scholarly journals Molecular characterization and immune protection of the 3-hydroxyacyl-CoA dehydrogenase gene in Echinococcus granulosus

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Jinwen Xian ◽  
Ning Wang ◽  
Pengpeng Zhao ◽  
Yanyan Zhang ◽  
Jimeng Meng ◽  
...  
Keyword(s):  
Echinococcus Granulosus ◽  
Interleukin 4 ◽  
Effective Vaccine ◽  
Control Group ◽  
Protective Effects ◽  
Immune Protection ◽  
Reading Frame ◽  
Reverse Transcription Pcr ◽  
Frame Sequence ◽  
Hydroxyacyl Coa Dehydrogenase

Abstract Background Cystic echinococcosis (CE) is a serious parasitic zoonosis caused by the larvae of the tapeworm Echinococcus granulosus. The development of an effective vaccine is one of the most promising strategies for controlling CE. Methods The E. granulosus 3-hydroxyacyl-CoA dehydrogenase (EgHCDH) gene was cloned and expressed in Escherichia coli. The distribution of EgHCDH in protoscoleces (PSCs) and adult worms was analyzed using immunofluorescence. The transcript levels of EgHCDH in PSCs and adult worms were analyzed using quantitative real-time reverse transcription PCR (RT-qPCR). The immune protective effects of the rEgHCDH were evaluated. Results The 924-bp open reading frame sequence of EgHCDH, which encodes a protein of approximately 34 kDa, was obtained. RT-qPCR analysis revealed that EgHCDH was expressed in both the PSCs and adult worms of E. granulosus. Immunofluorescence analysis showed that EgHCDH was mainly localized in the tegument of PSCs and adult worms. Western blot analysis showed that the recombinant protein was recognized by E. granulosus-infected dog sera. Animal challenge experiments demonstrated that dogs immunized with recombinant (r)EgHCDH had significantly higher serum IgG, interferon gamma and interleukin-4 concentrations than the phosphate-buffered saline (PBS) control group. The rEgHCDH vaccine was able to significantly reduce the number of E. granulosus and inhibit the segmental development of E. granulosus compared to the PBS control group. Conclusions The results suggest that rEgHCDH can induce partial immune protection against infection with E. granulosus and could be an effective candidate for the development of new vaccines. Graphical abstract

scholarly journals Molecular Characterization and Expression Analysis of the Gene Encoding 3-Hydroxyacyl-CoA Dehydrogenase (EGR-03347) from Echinococcus Granulosus and the Evaluation of the Immune Protection of the Definitive Hosts (dogs)

2021 ◽  
Author(s):  
Jinwen Xian ◽  
Ning Wang ◽  
Pengpeng Zhao ◽  
Yanyan Zhang ◽  
Jimeng Meng ◽  
...  
Keyword(s):  
Echinococcus Granulosus ◽  
Post Infection ◽  
Tricarboxylic Acid Cycle ◽  
Interleukin 4 ◽  
Effective Vaccine ◽  
Control Group ◽  
Gene Encoding ◽  
Transcript Levels ◽  
Key Enzyme ◽  
Hydroxyacyl Coa Dehydrogenase

Abstract Background: Cystic echinococcosis, a serious parasitic zoonosis, is caused by tapeworm (Echinococcus granulosus) larvae. The development of an effective vaccine is a promising strategy to control echinococcosis. E. granulosus has a complete tricarboxylic acid cycle pathway, in which 3-hydroxyacyl-CoA dehydrogenase (EGR-03347) is a key enzyme.Methods: In the present study, the cDNA encoding EGR-03347 in Echinococcus granulosus (rEGR-03347) was successfully cloned and the molecular and biochemical characterizations carried out. The immunoreactivity of recombinant EGR-03347 (rEGR-03347) was investigated using western blotting. The immunolocalization of EGR-03347 in different life stages of E. granulosus was determined using specific polyclonal antibody, quantitative real-time reverse transcription polymerase chain reaction was used to analyze their transcript levels in PSCs and 28-day strobilated worms stages. In addition, recombinant protein rEGR-03347 was mixed with the adjuvant Quil A for vaccinating dogs, after three vaccine injections, all the dogs were orally challenge-infected with 100000 protoscoleces of E. granulosus. After 28 days of infection, all the dogs were euthanized and necropsied for collecting and counting E. granulosus worms, post-infection the antibody and cytokine were measured for the immunogenicity analysis of this protein.Results: rEGR-03347 is a highly conserved protein, consisting of 308 amino acids. Recombinant EGR-03347 could be identifed in the sera of patients with CE and in mouse anti-rEGR-03347 sera. Immunofluorescence analysis showed that EGR-03347 mainly localized in the tegument of protoscoleces and adults, and their transcript levels were high in the 28-day strobilated worms. Furthermore, enzyme-linked immunosorbent assays post‑infection showed that IgG gradually increased after the first immunization with rEGR-03347 compared with the control group, rEGR-03347 changed the interferon gamma and interleukin 4 levels. We observed an 87.2 % reduction in E. granulosus numbers and 66.7 % inhibition of the segmental development of E. granulosus in the rEGR‑03347‑vaccinated dogs compared with the nonvaccinated controls.Conclusions: This is the first report characterizing a 3-hydroxyacyl-CoA dehydrogenase from the tapeworm E. granulosus. We have characterized the sequence, structure and location of EGR‑03347 and investigated the immunoreactivity, immunogenicity and serodiagnostic potential of rEGR‑03347 . The results demonstrated rEGR-03347 as a potential vaccine against E. granulosus infection in dogs.

