AIM: We designed two synthetic-core-specific peptides core 1 (C1) and core 2 (C2), and an E1-spec... more AIM: We designed two synthetic-core-specific peptides core 1 (C1) and core 2 (C2), and an E1-specific peptide (E1). We produced specific polyclonal antibodies against these peptides and used the antibodies for detection of HCV antigens on surface and within infected peripheral blood leukocytes. METHODS: Peripheral blood from a healthy individual who tested negative for HCV RNA was incubated with HCV type 4 infected serum for 1 h and 24 h at 37. Cells were stained by direct and indirect immunofluorescence and measured by flow cytometry. RESULTS: After 1 h of incubation, antibodies against C1, C2, and E1 detected HCV antigens on the surface of 27%, 26 % and 73 % of monocytes respectively, while 10%, 5%
Journal of Applied Sciences and Environmental Management, 2005
In a routine work to determine Hepatitis C Virus (HCV) molecules in human infected serum with bio... more In a routine work to determine Hepatitis C Virus (HCV) molecules in human infected serum with biotin/streptavidine enzyme linked immunosorbant assay (ELISA) technique, unexpected false positive was observed. No positive signals were noticed after changing all ELISA buffers. Subsequently, contaminated buffers were screened and analyzed for microbial contamination. Out of fifty-five, five randomly selected bacterial colonies were examined for biosynthesis of biotin using ELISA and/or Western blot binding biotin techniques in both supernatants and cell pellets. In compare to the E.coli reference strain a strong biotin signals in all examined isolates were recorded. All isolates were then examined for their genetic heterogeneity by PCR-RFLP technique of 23S rDNA genes. While, isolate BP(R2) which gave the highest biotin signal, was subjected for molecular identification. Comparative sequence analysis of the 16S rDNA gene (~1440 bp) revealed that this isolate is a member of the bacterial...
AIM To establish a cell culture system with long-term replication of hepatitis C virus (HCV) geno... more AIM To establish a cell culture system with long-term replication of hepatitis C virus (HCV) genome and expression of viral antigens in vitro. METHODS HepG2 cell line was tested for its susceptibility to HCV by incubation with a serum from a patient with chronic hepatitis C. Cells and supernatant were harvested at various time points during the culture. Culture supernatant was tested for its ability to infect naive cells. The presence of minus (antisense) RNA strand, and the detection of core and E1 antigens in cells were examined by RT-PCR and immunological techniques (flow cytometry and Western blot) respectively. RESULTS The intracellular HCV RNA was first detected on d 3 after infection and then could be consistently detected in both cells and supernatant over a period of at least three months. The fresh cells could be infected with supernatant from cultured infected cells. Flow cytometric analysis showed surface and intracellular HCV antigen expression using in house made polyc...
Abstract Visual discomfort could be a significant issue in open-plan spaces with skylight. Due to... more Abstract Visual discomfort could be a significant issue in open-plan spaces with skylight. Due to the glazing size and position in the field of view (FOV) are different from that in perimeter area within side-lit rooms, the evaluation method generated from where may not be referenceable. Post-occupancy evaluation (POE) survey including subjective feedback collection and instrument measurements are taken place in an open-plan office building with skylights to assess the visual discomfort evaluation method in the specific scenario. Based on the research findings, a simulation-based retrofitting plan of skylights and windows is proposed. The results of the glare rating experiment indicate that DGI outperforms DGP in core areas or deep spaces while the glazing appeared in the central area of FOV, the borderline thresholds of DGP shift to lower values as the stimulus range is lower than that in perimeter spaces (e.g. low vertical eye illuminance). Under the situation that the glare sources appeared around the upper edge of FOV, simple luminance metrics which defined within the region surrounding the glare perform better. Moreover, a field test also confirmed that the preferred light level on the workplane under skylight ranges from 150 lx to 1000 lx. The findings refers to glare rating and daylight retrofitting design process may provide insights into visual discomfort issue in open-plan spaces with skylights.
Intravenous Immunoglobulin G (IVIG) is one of the most important and widely used human plasma fra... more Intravenous Immunoglobulin G (IVIG) is one of the most important and widely used human plasma fractions. Although of this there are some precautions should be considered to use this product; e.g. biochemical, biological, immunological, and safety (virus transmission) for each produce patch. The aim of this study was to investigate four commercial IVIG preparations used in Egyptian hospitals. The results show that IVIG from Korean Green Cross Cooperation and Octapharma contained 100% intact IgG, while IVIG-VACSERA and VIGAM-S contained 98-99% intact IgG. IgG subclass distribution corresponded to the distribution in normal human plasma, except for IVIG-VACSERA, where IgG_2 slightly elevated. IVIG-VACSERA and VIAGM-S contained acceptable traces of IgA/IgM, while preparations from both Korean Green Cross Cooperation and Octapharma contained 130/150 and 100/80 microg/ml, respectively. Each preparation had a strong potential to react with respiratory and enteric bacterial LPS and tetanus toxoid antigen. Our results are in general agreement with previous status of worldwide diphtheria immunity status. The immunoreactivity with the other antigen will discuss.
