This paper describes the use of polynomial integer equations for high level model of digital circuits for property checking formal verification at this level. Most formal verification methods use low-level representations of a design like BDDs. BDD operations are not applicable to a large data path because of large CPU time and memory usage. In our method, a behavioral state machine is represented by a list of integer equations, and RT level properties are directly applied to this representation. Furthermore, this method is applied to circuits without having to separate their data and control sections. For this implementation, we use a canonical form of integer equations, which simplifies equations instead of solving them. This paper compares our results with those of the VIS verification tool that is a BDD based program.

Background: CD8+ T cells are thought to play an important role in protective immunity to tuberculosis. The major histocompatibility complex class I subtype HLA-A*0201 is one of the most prevalent class I alleles, with a frequency of over 30% in most populations. HLA-A*0201 transgenic, H-2Db/mouse beta2- microglobulin double-knockout mice (HHD) which express human HLA-A*0201 but no mouse class I, was shown to provide a powerful model for studying induction of HLA-A*0201-restricted immune responses in vivo. Methods: HHD mice were immunized with plasmid DNA encoding MPB51 by using a gene gun, and IFN-g production from the immune spleen cells was analyzed in response to a synthetic overlapping peptide library covering the mature MPB51 sequence. catatonic T lymphocytes (CTL) activity was measured using cytotoxicity assay and the three-color flowcytometry was used to reveal IFN-g-producing immune spleen cells. Results: Our findings were shown that only one peptide, p51-70, appeared to stimulate the immune splenocytes to produce IFN-g. Flow cytometric analysis with intracellular IFN-g and the T-cell phenotype revealed that the p51-70 peptide contains an immunodominant CD8+ T-cell epitope. Further analysis with computer-assisted algorithms permitted identification of a T-cell nona mer epitope, p54-62. Finally, we proved that the p54-62/HLA-A*0201 complex is strongly recognized by HLA class I-restricted CD8+ MPB51-specific CTL cells. Conclusion: These results suggest that vaccination with MPB51 gene elicited MPB51-specific CTL. In addition, the P54-62 epitope thus represent potential subunit component for the design of vaccines against tuberculosis.

Background and objectives: High mutation rate and immunosuppressive effect of some key proteins of hepatitis C virus (HCV) encouraged researchers to find conserved and small immunogenic units to induce immune responses. Thus epitope-based vaccines were introduced to find out any potential immunogens. However, the relatively weak immune responses against these vaccines require a creation of additional measures. This study aimed to construct plasmids carrying CTL epitope of hepatitis C virus using immunoinformatics modeling method and assessment of their preliminary immunogenicity.Materials and Methods: One H-2Dd and two HLA-A*0201-restricted CD8+ T-cell epitopes from E2, El and core regions of HCV were selected and immunoinformatically analyzed for optimum sequentiality. The multiepitope sequence was constructed by SOEing PCR and cloned in pcDNA3.1+ vector. PADRE, endoplasmic reticulum signal sequence (ERss) and hepatitis B surface antigen (HBsAg) immuno-enhancer sequences were fused to the 5' end of multiepitope constructs. In vitro, expression of each plasmid was analyzed by RT-PCR, dot-blot, Westem- blot and immunofluorescence techniques. Preliminary in vivo immunogenicity of the construct was assessed by delayed-type hypersensitivity (DTH) response in BALB/c mice. Results: In vitro, analyses confirmed the expression of plasmids. Preliminary, in vivo assessments indicated the processing and presentation of the H-2Dd epitope in BALB/c mice. Moreover, although the immunostimulation effects of ERss and PADRE were shown, fusion of HBsAg did not enhance the DTH response. Conclusion: This study shows the value of immunoinformatics prediction of hepatitis C virus epitopes as a multiepitope vaccine candidate and DTH assay for preliminary analysis using BALB/c mice. Data obtained, provide enough support for further evaluation of the designed constructs in HLA-A2 transgenic mice.

Background: Cervical cancer is the most prevalent tumor in developing countries and the second most frequent cancer among female population worldwide. Specific human papillomaviruses and, most notably, HPV types 16 and 18 are recognized as being caus-ally associated with cervical carcinomas. The early HPV type 16 genes, E6 and E7, di-rectly participate in the in vitro transformation of primary human keratinocytes and rep-resent an excellent target for immune therapy of HPV related disease.Objective: The aim of this study was the evaluation of the efficacy of a DNA vaccine containing human papillomaviruse type 16 E7 gene (Iranian isolate) in induction of CTL responses in an animal model.Methods: In this study, the expression vector containing HPV type 16 E7 gene was constructed and chosen as a model antigen in the development of a thera-peutic DNA vaccine in an animal model. CTL responses, cytokine assay, lymphocyte stimulation test, CD4 and CD8 staining and flowcytometry were done for evaluating of the immune responses.Results: Our findings indicate that the target DNA vaccine can induce an E7-specific CTL response, which is important in the lysis of infected tumor cells, compared to negative control (p<0.005) after in vivo immunization in the mouse system.Conclusion: The developed vaccine may be promising as an anti-cancer vac-cine.