scholarly journals Molecular Characterization of a Tetraspanin TSP11 Gene in Echinococcus granulosus and Evaluation Its Immunoprotection in Model Dogs

2021 ◽  
Vol 8 ◽  
Author(s):  
Jinwen Xian ◽  
Pengpeng Zhao ◽  
Ning Wang ◽  
Weiye Wang ◽  
Yanyan Zhang ◽  
...  
Keyword(s):  
Echinococcus Granulosus ◽  
Vaccine Development ◽  
Serum Levels ◽  
Disease Diagnosis ◽  
Vaccine Antigen ◽  
Interleukin 4 ◽  
Control Group ◽  
Potential Candidate ◽  
Intermediate Hosts ◽  
Wide Range

Cystic echinococcosis (CE) is a cosmopolitan zoonosis caused by the larval stage of Echinococcus granulosus, which affects humans and a wide range of mammalian intermediate hosts. Parasite tetraspanin proteins are crucial for host-parasite interactions, and therefore they may be useful for vaccine development or disease diagnosis. In the present study, the major antigen coding sequence of tetraspanin 11 (Eg-TSP11) from E. granulosus was determined. The results of immunolocalization showed that Eg-TSP11 was mainly located in the tegument of adult worms and protoscoleces. Western blotting analysis showed that the serum from dogs injected with recombinant Eg-TSP11 (rEg-TSP11) could recognize Eg-TSP11 among natural protoscolex proteins. Moreover, the serum from dogs with E. granulosus infection also recognized rEg-TSP11. Serum indirect enzyme-linked immunosorbent assays demonstrated that IgG levels gradually increased after the first immunization with rEg-TSP11 compared with those in the control group. Furthermore, the serum levels of interleukin 4, interleukin 5, and interferon gamma were significantly altered in the rEg-TSP11 group. Importantly, we found that vaccination with rEg-TSP11 significantly decreased worm burden and inhibited segment development in a dog model of E. granulosus infection. Based on these findings, we speculated that rEg-TSP11 might be a potential candidate vaccine antigen against E. granulosus infection in dogs.

scholarly journals Role of essential oils in antioxidant capacity and immunity in a rat model of mixed stress

2021 ◽  
Vol 51 (4) ◽  
pp. 426-436
Author(s):  
N. Seyidoglu ◽  
E. Koseli ◽  
R. Gurbanli ◽  
C. Aydin
Keyword(s):  
Essential Oils ◽  
Water Consumption ◽  
Rat Model ◽  
Animal Feed ◽  
Biological Effects ◽  
Interleukin 4 ◽  
Control Group ◽  
Protective Effects ◽  
Sprague Dawley ◽  
Stressed Group

Animal wellbeing is a balance between environmental stress and nutrition that regulates homeostasis. Augmentation of animal feed with essential oils can promote homeostasis. The present study was designed to observe the biochemical, immunological, and biological effects of daily administration of a mixture of essential oils (EOM) in a stressed rat model. Forty-eight adult male Sprague Dawley rats were  randomly allocated to four groups, namely a control group (C), a stressed group (S), a treated group (Tr), and a stressed group that  received the treatment (TrS). The treatment was applied by adding EOM to the water (0.2 ml/l) three days per week for 28 days. Two  chronic stressors (isolation and crowding) were applied to animals in groups S and TrS. Total oxidant status (TOS) increased in the S group  compared with C, whereas it decreased when fed with EOM. Although TOS was the same in S and C, it increased in Tr compared with C. There was a significant increase in interleukin 4 (IL-4) in S compared with C, and EOM reversed the IL-4 level. Nevertheless, an increase was seen in the weights of the liver, intestine, brain, and testes in TrS compared with S. The increase in water intake was a result of stress, but feeding with EOM decreased water consumption gradually. This study showed that 0.2 ml/l EOM had protective effects on antioxidant status, immunity and liver function, and decreased water consumption under stress conditions.