Hepatitis C virus is a major public health problem leading to cirrhosis and increased risk for de... more Hepatitis C virus is a major public health problem leading to cirrhosis and increased risk for development of hepatocellular carcinoma, both leading indications for liver transplantation. Egypt has the highest prevalence of hepatitis C of worldwide. Anti-HCV antibody is usually detected by enzyme immunoassay (EIA). Microarray analysis of 8 Anti-HCV antibody isotypes and 5 HCV peptides considered separately on 5 different matrixes was carried out on 50 hepatitis C Egyptian patients. The optimal substrate kind was chosen based on the greatest amount of antibody bonded with the minimal background. Mercaptosilane activated slides, agarose-slides and polyvinylidene difluoride membranes were the best substrates, while polyacrylamide and nitrocellulose membrane were less sensitive. IgM isotype gave the weakest signals in all patients whatever the substrate type used for antibody immobilization while IgG (total) and its subtypes IgG2, IgG3 and IgG(4) gave the strongest signals with most of the substrates follows by IgA(1), IgG(1) and total IgA, respectively. The results demonstrate that IgG2, IgG3 and IgG(4) are the dominant IgG subtypes, which may indicate that HCV patient immune response shift toward Th-2 immunity. The microarrays permitted the simultaneous serodetection of hepatitis C virus core and envelope peptides by using corresponding rabbit anti-peptides. Hepatitis C virus core and envelope peptides 1, 2, 3, 4 and 5 showed strong signals on all the used substrates except for polyacrylamide slides and nitrocellulose membranes.
Our aim was to determine the level of tetanus immunity in the general Egyptian population. The su... more Our aim was to determine the level of tetanus immunity in the general Egyptian population. The surveyed population consisted of 709 healthy subjects from five regions of Egypt, 2 month to 105 years of age. We used an ELISA assay to determine titers of anti-tetanus IgG. According to the widely used criteria, 31.7% was susceptible to tetanus (IgG level < 0.15 IU/ml), 15.7% protection (0.15-1.0 IU/ml), and 52.6% > or = 1.0 IU/ml). The results revealed that the majority (68.3%) population from 2 months to 50 years had a protective level of IgG against tetanus. The level of susceptibility increases with age until it reaches 90.3% Significantly, more males (23.7%) unprotected against tetanus. Our results suggest that monitoring immunization status and administering the tetanus vaccine as required are essential to insure adequate and long-lasting antibody levels.
The Egyptian journal of immunology / Egyptian Association of Immunologists, 2004
The aim of this study was to determine immune status to corynebacterium diphtheria by screening f... more The aim of this study was to determine immune status to corynebacterium diphtheria by screening for protective antibodies in a sample of Egyptian population. The study population consisted of 709 healthy subjects aged from 2 months to 105 years, inhabitants of 6 regions of Egypt. The study utilized Enzyme linked immunosorbent assay (ELISA) to measure serum levels IgG antibodies reactive with diphtheria toxoid. Levels of diphtheria toxoid antibody > or = 0.1 IU/ ml were defined as immune/protected, 23.9 % of the population were found to be susceptible to diphtheria (IgG level < 0.01 IU/ml), 43% had basic protection (0.01-0.09 IU/ml), and 33.1% were fully protected (0.1 IU/ml). The results revealed that serum levels of antitoxin antibodies decreased in old ages (< 60 y) with the females being more susceptible then males. These results recommend a booster immunization for the susceptible age groups.
Journal of microbiology and biotechnology, Jan 3, 2015
Escherichia coli is the most preferred microorganism to express heterologous protein for therapeu... more Escherichia coli is the most preferred microorganism to express heterologous protein for therapeutic use, as around 30% of the approved therapeutic proteins are currently being produced using it as a host. Due to its rapid growth, high yield of the product, cost-effectiveness and easy scale-up process, E. coli is an expression host of choice in Biotech industry for large scale production of proteins particularly non-glycosylated proteins, for therapeutic use. The availability of various E. coli expression vectors, strains, relatively easy protein folding mechanisms and bioprocess technologies, makes it very attractive for industrial applications. However, codon usage in E. coli and absence of post-translational modifications, such as glycosylation, phosphorylation and proteolytic processing limits its use for the production of slightly complex recombinant biopharmaceuticals. Several new technological advancements in the E. coli expression system to meet the Biotech industry requirem...
