فهرست مطالب حوریه سلیمانجاهی

In recent years, the collection of data from the monitoring of clinical trials in the form of electronic case report forms (eCRF) has been increasingly used, because avoiding mistakes, reducing the duration of clinical studies, and preventing increasing data collection costs can affect the success of the study. Accordingly, this research aimed to present the conceptual model and requirements of the proposed eCRF, which is based on NoSQL database to achieve complications caused by the COVID-19 vaccine.

In this study, using the descriptive-applicative method, the conceptual model of the eCRF system was first designed to collect the minimum set of data on the complications of the COVID-19 vaccine. Then, the functional and non-functional requirements of the proposed system were identified using the methods of observation, thinking and reflection, and questionnaire.

This research has two main parts: the proposed conceptual model and the functional and non-functional requirements of the eCRF system. A conceptual model was proposed based on the components, modules, and objectives of the eCRF core, which can be used by researchers and clinical trial specialists. Requirements including the ability to register, insert, delete, and edit the case report form, were also calculated for four groups of patients, health care workers, disease experts, and the executive manager.

The proposed system provides researchers and experts of clinical trials with classified and valid data regarding the registration of complications caused by the COVID-19 vaccine in an integrated manner. Therefore, some eCRF stakeholders such as patients, healthcare workers, disease experts, and executive managers and researchers can achieve a vaccine with acceptable safety and effectiveness by analyzing, interpreting, and sharing the recorded information.

Gene transfection is a powerful tool for changing cell function, which is used for treating various diseases. Nevertheless, various studies show that despite the variety of gene transfer methods, there are significant limitations in the application of each method. The present study aimed to compare two methods of gene transfer using PEI and Lipofectamine compounds in cancer cell.

In this study, Caco-2 and HCT-116 were used as target cells, and HEK-293 cells were used as control cells. For this purpose, initially, gene transfer was performed using PEI and Lipofectamine, and then their results were compared with each other based on the expression of GFP protein.

The results of this study showed that although gene transfer was carried out in HEK-293 cells at a favorable rate, this process was very insignificant in colorectal cells. In addition, in the meantime, some differences were observed between the amounts of gene transfer in colorectal cells, so that the amount of transfection in Caco-2 cells was lower than that of the HCT-116 cells.

The degree of acceptance of foreign genes in cells depends on several factors, including the type and origin of the cell. Since cancer cells have undergone many genetic changes, their genetic manipulation is associated with many limitations. For this reason, it is recommended to use other methods, such as cell selection by antibiotics and preparation of stable cell lines to reach a high level of gene transfer in these cells.

Exercise activities can act as an endogenous adjuvant, enhancing the efficiency of the host immune response after vaccination. However, the effect of repeated bouts of eccentric endurance exercise on cellular and humoral immune-related responses to the vaccine is unclear. The present study was performed to investigate the activation of type 1 and 2 (Th) helper lymphocytes following three bouts of endurance eccentric exercise as an adjuvant for DNA vaccination. Thirty-six female Balb-c mice (6-8 weeks, body mass mean=18g) were randomly divided into six groups: control, exercise, vaccine, PcDNA, exercise-vaccine and exercise-PcDNA (6 mice in each group). Different groups underwent DNA vaccine and PcDNA injection immediately after the last exercise, all stages of exercise and injections were repeated after 12 days according to the vaccine protocol, and 12 days after the second vaccine injection, spleen samples were collected. On-way variance analysis was used for statistical analysis and P<0.05 considered for significance. According to the results of the study, the ratio of Th1/Th2 after antigen stimulation was significantly different between the control group with vaccine and exercise-PcDNA (P <0.05). Also, the exercise-vaccine group was significantly different from all research groups (P <0.05). The results of the present study confirm the increase in the Th1/Th2 cell ratio following repeated bouts of eccentric endurance exercise. Stimulation of muscle damage through exercise before DNA vaccination may have been effective in invoking innate immune system cells and effectively stimulating acquired immunity.

