In the present study, for the first time, we released and assembled the particles of three major ... more In the present study, for the first time, we released and assembled the particles of three major structural proteins of velogenic NDV (M, HN, and F glycoproteins) as a NDV-VLPs. The ElISA result of the cytokines of splenocyte suspension cells showed that IL2, IL10, TNF-α, and IFN- ˠ titers were significantly higher (p ≤ 0.05) in mice that were immunized only with NDV-VLPs three times with a 10-day interval, in comparison to those that were immunized with NDV-VLPs twice in a 10-day interval and received a B1 live vaccine boost on the third interval. Flow cytometry results showed that CD8 + titers in the group that only received NDV-VLP was higher than other group. However, serum ELISA results did not show a significantly (p ≥ 0.05) higher NDV antibody titer in NDV-VLPs immunized mice compared to the boosted group. Besides, HI results of SPF chickens vaccinated with NDV-VLPs and boosted with B1 live vaccine were significantly (p ≤ 0.05) higher than those that only received NDV-VLPs. I...
Colorectal cancer is one of the most killing cancers and this has become a global problem. Curren... more Colorectal cancer is one of the most killing cancers and this has become a global problem. Current treatment and anticancer drugs cannot specifically target the cancerous cells, thus causing toxicity towards surrounding non-cancer cells. Hence, there is an urgent need to discover a more target-specific therapeutic agent to overcome this problem. Core-shell nanoparticles have emerged as good candidate for anticancer treatment. This study aimed to synthesize core-shell nanoparticles via green method which utilised crude peels extract of Garcinia mangostana as reducing and stabilising agents for drug delivery. Gold-silver core-shell nanoparticles (Au-AgNPs) were synthesized through seed germination process in which gold nanoparticles acted as the seed. A complete coating was observed through transmission electron microscopy (TEM) when the ratio of AuNPs and AgNPs was 1:9. The size of Au-AgNPs was 38.22 ± 8.41 nm and was mostly spherical in shape. Plant-based drug, protocatechuic acid (...
The combination of graphene-based materials and inorganic nanoparticles for the enhancement of th... more The combination of graphene-based materials and inorganic nanoparticles for the enhancement of the nanomaterial properties is extensively explored nowadays. In the present work, we used a sonochemical method to synthesize a copper/reduced graphene oxide (Cu/RGO) nanocomposite using Australian honey and vitamin C as capping and reducing agents, respectively. The honey-mediated copper/reduced graphene oxide (H/Cu/RGO) nanocomposite was then characterized through UV-visible, XRD, HRTEM, and FTIR analysis. The copper nanoparticles (Cu-NPs) in the nanocomposite formed uniform spherical shapes with a size of 2.20 ± 0.70 nm, which attached to the reduced graphene oxide (RGO) layers. The nanocomposite could suppress bacterial growth in both types of bacteria strains. However, in this study, the nanocomposite exhibited good bactericidal activity toward the Gram-positive bacteria than the Gram-negative bacteria. It also showed a cytotoxic effect on the cancer colorectal cell line HCT11, even ...
Journal of Materials Science: Materials in Medicine
Until now, a few studies have been conducted on the destructive effects of TiO2 NPs in living org... more Until now, a few studies have been conducted on the destructive effects of TiO2 NPs in living organisms, and studies on the toxicity of TiO2 NPs are still in the beginning phases. Because of the widespread use of TiO2 NPs in all areas of human life, it is essential to study their profound and fundamental toxic effects on each organ and body cell. Herein, we evaluate the effect of exposure to TiO2 NPs on in vitro models derived from the rat bone marrow and adipose tissues. Exposure to TiO2 NPs at 100 and 200 μg/ml exhibited cytotoxicity for the rat bone marrow mesenchymal stem cells (rBMSCs) and rat adipose mesenchymal stem cells (rATSC), respectively. Additionally, reduced rBMSCs and rATSCs frequencies in the S phase of the cell cycle. Moreover, TiO2 NPs enhanced the activity of cellular senescence-associated β-galactosidase in both model cells. Significantly higher relative expression of aging-related genes P53 and NF-kB (p < 0.05) and lower expression levels of anti-aging-relat...