Recombinant ferritin protects mice against challenge with Echinococcus granulosus ?

2009 ◽  
Vol 54 (4) ◽  
Author(s):  
Yana Wang ◽  
Zongji Li ◽  
Zhaoyu Li ◽  
Yang Bo ◽  
Wei Zhao
Keyword(s):  
Echinococcus Granulosus ◽  
Protective Efficacy ◽  
Effective Vaccine ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Intracellular Iron ◽  
Promising Candidate ◽  
Iron Storage ◽  
Iron Storage Protein

AbstractFerritin is well known as the main intracellular iron storage protein in both prokaryotes and eukaryotes, keeping it in a soluble and non-toxic form, though the role of ferritin as a vaccine candidate in echinococcosis has not yet been delineated. Through our study, ferritin was cloned from Echinococcus granulosus and expressed in Escherichia coli. The recombinant E. granulosus ferritin (rEgferritin) has a molecular weight of 19 kDa and could be recognized by anti-mice serum in Western blotting. The specific antibody was detected by enzyme-linked immunosorbent assay. Mice vaccinated with rEgferritin and challenged intraperitoneally with E. granulosus protoscoleces revealed significant protective efficacy up to 85.6%, compared with the control group. Thus, rEgferritin could be a promising candidate as an effective vaccine to prevent the infection of echinococcosis.

scholarly journals Characterization of p53 From the Marine Crab Portunus trituberculatus and Its Functions Under Low Salinity Conditions

2021 ◽  
Vol 12 ◽  
Author(s):  
Xianyun Ren ◽  
Lei Wang ◽  
Yao Xu ◽  
Qiong Wang ◽  
Jianjian Lv ◽  
...  
Keyword(s):  
Salinity Stress ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Portunus Trituberculatus ◽  
Control Group ◽  
Swimming Crab ◽  
Reading Frame ◽  
Protein Levels ◽  
Low Salinity ◽  
Reverse Transcription Pcr ◽  
The Guardian

Portunus trituberculatus, or the swimming crab, is tolerant of reduced salinity; however, the molecular mechanism of this tolerance is not clear. Cells can be damaged by hyperosmotic salinity. The protein p53, sometimes referred to as “the guardian of the genome,” displays versatile and important functions under changing environmental conditions. Herein, the P. trituberculatus p53 gene (designated as Ptp53) was cloned and studied. The full-length Ptp53 cDNA comprised 1,544bp, with a 1,314bp open reading frame, which encodes a putative polypeptide of 437 amino acids. Quantitative real-time reverse transcription PCR assays revealed ubiquitous expression of Ptp53 in all tissues examined, with the gills showing the highest expression level. Extensive apoptosis was detected under low salinity conditions using terminal deoxynucleotidyl transferase nick-end-labeling staining. Oxidative stress was induced under low salinity conditions, consequently leading to apoptosis. Low salinity stress caused significant upregulation of Ptp53 mRNA and protein levels in the gills. Moreover, compared with that in the control group, the mortality of Ptp53-silenced crabs under low salinity stress was enhanced significantly. Taken together, our findings suggest that Ptp53, via regulation of apoptosis and antioxidant defense, played important functions in the low salinity stress response of the swimming crab.

Levels interleukine-4 and interleukin-17 (IL-4, IL-17) of blood in patients with habitual recurrent pregnancy loss, which occurred in a loop in vitro fertilization

2016 ◽  
pp. 137-139
Author(s):  
K.P. Golovatyuk ◽  
Keyword(s):  
In Vitro Fertilization ◽  
Conditioned Medium ◽  
Mononuclear Cells ◽  
Recurrent Miscarriage ◽  
Interleukin 4 ◽  
Control Group ◽  
Interleukin 17 ◽  
Vitro Fertilization ◽  
Blood Mononuclear Cells

The objective: was to investigate the levels of cytokines IL-4 and IL-17 in serum and conditioned medium cultures of blood mononuclear cells (MNC) and evaluation association between their products and miscarriage, which occurred in IVF cycles. Patients and methods. We observed 240 patients with recurrent miscarriage, came in IVF cycles, and 100 apparently healthy fertile women in the control group. The concentrations of IL-4 and IL-17 in serum and conditioned medium of MNC cultures were determined. Results. The levels of IL-4 in the serum and conditioned medium in spontaneous and stimulated mitogen secretion was not significantly different from those in the control group, whereas IL-17 levels were higher than those in the control group serum, in conditioned media of stimulated and non-stimulated MNCs. Conclusion. Disregulation of activity of circulating blood mononuclear cells in women with recurrent miscarriage that followed IVF, is accompanied by increased secretion of IL-17 and almost constant production of IL-4 on the back of high stimulation index of production of these cytokines. Key words: in vitro fertilization, miscarriage, interleukin-4, interleukin-17, serum stimulated and non-stimulated mononuclear blood.

