our knowledge, it is the first time that A. capra and E. chaffeensis have been detected in yaks in China. Apart from that, we also found that co-infection of A. bovis and A. phagocytophilum is common in yaks (28.2%, 93/330). For triple co-infection, two yaks were infected with A. bovis, A. phagocytophilum, and A. capra, and two yaks were infected with A. bovis, A. phagocytophilum, and E. chaffeensis. Risk analysis shows that infection with A. bovis, A. phagocytophilum, and A. ovis was related to region and altitude. This study provides new data on the prevalence of Anaplasma spp. and E. chaffeensis in Qinghai, China, which may help to develop new strategies for active responding to these pathogens.

9.
Am J Transl Res ; 13(8): 9586-9592, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34540082

RESUMO

PURPOSE: To explore the effect of Etoricoxib on serum miR-214 expression level and inflammatory reaction in patients with knee osteoarthritis. METHODS: 96 patients with knee osteoarthritis admitted to our hospital (January 2019 to January 2020) were selected. 48 patients in the control group received Celecoxib and 48 patients in the observation group received Etoricoxib. The treatment effect, knee function, inflammatory factor level, immune function, and serum miR-214 expression level of the two groups were compared. 6 months after treatment, the incidence of complications (deformities, deep infections and severe pain) between the two groups was compared. RESULTS: (1) The observation group had a higher total effective rate (93.75%) in comparison to the control group (72.92%) (P<0.05). (2) Before treatment, the serum miR-214 expression level of the two groups was basically the same (P>0.05). After treatment, the serum miR-214 expression level of the two groups decreased significantly, with a more marked decrease in the observation group (P<0.05). (3) Before treatment, the levels of IL-1ß, TNF-α, and hs-CRP were not statistically different in the two groups (P>0.05). After treatment, IL-1ß, TNF-α, and hs-CRP in both groups decreased, and the decrease in the observation group was significantly greater (P<0.05). (4) Before treatment, the levels of CD3+CD8+ and CD3+ were basically the same in both groups (P>0.05). After treatment, the levels of CD3+CD8+ and CD3+ in the two groups increased, and for the observation group, were significantly greater P<0.05. (5) The Lysholm score was higher in the observation group than it was in the control group (inter-group effect: F = 58.070, P<0.001), and the Lysholm score of both groups tended to increase with time (time effect: F = 145.900, P<0.001). Grouping and time showed an interactive effect (interactive effect: F = 8.646, P<0.001). 6 months after treatment, observation group showed a lower complication rate when compared to the control groupt (P<0.05). CONCLUSION: Etoricoxib has a strong effect on patients with knee osteoarthritis. It can significantly reduce the expression of serum miR-214 and the level of inflammatory factors, and is worthy of clinical application.

11.
J AOAC Int ; 104(5): 1389-1393, 2021 Sep 27.
Artigo em Inglês | MEDLINE | ID: mdl-33769495

RESUMO

BACKGROUND: The diseases caused by the Capripoxvirus species have very similar symptoms and are difficult to distinguish clinically. According to a recent report, Capripoxvirus are not strictly host specific. OBJECTIVE: This study aimed to identify the viruses from ovine (include sheep and goat) or bovine, which will assist in selecting the appropriate vaccine and correct measures to control diseases. METHOD: Universal primers for all Capripoxvirus and specific probes for lumpy skin disease virus, sheeppox virus, and goatpox virus were designed and analyzed to identify the viruses from ovine (including sheep and goats) or bovine species. The parameters of the system, such as the annealing temperatures and the quantities of primers and probes used, were optimized. The sensitivity, specificity, and reproducibility were tested. RESULTS: Each probe showed a specific fluorescent signal, with no cross reaction with other pathogens that cause symptoms similar to those of the poxviruses. The LOD was 102 copies of the target genome DNA. The 557 local clinical samples and samples from Ethiopia were successfully detected and the results were consistent with a restriction fragment length polymorphism PCR analysis of the P32 and RPO30 genes and gene sequencing. CONCLUSIONS: This optimized real-time PCR detection system has good diagnostic sensitivity and specificity and can be used for the rapid and effective differential diagnosis of these diseases in goats, sheep, and cattle. HIGHLIGHTS: It is a rapid detection method to distinguish the viruses from ovine (include sheep and goat) or bovine.


