The need for an intensive care protocol, sometimes weekly or biweekly, has led to a significant i... more The need for an intensive care protocol, sometimes weekly or biweekly, has led to a significant increase in laboratory costs for kidney recipients. In the present study, an inhouse tetraplex nested PCR assay was developed and validated for the specific detection of BKV, JCV, HCMV and EBV in clinical samples. We determined the Limit of Detection (LOD) and analytical specificity. To demonstrate the diagnostic performance of the assay, a total of 102 archival plasma samples were tested and compared with a commercial uniplex real-time PCR kits. The analytical sensitivity of the in-house tetraplex nested PCR assay was 173 copies/ml, when all four viruses were present in the specimens. These values were 79.2, 58.7, 87.6 and 96.1 copies/ml when only, BKV, JCV, HCMV and EBV respectively, were present. The cross-reactivity assays were shown no detectable signal in the tetraplex PCR results. The estimated diagnostic sensitivities were 92.6% for BKV, 92.3% for JCV and 100% for both HCMV and EBV as compared with commercial kits. Regarding the sensitivity and specificity, it seems that the developed Multiplex Nested PCR assay could be used as a reliable virusassociated renal rejection (VRR) panel in post renal transplant surveillance.
Burkholderia mallei is the etiologic agent of glanders. It is difficult to diagnose this zoonotic... more Burkholderia mallei is the etiologic agent of glanders. It is difficult to diagnose this zoonotic disease in its early stages. Some methods such as the complement fixation test (CFT) cause some problems for veterinary authorities and financial losses to animal owners due to false-positive results. The mallein test requires appropriate laboratory equipment and skilled personnel. To quickly and accurately diagnose the disease, especially in areas where animals cannot be kept, new methods (such as the Western blot test [WBT]) should be used to identify the disease. This study designed and optimized the Western blot (immunoblot) test using sera from 84 glanderous equids, and the sensitivity and specificity of ELISA and CFT were compared with the WBT. ELISA tests are based on B. mallei antigens whereas a purified lipopolysaccharide-containing B. mallei antigen is used in the WBT. The sensitivity and specificity of the tests were estimated using the cut-off values recommended by the test developers. The WBT and ELISA were significantly more specific than the CFT. The ELISA based on B. mallei antigens was significantly less sensitive than the CFT. Given their comparable sensitivities and specificities, the CFT (95.7%, 98.5%), the WBT (95%, 100%) and the ELISA (85%, 100%) should be further developed. The CFT is still the prescribed technique for serological investigation of equids for trade purposes to certify individual animals without glanders. Therefore, more efforts should be made to further improve and optimize the WBT and ELISA tests.
The S-layer is the surface cell wall layer in cyanobacteria. S-layers have a 5-10 nm thickness an... more The S-layer is the surface cell wall layer in cyanobacteria. S-layers have a 5-10 nm thickness and identical, regularly arranged pores with diameters of 2-8 nm. The self- assembly process can occur in the air-water interface. Drug targeting is the selective delivery of drugs to particular tissues and cells and it can increase the efficiency of medical treatment, reduce the required dose and decrease the side effects of a drug. The mechanisms of attaching drugs to drug targeting materials vary according to the physicochemical properties of these materials. In this study, the S-layer of Spirulina ISC6 cyanobacteria was used to deliver the anti-blood cancer drug L-Asparaginase to cancer cell lines. In this study the use of nano particles of the Spirulina ISC6 S-layer for the delivery of L- Asparaginase to cancer cell lines was investigated. Spirulina ISC6 was cultured in zarrouk media and the S- layer was extracted from the outer membrane of cyanobacteria using the standard method. Quantification and qualification of the extracted S-layer was done and it was sterilized by filtration. Different concentrations of S- layer and L-Asparaginase were tested on cancer cells. The cell mortality was compared with that of untreated cells, S-layer alone and L-Asparaginase treated cells alone. Results showed that cancer cells treated with S-layer and L-Asparaginase decreased better than cancer cells treated with other compounds.