Viral fusion protein through protected areas called fusion peptide are essential for entry of the virus into the host cell by membrane integration. Fusion phenomenon occurs at the host cell surface or in the cytoplasmic compounds and in cytoplasmic components. The fusion proteins are divided into three categories according to variations that are likely during the fusion. In the first category, which includes viruses such as orthomixo, paramyxo, filo, corona, and retroviruses, definite domains of fusion proteins are cleaved and removed and they become mature and functional. The second group of fusion proteins is alpha and flaviviruses become their functional form without cleavage. The third category also includes viruses such as vesicular stomatitis, herpes simplex, and baculovirus, they have common features of the first and second categories.The changes in fusion protein in the levels before and after fusion, description of fusion proteins in viruses such as influenza, filo, and reoviruses as a prototype of fusion protein viruses and their therapeutic applications of fusion protein as potential drugs such as Lactoferrin and Enfuvertide in preventing the occurrence of fusion phenomenon is an important issue for consideration about multiplication and virus entry.

The Coronaviridae family includes viruses that are considered the causative agents of respiratory infections, and among human RNA viruses have the largest genome. Coronaviruses undergo elusive genetic changes through mutation during replication. A gene mutation is a permanent alteration in the nucleic acid sequence; if it occurs in large numbers, it causes changes in the biological features of a species. Fundamentally, viruses adapt to the human body during replication. Several studies have shown that most mutations do not have much effect on pathogenicity. Sequence diversity in new coronaviruses is very low. However, antigen drift has been observed among some coronaviruses. Most coronavirus mutations occur intermittently in Iran and other countries and have little effect on the pathogenicity of the virus but have increased its rate of transmission. In mutated viruses, deletion of nucleotide sequences has been observed relatively in some reading frames extensively. Studies have shown that the host protein induced mutagenesis through interaction via viral proteins. The most important mutation in SARS-CoV2 compared to the original Wuhan virus was the spike D614G mutation and the lineage of B.1.1.7, 20I/501Y.V1become the dominant and exhibit greater virus spread but did not associate with higher viral loads and morbidity. However, it may affect the effectiveness of the vaccines and mortality rate.

SARS-COV-2, the latest member of Coronaviridea family as the cause of the recent global epidemic, has inflicted sever damages in different fields on most governments. Appropriate strategies such as identifying infected individuals and isolating them, trying to produce an effective vaccine, finding efficient treatment and making correct decisions in the social, economic and health fields are the current priorities of the world. Serological tests provide useful information in all of the above areas. In this article we will discuss the importance of performing SARS-COV-2 serological tests in diagnosis, vaccination, treatment and proper planning in the society.

Autophagy has been introduced as a protective mechanisms in cell in recent years. Regarding to the changes induced in autophagy factors following an acute physical exercise. The purpose of this study was to investigate the changes in autophagy factors in time courses of acute exhaustive endurance exercise in Tibialis Anterior skeletal muscle of BALB/c mice.

Regarding the purpose of the study, 12 BALB/c mice were divided into two groups of exercise and control. Subjects were anatomized immediately and 12 hours after the end of the exercise session. The Real Time-PCR method was used to determine the expression of autophagy factors.

Independent t-test showed significant increases in autophagy activation immediately after exercise and significant decreases 12 hours following acute exhaustive endurance exercise (P≤0.05).

  Our results proposed that significant increases induced in autophagy factors immediately after acute exhaustive exercise. Possible correlations between autophagy activation and changes in oxidative stress, some immunological factors and metabolic responses could be some of mechanisms of autophagy activation following acute exercise.

Colorectal Cancer (CRC) is the third most common cancer and the second leading cause of cancer-related death all around the world with an estimated 1.2 million new cases and a half million deaths each year. Although some curing methods such as surgery and chemotherapy are used in the early stage of CRC, targeted therapy is one of the principal modes of cancer treatment that could be a reliable and noninvasive strategy. It seems that Mesenchymal Stem Cells (MSCs) which are undifferentiated and multipotent with the ability to self-renew and differentiate into several distinct cell lineages, are emerging as a targeted anticancer agent and exert their biological and therapeutical activity through immune system modulation and release of cytokines and chemokines to the tumor site. These cells also play a significant role as a carrier to delivery drug to the primary site or metastasis of tumors. There are some studies on MSC`s potential to destroy tumors by inhibition of proliferation and induction of apoptosis. In contrast, tumor tendency is one of the most important characteristics of MSCs in which Adipose Derived MSCs (ADSCs) are the cause of tumor growth and metastasis. Furthermore, obesity indicated that increase of adipose tissue and ADSCs are in association with colon and breast cancers. Therefore, the double-edge sword role of MSCs is revealed as a therapeutic agent.