IntroductionThe control of Newcastle disease virus (NDV) infection depends solely on vaccination ... more IntroductionThe control of Newcastle disease virus (NDV) infection depends solely on vaccination which in most cases is not sufficient to restrain the consequences of such a highly evolving viral disease. Finding out substances for preparing an efficient anti-ND drug would be of high value. n-Docosanol is a saturated fatty alcohol with an inhibitory effect against many enveloped viruses. In this study, we evaluated the therapeutic effect of n-docosanol on NDV infection and shedding in chickens.MethodsChickens infected with a highly virulent NDV were treated with low to high concentrations of n-docosanol (20, 40, and 60 mg/kg body weight) for 4-successive days, once they showed the disease symptoms. Survival and curative rates, virus load, histopathological scoring, and virus shedding were defined.ResultsSymptoms development was found to discontinue 24–72 hours post-treatment. Survival rate in the NDV-infected chickens raised 37.4–53.2% after the treatment. n-Docosanol treatment was ...
Human norovirus (HuNoV) is the leading cause of nonbacterial gastroenteritis worldwide with the G... more Human norovirus (HuNoV) is the leading cause of nonbacterial gastroenteritis worldwide with the GII.4 genotype accounting for over 80% of infections. The major capsid protein of GII.4 variants is evolving rapidly, resulting in new epidemic variants with altered antigenic potentials that must be considered for the development of an effective vaccine. In this study, we identify and characterize linear blockade B-cell epitopes in HuNoV GII.4. Five unique linear B-cell epitopes, namely P2A, P2B, P2C, P2D, and P2E, were predicted on the surface-exposed regions of the capsid protein. Evolving of the surface-exposed epitopes over time was found to correlate with the emergence of new GII.4 outbreak variants. Molecular dynamic simulation (MD) analysis and molecular docking revealed that amino acid substitutions in the putative epitopes P2B, P2C, and P2D could be associated with immune escape and the appearance of new GII.4 variants by affecting solvent accessibility and flexibility of the an...
This study was aimed to evaluate the immunogenicity of recombinant plasmid deoxyribonucleic acid ... more This study was aimed to evaluate the immunogenicity of recombinant plasmid deoxyribonucleic acid (DNA), pBud-H5-green fluorescent protein (GFP)-interferon-regulatory factor (IRF)3 following delivery using polyamidoamine (PAMAM) dendrimer and transactivator of transcription (TAT)-conjugated PAMAM dendrimer as well as the effect of IRF3 as the genetic adjuvant. BALB/c mice were vaccinated transdermally with pBud-H5-GFP, PAMAM/pBud-H5-GFP, TAT-PAMAM/pBud-H5-GFP, and TAT-PAMAM/pBud-H5-GFP-IRF3. The expression analysis of H5 gene from the blood by using quantitative real-time reverse transcriptase polymerase chain reaction confirmed the ability of PAMAM dendrimer as a carrier for gene delivery, as well as the ability of TAT peptide to enhance the delivery efficiency of PAMAM dendrimer. Mice immunized with modified PAMAM by TAT peptide showed higher hemagglutination inhibition titer, and larger CD3/CD4 T cells and CD3/CD8 T cells population, as well as the production of cytokines, namely,...
Journal of Pharmacy & Pharmaceutical Sciences, 2016
PURPOSE: In order to enhance cellular uptake and to facilitate transdermal delivery of DNA vaccin... more PURPOSE: In order to enhance cellular uptake and to facilitate transdermal delivery of DNA vaccine, polyamidoamine (PAMAM) dendrimers conjugated with HIV transactivator of transcription (TAT) was developed. METHODS: First, the plasmid DNA (pIRES-H5/GFP) nanoparticle was formulated using PAMAM dendrimer and TAT peptide and then characterized for surface charge, particle size, DNA encapsulation and protection of the pIRES-H5/GFP DNA plasmid to enzymatic digestion. Subsequently, the potency of the TAT-conjugated dendrimer for gene delivery was evaluated through in vitro transfection into Vero cells followed by gene expression analysis including western blotting, fluorescent microscopy and PCR. The effect of the TAT peptide on cellular uptake of DNA vaccine was studied by qRT-PCR and flow cytometry. Finally, the ability of TAT-conjugated PAMAM dendrimer for transdermal delivery of the DNA plasmid was assessed through artificial membranes followed by qRT-PCR and flow cytometry. RESULTS: ...
