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Background:Accumulative research is in progress to clarify clinical aspects of GBV-C. The possibility of interaction between HCV and GBV-C as well as its consequence on development of liver diseases is the most important clinical aspect which encourages researchers to develop a rapid and cost effective technique for simultaneous detection of both viruses. Methods: In this study, a SYBR Green real time multiplex RT-PCR technique as a new economical and sensitive method was designed and validated for simultaneous detection of HCV/GBV-C in HCV positive plasma samples. SYBR green real time RT-PCR technique optimization was performed separately for each virus. Multiplex PCR was established next. Standard sera with known concentrations of HCV RNA and dual HCV/GBV-C positive control samples along with negative control samples were used to validate the assay. Results and Conclusions:  Fifty six non cirrhotic HCV positive plasma samples [29 of genotype 3a and 27 of genotype 1a] were collected from patients before receiving treatment. 20.6% of genotype 3a and 18.7% of genotype 1a showed HCV/GBV-C co-infection. As a result, 19.6% of 56 samples had HCV/GBV-C co-infection that was compatible with other results from all over the world. SYBR Green real time multiplex RT-PCR technique can be used to detect HCV/GBV-C co-infection in plasma samples. Furthermore, with application of this method more time and cost could be saved in clinical-research settings.

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The purpose is examining factors that promote satisfaction for student in the school. This research is fundamental both quantitative and qualitative in nature. In this study, two methods have been used: a) Survey research methods (survey) b) Correlation method. For a multistage cluster sampling method was used. In order to assess the individual's perspective, creating table of contents based on studies and interviews with architects and experts, will be discussed. According to this table question naive was designed and distributed among the population. After classifying data using the software SPSS, the analysis is discussed. Five factors were extracted on patient satisfaction. These factors include: physical comfort, perceived environmental, psychological security, environmental attractiveness, sense of belonging. Operating resultant T-test analysis was located. Finally, it was found that 95% of the sample mean is greater than from average. At least 70% percent of the population had an agreement with the agent.

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The effects of four insecticides, abamectin (1500 and 750mgl^-1), emamectin benzoate (1000 and 500mgl^-1), acetamiprid (500 and 250mgl^-1), and flubendiamide (500 and 250mgl^-1), were studied on different preimaginal stages of T. brassicae and T. evanescens, the egg parasitoids of tomato leaf miner Tuta absoluta (Lepidoptera: Gelechiidae). Parasitized eggs of the Angoumois grain moth Sitotroga cerealella (Lepidoptera: Gelechiidae) were treated by the dipping method at the larval, prepupal, and pupal stages of the parasitoid. For persistence evaluation, the insecticides were applied at the recommended concentration on tomato plants by a hand sprayer till runoff point. Plants were maintained under a transparent polyethylene rain cover in the field. Leaves of the treated tomato plants were sampled and transferred to the laboratory at time intervals of 3, 5, 16, and 31 days after application. Based on our study, abamectin was the most harmful insecticide for immature stages of both parasitoids T. brassicae and T. evanescence. Treatment by abamectin at the pupal stage had more adverse effects compared to prepupal or larval stages. Acetamiprid with 30.5% and 31.6% mortality in less than five days was classified as the short-lived insecticide for T. brassicae and T. evanescens, respectively. The same result was obtained in flubendiamide treatment which caused 27.2% and 26.1% mortality to the parasitoids, respectively. Abamectin with 16.1% and 13.8% mortality in less than 16 days was slightly persistent. However, emamectin benzoate with 13.3% and 15.5% mortality in less than 30 days was classified as moderately persistent for those two species, respectively. Therefore, flubendiamide and acetamiprid were non-harmful to both T. brassicae and T. evanescence wasps and are good candidates to be incorporated into IPM programs in combination with biological agents for the control of tomato leaf miner T. absoluta. By contrast, emamectin and abamectin should be used with greater care as a part of an IPM procedure.