Purpose The prevalence of HCV infection has increased during recent years and the incidence reach... more Purpose The prevalence of HCV infection has increased during recent years and the incidence reach 3% of the world's population, and in some countries like Egypt, may around 20%. The developments of effective and preventive agents are critical to control the current public health burden imposed by HCV infection. Lactoferrin in general and camel lactoferrin specifically has been shown to have a compatitive anti-viral activity against hepatitis C virus (HCV). The purpose of this study was to examine and compare the anti-infectivity of native human, camel, bovine and sheep lactoferrin on continuous of HCV infection in HepG2 cells. Material and methods Used Lfs were purified by Mono S 5/50 GL column and Superdex 200 5/150 column. The purified Lfs were evaluated in two ways; 1. the pre-infected cells were treated with the Lfs to inhibit intracellular replication at different concentrations and time intervals, 2. Lfs were directly incubated with the virus molecules then used to cells i...
Purpose To extend the study of the camel milk proteins which have antiviral activity against HCV,... more Purpose To extend the study of the camel milk proteins which have antiviral activity against HCV, camel naïve polyclonal IgGs, α-lactalbumin were purified from camel milk and their anti-HCV effect was examined using PBMCs and Huh7.5 cell-lines. They were compared with the activity of human polyclonal IgGs and camel lactoferrin and casein. Material and methods Three types of experiments were performed on PBMCs and HuH7.5 cell. HCV was directly incubated with the purified proteins and then mixed with both cell types, or the proteins were incubated with the cells and then exposed to HCV, or the HCV pre-infected cells were treated with the proteins to inhibit intracellular replication. The proteins were added to cells or virus at different concentrations and time intervals. Results Pretreated PBMCs and Huh7.5 cells with milk proteins were not protected when exposed to HCV infection. The direct interaction between HCV and camel IgGs and camel lactoferrin (cLf) led to a complete inhibitio...
AIM: We designed two synthetic-core-specific peptides core 1 (C1) and core 2 (C2), and an E1-spec... more AIM: We designed two synthetic-core-specific peptides core 1 (C1) and core 2 (C2), and an E1-specific peptide (E1). We produced specific polyclonal antibodies against these peptides and used the antibodies for detection of HCV antigens on surface and within infected peripheral blood leukocytes. METHODS: Peripheral blood from a healthy individual who tested negative for HCV RNA was incubated with HCV type 4 infected serum for 1 h and 24 h at 37. Cells were stained by direct and indirect immunofluorescence and measured by flow cytometry. RESULTS: After 1 h of incubation, antibodies against C1, C2, and E1 detected HCV antigens on the surface of 27%, 26 % and 73 % of monocytes respectively, while 10%, 5%
Journal of Applied Sciences and Environmental Management, 2005
In a routine work to determine Hepatitis C Virus (HCV) molecules in human infected serum with bio... more In a routine work to determine Hepatitis C Virus (HCV) molecules in human infected serum with biotin/streptavidine enzyme linked immunosorbant assay (ELISA) technique, unexpected false positive was observed. No positive signals were noticed after changing all ELISA buffers. Subsequently, contaminated buffers were screened and analyzed for microbial contamination. Out of fifty-five, five randomly selected bacterial colonies were examined for biosynthesis of biotin using ELISA and/or Western blot binding biotin techniques in both supernatants and cell pellets. In compare to the E.coli reference strain a strong biotin signals in all examined isolates were recorded. All isolates were then examined for their genetic heterogeneity by PCR-RFLP technique of 23S rDNA genes. While, isolate BP(R2) which gave the highest biotin signal, was subjected for molecular identification. Comparative sequence analysis of the 16S rDNA gene (~1440 bp) revealed that this isolate is a member of the bacterial...
AIM To establish a cell culture system with long-term replication of hepatitis C virus (HCV) geno... more AIM To establish a cell culture system with long-term replication of hepatitis C virus (HCV) genome and expression of viral antigens in vitro. METHODS HepG2 cell line was tested for its susceptibility to HCV by incubation with a serum from a patient with chronic hepatitis C. Cells and supernatant were harvested at various time points during the culture. Culture supernatant was tested for its ability to infect naive cells. The presence of minus (antisense) RNA strand, and the detection of core and E1 antigens in cells were examined by RT-PCR and immunological techniques (flow cytometry and Western blot) respectively. RESULTS The intracellular HCV RNA was first detected on d 3 after infection and then could be consistently detected in both cells and supernatant over a period of at least three months. The fresh cells could be infected with supernatant from cultured infected cells. Flow cytometric analysis showed surface and intracellular HCV antigen expression using in house made polyc...