Oncolytic viruses (OVs) are a new approach in treatment of cancer. Antitumor efficacy of OVs were limited due to insufficient and non-specific viral delivery to tumor sites. To overcome this issue, mesenchymal stem cells (MSCs) were used for their ability to specifically homing into tumors. The main aim of this study was to use MSCs as carriers and investigate the effect of oncolytic reovirus infection in MSCs, induction of apoptosis, nitric oxide (NO) secretion and their effects for selectively killing tumor cells, to use in future. MSCs isolated from mice adipose tissue and confirmed. Then, the ability of the virus to infect MSCs and the effect of reovirus infection in induction of apoptosis and NO secretion in MSCs were evaluated. The results demonstrate that reovirus could replicate on MSCs. The finding indicated that the NO production significantly was higher at 72 h post infection with different MOI in comparison to the control cells. Also, reovirus induced high level of apoptosis in the MSCs at 48 h post infection compared with the control cells. Based on observed results, reovirus increased the secretion of iNOS (inducible nitric oxide) in the infected MSCs at 48 h post infection; therefore, high amounts of NO and reovirus replication were found to trigger apoptosis at 48 h post infection. Therefore, by optimizing the replication time of virus in the MSCs, specific viral delivery to tumor sites are available and causes cancer cells’ death.

Human papillomavirus (HPV) oncoproteins, including E6 and E7 are constitutively expressed in cervical cancer cells. These proteins are ideal targets to be used for developing therapeutic vaccines against existing HPV-associated carcinomas. The aim of this study was to measure the proliferation response rate of splenic lymphocytes derived from E7-HPV16 encoding plasmid injection on the tumor mouse model of papillomavirus.
C57BL/6 mice were inoculated subcutaneous with 5× 10⁵ TC-1 cells in three times with two weeks intervals and then immunized with HPV-16 E7 DNA vaccine. The proliferation response of splenic cells was measured by MTT assay. IL12 cytokine was measured by ELISA assay and the mass of tumor was calculated with caliper for six weeks.
Following the application of DNA vaccines containing E7 therapeutic gene, the proliferative response of splenic cells was provoked significanltly higher than the stimulation in control group (P Our findings revealed that the application of DNA vaccine containing E7 gene in a tumor mouse model may induce anti-tumor cellular immune responses.

More than 99% of cervical cancers contain human papillomavirus (HPV), particularly the high-risk HPV type 16 (HPV-16). Among therapeutic HPV vaccines, DNA vaccines have emerged as a potentially promising approach. The main problem with DNA vaccination is the efficient delivery of the genes. A different delivery system has been used to bypass this problem. Archaeosomes have shown high stability during oxidative stress. In this study, we prepared the archaeosome Halobacterium salinarum polar lipid and used it as a delivery system and adjuvants for formulation with the E6/E7/L1 chimeric plasmid as an HPV vaccine candidate.
The recombinant pIRES2-plasmid that contained an E6/E7/L1 chimeric gene of HPV were purified after extraction. Halobacterium salinarum total polar lipids were prepared according to a method by Bligh and Dyer. The archaeosome-pDNA complex was prepared by the addition of plasmid DNA to an archaeal lipid solution and the mixture kept at room temperature to allow for complex formation. Particle sizes and zeta potential of the samples were measured using dynamic light scattering. We measured the relative tumor volume after administration of TC-1 cells to C57BL/6 mice.
Zeta potential of the anionic archaeosomes was -6.84mV while archaeosome-pDNA complexes were -29 mV. The highly negative zeta potential of archaeosome-pDNA complexes demonstrated excellent loading of the plasmid on the nanoparticle surface and electrostatic stability. The results showed that the archaeosome-containing E6/E7/L1 chimeric gene significantly inhibited the rate of tumor growth in comparison with the control groups.
Archaeosomes are easy and cost-economic to prepare and highly stable. They may hold tremendous promise as vaccine delivery vehicles