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During the last decade, plant and microbial-derived metabolites have received growing attention as potential tools for pest management in agriculture. Pederin (C₂₅H₄₅NO₉) is a vesicant toxin produced by Pseudomonas-like bacterial symbionts of rove beetles within the genus Paederus (Col: Staphylinidae). In this study, the toxicity of pederin to two stored product pests, Ephestia kuehniella Zeller (Lep: Pyralidae) and Tribolium confusum Jacquelin du Val (Col: Tenebrionidae) was evaluated using laboratory bioassays. Probit analysis estimated the median lethal concentrations of pederin as 1311.96 and 596.36ppm for E. kuehniella fourth larval instar and T. confusum adults, respectively. We also measured the activity of two major digestive enzymes (amylases and proteases) as well as three major detoxifying enzymes (P_450s monooxygenases, glutathione S-transferases, and carboxyl esterases) in insects treated orally with pederin. Feeding on pederin resulted in significant decrease in the activity of amylolytic, proteolytic, and carboxyl esterase enzymes, but significant increase in the activity of P_450s and glutathione S-transferases. Results of this study may highlight pederin as a novel source of pesticides with unique mode of action for use in pest management programs.

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Introduction: Hepatitis C virus (HCV) is the major cause of acute and chronic hepatitis, cirrhosis, and hepatocellular carcinoma worldwide. Because of increasing number of infected patients throughout the world, various studies are concentrated on development of high quality diagnostic tests and genotyping in various regions of the world. Methods: In this study, we used RT-PCR to isolate and amplify the core gene sequence of the hepatitis C virus that is very conserved among various genotypes, from an isolate derived from an Iranian patient with chronic hepatitis. Then it was cloned in pUC18 vector and sequenced with universal primers. In order to produce recombinant core protein, the core gene was cloned in PET28A expression vector and the recombinant plasmid was transformed into BL21 (DE3) bacteria. Results and Discussion: The results of sequencing showed that the core gene of the hepatitis C virus from an isolate derived from an Iranian patient with chronic hepatitis has a higher similarity with 1a (96%) and 1b (95%) strains of the virus. This result is similar to those obtained by previous studies. The presence of a 23.5 KD band in SDS-PAGE and Western blot using monoclonal antibody, proved the expression of Core protein in PET28A vector. Mass production of this protein could lead to its use in detection of anti-HCV antibodies in infected patients by immunoassays.

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Objectives: Human Immunodeficiency Virus type 1 is the causative agent of Acquired Immunodeficiency syndrome “AIDS” in human and demonstration of HIV-1 genome in samples is accepted as evidence of infection. Transmission of Infection during window period in blood transfusion settings is a world wide concern. Also there is a need for a rapid, sensitive and accurate technique to detect HIV-1 infection prior to antibody appearance in patients and new borns. Material and Method: A rapid Visual DNA Chip based on RT-Nested PCR and Enzyme-Substrate detection system was developed. At first a specific RT-Nested PCR was developed and the products were confirmed in gel electrophoresis and the products were labeled with DIG (Digoxigenin). The labeled products were then hybridized with the pre-prepared chip with an anchored specific probe. After the washing procedure an antibody against DIG conjugated with alkaline phosphates enzyme was used. After the second washing procedure the BCIP/NBT substrate was used and development of color was interpreted as positive while the negative samples developed no color. Results: 35 sera samples from different stages of HIV infection (AIDS, Asymptomatic and Symptomatic Infection) as well as 20 confirmed negative sera samples were collected and checked with the developed assay. All the positive samples developed reaction while the negative samples had no reaction. Conclusion: In the current study the developed assay showed high sensitivity and specificity to detect HIV-1 infection. It seems that the viral genome could be detected prior to antibody appearance and hence the window period could be shortened. Also the assay could be used to detect infection in new borns from infected mothers, because the maternal antibody could pass the placenta and antibody based assay have false positive results. Because of the high sensitivity, the developed assay could also detect infection in very low viral load conditions.

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Objective: Hepatitis C virus is the major cause of viral hepatitis and its diagnosis in suspected specimens is of great importance. The risk of transfusion- transmitted virus infection is primarily the result of failure in serological screening tests to detect recently infected donors in the pre-seroconversion window period of infection. Therefore, sensitive and accurate diagnosis of HCV prior to antibody production to reduce window period is necessary. Materials and Methods: In the present study, a sensitive and specific RT-Nested PCR method for detection of a conserved HCV 5'UTR sequence was developed. Two pairs of primers for amplification of the target sequence in two rounds of PCR were selected. The developed RT-Nested PCR assay was performed on HCV-antibody confirmed positive samples as well as negative controls and standard samples. In order to compare the results, One Step RT-PCR kit was used in this study. Results: 25 HCV-positive plasma samples whose positivity were confirmed by ELISA and Western Blot tests, also as well as 10 fold dilutions of a high viral load plasma sample obtained from a HCV-positive patient as standard samples and 25 negative control plasmas from healthy blood donors were collected and tested by this assay. In all of positive samples a 175bp band was observed on agarose gel electrophoresis, but no band could be detected in negative control plasma. Results from developed RT-PCR assay and One Step RT-PCR kit showed a good correlation. Conclusion: According to the results of this study, the developed RT-Nested PCR assay has a good sensitivity and specificity for diagnosis of HCV infection. It has the advantage of viral genome detection prior to seroconversion and can be used to detect HCV infection during window period