Plasmid DNA (pDNA)-based vaccines have emerged as effective subunit vaccines against viral and ba... more Plasmid DNA (pDNA)-based vaccines have emerged as effective subunit vaccines against viral and bacterial pathogens. In this study, a DNA vaccine, namely plasmid internal ribosome entry site-HN/F, was applied in ovo against Newcastle disease (ND). Vaccination was carried out using the DNA vaccine alone or as a mixture of the pDNA and dextran-spermine (D-SPM), a nanoparticle used for pDNA delivery. The results showed that in ovo vaccination with 40 μg pDNA/egg alone induced high levels of antibody titer (P&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt;0.05) in specific pathogen-free (SPF) chickens at 3 and 4 weeks postvaccination compared to 2 weeks postvaccination. Hemagglutination inhibition (HI) titer was not significantly different between groups injected with 40 μg pDNA + 64 μg D-SPM and 40 μg pDNA at 4 weeks postvaccination (P&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;gt;0.05). Higher antibody titer was observed in the group immunized with 40 μg pDNA/egg at 4 weeks postvaccination. The findings also showed that vaccination with 40 μg pDNA/egg alone was able to confer protection against Newcastle disease virus strain NDIBS002 in two out of seven SPF chickens. Although the chickens produced antibody titers 3 weeks after in ovo vaccination, it was not sufficient to provide complete protection to the chickens from lethal viral challenge. In addition, vaccination with pDNA/D-SPM complex did not induce high antibody titer when compared with naked pDNA. Therefore, it was concluded that DNA vaccination with plasmid internal ribosome entry site-HN/F can be suitable for in ovo application against ND, whereas D-SPM is not recommended for in ovo gene delivery.
The study describes the development of DNA vaccines by using the HN and F genes from AF2240 NDV s... more The study describes the development of DNA vaccines by using the HN and F genes from AF2240 NDV strain, namely pIRES/HN, pIRES/F and pIRES-F/HN. Transient expression analysis of the constructs in Vero cells showed the successful expression of the gene inserts in vitro. In vivo experimental showed that single vaccination by constructed plasmid DNA (pDNA) and then boosted with inactivated vaccine, showed a significant difference in ELISA antibody levels (p<0.05) elicited by either pIRES/F, pIRES/F+ pIRES/HN or pIRES-F/HN at one week after the booster in specific pathogen free (SPF) chickens, compared with the inactivated vaccine alone. It was concluded that recombinant pDNA can be used to increase the efficacy of the inactivated vaccine immunization procedure.
In the present study, for the first time, we released and assembled the particles of three major ... more In the present study, for the first time, we released and assembled the particles of three major structural proteins of velogenic NDV (M, HN, and F glycoproteins) as a NDV-VLPs. The ElISA result of the cytokines of splenocyte suspension cells showed that IL2, IL10, TNF-α, and IFN- ˠ titers were significantly higher (p ≤ 0.05) in mice that were immunized only with NDV-VLPs three times with a 10-day interval, in comparison to those that were immunized with NDV-VLPs twice in a 10-day interval and received a B1 live vaccine boost on the third interval. Flow cytometry results showed that CD8 + titers in the group that only received NDV-VLP was higher than other group. However, serum ELISA results did not show a significantly (p ≥ 0.05) higher NDV antibody titer in NDV-VLPs immunized mice compared to the boosted group. Besides, HI results of SPF chickens vaccinated with NDV-VLPs and boosted with B1 live vaccine were significantly (p ≤ 0.05) higher than those that only received NDV-VLPs. I...
Colorectal cancer is one of the most killing cancers and this has become a global problem. Curren... more Colorectal cancer is one of the most killing cancers and this has become a global problem. Current treatment and anticancer drugs cannot specifically target the cancerous cells, thus causing toxicity towards surrounding non-cancer cells. Hence, there is an urgent need to discover a more target-specific therapeutic agent to overcome this problem. Core-shell nanoparticles have emerged as good candidate for anticancer treatment. This study aimed to synthesize core-shell nanoparticles via green method which utilised crude peels extract of Garcinia mangostana as reducing and stabilising agents for drug delivery. Gold-silver core-shell nanoparticles (Au-AgNPs) were synthesized through seed germination process in which gold nanoparticles acted as the seed. A complete coating was observed through transmission electron microscopy (TEM) when the ratio of AuNPs and AgNPs was 1:9. The size of Au-AgNPs was 38.22 ± 8.41 nm and was mostly spherical in shape. Plant-based drug, protocatechuic acid (...