Assuntos
Capripoxvirus , Doenças das Cabras , Vírus da Doença Nodular Cutânea , Infecções por Poxviridae , Doenças dos Ovinos , Animais , Capripoxvirus/genética , Bovinos , Doenças das Cabras/diagnóstico , Cabras , Vírus da Doença Nodular Cutânea/genética , Infecções por Poxviridae/diagnóstico , Infecções por Poxviridae/veterinária , Reação em Cadeia da Polimerase em Tempo Real , Reprodutibilidade dos Testes , Ovinos , Doenças dos Ovinos/diagnóstico
12.
Pathogens ; 10(2)2021 Feb 22.
Artigo em Inglês | MEDLINE | ID: mdl-33671750

RESUMO

Melophagus ovinus (sheep ked) is a hematophagous ectoparasite that mainly parasitizes sheep. In addition to causing inflammation, wool loss, and skin damage to the animal hosts, M. ovinus also serves as a vector for a variety of pathogens and is highly likely to participate in the life and transmission cycle of pathogenic organisms. Herein, we investigated the presence and molecular characterization of vector-borne pathogens in M. ovinus from Qinghai-Tibet Plateau, China. A total of 92 M. ovinus pools collected from the Qinghai province of China were screened for the presence of selected vector-borne pathogens. The overall positive rate of A. ovis, A. bovis, A. phagocytophilum, and T. ovis in M. ovinus was 39.1%, 17.4%, 9.8%, and 89.1%, respectively. All of the samples were negative for Border disease virus (BDV), other Anaplasma species, Babesia spp., Rickettsia spp., and Borrelia spp. Co-infection of different Anaplasma species and T. ovis occurred in 51.2% of all samples with T. ovis. The positive rates of A. ovis, A. bovis, and A. phagocytophilum in different regions and altitudes of the sampling sites were significantly different. Sequence and phylogenetic analysis of target genes confirmed their identity with corresponding pathogens. Our results elucidate the occurrence and molecular characterization of Anaplasma spp. and Theileria spp. in M. ovinus, which could act as potential zoonotic reservoirs. To the best of our knowledge, this is the first report of the detection of A. bovis and A. phagocytophilum DNA in M. ovinus. This study gives the first extensive molecular survey of vector-borne pathogens with veterinary and public health significance in M. ovinus from the Qinghai-Tibet Plateau, China.

13.
BMC Vet Res ; 17(1): 48, 2021 Jan 23.
Artigo em Inglês | MEDLINE | ID: mdl-33485335

RESUMO

BACKGROUND: Brucellosis is a serious zoonosis disease that frequently causes significant economic loss in animal husbandry and threatens human health. Therefore, we established a rapid, accurate, simple and sensitive fluorescent immunochromatographic strip test (ICST) based on quantum dots (QDs) for detection the antibodies of Brucella infection animals serum. RESULTS: The test strips were successfully prepared by quantum dot fluorescent microspheres (QDFM) as tracers, which were covalently coupled to an outer membrane protein of Brucella OMP22. The outer membrane protein OMP28 and monoclonal antibodies of OMP22 were separately dispensed onto a nitrocellulose membrane as test and quality control lines, respectively. The critical threshold for determining negative or positive through the ratio of the fluorescent signal of the test line and the control line (HT / HC) is 0.0492. The repeatability was excellent with an overall average CV of 8.78%. Under optimum conditions, the limit of detection was 1.05 ng/mL (1:512 dilution). With regard to the detection of brucellosis in 150 clinical samples, the total coincidence rate of ICST and Rose Bengal plate test (RBPT) was 97.3%, the coincidence rate of positive samples was 98.8%, the coincidence rate of negative samples was 95.3%, the sensitivity of RBPT is 1:32, and no cross reaction with the sera of other related diseases was observed. CONCLUSION: In our present study, the QDFM has promising application for on-site screening of brucellosis owing to its high detection speed, high sensitivity, high specificity and low cost.