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Objective: In this study, a SYBR Green real-time RT-PCR assay for quantification of HIV-1 viral RNA was developed. Materials and Methods: This assay was performed based on amplification of the pol region of HIV-1 and product analysis by an ABI 7500 system. We quantified HIV-1 viral load in 26 seropositive patients by this system and the data were subsequently compared with results obtained with a reference technique represented by COBAS AMPLICOR HIV-1 Monitor test. Results: The results demonstrated that this technique could detecte up to 500 HIV-1 RNA copies/ml of plasma. The linearity of this approach was conserved over a wide range of HIV-1 copy numbers (5×102-5×109). Since no positive signal was observed in seronegative volunteers, the specificity of the test was calculated as 100%. Comparison of the results with those obtained by the reference quantification method, revealed a significant correlation between the results (R2= 0.95). Conclusion: On the basis of the most recent recorded cases for HIV-1 infection and AIDS in Iran, the prevalence of this disease is rising rapidly and the situation has been called to be alarming by national health representatives. Determination of HIV-1 viral load in plasma has been considered as the most effective single prediction tool of clinical outcome. Indeed, the development and stabilization of HIV-1 RNA assays have given physicians a unique tool for monitoring HIV-1 patients treated with antiviral drugs. In this study, we have developed a SYBR-Green Real Time RT-PCR assay for quantitative analysis of HIV-1 in infected patients. Since a synthetic RNA standard was used in this assay, the upper limit of detection was detected to be higher than the standard test (5×10 9 versus 7.5×10 5). This can be important in patients with acute high viral load infections. Reproducibility was assessed by Intra assay and Inter assay analysis. Coefficient of variations Ct, in reproducibility tests for Intra assay and Inter assay variability were less than

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Objective: The global HIV epidemic continues to expand and exceeding previous predictions. An effective vaccine represents the best hope to curtail the HIV epidemic. DNA vaccines induce humoral and cellular responses and mimic live vaccines without their pathogenic potential. The importance of CD8+ CTL responses in controlling HIV and SIV viremia has led to production of a series of vaccine candidates that effectively induce these responses. It is now widely believed that an HIV vaccine strategy must stimulate both a strong humoral (antibody) as well as cell-mediated (CTL) immune response.The p24 and gp41 play many important roles in host-virus interaction and pathogenesis. These proteins are considered as attractive vaccine candidate in which their immunogenecity and immunomodulatory effects have been confirmed. Materials and Methods: In this study, a construct, pcDNA3.1Hygro- (p24-gp41), was evaluated as a DNA vaccine candidate in Balb/C mice for generation of effective cellular immune responses. For immunizing, we used dendrosome, a novel family of vehicles for transfection and therapy. IFN-γ cytokine production and total antibody were detected by ELISA. Lymphoprolifration assay was performed by MTT test. Results: ELISA and MTT assays confirmed that the cited p24-gp41 fusion gene is able to enhance immune responses in mice. Conclusion: The construct that was used in this research can be a good candidate for DNA vaccine against HIV-1, if the future complementary tests demonstrate the same trends of immunogenic responses shown in this study.

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Aims: Electronic health can lead to health preservation and promotion using information and communication technologies to receive and record accurate data, appropriate storage, and retrieval, as well as the health information management approach. The present systematic review aimed to assess the E-health application during the COVID-19.
Information & Methods: The present systematic review was done based on PRISMA protocols. The study data were retrieved using the E-health and COVID-19 keywords in the related studies from August 4, 2021, in PubMed, Scopus, Magiran, and Sid databases. Moreover, the inclusion criteria were original research studies that used E-health to manage patients with COVID-19.
Findings: A total of 10 articles were included in the study, 40% of which focused on the impact of E-health on reducing fear and anxiety caused by COVID-19, 30% on the E-health in early diagnosis and progression of the disease, 10% on the E-health application in the field of prevention, 10% on E-health in the field of disease control, and 10% on E-health for quick investigations of the disease process and access new medical information. The used technologies included virtual training through WhatsApp video calling, Instagram, iGap and Telegram voice and text messaging, artificial intelligence, and data mining techniques.
Conclusion: E-health tools played a prominent role during the COVID-19 in the prevention, diagnosis, control, and fear reduction of coronavirus disease. Various practical strategies such as financing, implementation and legal requirements can be considered to effectively use the capabilities of eHealth tools in disease management.
 