9 : JOURNAL OF VETERINARY RESEARCH 2002; 57(1):47-50. EVALUATION OF THE HALF LIFE OF MATERNAL ANT... more 9 : JOURNAL OF VETERINARY RESEARCH 2002; 57(1):47-50. EVALUATION OF THE HALF LIFE OF MATERNAL ANTIBODIES AGAINST AVIAN INFLUENZA (AI) IN BROILER AND LAYER CHICKS IN MASHHAD KALIDARI GH.A.,HARZANDI N.,DIANAT MOGHADAM SR ...
Evidence-Based Complementary and Alternative Medicine
Background. Drug resistance is currently possible anywhere in the world. Due to the discovery of ... more Background. Drug resistance is currently possible anywhere in the world. Due to the discovery of antimicrobials, medicine, and health have made tremendous advances over the past several decades. Aim. This research evaluated the antimicrobial and cytotoxicity effects of green synthesis of copper oxide nanoparticles (CuO NPs) on Lactobacillus acidophilus and human embryonic kidney 293 cells (HEK). Method and Materials. Propolis was sampled and extracted. Green synthesis of CuO NPs was synthesized and characterized using SEM, TEM, DLS, BET, and zeta potential methods. L. acidophilus (ATCC 4356) was used, and the antimicrobial tests were carried out at different concentrations (10≥ mg/ml). Moreover, the cytotoxicity was evaluated using an MTT assay on human embryonic kidney 293 cells (HEK). Results. Synthesized CuO NPs using propolis extracts from Khalkhal (sample 1) and Gillan (sample 2) showed −13.2 and −14.4 mV, respectively. The hydrodynamic sizes of well-dispersed samples 1 and 2 w...
Background & Objectives: The identification of Brucella spp. antigens with the capacity to el... more Background & Objectives: The identification of Brucella spp. antigens with the capacity to elicit a protective immune response is of the great interest for the researchers. So, characterization and assessment of diverse antigens of Brucella need to be evaluated. In this study, we report the cloning and expression of the gene coding for 31 KDa OMP (OMP31) of Brucella melitensis 16M. Methods: Brucella melitensis Omp31 gene was amplified with specific primers, cloned into pJET1/2 and subsequently subcloned in pET28a (+) vector. Both these recombinant plasmids were sequenced and then after, expression of recombinant protein was induced by 1mM IPTG. Western blot analysis was also performed by polyclonal rabbit antiserum. Results: Omp31 successfully was cloned in both plasmid vectors. The recombinant Omp31 was expressed in E.coli host and purified with significant yield. Western blot results along with those of sequencing ensured accurate production of recombinant omp31 and retaining of its partial epitopes. Conclusion: Our results show that, an expression host such as E. coli is suitable for omp31 production.
World Academy of Science, Engineering and Technology, International Journal of Medical and Health Sciences, 2014
Authors : F. Motamedi Sedeh, Sh. Chahardoli, H. Mahravani, N. Harzandi, M. Sotoodeh, S. K. Shafae... more Authors : F. Motamedi Sedeh, Sh. Chahardoli, H. Mahravani, N. Harzandi, M. Sotoodeh, S. K. Shafaei Abstract : Foot-and-mouth disease virus (FMDV) is the most economically important disease of livestock. The aim of the study is inactivation of FMD virus type O/IRN/2010 by electron beam without antigenic changes as electron radio vaccine. The BALB/C mice were divided into three groups, each group containing five mice. Three groups of mice were inoculated with conventional vaccine and electron beam irradiated vaccine FMDV type O/IRN/2010 subcutaneously three weeks interval, the final group as negative control. The sera were separated from the blood samples of mice 14 days after last vaccination and tested for the presence of antibodies against FMDV type O/IRN/2010 by serum neutralization test. The Serum Neutralization Test (SNT) was carried out and antibody titration was calculated according to the Kraber protocol. The results of the SNT in three groups of mice showed the titration of ...