Assuntos
Brucella/imunologia , Brucelose/veterinária , Pontos Quânticos/química , Animais , Anticorpos Antibacterianos/química , Anticorpos Antibacterianos/imunologia , Anticorpos Monoclonais/química , Anticorpos Monoclonais/imunologia , Brucelose/diagnóstico , Brucelose/imunologia , Bovinos , Cromatografia de Afinidade/métodos , Cromatografia de Afinidade/veterinária , Cabras , Microesferas , Fitas Reagentes , Sensibilidade e Especificidade , Ovinos
14.
Bioorg Chem ; 100: 103886, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32371249

RESUMO

Chemical modifications on the A ring of limonin (1) and deoxylimonin (2) afforded 28 structural characterized derivatives, which were firstly subjected to preliminary in vivo analgesic and anti-inflammatory screen by mice model. The most promising candidate, deoxylimonin analog II-B-2 (70 mg/kg) with 3,4-dimethoxyphenylethyl moiety substitued δ-lactam in the A ring, exhibited better analgesic activity than aspirin (200 mg/kg) and stronger anti-inflammatory efficacy than naproxen (150 mg/kg). Further in vivo evaluation confirmed its advantage over limonin and showed dose-response dependent manner, and follow-up research suggested that the anti-inflammatory effect of compound II-B-2 may be attributed to the downregulation of cyclooxygenase 2 expression and the suppression of prostaglandin E2 formation.


Assuntos
Analgésicos/química , Analgésicos/farmacologia , Anti-Inflamatórios não Esteroides/química , Anti-Inflamatórios não Esteroides/farmacologia , Limoninas/química , Limoninas/farmacologia , Analgésicos/uso terapêutico , Animais , Anti-Inflamatórios não Esteroides/uso terapêutico , Ciclo-Oxigenase 2/metabolismo , Dinoprostona/metabolismo , Modelos Animais de Doenças , Edema/tratamento farmacológico , Edema/metabolismo , Feminino , Lactamas/química , Lactamas/farmacologia , Lactamas/uso terapêutico , Limoninas/uso terapêutico , Masculino , Camundongos , Camundongos Endogâmicos ICR , Dor/tratamento farmacológico , Dor/metabolismo , Ratos Endogâmicos Lew
15.
Microorganisms ; 8(4)2020 Apr 04.
Artigo em Inglês | MEDLINE | ID: mdl-32260465

RESUMO

Edwardsiella piscicida, a facultative aerobic pathogen belonging to the Enterobacteriaceae family, is the etiological agent of edwardsiellosis that causes significant economic loses in the aquaculture industry. cAMP receptor protein (CRP) is one of the most important transcriptional regulators, which can regulate large quantities of operons in different bacteria. Here we characterize the crp gene and report the effect of a crp deletion in E. piscicida. The crp-deficient mutant lost the capacity to utilize maltose, and showed significantly reduced motility due to the lack of flagella synthesis. We further constructed a ΔPcrp mutant to support that the phenotype above was caused by the crp deletion. Evidence obtained in fish serum killing assay and competitive infection assay strongly indicated that the inactivation of crp impaired the ability of E. piscicida to evade host immune clearance. More importantly, the virulence of the crp mutant was attenuated in both zebrafish and channel catfish, with reductions in mortality rates. In the end, we found that crp mutant could confer immune protection against E. piscicida infection to zebrafish and channel catfish, indicating its potential as a live attenuated vaccine.

16.
Korean J Parasitol ; 58(1): 37-46, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-32145725

RESUMO

Livestock husbandry is vital to economy of the Tarim Basin, Xinjiang Autonomous Region, China. However, there have been few surveys of the distribution of ixodid ticks (Acari: Ixodidae) and tick-borne pathogens affecting domestic animals at these locations. In this study, 3,916 adult ixodid ticks infesting domestic animals were collected from 23 sampling sites during 2012-2016. Ticks were identified to species based on morphology, and the identification was confirmed based on mitochondrial 16S and 12S rRNA sequences. Ten tick species belonging to 4 genera were identified, including Rhipicephalus turanicus, Hyalomma anatolicum, Rh. bursa, H. asiaticum asiaticum, and Rh. sanguineus. DNA sequences of Rickettsia spp. (spotted fever group) and Anaplasma spp. were detected in these ticks. Phylogenetic analyses revealed possible existence of undescribed Babesia spp. and Borrelia spp. This study illustrates potential threat to domestic animals and humans from tick-borne pathogens.


