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1.
Biotechnol Appl Biochem ; 64(3): 406-414, 2017 May.
Article in English | MEDLINE | ID: mdl-26970530

ABSTRACT

Porcine circovirus type 2 (PCV2) still represents a major problem to the swine industry worldwide, causing high mortality rates in infected animals. Virus-like particles (VLPs) have gained attention for vaccine development, serving both as scaffolds for epitope expression and immune response enhancers. The commercial subunit vaccines against PCV2 consist of VLPs formed by the self-assembly of PCV2 capsid protein (CP) expressed in the baculovirus vector system. In this work, a PCV2 protective epitope was inserted into three different regions of papaya ringspot virus (PRSV) CP, namely, the N- and C-termini and a predicted antigenic region located near the N-terminus. Wild-type and chimeric CPs were modeled in silico, expressed in Escherichia coli, purified, and visualized by transmission electron microscopy. This is the first report that shows the formation of chimeric VLPs using PRSV as epitope-presentation scaffold. Moreover, it was found that PCV2 epitope localization strongly influences VLP length. Also, the estimated yields of the chimeric VLPs at a small-scale level ranged between 65 and 80 mg/L of culture medium. Finally, the three chimeric VLPs induced high levels of immunoglobulin G against the PCV2 epitope in immunized BALB/c mice, suggesting that these chimeric VLPs can be used for swine immunoprophylaxis against PCV2.


Subject(s)
Capsid Proteins , Circovirus , Epitopes , Gene Expression , Plant Viruses , Viral Vaccines , Animals , Capsid Proteins/biosynthesis , Capsid Proteins/genetics , Capsid Proteins/immunology , Circovirus/genetics , Circovirus/immunology , Epitopes/biosynthesis , Epitopes/genetics , Epitopes/immunology , Mice , Plant Viruses/genetics , Plant Viruses/immunology , Plant Viruses/metabolism , Porcine Postweaning Multisystemic Wasting Syndrome/immunology , Porcine Postweaning Multisystemic Wasting Syndrome/prevention & control , Swine , Viral Vaccines/biosynthesis , Viral Vaccines/genetics , Viral Vaccines/immunology
2.
Toxicol Pathol ; 41(6): 857-65, 2013 Aug.
Article in English | MEDLINE | ID: mdl-23197198

ABSTRACT

In this study of chemoprevention in the rat modified resistant hepatocyte model, preneoplastic cells were diminished by >60% with quercetin pretreatment compared with those rats treated with N-Diethylnitrosamine (DEN) to induce liver cancer. This decrease occurred associated with an abolished DEN-induced lipid peroxidation as well as activation of caspase 9 and increased caspase 3, as determined by increased expression of cleaved caspase 3 and 9, but not cleaved caspase 8 and increased fragmentation of Poly (ADP-ribose) polymerase (PARP) inducing apoptosis of presumed genetically injured cells, when quercetin was administered before the initiation agent.


Subject(s)
Apoptosis/drug effects , Liver Neoplasms, Experimental/pathology , Liver Neoplasms, Experimental/prevention & control , Precancerous Conditions/drug therapy , Quercetin/pharmacology , Animals , Anticarcinogenic Agents/pharmacology , Antineoplastic Agents/pharmacology , Carcinogenesis/drug effects , Caspases/metabolism , Chemoprevention/methods , Liver/chemistry , Liver/drug effects , Liver/pathology , Liver Neoplasms, Experimental/chemistry , Liver Neoplasms, Experimental/metabolism , Male , Oxidative Stress/drug effects , Precancerous Conditions/chemistry , Precancerous Conditions/metabolism , Precancerous Conditions/pathology , Rats , Rats, Inbred F344
3.
BMC Cancer ; 10: 159, 2010 Apr 24.
Article in English | MEDLINE | ID: mdl-20416104

ABSTRACT

BACKGROUND: Recently, extremely low frequency electromagnetic fields (ELF-EMF) have been studied with great interest due to their possible effects on human health. In this study, we evaluated the effect of 4.5 mT-120 Hz ELF-EMF on the development of preneoplastic lesions in experimental hepatocarcinogenesis. METHODS: Male Fischer-344 rats were subjected to the modified resistant hepatocyte model and were exposed to 4.5 mT - 120 Hz ELF-EMF. The effects of the ELF-EMF on hepatocarcinogenesis, apoptosis, proliferation and cell cycle progression were evaluated by histochemical, TUNEL assay, caspase 3 levels, immunohistochemical and western blot analyses. RESULTS: The application of the ELF-EMF resulted in a decrease of more than 50% of the number and the area of gamma-glutamyl transpeptidase-positive preneoplastic lesions (P = 0.01 and P = 0.03, respectively) and glutathione S-transferase placental expression (P = 0.01). The number of TUNEL-positive cells and the cleaved caspase 3 levels were unaffected; however, the proliferating cell nuclear antigen, Ki-67, and cyclin D1 expression decreased significantly (P < or = 0.03), as compared to the sham-exposure group. CONCLUSION: The application of 4.5 mT-120 Hz ELF-EMF inhibits preneoplastic lesions chemically induced in the rat liver through the reduction of cell proliferation, without altering the apoptosis process.


Subject(s)
Cell Proliferation/radiation effects , Electromagnetic Fields , Liver Neoplasms/prevention & control , Liver/radiation effects , Precancerous Conditions/prevention & control , Animals , Apoptosis/radiation effects , Blotting, Western , Caspase 3/metabolism , Cell Cycle/radiation effects , Cyclin D1/metabolism , Glutathione S-Transferase pi/metabolism , Immunohistochemistry , In Situ Nick-End Labeling , Ki-67 Antigen/metabolism , Liver/metabolism , Liver/pathology , Liver Neoplasms/chemically induced , Liver Neoplasms/metabolism , Liver Neoplasms/pathology , Male , Precancerous Conditions/chemically induced , Precancerous Conditions/metabolism , Precancerous Conditions/pathology , Rats , Rats, Inbred F344 , Time Factors , gamma-Glutamyltransferase/metabolism
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