Withdrawn: Hepatoprotective effects of Tribulus terrestris L. (Zygophyllales: Zygophyllaceae) hydro-alcholic extract on non-alcoholic fatty liver-induced rats

2016 ◽  
Vol 3 (5) ◽  
pp. 143
Author(s):  
Fatemeh Almasi ◽  
Mozafar Khazaei ◽  
Shima Chehrei ◽  
Ali Ghanbari
Keyword(s):  
Fatty Liver ◽  
Liver Tissue ◽  
Serum Lipid ◽  
Histopathological Examination ◽  
Serum Levels ◽  
Lipid Profiles ◽  
Control Group ◽  
Tribulus Terrestris ◽  
Protective Effects ◽  
Alcoholic Fatty Liver

Non-alcoholic fatty liver induces many complications to the liver tissue and also serum related parameters. Medicinal plants are the safe therapeutic strategy for the treatment of diseases. In this regards, the present study was conducted to evaluate the effect of Tribulus terrestris L. (Zygophyllales: Zygophyllaceae) extract on non-alcoholic fatty liver in rats. In this experimental study, thirty male Wistar rats were divided into five groups (n = 6). Animals in experimental groups were received high fructose diet (70%) (HDF) daily alone or in combined with daily intraperitoneal injection of 500, 700 and 1,000 mg/kg extract of T. terrestris. Control group of rats was feed with standard chow. The serum levels of biomarkers of liver and serum lipid profiles were assessed, also histopathological examination of liver tissue done. Data were analyzed using One-way ANOVA method followed by Tukey’s post-hoc multiple comparison test and P < 0.05 was considered statistically significant. There were significant improvements for biomarkers of liver tissue (P < 0.05) and serum lipid profiles (P < 0.01) in the HFD-fed rats that were treated with T. terrestris extract compare to HFD-fed group. In addition, accumulation of lipids in hepatocytes was significantly reduced in the HFD-fed + extract administrated groups in comparison to HFD-fed rats (P < 0.01). T. terrestris extract has protective effects against non-alcoholic fatty liver by changing biomarkers of liver tissue, serum lipid profiles and histopathological anomalies of liver tissue, to normal range.

scholarly journals Newly Developed Recombinant Antithrombin Protects the Endothelial Glycocalyx in an Endotoxin-Induced Rat Model of Sepsis

2020 ◽  
Vol 22 (1) ◽  
pp. 176
Author(s):  
Toshiaki Iba ◽  
Jerrold H. Levy ◽  
Koichiro Aihara ◽  
Katsuhiko Kadota ◽  
Hiroshi Tanaka ◽  
...  
Keyword(s):  
Dose Group ◽  
Low Dose ◽  
Leukocyte Adhesion ◽  
Endothelial Glycocalyx ◽  
High Dose Group ◽  
Control Group ◽  
High Dose ◽  
Protective Effects ◽  
Circulating Levels

(1) Background: The endothelial glycocalyx is a primary target during the early phase of sepsis. We previously reported a newly developed recombinant non-fucosylated antithrombin has protective effects in vitro. We further evaluated the effects of this recombinant antithrombin on the glycocalyx damage in an animal model of sepsis. (2) Methods: Following endotoxin injection, in Wistar rats, circulating levels of hyaluronan, syndecan-1 and other biomarkers were evaluated in low-dose or high-dose recombinant antithrombin-treated animals and a control group (n = 7 per group). Leukocyte adhesion and blood flow were evaluated with intravital microscopy. The glycocalyx was also examined using side-stream dark-field imaging. (3) Results: The activation of coagulation was inhibited by recombinant antithrombin, leukocyte adhesion was significantly decreased, and flow was better maintained in the high-dose group (both p < 0.05). Circulating levels of syndecan-1 (p < 0.01, high-dose group) and hyaluronan (p < 0.05, low-dose group; p < 0.01, high-dose group) were significantly reduced by recombinant antithrombin treatment. Increases in lactate and decreases in albumin levels were significantly attenuated in the high-dose group (p < 0.05, respectively). The glycocalyx thickness was reduced over time in control animals, but the derangement was attenuated and microvascular perfusion was better maintained in the high-dose group recombinant antithrombin group (p < 0.05). (4) Conclusions: Recombinant antithrombin maintained vascular integrity and the microcirculation by preserving the glycocalyx in this sepsis model, effects that were more prominent with high-dose therapy.

