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 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 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.

Infectivity of Echinococcus granulosus protoscolices under different conditions of temperature and humidity

2008 ◽  
Vol 82 (4) ◽  
pp. 297-300 ◽  
Author(s):  
A.I. Diker ◽  
R. Tinar ◽  
B. Senlik
Keyword(s):  
Relative Humidity ◽  
Echinococcus Granulosus ◽  
Cold Storage ◽  
Post Infection ◽  
Control Group ◽  
Fourth Group ◽  
Larval Stages ◽  
The Third ◽  
Control Programmes ◽  
Different Temperatures

AbstractThe aim of this study was to examine the effect of different temperatures and humidities on the infectivity of Echinococcus granulosus protoscolices. Eighteen dogs (6 groups, n = 3 each) were fed with offal mince harbouring approximately 20,000 protoscolices of E. granulosus of different viabilities. Dogs were infected with E. granulosus protoscolices of: (1) 5% viability at − 10°C and 50% relative humidity (RH); (2) 30% viability at 0°C and 60% RH; (3) 20% viability at +10°C and 65% RH; (4) 15% viability at +30°C and 75% RH; (5) 11% viability at +40°C and 80% RH; (6) 68% viability (control group). Dogs in each group were necropsied at 29–49 days post-infection. Mean intensities of E. granulosus recovered from dogs were 256.7 ± 60.3 in the second group; 32.7 ± 7.1 in the third group; 40.3 ± 15.5 in the fourth group and 1533 ± 513 in the control group. However, no parasites were recovered from the first and fifth groups. Results obtained in the present study show that larval stages could be infective for 1 to 4 weeks during spring, autumn or winter months when maximal temperatures are approximately 0–10°C. In conclusion, cold-storage depots in slaughterhouses and abattoirs where sheep carcasses might be discarded should be kept at − 20°C for 2–3 days, dogs should be properly controlled and adequate control programmes must be established in areas where the disease is endemic.

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.

Tryptophan-Niacin Metabolism in Rat with Puromycin Aminonucleoside-Induced Nephrosis

2006 ◽  
Vol 76 (1) ◽  
pp. 28-33 ◽  
Author(s):  
Yukari Egashira ◽  
Shin Nagaki ◽  
Hiroo Sanada
Keyword(s):  
Body Weight ◽  
Urinary Excretion ◽  
Enzyme Activities ◽  
Treated Group ◽  
Control Group ◽  
Puromycin Aminonucleoside ◽  
Low Level ◽  
Key Enzyme

We investigated the change of tryptophan-niacin metabolism in rats with puromycin aminonucleoside PAN-induced nephrosis, the mechanisms responsible for their change of urinary excretion of nicotinamide and its metabolites, and the role of the kidney in tryptophan-niacin conversion. PAN-treated rats were intraperitoneally injected once with a 1.0% (w/v) solution of PAN at a dose of 100 mg/kg body weight. The collection of 24-hour urine was conducted 8 days after PAN injection. Daily urinary excretion of nicotinamide and its metabolites, liver and blood NAD, and key enzyme activities of tryptophan-niacin metabolism were determined. In PAN-treated rats, the sum of urinary excretion of nicotinamide and its metabolites was significantly lower compared with controls. The kidneyα-amino-β-carboxymuconate-ε-semialdehyde decarboxylase (ACMSD) activity in the PAN-treated group was significantly decreased by 50%, compared with the control group. Although kidney ACMSD activity was reduced, the conversion of tryptophan to niacin tended to be lower in the PAN-treated rats. A decrease in urinary excretion of niacin and the conversion of tryptophan to niacin in nephrotic rats may contribute to a low level of blood tryptophan. The role of kidney ACMSD activity may be minimal concerning tryptophan-niacin conversion under this experimental condition.