9 : JOURNAL OF VETERINARY RESEARCH 2002; 57(1):47-50. EVALUATION OF THE HALF LIFE OF MATERNAL ANT... more 9 : JOURNAL OF VETERINARY RESEARCH 2002; 57(1):47-50. EVALUATION OF THE HALF LIFE OF MATERNAL ANTIBODIES AGAINST AVIAN INFLUENZA (AI) IN BROILER AND LAYER CHICKS IN MASHHAD KALIDARI GH.A.,HARZANDI N.,DIANAT MOGHADAM SR ...
The need for an intensive care protocol, sometimes weekly or biweekly, has led to a significant i... more The need for an intensive care protocol, sometimes weekly or biweekly, has led to a significant increase in laboratory costs for kidney recipients. In the present study, an inhouse tetraplex nested PCR assay was developed and validated for the specific detection of BKV, JCV, HCMV and EBV in clinical samples. We determined the Limit of Detection (LOD) and analytical specificity. To demonstrate the diagnostic performance of the assay, a total of 102 archival plasma samples were tested and compared with a commercial uniplex real-time PCR kits. The analytical sensitivity of the in-house tetraplex nested PCR assay was 173 copies/ml, when all four viruses were present in the specimens. These values were 79.2, 58.7, 87.6 and 96.1 copies/ml when only, BKV, JCV, HCMV and EBV respectively, were present. The cross-reactivity assays were shown no detectable signal in the tetraplex PCR results. The estimated diagnostic sensitivities were 92.6% for BKV, 92.3% for JCV and 100% for both HCMV and EB...
The propolis produced by bees is used by them to protect their hives. The cavity inside the hive’... more The propolis produced by bees is used by them to protect their hives. The cavity inside the hive’s walls is filled in during cold days to reduce entry points and mummify any intruders to ensure their survival. A current focus in nanotechnology and nanoscience is the green biosynthesis of nanoparticles (NPs) using biomaterials. Research on green methods for making metal oxide NPs is gaining momentum to safeguard the environment from the potential dangers associated with toxic chemicals. This study aimed to synthesize copper NPs (CuNPs) via propolis extraction, a novel application of nanoscience. The study was conducted under a range of pH, time conditions, and concentration ratios, and its properties were characterized by UV-Vis absorption spectra, XRD, and FTIR. An FTIR analysis revealed that compounds found in propolis extract could have an effect on the surface modification of the synthesized NPs. The propolis (Khalkhal) extract spectrum exhibited a sharp peak at 3422 cm−1, caused...
OBJECTIVE(S) BK polyomavirus virus primarily infects humans in their early life stages, and in la... more OBJECTIVE(S) BK polyomavirus virus primarily infects humans in their early life stages, and in later life stages, immunosuppressed patients may develop asymptomatic infections. The nucleotides 1744-1812 in the VP1 gene are traditionally used to determine this virus's genotype. MATERIALS AND METHODS The complete genome of the BKV samples from patients referred to Masih Daneshvari Hospital's virology research center was amplified by previously known primer sets. The phylogenetic diversity of the whole genome, different genomic sections, and the non-coding control region of BK virus samples were investigated. Using software Mega X and references, the samples' genotype was determined in separate genomic fragments and the whole genome. RESULTS The samples were classified into two genotypes (I and IV) and five subtypes (Ia, Ib-2, IVc-1, and IVc-2), but none of the isolates belonged to genotype II, III, V, or VI. The Large T antigen-based phylogenetic tree provided 100% bootstrap values for these divisions, which were superior to those (96-100%) used in the VP1 sequence. Among the genomic segments, LTag and VP1 had the most mutations. The non-coding control area contained mutations at the O41 position in the granulocyte/macrophage stimulus gene and the P31 position in the NF-1 gene. CONCLUSION The validity of the phylogenetic analysis was supported by sequence analysis, which found SNPs that could be useful for sub-classifying isolates. More research with a large number of samples and in the wider geographical areas is needed to understand the genetic diversity of the BKV in Iran and also to determine these SNPs' clinical significance in terms of patient outcome and viral load dynamics. This article is protected by copyright. All rights reserved.