Abstract Visual discomfort could be a significant issue in open-plan spaces with skylight. Due to... more Abstract Visual discomfort could be a significant issue in open-plan spaces with skylight. Due to the glazing size and position in the field of view (FOV) are different from that in perimeter area within side-lit rooms, the evaluation method generated from where may not be referenceable. Post-occupancy evaluation (POE) survey including subjective feedback collection and instrument measurements are taken place in an open-plan office building with skylights to assess the visual discomfort evaluation method in the specific scenario. Based on the research findings, a simulation-based retrofitting plan of skylights and windows is proposed. The results of the glare rating experiment indicate that DGI outperforms DGP in core areas or deep spaces while the glazing appeared in the central area of FOV, the borderline thresholds of DGP shift to lower values as the stimulus range is lower than that in perimeter spaces (e.g. low vertical eye illuminance). Under the situation that the glare sources appeared around the upper edge of FOV, simple luminance metrics which defined within the region surrounding the glare perform better. Moreover, a field test also confirmed that the preferred light level on the workplane under skylight ranges from 150 lx to 1000 lx. The findings refers to glare rating and daylight retrofitting design process may provide insights into visual discomfort issue in open-plan spaces with skylights.
Intravenous Immunoglobulin G (IVIG) is one of the most important and widely used human plasma fra... more Intravenous Immunoglobulin G (IVIG) is one of the most important and widely used human plasma fractions. Although of this there are some precautions should be considered to use this product; e.g. biochemical, biological, immunological, and safety (virus transmission) for each produce patch. The aim of this study was to investigate four commercial IVIG preparations used in Egyptian hospitals. The results show that IVIG from Korean Green Cross Cooperation and Octapharma contained 100% intact IgG, while IVIG-VACSERA and VIGAM-S contained 98-99% intact IgG. IgG subclass distribution corresponded to the distribution in normal human plasma, except for IVIG-VACSERA, where IgG_2 slightly elevated. IVIG-VACSERA and VIAGM-S contained acceptable traces of IgA/IgM, while preparations from both Korean Green Cross Cooperation and Octapharma contained 130/150 and 100/80 microg/ml, respectively. Each preparation had a strong potential to react with respiratory and enteric bacterial LPS and tetanus toxoid antigen. Our results are in general agreement with previous status of worldwide diphtheria immunity status. The immunoreactivity with the other antigen will discuss.
Hepatitis C virus is a major public health problem leading to cirrhosis and increased risk for de... more Hepatitis C virus is a major public health problem leading to cirrhosis and increased risk for development of hepatocellular carcinoma, both leading indications for liver transplantation. Egypt has the highest prevalence of hepatitis C of worldwide. Anti-HCV antibody is usually detected by enzyme immunoassay (EIA). Microarray analysis of 8 Anti-HCV antibody isotypes and 5 HCV peptides considered separately on 5 different matrixes was carried out on 50 hepatitis C Egyptian patients. The optimal substrate kind was chosen based on the greatest amount of antibody bonded with the minimal background. Mercaptosilane activated slides, agarose-slides and polyvinylidene difluoride membranes were the best substrates, while polyacrylamide and nitrocellulose membrane were less sensitive. IgM isotype gave the weakest signals in all patients whatever the substrate type used for antibody immobilization while IgG (total) and its subtypes IgG2, IgG3 and IgG(4) gave the strongest signals with most of the substrates follows by IgA(1), IgG(1) and total IgA, respectively. The results demonstrate that IgG2, IgG3 and IgG(4) are the dominant IgG subtypes, which may indicate that HCV patient immune response shift toward Th-2 immunity. The microarrays permitted the simultaneous serodetection of hepatitis C virus core and envelope peptides by using corresponding rabbit anti-peptides. Hepatitis C virus core and envelope peptides 1, 2, 3, 4 and 5 showed strong signals on all the used substrates except for polyacrylamide slides and nitrocellulose membranes.