Assuntos
Ácaros e Carrapatos/genética , Ácaros e Carrapatos/patogenicidade , Animais Domésticos/parasitologia , Análise de Sequência de DNA , Doenças Transmitidas por Carrapatos/parasitologia , Animais , China , RNA Ribossômico , RNA Ribossômico 16S
17.
RSC Med Chem ; 11(7): 843-847, 2020 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-33479680

RESUMO

A practical synthetic route, consisting of 5 steps, has been developed and applied successfully for converting limonin/deoxylimonin into the corresponding amino derivatives I- 5a-I- 5e and II- 5a-II- 5e. Deoxylimonin analogs II- 5a and II- 5b bearing an open A ring structure underwent ring closure reaction by employing the Mitsunobu procedure to afford II- 6a and II- 6b. All of the 12 newly synthesized compounds were subjected to preliminary screening for evaluating their inflammation and nociception inhibition efficacy. The most promising compounds, I- 5b and II- 5d, were selected for further investigation by a carrageenan-induced mouse paw edema model, both of which displayed a dose-response dependent manner and demonstrated superior anti inflammation capacity to that of indomethacin in the first 2 hours.

18.
Fish Shellfish Immunol ; 97: 135-145, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31846774

RESUMO

Interleukin-1 receptor-associated kinase 1 (IRAK1) and IRAK4 are critical signalling mediators and play pivotal roles in the innate immune and inflammatory responses mediated by TLR/IL-1R. In the present study, two IRAK family members, OnIRAK1 and OnIRAK4, were identified in the Nile tilapia Oreochromis niloticus with a conserved N-terminal death domain and a protein kinase domain, similar to those of other fishes and mammals. The gene structures of OnIRAK1 and OnIRAK4 are organized into fifteen exons split by fourteen introns and ten exons split by nine introns. OnIRAK1 and OnIRAK4 were broadly expressed in all of the tissues tested, with the highest expression levels being observed in the blood and the lowest expression levels being observed in the liver. Both genes could be detected from 2 d post-fertilization (dpf) to 8 dpf during embryonic development. Moreover, the expression levels of OnIRAK1 and OnIRAK4 were clearly altered in all five tissues after Streptococcus agalactiae infection in vivo and could be induced by LPS, Poly I: C, S. agalactiae WC1535 and △CPS in Nile tilapia macrophages. The overexpression of OnIRAK1 and OnIRAK4 in 293T cells showed that they were both distributed in the cytoplasm and could significantly increase NF-κB activation. Interestingly, after transfection, OnIRAK1 significantly upregulated OnMyd88-induced NF-κB activation, while OnIRAK4 had no effect on OnMyd88-induced NF-κB activation. Co-immunoprecipitation (Co-IP) assays showed that OnMyd88 did not interact with either OnIRAK1 or OnIRAK4 and that OnIRAK1 did not interact with OnIRAK4. Taken together, these findings suggest that OnIRAK1 and OnIRAK4 could play important roles in TLR/IL-1R signalling pathways and the immune response to pathogen invasion.


Assuntos
Ciclídeos/genética , Ciclídeos/imunologia , Proteínas de Peixes/genética , Quinases Associadas a Receptores de Interleucina-1/genética , Quinases Associadas a Receptores de Interleucina-1/metabolismo , Infecções Estreptocócicas/veterinária , Animais , Clonagem Molecular , Regulação para Baixo , Doenças dos Peixes/imunologia , Doenças dos Peixes/microbiologia , Proteínas de Peixes/imunologia , Regulação da Expressão Gênica , Imunidade Inata , Filogenia , Alinhamento de Sequência , Transdução de Sinais/imunologia , Infecções Estreptocócicas/imunologia , Streptococcus agalactiae , Regulação para Cima
19.
J Fish Biol ; 96(1): 185-193, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31721203