The need for an intensive care protocol, sometimes weekly or biweekly, has led to a significant i... more The need for an intensive care protocol, sometimes weekly or biweekly, has led to a significant increase in laboratory costs for kidney recipients. In the present study, an inhouse tetraplex nested PCR assay was developed and validated for the specific detection of BKV, JCV, HCMV and EBV in clinical samples. We determined the Limit of Detection (LOD) and analytical specificity. To demonstrate the diagnostic performance of the assay, a total of 102 archival plasma samples were tested and compared with a commercial uniplex real-time PCR kits. The analytical sensitivity of the in-house tetraplex nested PCR assay was 173 copies/ml, when all four viruses were present in the specimens. These values were 79.2, 58.7, 87.6 and 96.1 copies/ml when only, BKV, JCV, HCMV and EBV respectively, were present. The cross-reactivity assays were shown no detectable signal in the tetraplex PCR results. The estimated diagnostic sensitivities were 92.6% for BKV, 92.3% for JCV and 100% for both HCMV and EBV as compared with commercial kits. Regarding the sensitivity and specificity, it seems that the developed Multiplex Nested PCR assay could be used as a reliable virusassociated renal rejection (VRR) panel in post renal transplant surveillance.
Burkholderia mallei is the etiologic agent of glanders. It is difficult to diagnose this zoonotic... more Burkholderia mallei is the etiologic agent of glanders. It is difficult to diagnose this zoonotic disease in its early stages. Some methods such as the complement fixation test (CFT) cause some problems for veterinary authorities and financial losses to animal owners due to false-positive results. The mallein test requires appropriate laboratory equipment and skilled personnel. To quickly and accurately diagnose the disease, especially in areas where animals cannot be kept, new methods (such as the Western blot test [WBT]) should be used to identify the disease. This study designed and optimized the Western blot (immunoblot) test using sera from 84 glanderous equids, and the sensitivity and specificity of ELISA and CFT were compared with the WBT. ELISA tests are based on B. mallei antigens whereas a purified lipopolysaccharide-containing B. mallei antigen is used in the WBT. The sensitivity and specificity of the tests were estimated using the cut-off values recommended by the test developers. The WBT and ELISA were significantly more specific than the CFT. The ELISA based on B. mallei antigens was significantly less sensitive than the CFT. Given their comparable sensitivities and specificities, the CFT (95.7%, 98.5%), the WBT (95%, 100%) and the ELISA (85%, 100%) should be further developed. The CFT is still the prescribed technique for serological investigation of equids for trade purposes to certify individual animals without glanders. Therefore, more efforts should be made to further improve and optimize the WBT and ELISA tests.
The S-layer is the surface cell wall layer in cyanobacteria. S-layers have a 5-10 nm thickness an... more The S-layer is the surface cell wall layer in cyanobacteria. S-layers have a 5-10 nm thickness and identical, regularly arranged pores with diameters of 2-8 nm. The self- assembly process can occur in the air-water interface. Drug targeting is the selective delivery of drugs to particular tissues and cells and it can increase the efficiency of medical treatment, reduce the required dose and decrease the side effects of a drug. The mechanisms of attaching drugs to drug targeting materials vary according to the physicochemical properties of these materials. In this study, the S-layer of Spirulina ISC6 cyanobacteria was used to deliver the anti-blood cancer drug L-Asparaginase to cancer cell lines. In this study the use of nano particles of the Spirulina ISC6 S-layer for the delivery of L- Asparaginase to cancer cell lines was investigated. Spirulina ISC6 was cultured in zarrouk media and the S- layer was extracted from the outer membrane of cyanobacteria using the standard method. Quantification and qualification of the extracted S-layer was done and it was sterilized by filtration. Different concentrations of S- layer and L-Asparaginase were tested on cancer cells. The cell mortality was compared with that of untreated cells, S-layer alone and L-Asparaginase treated cells alone. Results showed that cancer cells treated with S-layer and L-Asparaginase decreased better than cancer cells treated with other compounds.