Foot-and-mouth disease is an important viral disease of cloven-hoofed animals. Inactivated whole particle virus vaccines are still widely used in prophylactic vaccination campaigns. The choice of adjuvant is a very important factor in enhancing immune responses and the efficacy of inactivated vaccines. Montanide ISA 61 VG is a new ready-to-use mineral oil-based adjuvant developed by SEPPIC Inc. (SEPPIC, France) with high-potential immune responses needed for clinical protection against FMD infection. In this study, we compared the efficacy of two FMD vaccines either formulated with the new oil-based adjuvant ISA 61 VG and saponin, or with aluminum hydroxide gel and saponin. Both vaccines contained the same antigen payloads of O2010/IR. Two groups of 15 naive cattle received a single vaccination with different doses (full dose, 1/3 dose and 1/9 dose) to calculate their PD50 (50% protective dose) after being challenged with the homologous virulent virus. The mean neutralizing antibody titer was determined at 0, 7, 14 and 21 days after vaccination, measured by a micro neutralization test. The new vaccine improved humoral immune responses by 19%, while inducing a higher geometric mean. The titer for neutralizing antibodies was 2.91 log10 compared to the alum-gel based adjuvant vaccine which was 2.44 log10 (P-value=0.1782). The new vaccine showed a PD50 value of 10.05 as compared to a PD50 value of 4.171, respectively. According to the results, the FMD vaccine formulated with the new oil adjuvant, ISA 61 VG, shows potential as an alternative vaccine for routine and emergency vaccinations in the FMD enzootic region.

The rotavirus nonstructural protein 4 (NSP4)، is a multi functional protein that play key role in both viral morphogenesis and cytopathic effect associated with cell death. However، the complete biological effect of NSP4 remains to be clarified. Since to obtain further knowledge about this protein there is a need for recognizing antibody and there is no commercial antibody against this protein، this study was designed to produce polyclonal antibodies against a synthetic immunogenic peptide of RF rotavirus NSP4 protein in order to be used as diagnostic and research tool. In the present study a peptide sequences corresponding to NSP4 114-135 conjugated to Bovine Serum Albumin (BSA) by glutaraldehyde through a single-step coupling protocol. The conjugated peptides were extensively dialyzed and injected to New Zealand white rabbits. The NSP4 114-135 peptide-specific antiserum was confirmed by both IF and Western blot analysis. The result indicated successful production of polyclonal antibody raised against native NSP4 protein. In conclusion، since NSP4 is a toxic protein for the cells، it is impracticable to express full length of NSP4 protein in expression systems. Therefore، the introducing immunogenic peptide in appropriate animal may be the best approach to produce its specific antibody.

Rotaviruses are the members of the Reoviridae family containing double-stranded RNA (dsRNA) genome which are the main cause of gastroinentritis particularly in children less than three years. This study was designed to evaluate the detection of rotavirus genome by new silver staining method using polyacrylamide gel electrophoresis (PAGE). In this descriptive study, the samples were collected from infected MA-104 cell culture and the RNA electrophoresis was performed in 10% polyacrylamide slab gels after RNA extraction. According to polyacrylamide gel electrophoresis and sensitive staining analysis, rotavirus RNA segments were divided into 4 groups and single-nucleotides differences were clearly detected rapidly. New silver staining method using polyacrylamide gel electrophoresis has the capacity to detect the rotavirus electeropherotype within a few minutes even in small DNA/RNA pieces up to 7 picograms.

The aim of the present study was to identify a suitable cell line for studies of recombinant protein production in eukaryotic system. After transfection, altered expression levels of RNA and its target protein were analyzed. To investigate the in vitro expression of E6 protein of human papillomavirus type 16 in cell culture, the plasmid pcDNA3-E6, and two different cell lines MCF7 and CHO were used. Following transfection, RNA expression was determined by RT- PCR and its protein target was evaluated by Western blot using SDS-PAGE gel and Anti-E6 monoclonal antibody. In this study due to the low concentration of proteins in MCF7cells, RT-PCR was positive and Western blotting was negative, while RT-PCR and Western blotting were positive for CHO cell lines. Regarding the results, the use of CHO expression system as a tool for efficient transfection of recombinant protein production is suggested.