scholarly journals Lateral rectus muscle differentiation potential in paralytic esotropia patients

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Qing Xia ◽  
Xiangtian Ling ◽  
Zhonghao Wang ◽  
Tao Shen ◽  
Minghao Chen ◽  
...  
Keyword(s):  
Gene Expression ◽  
Expression Profiles ◽  
Gene Expression Profiles ◽  
Rectus Muscle ◽  
Lateral Rectus ◽  
Control Group ◽  
Differentiation Potential ◽  
Partial Restoration ◽  
Reverse Transcription Pcr ◽  
Lateral Rectus Muscle

Abstract Purpose and background Recently, we found that maximal medial rectus recession and lateral rectus resection in patients with complete lateral rectus paralysis resulted in a partial restoration of abduction. In an attempt to understand some of the mechanisms involved with this effect we examined gene expression profiles of lateral recti from these patients, with our focus being directed to genes related to myogenesis. Materials and methods Lateral recti resected from patients with complete lateral rectus paralysis and those from concomitant esotropia (controls) were collected. Differences in gene expression profiles between these two groups were examined using microarray analysis and quantitative Reverse-transcription PCR (qRT-PCR). Results A total of 3056 differentially expressed genes (DEGs) were identified between these two groups. Within the paralytic esotropia group, 2081 genes were up-regulated and 975 down-regulated. The results of RT-PCR revealed that PAX7, MYOG, PITX1, SIX1 and SIX4 showed higher levels of expression, while that of MYOD a lower level of expression within the paralytic esotropia group as compared with that in the control group (p < 0.05). Conclusion The decreased expression of MYOD in the paralytic esotropia group suggested that extraocular muscle satellite cell (EOMSCs) differentiation processes were inhibited. Whereas the high expression levels of PAX7, SIX1/4 and MYOG, suggested that the EOMSCs were showing an effective potential for differentiation. The stimulation resulting from muscle surgery may induce EOMSCs to differentiate and thus restore abduction function.

scholarly journals Hepatoprotective effect of galangin on carbon tetrachloride-induced hepatotoxicity via the LKB1/AMPK pathway

2021 ◽  
Vol 19 ◽  
pp. 205873922110008
Author(s):  
Meng Chen ◽  
Xinyan Song ◽  
Jifang Jiang ◽  
Lei Xing ◽  
Pengfei Wang
Keyword(s):  
Carbon Tetrachloride ◽  
Liver Toxicity ◽  
Corn Oil ◽  
Histopathological Examination ◽  
Hepatoprotective Effect ◽  
Control Group ◽  
Group Model ◽  
Protective Effects ◽  
Model Group ◽  
Expression Levels

To investigate the protective effects of galangin on liver toxicity induced by carbon tetrachloride (CCl4) in mice. Mouse hepatotoxicity model was established by intraperitoneal injection (i.p.) of 10 ml/kg body weight CCl4 that diluted with corn oil to a proportion of 1:500 on Kunming mice. The mice were randomly divided into five groups named control group, model group, and 1, 5, and 10 mg/kg galangin group. The levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were analyzed by ELISA. Liver histopathological examination was observed via optical microscopy. The levels of superoxide dismutase (SOD), malondialdehyde (MDA), glutathione (GSH), and glutathion (GSSG) were analyzed to assess oxidative stress. Finally, western blot assay was carried out to analyse the expression levels of total AMP-activated protein kinase (AMPK), phospho-AMPK (p-AMPK), total liver kinase B1 (LKB1), and phospho-LKB1 (p-LKB1). Compared with the control group, in the model group, the levels of AST, ALT, MDA, and GSSG increased significantly ( p < 0.01); the activity of SOD and GSH decreased significantly ( p < 0.01); and the histopathological examination revealed liver necrosis. However, treatment with galangin (5 and 10 mg/kg) significantly reversed these CCl4-induced liver damage indicators. Furthermore, treatment with galangin (10 mg/kg) significantly increased the p-AMPK and p-LKB1 expression levels ( p < 0.01). This study supports the hepatoprotective effect of galangin against hepatotoxicity, perhaps occurring mainly through the LKB1/AMPK-mediated pathway.

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