9 : JOURNAL OF VETERINARY RESEARCH 2002; 57(1):47-50. EVALUATION OF THE HALF LIFE OF MATERNAL ANT... more 9 : JOURNAL OF VETERINARY RESEARCH 2002; 57(1):47-50. EVALUATION OF THE HALF LIFE OF MATERNAL ANTIBODIES AGAINST AVIAN INFLUENZA (AI) IN BROILER AND LAYER CHICKS IN MASHHAD KALIDARI GH.A.,HARZANDI N.,DIANAT MOGHADAM SR ...
Evidence-Based Complementary and Alternative Medicine
Background. Drug resistance is currently possible anywhere in the world. Due to the discovery of ... more Background. Drug resistance is currently possible anywhere in the world. Due to the discovery of antimicrobials, medicine, and health have made tremendous advances over the past several decades. Aim. This research evaluated the antimicrobial and cytotoxicity effects of green synthesis of copper oxide nanoparticles (CuO NPs) on Lactobacillus acidophilus and human embryonic kidney 293 cells (HEK). Method and Materials. Propolis was sampled and extracted. Green synthesis of CuO NPs was synthesized and characterized using SEM, TEM, DLS, BET, and zeta potential methods. L. acidophilus (ATCC 4356) was used, and the antimicrobial tests were carried out at different concentrations (10≥ mg/ml). Moreover, the cytotoxicity was evaluated using an MTT assay on human embryonic kidney 293 cells (HEK). Results. Synthesized CuO NPs using propolis extracts from Khalkhal (sample 1) and Gillan (sample 2) showed −13.2 and −14.4 mV, respectively. The hydrodynamic sizes of well-dispersed samples 1 and 2 w...
Background & Objectives: The identification of Brucella spp. antigens with the capacity to el... more Background & Objectives: The identification of Brucella spp. antigens with the capacity to elicit a protective immune response is of the great interest for the researchers. So, characterization and assessment of diverse antigens of Brucella need to be evaluated. In this study, we report the cloning and expression of the gene coding for 31 KDa OMP (OMP31) of Brucella melitensis 16M. Methods: Brucella melitensis Omp31 gene was amplified with specific primers, cloned into pJET1/2 and subsequently subcloned in pET28a (+) vector. Both these recombinant plasmids were sequenced and then after, expression of recombinant protein was induced by 1mM IPTG. Western blot analysis was also performed by polyclonal rabbit antiserum. Results: Omp31 successfully was cloned in both plasmid vectors. The recombinant Omp31 was expressed in E.coli host and purified with significant yield. Western blot results along with those of sequencing ensured accurate production of recombinant omp31 and retaining of its partial epitopes. Conclusion: Our results show that, an expression host such as E. coli is suitable for omp31 production.
World Academy of Science, Engineering and Technology, International Journal of Medical and Health Sciences, 2014
Authors : F. Motamedi Sedeh, Sh. Chahardoli, H. Mahravani, N. Harzandi, M. Sotoodeh, S. K. Shafae... more Authors : F. Motamedi Sedeh, Sh. Chahardoli, H. Mahravani, N. Harzandi, M. Sotoodeh, S. K. Shafaei Abstract : Foot-and-mouth disease virus (FMDV) is the most economically important disease of livestock. The aim of the study is inactivation of FMD virus type O/IRN/2010 by electron beam without antigenic changes as electron radio vaccine. The BALB/C mice were divided into three groups, each group containing five mice. Three groups of mice were inoculated with conventional vaccine and electron beam irradiated vaccine FMDV type O/IRN/2010 subcutaneously three weeks interval, the final group as negative control. The sera were separated from the blood samples of mice 14 days after last vaccination and tested for the presence of antibodies against FMDV type O/IRN/2010 by serum neutralization test. The Serum Neutralization Test (SNT) was carried out and antibody titration was calculated according to the Kraber protocol. The results of the SNT in three groups of mice showed the titration of ...