Apoptosis or programmed cell death is known as a conserved gene-directed mechanism for the specific elimination of unnecessary or unwanted cells from an organism and is involved in many immune system mechanisms and diseases. The main differences of this pathway with cell necrosis as two main pathways for elimination of unwanted cells are the absence of inflammation and the restricted effect on target cells. Apoptosis has an important role in biological processes such as normal development، tissue homeostasis، elimination of destroyed cells or virus infected cells and remove of self antigen-activated immune cells. A poptosis is important for regulation of cell growth and proliferation، development and body health and many autoimmune diseases، cancers and viral infections are the result of impaired or inhibited programmed cell death. Therefore the main objective of apoptosis research is to identify molecular components and regulatory mechanisms specially Bcl-2 and IAP family as the most important regulators of apoptosis and this information may lead to application of therapeutic agents that can modulate this process in the treatment of neurodegenerative disorders and proliferative diseases such as cancer.

While hepatitis C virus (HCV) is the major cause of acute and chronic hepatitis, cirrhosis, and hepatocellular carcinoma worldwide, no vaccine against this infection has been proved to date. Cellular immune responses play an important role for eradicating persistent viral infections. Among different vaccine strategies, the use of DNA vaccine has been shown to be a promising approach for enhancing cellular immune responses. In spite of their advantages, DNA-based vaccines might induce weaker antibody and cytotoxic T-lymphocyte responses compared to protein immunization. To overcome this obstacle, several methods such as different immunization regimens, fusion of particle forming units [like hepatitis B surface antigen (HBsAg)] and co-expressing cytokines have been tested. The aim of this study was to design, construct, and evaluate an HBsAg-fused core-based DNA vaccine against HCV infection. The HCV core gene was amplified by polymerase chain reaction (PCR) and cloned in BamHI/EcoRV sites of pcDNA3.1 containing HBsAg. The constructed plasmid (pCHCORE) was analyzed by restriction enzyme and sequencing analyses and evaluated for the protein expression in HEK293T cell line by western blot analysis with anti-HBsAg polyclonal antibody. The correctness of the constructed DNA vaccine was shown by restriction enzyme analysis and sequencing. Western blotting results confirmed the expression of HBsAg-HCV fusion protein with an expected molecular weight in HEK239T cell line. In accordance with previous studies, the constructed vector (pCHCORE) compromising the fusion of HBsAg to HCV core in pCDNA3 plasmid might be used as a HCV DNA vaccine (due to proper expression in cell lines) to induce augmented cellular immune responses.

Herpes simplex encephalitis (HSE) is a rare but very serious disorder caused by herpes simplex type 1 virus (HSV-1). Following primary HSV-1 infection, viral latency develops in the trigeminal ganglia (TG). Therefore, prevention from primary HSV -1 infection and subsequent latency establishment is an important feature for vaccine development.
We evaluate efficacy of DNA vaccine encoding Glycoprotein D-1 (gD-1) gene of HSV-1 against lethal ocularly challenge of TCID 5 × 105 plaque-forming units (pfu) per eye of wild HSV-1 versus negative control groups.
The data demonstrated protective effects of DNA vaccine encoding Herpes simplex virus Glycoprotein D gene on latency rate in TG and mortality rate.
These results indicated that immunization with g D-1 DNA vaccine is a promising approach for eliciting a protective immunity against a HSV-1 lethal ocular challenge.