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Objective: The use of bacterial plasmids carrying specific genes of pathogens as genetic vaccines is a relatively new technique for induction of cellular immune responses against microbial pathogens. Mechanisms of production of specific immune responses against these vaccines are not still completely understood. Therefore, it is necessary to examine various routes of inoculation to find the best way of immunization for specific antigens. In this research, intramuscular method of inoculation of influenza vaccine nucleoprotein (NP) encoding vector was compared with that of intra-dermal method. Materials and Methods: In this study, the ability of two different methods of immunization (intramuscular and intra-dermal) in induction of CTL responses as well as their efficiency in clearance of influenza virus from the lung of BALB/c mice was compared. Female BALB/c mice were immunized with influenza virus NP expressing plasmids on days 0, 14 and 28. CTL activity of mice was evaluated by lactate dehydrogenase technique two weeks after the last inoculation. In addition, the mice were challenged by live influenza virus and the viral titer was measured 4 days post-challenge in the lungs of animals. The results of experiments demonstrated that intramuscular immunization of mice induces a stronger CTL response. Mice immunized by intramuscular route also showed a higher ability in virus clearance from the lungs. Conclusion: Results of this study showed that different routes of immunization of influenza NP genetic vaccine induce different levels of cell-mediated immune responses and protection from the live virus.

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Objective: In this project, our aim was to construct a novel expressing vector harboring a new sequence from overlapping region of NS3 gene of HCV from infected Iranian patient. Materials and Methods: The partial NS3 (pNS3) gene was amplified by Nested-RT-PCR method using sera of HCV infected patients harboring genotype 1a. After purification and cloning the pNS3 into TA-cloning vector, the best colony was selected based on Blue/White screening and colony-PCR following by confirmation with sequencing and restriction digestion with BglII. The sequenced gene was compared with other reference sequences using alignment softwares. The resultant pNS3 gene subcloned into the expression vector, IRES vector, followed by selection the suitable clones by 2 different colony-PCRs. The gene expression was evaluated using GFP detection, RT-PCR and western blotting techniques after transfection of the IRES-pNS3 vector into the 293 cell line. Results: After pNS3 sequence amplification by RT-PCR, sequencing results showed high homology among the sequences with other reference sequences. This result also showed that it belonged to genotype 1 of HCV. Colony-PCR showed the insertion of gene into expressing vector with the right orientation. GFP expression, RT-PCR and western blotting confirmed transfection of vector, expression of pNS3 gene and production of its protein in 293 cells respectively. Conclusion: This novel expressing vector harboring partial region of NS3 gene in compare to full NS3 gene maybe more useful in immune induction by antigen presenting cells due to absence of genes responsible for protease activity of the protein in the setting of HCV vaccine.

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Objective: In this study, two conserved genes (M1 and NP) of influenza virus were expressed in a bicistronic vector in order to develop a universal gene based vaccine. Materials and Methods: Plasmids M1-pIRES2-EGFP, pIRES2-NP were constructed by cloning the PCR products of M1 and NP genes which were amplified from the A/Peurto Rico/8/34 (H1N1) influenza virus strain into the plasmid expression vector pIRES2-EGFP, respectively. For construction of M1-pIRES2-NP bicistronic plasmid, M1 gene was extracted from M1-pIRES-EGFP plasmid and sub-cloned into pIRES2-NP construct. Finally, simultaneous expression of both genes was assessed by transient transfection of bicistronic plasmid into BHK-21 cell lines and subsequent immunofluorescence staining. Results: The results of enzymatic double digestions on the constructed plasmids and sequencing demonstrated the success of cloning processes of above mentioned genes. Correct expression of these genes was confirmed by M1-pIRES2-NP plasmid expression in BHK-21 cell lines confirmed by immunofluoresence microscopy. Conclusion: Simultaneous expression of influenza M1 and NP genes from a bicistronic plasmid containing “IRES” sequence is achievable.