9 : JOURNAL OF VETERINARY RESEARCH 2002; 57(1):47-50. EVALUATION OF THE HALF LIFE OF MATERNAL ANT... more 9 : JOURNAL OF VETERINARY RESEARCH 2002; 57(1):47-50. EVALUATION OF THE HALF LIFE OF MATERNAL ANTIBODIES AGAINST AVIAN INFLUENZA (AI) IN BROILER AND LAYER CHICKS IN MASHHAD KALIDARI GH.A.,HARZANDI N.,DIANAT MOGHADAM SR ...
The need for an intensive care protocol, sometimes weekly or biweekly, has led to a significant i... more The need for an intensive care protocol, sometimes weekly or biweekly, has led to a significant increase in laboratory costs for kidney recipients. In the present study, an inhouse tetraplex nested PCR assay was developed and validated for the specific detection of BKV, JCV, HCMV and EBV in clinical samples. We determined the Limit of Detection (LOD) and analytical specificity. To demonstrate the diagnostic performance of the assay, a total of 102 archival plasma samples were tested and compared with a commercial uniplex real-time PCR kits. The analytical sensitivity of the in-house tetraplex nested PCR assay was 173 copies/ml, when all four viruses were present in the specimens. These values were 79.2, 58.7, 87.6 and 96.1 copies/ml when only, BKV, JCV, HCMV and EBV respectively, were present. The cross-reactivity assays were shown no detectable signal in the tetraplex PCR results. The estimated diagnostic sensitivities were 92.6% for BKV, 92.3% for JCV and 100% for both HCMV and EB...
The propolis produced by bees is used by them to protect their hives. The cavity inside the hive’... more The propolis produced by bees is used by them to protect their hives. The cavity inside the hive’s walls is filled in during cold days to reduce entry points and mummify any intruders to ensure their survival. A current focus in nanotechnology and nanoscience is the green biosynthesis of nanoparticles (NPs) using biomaterials. Research on green methods for making metal oxide NPs is gaining momentum to safeguard the environment from the potential dangers associated with toxic chemicals. This study aimed to synthesize copper NPs (CuNPs) via propolis extraction, a novel application of nanoscience. The study was conducted under a range of pH, time conditions, and concentration ratios, and its properties were characterized by UV-Vis absorption spectra, XRD, and FTIR. An FTIR analysis revealed that compounds found in propolis extract could have an effect on the surface modification of the synthesized NPs. The propolis (Khalkhal) extract spectrum exhibited a sharp peak at 3422 cm−1, caused...
OBJECTIVE(S) BK polyomavirus virus primarily infects humans in their early life stages, and in la... more OBJECTIVE(S) BK polyomavirus virus primarily infects humans in their early life stages, and in later life stages, immunosuppressed patients may develop asymptomatic infections. The nucleotides 1744-1812 in the VP1 gene are traditionally used to determine this virus's genotype. MATERIALS AND METHODS The complete genome of the BKV samples from patients referred to Masih Daneshvari Hospital's virology research center was amplified by previously known primer sets. The phylogenetic diversity of the whole genome, different genomic sections, and the non-coding control region of BK virus samples were investigated. Using software Mega X and references, the samples' genotype was determined in separate genomic fragments and the whole genome. RESULTS The samples were classified into two genotypes (I and IV) and five subtypes (Ia, Ib-2, IVc-1, and IVc-2), but none of the isolates belonged to genotype II, III, V, or VI. The Large T antigen-based phylogenetic tree provided 100% bootstrap values for these divisions, which were superior to those (96-100%) used in the VP1 sequence. Among the genomic segments, LTag and VP1 had the most mutations. The non-coding control area contained mutations at the O41 position in the granulocyte/macrophage stimulus gene and the P31 position in the NF-1 gene. CONCLUSION The validity of the phylogenetic analysis was supported by sequence analysis, which found SNPs that could be useful for sub-classifying isolates. More research with a large number of samples and in the wider geographical areas is needed to understand the genetic diversity of the BKV in Iran and also to determine these SNPs' clinical significance in terms of patient outcome and viral load dynamics. This article is protected by copyright. All rights reserved.
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