RESUMO

In this study, we cloned the complementary (c)DNA sequences of tumour necrosis factor receptor (TNFR)-associated factor 3 (traf3) in Nile tilapia, Oreochromis niloticus. The expression patterns of the traf3 gene were investigated and preliminary functional analyses were performed. In healthy fish, traf3 transcript was broadly expressed in all examined tissues, with the highest expression level in the blood and the lowest in the liver. The traf3 gene reached its highest expression at 8 days post-fertilisation (dpf) during embryonic development. Moreover, we found that expression of traf3 was clearly altered following stimulation with Streptococcus agalactiae in vivo and that traf3 could be induced by lipopolysaccharides (LPS), Poly I: C and S. agalactiae WC1535 in Nile tilapia macrophages. Overexpression in 293T cells showed that Traf3 protein was mainly distributed in the cytoplasm and could significantly increase nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) activation. Taken together, these results implied that traf3 could play important roles in the immune response to pathogen invasion.


Assuntos
Ciclídeos/imunologia , NF-kappa B/metabolismo , Fator 3 Associado a Receptor de TNF , Animais , Técnicas de Cultura de Células , Ciclídeos/genética , Doenças dos Peixes/imunologia , Proteínas de Peixes/genética , Expressão Gênica , Lipopolissacarídeos/imunologia , Macrófagos/imunologia , Macrófagos/metabolismo , Macrófagos/microbiologia , NF-kappa B/imunologia , Streptococcus agalactiae/imunologia , Fator 3 Associado a Receptor de TNF/genética , Fator 3 Associado a Receptor de TNF/imunologia , Fator 3 Associado a Receptor de TNF/metabolismo
20.
Appl Microbiol Biotechnol ; 103(21-22): 9023-9035, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31654082

RESUMO

Streptococcus agalactiae is a major pathogen causing streptococcosis. To prevent and control this bacterial disease, antagonistic bacteria have become a new research hotspot. This study evaluated the probiotic potential of Bacillus velezensis LF01 strain, which is antagonistic to S. agalactiae. The active compounds produced by LF01 showed antimicrobial activity against a broad spectrum of fish pathogens, including S. agalactiae, Streptococcus iniae, Aeromonas hydrophila, Edwardsiella tarda, Edwardsiella ictaluri, Aeromonas schubertii, Aeromonas veronii, Aeromonas jandaei, and Vibrio harveyi. The antimicrobial compounds were heat stable, pH stable, UV stable, resistant to proteases, and could be stored for a long time. To evaluate the probiotic function of LF01 in Nile tilapia, juveniles were divided into three treatment groups: a control group, an interval feeding group, and a continuous feeding group. Tilapia fed with LF01-supplemented diets (1.0 × 109 CFU/g) showed significantly better growth performances than those of the control group (P < 0.05). Tilapia fed with LF01-supplemented diets significantly increased lysozyme (LZY) and superoxide dismutase (SOD) activities. The expression of three immune-related genes (C3, lyzc, and MHC-IIß) was higher in the intestine, head kidney, and gill of tilapia from the continuous feeding group than in those from the control group (P < 0.05). Tilapia fed with LF01-supplemented diets showed remarkably improved survival rates after S. agalactiae infection, and analysis of their intestinal tract pathogens revealed that the abundance of Edwardsiella and Plesiomonas had significantly decreased compared with the control group. Our findings demonstrate that LF01 is an effective antagonist against various fish pathogens and has potential for controlling infections by Streptococcus spp. and other pathogens in tilapia.


Assuntos
Antibiose/fisiologia , Bacillus/fisiologia , Agentes de Controle Biológico/farmacologia , Ciclídeos/microbiologia , Infecções Estreptocócicas/prevenção & controle , Streptococcus agalactiae/fisiologia , Animais , Doenças dos Peixes/microbiologia , Doenças dos Peixes/prevenção & controle , Sequenciamento de Nucleotídeos em Larga Escala , Probióticos/farmacologia , Infecções Estreptocócicas/veterinária
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