Group A rotaviruses (GARV) are responsible for the vast majority of severe diarrhea worldwide that kills an estimated 600،000-870،000 children annually. Since infantile gastroenteritis is a main health problem، therefore diagnosis and treatment of this disease is crucial. Gene rearrangements have been detected in vitro during serial passages of the virus at a high multiplicity of infection (MOI) in cell culture، as well as in chronically infected immunodeficient individuals. In this study، we developed an RT-PCR method to detect and diagnose the standard and gene rearranged bovine rotavirus. Rotavirus RNA was extracted from confluent monolayers of infected MA-104 cells، stained with silver nitrate، and then electrophoresed in a 10% polyacrylamide gel. The full-length gene products that encoded the NSP1، 2، and 3 genes of the standard and rearranged rotavirus were amplified by RT-PCR using specific primers. We observed rearranged NSP1 and NSP3 genes that had different migration patterns seen with polyacrylamide gel electrophoresis. NSP1، 2، and 3 gene segments from standard and rearranged rotaviruses were amplified by RT-PCR، then the complete nucleotide sequence of each gene was subjected to sequencing. The results showed the generation of gene rearrangement through serial passages of the bovine rotavirus RF strain. Serial passage of rotavirus in cell culture at a high MOI and chronic infection in immunodeficient target groups might alter rotavirus evolution. The methods utilized for detection and characterization of rotaviruses are continually evolving and being refined. Data collection is necessary to understand the molecular and antigenic features of the rotavirus in order to have a successful implementation of rotavirus studies and the development of a rotavirus vaccine. This study shows the importance of genetic variation and can provide valuable information about the amplification، diversity، biology، and evolution of rotaviruses.

Titration of viruses is important to determine the quantity of virus in vaccine development، master virus seed stock preparation، viral vector studies and virus replication. In this study، we compared the CCID50% and plaque assay as a standard titration method for rotavirus (RF) and HSV-1. The MA104 and Vero cells were inoculated by RF and HSV-1 in 6- and 96-well plates. Following infection and adsorption، the optimal time for the cytopathic effect caused by the viruses was noted and the results compared. The CPE (Cytopathic Effect) of RF was observed in less than 18 hours، which increased until 72 hours after inoculation. In HSV-1، the CPE was observed 24 and 72 hours after inoculation. The virus titration in the plaque assay was monitored at 96 hours post-infection for RF and at 72 hours post-infection for HSV-1. In both viruses the plaque titer method was lower than the CCID50 method، since the results indicated that 1 CCID50% was equal to 0. 7 PFU. The plaque assay is one of the most accurate methods for viral titration. For the plaque assay، individual lesions may be isolated، which the plaques can be counted. The CCID50% method is not applicable for purification of homogenous viruses، nor is this technique reproducible.

Human cytomegalovirus (CMV) is a major life-threatening pathogen for hematopoietic stem cell transplant recipients. Specific tests are used for the diagnosis and monitoring of CMV infection in transplant patients. This study evaluates the performance of pp65 antigenemia and qualitative PCR assays for monitoring CMV in such patients. We analyzed 179 clinical samples from 41 patients by using a validated home-brewed qualitative PCR and a commercial antigenemia assay. The obtained results were evaluated using quantitative real-time PCR as the gold standard. CMV was observed in 26.8% of samples analyzed by the antigenemia assay and in 42.6% of the samples by qualitative PCR. Among 179 clinical samples, 50.8% were negative and 21.2% were positive by both assays. On the other hand, 26.3% were only positive by qualitative PCR whereas 1.7% were positive by the antigenemia assay. A comparison of the results with real-time PCR showed that qualitative PCR has a higher sensitivity than the antigenemia assay (98.7% vs. 45.7%). The specificity of both assays was equal (96.8%). Quantitative results of the antigenemia assay showed good correlation with real-time PCR (r=0.715; p<0.001). Both the qualitative PCR and antigenemia assays have special deficiencies for efficient diagnosis of CMV infection. Therefore, effective management of CMV infection in transplant patients requires the use of other sensitive quantitative methods such as qPCR.