Our aim was to determine the level of tetanus immunity in the general Egyptian population. The su... more Our aim was to determine the level of tetanus immunity in the general Egyptian population. The surveyed population consisted of 709 healthy subjects from five regions of Egypt, 2 month to 105 years of age. We used an ELISA assay to determine titers of anti-tetanus IgG. According to the widely used criteria, 31.7% was susceptible to tetanus (IgG level &amp;amp;lt; 0.15 IU/ml), 15.7% protection (0.15-1.0 IU/ml), and 52.6% &amp;amp;gt; or = 1.0 IU/ml). The results revealed that the majority (68.3%) population from 2 months to 50 years had a protective level of IgG against tetanus. The level of susceptibility increases with age until it reaches 90.3% Significantly, more males (23.7%) unprotected against tetanus. Our results suggest that monitoring immunization status and administering the tetanus vaccine as required are essential to insure adequate and long-lasting antibody levels.
The Egyptian journal of immunology / Egyptian Association of Immunologists, 2004
The aim of this study was to determine immune status to corynebacterium diphtheria by screening f... more The aim of this study was to determine immune status to corynebacterium diphtheria by screening for protective antibodies in a sample of Egyptian population. The study population consisted of 709 healthy subjects aged from 2 months to 105 years, inhabitants of 6 regions of Egypt. The study utilized Enzyme linked immunosorbent assay (ELISA) to measure serum levels IgG antibodies reactive with diphtheria toxoid. Levels of diphtheria toxoid antibody > or = 0.1 IU/ ml were defined as immune/protected, 23.9 % of the population were found to be susceptible to diphtheria (IgG level < 0.01 IU/ml), 43% had basic protection (0.01-0.09 IU/ml), and 33.1% were fully protected (0.1 IU/ml). The results revealed that serum levels of antitoxin antibodies decreased in old ages (< 60 y) with the females being more susceptible then males. These results recommend a booster immunization for the susceptible age groups.
Journal of microbiology and biotechnology, Jan 3, 2015
Escherichia coli is the most preferred microorganism to express heterologous protein for therapeu... more Escherichia coli is the most preferred microorganism to express heterologous protein for therapeutic use, as around 30% of the approved therapeutic proteins are currently being produced using it as a host. Due to its rapid growth, high yield of the product, cost-effectiveness and easy scale-up process, E. coli is an expression host of choice in Biotech industry for large scale production of proteins particularly non-glycosylated proteins, for therapeutic use. The availability of various E. coli expression vectors, strains, relatively easy protein folding mechanisms and bioprocess technologies, makes it very attractive for industrial applications. However, codon usage in E. coli and absence of post-translational modifications, such as glycosylation, phosphorylation and proteolytic processing limits its use for the production of slightly complex recombinant biopharmaceuticals. Several new technological advancements in the E. coli expression system to meet the Biotech industry requirem...
Purpose The prevalence of HCV infection has increased during recent years and the incidence reach... more Purpose The prevalence of HCV infection has increased during recent years and the incidence reach 3% of the world's population, and in some countries like Egypt, may around 20%. The developments of effective and preventive agents are critical to control the current public health burden imposed by HCV infection. Lactoferrin in general and camel lactoferrin specifically has been shown to have a compatitive anti-viral activity against hepatitis C virus (HCV). The purpose of this study was to examine and compare the anti-infectivity of native human, camel, bovine and sheep lactoferrin on continuous of HCV infection in HepG2 cells. Material and methods Used Lfs were purified by Mono S 5/50 GL column and Superdex 200 5/150 column. The purified Lfs were evaluated in two ways; 1. the pre-infected cells were treated with the Lfs to inhibit intracellular replication at different concentrations and time intervals, 2. Lfs were directly incubated with the virus molecules then used to cells i...
Purpose To extend the study of the camel milk proteins which have antiviral activity against HCV,... more Purpose To extend the study of the camel milk proteins which have antiviral activity against HCV, camel naïve polyclonal IgGs, α-lactalbumin were purified from camel milk and their anti-HCV effect was examined using PBMCs and Huh7.5 cell-lines. They were compared with the activity of human polyclonal IgGs and camel lactoferrin and casein. Material and methods Three types of experiments were performed on PBMCs and HuH7.5 cell. HCV was directly incubated with the purified proteins and then mixed with both cell types, or the proteins were incubated with the cells and then exposed to HCV, or the HCV pre-infected cells were treated with the proteins to inhibit intracellular replication. The proteins were added to cells or virus at different concentrations and time intervals. Results Pretreated PBMCs and Huh7.5 cells with milk proteins were not protected when exposed to HCV infection. The direct interaction between HCV and camel IgGs and camel lactoferrin (cLf) led to a complete inhibitio...
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