The combination of graphene-based materials and inorganic nanoparticles for the enhancement of th... more The combination of graphene-based materials and inorganic nanoparticles for the enhancement of the nanomaterial properties is extensively explored nowadays. In the present work, we used a sonochemical method to synthesize a copper/reduced graphene oxide (Cu/RGO) nanocomposite using Australian honey and vitamin C as capping and reducing agents, respectively. The honey-mediated copper/reduced graphene oxide (H/Cu/RGO) nanocomposite was then characterized through UV-visible, XRD, HRTEM, and FTIR analysis. The copper nanoparticles (Cu-NPs) in the nanocomposite formed uniform spherical shapes with a size of 2.20 ± 0.70 nm, which attached to the reduced graphene oxide (RGO) layers. The nanocomposite could suppress bacterial growth in both types of bacteria strains. However, in this study, the nanocomposite exhibited good bactericidal activity toward the Gram-positive bacteria than the Gram-negative bacteria. It also showed a cytotoxic effect on the cancer colorectal cell line HCT11, even ...
Journal of Materials Science: Materials in Medicine
Until now, a few studies have been conducted on the destructive effects of TiO2 NPs in living org... more Until now, a few studies have been conducted on the destructive effects of TiO2 NPs in living organisms, and studies on the toxicity of TiO2 NPs are still in the beginning phases. Because of the widespread use of TiO2 NPs in all areas of human life, it is essential to study their profound and fundamental toxic effects on each organ and body cell. Herein, we evaluate the effect of exposure to TiO2 NPs on in vitro models derived from the rat bone marrow and adipose tissues. Exposure to TiO2 NPs at 100 and 200 μg/ml exhibited cytotoxicity for the rat bone marrow mesenchymal stem cells (rBMSCs) and rat adipose mesenchymal stem cells (rATSC), respectively. Additionally, reduced rBMSCs and rATSCs frequencies in the S phase of the cell cycle. Moreover, TiO2 NPs enhanced the activity of cellular senescence-associated β-galactosidase in both model cells. Significantly higher relative expression of aging-related genes P53 and NF-kB (p < 0.05) and lower expression levels of anti-aging-relat...
IntroductionThe control of Newcastle disease virus (NDV) infection depends solely on vaccination ... more IntroductionThe control of Newcastle disease virus (NDV) infection depends solely on vaccination which in most cases is not sufficient to restrain the consequences of such a highly evolving viral disease. Finding out substances for preparing an efficient anti-ND drug would be of high value. n-Docosanol is a saturated fatty alcohol with an inhibitory effect against many enveloped viruses. In this study, we evaluated the therapeutic effect of n-docosanol on NDV infection and shedding in chickens.MethodsChickens infected with a highly virulent NDV were treated with low to high concentrations of n-docosanol (20, 40, and 60 mg/kg body weight) for 4-successive days, once they showed the disease symptoms. Survival and curative rates, virus load, histopathological scoring, and virus shedding were defined.ResultsSymptoms development was found to discontinue 24–72 hours post-treatment. Survival rate in the NDV-infected chickens raised 37.4–53.2% after the treatment. n-Docosanol treatment was ...
Human norovirus (HuNoV) is the leading cause of nonbacterial gastroenteritis worldwide with the G... more Human norovirus (HuNoV) is the leading cause of nonbacterial gastroenteritis worldwide with the GII.4 genotype accounting for over 80% of infections. The major capsid protein of GII.4 variants is evolving rapidly, resulting in new epidemic variants with altered antigenic potentials that must be considered for the development of an effective vaccine. In this study, we identify and characterize linear blockade B-cell epitopes in HuNoV GII.4. Five unique linear B-cell epitopes, namely P2A, P2B, P2C, P2D, and P2E, were predicted on the surface-exposed regions of the capsid protein. Evolving of the surface-exposed epitopes over time was found to correlate with the emergence of new GII.4 outbreak variants. Molecular dynamic simulation (MD) analysis and molecular docking revealed that amino acid substitutions in the putative epitopes P2B, P2C, and P2D could be associated with immune escape and the appearance of new GII.4 variants by affecting solvent accessibility and flexibility of the an...