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.

scholarly journals Impact of MASP2 gene polymorphism and gene-tea drinking interaction on susceptibility to tuberculosis

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Zihao Li ◽  
Mian Wang ◽  
Hua Zhong ◽  
Xin Huang ◽  
Xinyin Wu ◽  
...  
Keyword(s):  
Additive Model ◽  
Lectin Pathway ◽  
Control Group ◽  
Single Nucleotide ◽  
Key Enzyme ◽  
Healthy Control ◽  
Tea Drinking ◽  
Mannan Binding Lectin ◽  
The Complement System ◽  
Negative Interactions

AbstractMannan-binding lectin-associated serine protease-2 (MASP-2) has been reported to play an important role as a key enzyme in the lectin pathway of the complement system. The objectives of our study were to determine whether the single-nucleotide polymorphism (SNPs) of MASP2 and the gene-tea drinking interaction were associated with the susceptibility to TB. In total, 503 patients and 494 healthy controls were contained. Three SNPs (rs12142107, rs12711521, and rs7548659) were genotyped. The association between the SNPs and susceptibility to TB were investigated by conducting multivariate unconditional logistic regression analysis. The gene-tea drinking interactions were analyzed by the additive model of marginal structural linear odds models. Both genotype AC + AA at rs12711521 of MASP2 genes and genotype GT + GG at rs7548659 of MASP2 genes were more prevalent in the TB patient group than the healthy control group (OR: 1.423 and 1.439, respectively, P < 0.05). In addition, The relative excess risk of interaction (RERI) between tea drinking and rs12142107, rs12711521, and rs7548659 of MASP2 genes was found to suggest negative interactions, which reached − 0.2311 (95% confidence interval (CI): − 0.4736, − 0.0113), − 0.7080 (95% CI − 1.3998, − 0.0163), and − 0.5140 (95% CI − 0.8988, − 0.1291), respectively (P < 0.05). Our finding indicated that the SNPs (rs12711521 and rs7548659) of MASP2 were associated with the susceptibility to TB. Furthermore, there were negative interactions between tea drinking and rs12142107, rs12711521, and rs75548659 of MASP2 gene, respectively. Our research provides a basis for studying the pathogenesis and prevention of tuberculosis.

Effect of Theaflavin on Inflammatory and Remolding of Airway in the Asthma Mice

2021 ◽  
Vol 11 (6) ◽  
pp. 1091-1098
Author(s):  
Jingju Hu ◽  
Jing Yang ◽  
Hua Guo ◽  
Xuesong Yao ◽  
Haiyan Qiu ◽  
...  
Keyword(s):  
Interleukin 4 ◽  
Control Group ◽  
Model Group ◽  
Interleukin 5 ◽  
Treatment Groups ◽  
Interleukin 13 ◽  
Asthma Model ◽  
Lung Resistance ◽  
Neutrophile Granulocytes ◽  
Number Of Cells

To study the effect of theaflavin on the airway’s inflammation and remodeling in mice with asthma. The mice were divided into the control, asthma model, and the theaflavin treatment groups to analyze the changes in pulmonary compliance and lung resistance of the mice with asthma to theaflavin treatment. The theaflavin treatment groups consisted of the low-dose (15 mg/kg theaflavin-intragastric administration), medium-dose (30 mg/kg), and high-dose (60 mg/kg) groups. Alveoli lavage liquid was gathered from the mice to count the number of inflammatory cells, and the levels of interleukin 4 (IL-4), interleukin 5 (IL-5), interleukin 13 (IL-13), and eotaxin were detected by ELISA. The levels of proteins, such as transforming growth factor-1 (TGF-1), alpha-smooth muscle actin (α-SMA), CyclinD1,CyclinD2, Toll-like receptors-4 (TLR4), myeloid differentiation factor 88 (MyD88), and NF-κBp65, which showed the performance of lung tissue was tested by Western blotting. Compared to the control group, the lung resistance of the asthma model mice was increased, and compliance was decreased after increasing concentrations of acetylcholine (Mch) stimulation. Compared to the asthma model group, the pulmonary resistance was decreased, and pulmonar compliance was increased according to the rising concentration of Mch in theaflavin-L, theaflavin-M and theaflavin-H mice. Compared to the control group, the number of cells, macrophages, acidophilic cells, lymph, and neutrophile granulocytes increased in the alveolar perfusion fluid of asthmatic mice. The level of interleukin 4, interleukin 5, interleukin 13, and eotaxin, TGF-β1, α-SMA, Cyclin D1, MyD88, TLR4, Cyclin D2, and NF-κBp65 proteins of the lung was also increased. Compared to the model group, the number of cells, macrophages, acidophilic cells, lymph, and neutrophile granulocytes were decreased successively in the alveolar lavage fluid in the theaflavin-L, theaflavin-M, and theaflavin-H mice. Meanwhile, the content of interleukin 4, interleukin 5, interleukin 13, and eotaxin were decreased successively, and the level of TGF-β1, α-SMA, Cyclin D1, MyD88, TLR4, Cyclin D2, and NF-Bp65 protein increased successively in the theaflavin-L, theaflavin-M, and theaflavin-H mice. Theaflavin has been found to reduce airway inflammation, impede airway remodeling, and decrease the TLR 4/MyD88/NF-B signaling in asthmatic mice.