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Objective: Human cytomegalovirus (HCMV) is a beta-herpesvirus that causes persistent infection in humans, as well as severe disease in fetuses and immunocompromised individuals. Although HCMV is not currently causally implicated in human cancer, emerging evidence suggests that HCMV infection may be specifically associated with malignancies such as gliomas. Gliomas are one of the most common brain tumors that affect humans. It is classified into four grades. In this study, we have developed and used a real-time PCR method for the detection and diagnosis of HCMV infection in glioma brain tumor samples. Methods: Paraffin-embedded tumor samples were chosen from patients who referred to Imam Khomeini Hospital Neurosurgery Ward. DNA was extracted from paraffin-embedded tissues by a DNA extraction kit. After designing specific primers for the HCMV US28 region, a real-time PCR method was developed for detection of HCMV US28. Results: The results of qualitative real-time PCR on 4/18 patients (22.2%) were positive. Two patients with positive HCMV results died. Conclusion: This is the first study that has monitored HCMV genes in samples from glioma patients in Iran. Considering the results of this study and controversies associated with other studies, a more comprehensive study using this and other diagnostic methods is suggested.

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Objective: Despite availability of an effective vaccine against hepatitis B virus (HBV), the global prevalence of this virus infection has not diminished significantly. Contrary to numerous other human viruses, HBV does not have the ability to propagate in cell culture. However, infectious virus has been produced by transfection of human hepatoma cells with plasmids that contain full length HBV genome. Generation and optimization of appropriate cell culture systems can help us in demonstrating the quality of genome replication by PCR as well as expression of surface antigen secretion. Interferon stimulating genes (ISGs) are usually produced in response to interferon and can be determined as a measure of response to IFN-therapy. Therefore, in pharmacological studies, in addition to assessing the effects of a medicine on viral determinants of replication, its’ effects on stimulation of various ISGs, as indicators of innate immune responses, can be achieved. Methods: In this study, we transfected the Huh-7 hepatoma cell line with pCH-9/3091. HBsAg production and viral mRNA transcription were subsequently evaluated. In this system, by using ISGs-specific primers, the ISG mRNAs recognition method was optimized and utilized. Results: Huh-7 cells supported HBV replication. The peak HBsAg secretion was observed at 72 h post-transfection. By using designed primers for the S and pg/pC regions, transcription and genome replication of the virus was shown. RT-PCR results for ISG production by transfected cells showed no role for HBV in enhancement of ISGs levels in Huh-7 cells. Conclusion: The results indicated that this system can be used for functional studies of HBV-specific genes as well as assessment of the effects of new drugs or new vaccines. In addition, it may be used to study the mechanisms of drug resistance that have resulted in difficulties in response to HBV antivirals, including IFN-α.

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Quick and authentic identification of exotic and potentially invasive taxa with capability of causing high economic losses or detriments is essential prerequisite for effective plant quarantine and biological control initiatives. The order Thysanoptera includes several agricultural pest species that, not only because of their minute size but also due to their cryptic behavior, incline to undetected transport through international trade of plants. Identification of thrips, particularly at species level, is pretty demanding and requires expertise in knowledge about Thysanoptera. Moreover, in most cases, identification of larval Thysanoptera to species is impossible without presence of adults. Hence, there is a great desire for a facile, accurate, and highly reliable technique for thrips identification. The present study describes species-specific primers for four pest thrips species, and the use of a multiplex PCR assay to detect and to distinguish between the four target species. Five primers were used to simultaneously amplify a specific region of the mitochondrial DNA and produce species-specific fragments. Results indicated that the primers were capable of detecting these four species and amplifying uniquely sized, species-specific PCR products. Furthermore, using a multiplex PCR assay, the primers maintained specificity and sensitivity, and allowed detection of each of the four species in a single reaction. The stringency of the method was tested using specimens of different developmental stages and consistent results were obtained for all of the examined samples. This method is simple enough to be implemented by non-experts and also can be extended to any organism for which quick and reliable identification is needed.

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Formation of institutions depends on leaders and elite’s beliefs. Some factors influence the beliefs of people. The aim of this study is to identify beliefs affecting the formation of efficient institutions. In this research, the knowledge has been generated by the method of inductive qualitative content analysis, based on the lives of the six persons who have been introduced the experts as influential people in the formation of successful economic institutions. Based on the content analysis of the interviews, the self-efficacy beliefs have positive effects on the formation of efficient institutions, and skills and experiences affect the beliefs. Of two factors influencing self-efficacy belief, the experience of domination (impact of life circumference, and experiences of childhood and teenage) is of high scores in the first three persons than three next ones; however the substitution experience (impact of parents) is similar among sample under study. All six experts had good socioeconomic positions.