Introduction: Effect of Aliado fungicide (Masio, Spain) with active ingredient of Mancoezbe 40%, chlorothalonil 20% and simoxanil 5% as a 65% WP formulation for control of citrus gummosis disease in laboratory, greenhouse and orchard conditions was compared with common fungicides (copper oxychloride and Bordeaux mixture in ratios of 100 and 50 g/l respectively). The growth of Phytophthora parasitica and P. citrophthora was controlled in the culture media containing the recommended doses of 15, 20, 25, 30 and 35 g/l. Results of greenhouse experiment with one-year-old sweet orange seedlings of Siavaraze (a local variety) indicated that Aliado is effective on prevention of the wound development at the doses of 25 and 35 g/l for P. parasitica and 20 and 35 g/l for P. citrophthora. The best result was obtained with painting of 15 grams per liter of the fungicide twice a year (spring and autumn) in the field assessment. Materials and Methods: In this study, P. parasitica and P. citrophthora were isolated from the infected soil. The fungi were stored after purification and cultivation in the test tube for the next steps. To evaluate the effect of Aliado in laboratory conditions, different doses of the fungicide containing 0, 15, 20, 25, 30 and 35 grams per liter were added to the CMA medium and distributed in petri dishes. Then, 5 mm discs were prepared from the 7-day-old pragmatic margin of the fungus-like and placed in the center of Petri dishes. After maintaining them at 24 ° C for 4 days, the radial growth of mycelium was measured. In order to investigate the effect of the fungicide in greenhouse condition, seeds of Siavaraze local sweet orange as a sensitive variety was prepared and treated with 0. 5 percent sodium hypochlorite solution and cultivated in a soil composition disinfected with water vapor in a controlled greenhouse of Citrus and subtropical Fruit Research center. Evaluation of the effect of fungicides was carried out after seedlings reached about 20 centimeters. For staining treatments, 24 one year old seedlings of Siavaraze were used. First, at 15 cm above the surface of the soil, a part of the bark was removed from the trunk and discs of the same size from the fungi-like colonies were placed instead. In the control plants, the culture medium without mycelium was placed on the surface of the wood. To evaluate the treated seedlings, about 40 days after application of the fungicides, the size of the new wound was calculated. To study the effect of Aliado in orchard condition, 24 trees of 12 years old with Bakraee and Lime rootstocks infected with trunk gummosis in Kerman (Jiroft) and Fars (Jahrom) provinces were used in a randomized complete block design with 8 treatments in 3 replications. The wound of the trunk marked and bark of the infected area was gently scratched and painted by a brush with the same treatments and doses of the laboratory test. Six months after application, the size of the new wound was recorded. The analysis of variance and the mean comparison were performed based on MSTATC statistical program and the Tukey's test. Results and Discussion: Laboratory studies showed that Aliado fungicide completely prevented the growth of fungi-like in the recomended doses of the company. Under greenhouse conditions, painting of Aliado with 25 and 35 grams per liter for P. parasitica and 20 and 35 grams per liter for P. citrophthora have been effective in stopping the development of stem wounds. This research was also conducted in field conditions in the orchards with gummosis symptoms in Kerman and Fars provinces. In the orchard condition, two times application of all treatments containing Aliado fungicide (even a dose of at least 15 g/l) was effective. The results of the experiment are in agreement with those of the world's researchers, which have reported the use of fungicide as an effective method for controlling phytophthora fungus-like of gummosis. Therefore, in citrus cultivation areas of the country, especially the southern provinces, where the climatic conditions are favorable for the activity of the fungi-like, treatment of the crowns of citrus trees with Aliado in the spring and the autumn with the minimum proposed dose of the fungicide is recommended. Observations showed that all treatments of Aliado are effective by drying the gum in controlling disease progression through disinfection. In some treatments, the emergence of gum in the new places was observed with a little distance from the previous wounds. It can be because the effect of fungicide has decreased after a few months. In climatic conditions of the southern regions of the country the fungi-like are active throughout the year. So the use of fungicide in spring and autumn is recommended. The use of Aliet fungicide in two doses of 160 and 200 g in 100 liters of water had a protective effect less than one year.