The Action of Shenmai Injection on the Inflammation and Proliferation of Smooth Muscle Cells of the Airway in Asthmatic Rats

2021 ◽  
Vol 11 (3) ◽  
pp. 386-391
Author(s):  
He Zhu ◽  
Yali Guo ◽  
Xiaoli Wang ◽  
Min Zhu ◽  
Jiahui Lei ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Transient Receptor Potential ◽  
Receptor Potential ◽  
Muscle Cells ◽  
Interleukin 4 ◽  
Nuclear Antigen ◽  
Control Group ◽  
Airway Smooth Muscle Cells ◽  
Shenmai Injection

To observe the effect of transient receptor potential ankyrin 1 (TRPA1) channel on the proliferation and inflammation of airway smooth muscle cells (SMC) in asthmatic rats, the rats were randomly allocated into three treatment groups: control, asthma, and Shenmai injection (SMI), with 15 rats in each group. Asthmatic rat models were induced by ovalbumin (OVA) inhalation. Rats in the control and asthma groups were intraperitoneally injected 2 mL NS daily, whereas rats in the SMI treatment group were intraperitoneally injected with 2 mL SMI daily. RT-qPCR and western blotting were used to test for TRPA1 and proliferating cell nuclear antigen (PCNA) mRNA and protein expression. ELISA was used to test the expression of interleukin-4 (IL-4), interleukin-5 (IL-5), and interleukin-13 (IL-13) in the serum. Compared with the control group, there were significantly higher levels of TRPA1 and PCNA mRNA and protein, as well as of IL-4, IL-5, and IL-13 in asthmatic rats (P< 0.05). After SMI treatment, there was significantly lower expression of TRPA1, PCNA, IL-4, IL-5, and IL-13 compared to the levels in asthmatic rats (P < 0.05). TRPA1, IL-4, IL-5, and IL-13 were highly expressed in the tracheal SMC of asthmatic rats. Inhibiting TRPA1, IL-4, IL-5, and IL-13 using SMI may be one of the mechanisms that can intervene chronic airway inflammation and asthma proliferation.

Saccharomyces cerevisiae contains two functional citrate synthase genes

1986 ◽  
Vol 6 (6) ◽  
pp. 1936-1942
Author(s):  
K S Kim ◽  
M S Rosenkrantz ◽  
L Guarente
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Carbon Source ◽  
Citrate Synthase ◽  
Tricarboxylic Acid Cycle ◽  
Nuclear Gene ◽  
Tricarboxylic Acid ◽  
Yeast Genome ◽  
Gene Encoding ◽  
Acid Cycle ◽  
Nonfermentable Carbon Source

The tricarboxylic acid cycle occurs within the mitochondria of the yeast Saccharomyces cerevisiae. A nuclear gene encoding the tricarboxylic acid cycle enzyme citrate synthase has previously been isolated (M. Suissa, K. Suda, and G. Schatz, EMBO J. 3:1773-1781, 1984) and is referred to here as CIT1. We report here the isolation, by an immunological method, of a second nuclear gene encoding citrate synthase (CIT2). Disruption of both genes in the yeast genome was necessary to produce classical citrate synthase-deficient phenotypes: glutamate auxotrophy and poor growth on rich medium containing lactate, a nonfermentable carbon source. Therefore, the citrate synthase produced from either gene was sufficient for these metabolic roles. Transcription of both genes was maximally repressed in medium containing both glucose and glutamate. However, transcription of CIT1 but not of CIT2 was derepressed in medium containing a nonfermentable carbon source. The significance of the presence of two genes encoding citrate synthase in S. cerevisiae is discussed.

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