The aim of this study was to construct a pcDNA3.1+ vector containing FMDV type O/IRN/1/2007-VP1 gene, protein expression in BHKT7 cells and evaluation of immune response in BALB/c mice. FMDV type O/IRN/1/2007 was isolated from a cattle in Ray in 2007 and serotyped. The purified VP1 gene was sub-cloned into the PTZ57R/T vector and pcDNA3.1+ expression vector. The PCR product of Vp1 gene without stop codon was sub-cloned upstream of EGFP gene into the pEGFP-N1 vector to evaluate VP1-GFP fusion protein expression. The pcDNA3.1-VP1 and pEGFP-VP1 vectors were transfected into BHKT7 cell line. The expression of VP1 protein was evaluated by SDS-PAGE, western blotting and florescent analysis of VP1-GFP fusion protein. The mice were injected subcutaneously by pcDNA3.1-VP1 vector as DNA vaccine and titration of neutralizing antiserum and T cell proliferation assay were done to evaluate the immune response. Insertion of VP1 gene was confirmed by double digestion of sub-cloned PTZ57R/T, pcDNA3.1+ and pEGFP-N1 vectors. The specific band in western blotting was also confirmed the VP1 protein expression in BHKT7 cells. The expression of VP1-GFP fusion protein was observed under the immune-florescent inverted microscopy as more green florescent spots versus expression of GFP protein, alone. The neutralizing antiserum titer and T cell proliferation increased significantly in the group of mice vaccinated with pcDNA3.1+-VP1 vector verses control groups (P<0.05). The results showed that the target gene was amplified, cloned in the cloning and expression vectors and protein expression was confirmed successfully. According to the confirmed VP1 protein expression and increasing neutralizing antiserum titer and T cell proliferation by pcDNA3.1+-VP1 vector (P<0.05), it can be used as DNA vaccine against FMDV type O/IRN/2007.

Cytomegalovirus is one of the most important viruses that its presence in transplant recipients has been surveyed. This virus can led to the severe disease or death. The prevalence of CMV infections in transplant recipients varies from one country to another. Indirect immunofluorescent method represents the virus multiplication and active infection, while PCR technique shows the presence ofvirus. The aim of this study was to determine the prevalence of active CMV infection in kidney transplant recipients using PCR and indirect immunofluorescent methods based on the season, age, and sex variables. samples from kidney recipients admitted to Gholhak laboratory during year 2006 and 2007 from all over Iran. This study was performed using PCR technique and indirect immunofluorescent method (PP65 antigenemia assay). The results were analyzed by Chi2 and lojestic regression. In total 923 samples were investigated. Seventy one (7.7%) and 323(35%) of samples were positive by antigenemia and PCR respectively. There was no correlation between CMV infection and age or sex of the patients (p >0.05). However there was a correlation between presence of virus and season in either used methods of study, which was statistically significant in the spring. The spring has meaningfull (p =0.01). Positive results obtained by PCR were more than antigenemia and this shows the higher sensitivity of this method to detect viral infection. we found no correlation between viral infection and sex or age by tested methods, but the infection rate had significant increase during spring.

Bacteriophage vectors recently have been considered as a gene transfer and vaccine delivery vehicles chiefly due to their low cost, safety, and physical stability. Since, little is known about phage mediated gene transfer in mammalian hosts, A group of invitro experiments were performed to ascertain gene transfercapability of these vehicles.

For this purpose, different genes eg. (EGFP as positive control and PBR322 as negative control) were sub cloned into the vehicle Lambda ZAP cassettePossessing expression capability in animal cells.New Recombinant-phage constructs using packaged using bacteriophage lysates. In order to evaluate functionality of the produced bacterio phages EGFP bacteriophage particles were added to eukaryotic cell cultures and after 36 hours, amplified and purified phage particles were assessed for gene expression criteria in cell culture models. Results and

The observation of fluorescent signals arising from GFP(green fluorescent protein) means vehicle expression in eukaryotic systems.stability of the recombinant phages as oral vaccine vehicles were checkedIn similar conditionsas in gastrointestinal tract. (Ph &destructing enzymes) The results of stability assay showed that the phages have potential for employing as oral vaccine vehicles.

Herpes simplex virus (HSVs) is a widespread human infectious agent, responsible for persistent and latent infections. Herpes simplex virus infections are usually continually recurrent in the normal population and represent a significant cause of complications in immunocompromised patients. In this study HSVs were propagated in BK cells and more than 502 samples were taken and analyzed for HSV IgG antibodies using Virus Neutralization Test (VNT) as golden standard test for evaluating in house Enzyme Linked Immunosorbent Assay (ELISA). Based on the results 80.48 % and 81.67% were positive (1.8) in VNT and ELISA respectively. There was a significant correlation between the VNT and ELISA tests in the tested samples (Pearson’s r = 0.96). Our data showed that the in house ELISA can be used for screening and determination of the prevalence of HSV IgG antibodies, which can facilitates patient management using suitable and cost effective laboratory diagnostic tests.