This study was aimed to evaluate the immunogenicity of recombinant plasmid deoxyribonucleic acid ... more This study was aimed to evaluate the immunogenicity of recombinant plasmid deoxyribonucleic acid (DNA), pBud-H5-green fluorescent protein (GFP)-interferon-regulatory factor (IRF)3 following delivery using polyamidoamine (PAMAM) dendrimer and transactivator of transcription (TAT)-conjugated PAMAM dendrimer as well as the effect of IRF3 as the genetic adjuvant. BALB/c mice were vaccinated transdermally with pBud-H5-GFP, PAMAM/pBud-H5-GFP, TAT-PAMAM/pBud-H5-GFP, and TAT-PAMAM/pBud-H5-GFP-IRF3. The expression analysis of H5 gene from the blood by using quantitative real-time reverse transcriptase polymerase chain reaction confirmed the ability of PAMAM dendrimer as a carrier for gene delivery, as well as the ability of TAT peptide to enhance the delivery efficiency of PAMAM dendrimer. Mice immunized with modified PAMAM by TAT peptide showed higher hemagglutination inhibition titer, and larger CD3/CD4 T cells and CD3/CD8 T cells population, as well as the production of cytokines, namely,...
Journal of Pharmacy & Pharmaceutical Sciences, 2016
PURPOSE: In order to enhance cellular uptake and to facilitate transdermal delivery of DNA vaccin... more PURPOSE: In order to enhance cellular uptake and to facilitate transdermal delivery of DNA vaccine, polyamidoamine (PAMAM) dendrimers conjugated with HIV transactivator of transcription (TAT) was developed. METHODS: First, the plasmid DNA (pIRES-H5/GFP) nanoparticle was formulated using PAMAM dendrimer and TAT peptide and then characterized for surface charge, particle size, DNA encapsulation and protection of the pIRES-H5/GFP DNA plasmid to enzymatic digestion. Subsequently, the potency of the TAT-conjugated dendrimer for gene delivery was evaluated through in vitro transfection into Vero cells followed by gene expression analysis including western blotting, fluorescent microscopy and PCR. The effect of the TAT peptide on cellular uptake of DNA vaccine was studied by qRT-PCR and flow cytometry. Finally, the ability of TAT-conjugated PAMAM dendrimer for transdermal delivery of the DNA plasmid was assessed through artificial membranes followed by qRT-PCR and flow cytometry. RESULTS: ...
Plasmid DNA (pDNA)-based vaccines have emerged as effective subunit vaccines against viral and ba... more Plasmid DNA (pDNA)-based vaccines have emerged as effective subunit vaccines against viral and bacterial pathogens. In this study, a DNA vaccine, namely plasmid internal ribosome entry site-HN/F, was applied in ovo against Newcastle disease (ND). Vaccination was carried out using the DNA vaccine alone or as a mixture of the pDNA and dextran-spermine (D-SPM), a nanoparticle used for pDNA delivery. The results showed that in ovo vaccination with 40 μg pDNA/egg alone induced high levels of antibody titer (P&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt;0.05) in specific pathogen-free (SPF) chickens at 3 and 4 weeks postvaccination compared to 2 weeks postvaccination. Hemagglutination inhibition (HI) titer was not significantly different between groups injected with 40 μg pDNA + 64 μg D-SPM and 40 μg pDNA at 4 weeks postvaccination (P&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;gt;0.05). Higher antibody titer was observed in the group immunized with 40 μg pDNA/egg at 4 weeks postvaccination. The findings also showed that vaccination with 40 μg pDNA/egg alone was able to confer protection against Newcastle disease virus strain NDIBS002 in two out of seven SPF chickens. Although the chickens produced antibody titers 3 weeks after in ovo vaccination, it was not sufficient to provide complete protection to the chickens from lethal viral challenge. In addition, vaccination with pDNA/D-SPM complex did not induce high antibody titer when compared with naked pDNA. Therefore, it was concluded that DNA vaccination with plasmid internal ribosome entry site-HN/F can be suitable for in ovo application against ND, whereas D-SPM is not recommended for in ovo gene delivery.
The study describes the development of DNA vaccines by using the HN and F genes from AF2240 NDV s... more The study describes the development of DNA vaccines by using the HN and F genes from AF2240 NDV strain, namely pIRES/HN, pIRES/F and pIRES-F/HN. Transient expression analysis of the constructs in Vero cells showed the successful expression of the gene inserts in vitro. In vivo experimental showed that single vaccination by constructed plasmid DNA (pDNA) and then boosted with inactivated vaccine, showed a significant difference in ELISA antibody levels (p<0.05) elicited by either pIRES/F, pIRES/F+ pIRES/HN or pIRES-F/HN at one week after the booster in specific pathogen free (SPF) chickens, compared with the inactivated vaccine alone. It was concluded that recombinant pDNA can be used to increase the efficacy of the inactivated vaccine immunization procedure.
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