Molecular cloning of S1 glycoprotein gene of infectious bronchitis virus (IBV) serotype 793/B in secretory Pichia pastoris vector

In vitro protein expression is an important method of obtaining large amounts of viral proteins to investigate their biological properties. The S1 glycoprotein of infectious bronchitis virus, due to its effective immune-dominant role is an appropriate candidate for production of recombinant vaccine against infectious bronchitis disease. In this study, the S1 gene fragment of infectious bronchitis virus strain793/B was amplified using reverse transcriptase-polymerase chain reaction (RT-PCR) and purified. Itwas then cloned into pPICZαA a secretory expression vector of Pichia pastoris. The insertion was proved by PCR analysis and isolation of gene from construct by restriction enzymes and finally, it was sequenced. After the expression of S1 gene in P. pastoris expression system, it was found that it could be used in the production of recombinant vaccines against infectious bronchitis disease. Keywords: Infectious bronchitis, S1 glycoprotein, cloning, Pichia pastori

A path walkability assessment index model for evaluating and facilitating retail walking using decision-tree-making (DTM) method

Transportation is the major contributor of ever-increasing CO2 and Greenhouse Gas emissions in cities. The ever-increasing hazardous emissions of transportation and energy consumption have persuaded transportation and urban planners to motivate people to nonmotorized mode of travel, especially walking. Currently, there are several urban walkability assessment models; however, coping with a limited range of walkability assessment variables make these models not fully able to promote inclusive walkable urban neighborhoods. In this regard, this study develops the path walkability assessment (PWA) index model which evaluates and analyzes path walkability in association with the pedestrian's decision-tree-making (DTM). The model converts the pedestrian's DTM qualitative data to quantifiable values. This model involves ninetytwo (92) physical and environmental walkability assessment variables clustered into three layers of DTM (Layer 1: features; Layer 2: Criteria; and Layer 3: Sub-Criteria), and scoped to shopping and retail type of walking. The PWA model as a global decision support tool can be applied in any neighborhood in the world, and this study implements it at Taman Universiti neighborhood in Skudai, Malaysia. The PWA model has established the walkability score index which determines the grading rate of walkability accomplishment for each walkability variable of the under-survey neighborhood. Using the PWA grading index enables urban designers to manage properly the financial resource allocation for inspiring walkability in the targeted neighborhood.Scopu

A path walkability assessment index model for evaluating and facilitating retail walking using decision-tree-making (DTM) method

Transportation is the major contributor of ever-increasing CO2 and Greenhouse Gas emissions in cities. The ever-increasing hazardous emissions of transportation and energy consumption have persuaded transportation and urban planners to motivate people to nonmotorized mode of travel, especially walking. Currently, there are several urban walkability assessment models; however, coping with a limited range of walkability assessment variables make these models not fully able to promote inclusive walkable urban neighborhoods. In this regard, this study develops the path walkability assessment (PWA) index model which evaluates and analyzes path walkability in association with the pedestrian's decision-tree-making (DTM). The model converts the pedestrian's DTM qualitative data to quantifiable values. This model involves ninetytwo (92) physical and environmental walkability assessment variables clustered into three layers of DTM (Layer 1: features; Layer 2: Criteria; and Layer 3: Sub-Criteria), and scoped to shopping and retail type of walking. The PWA model as a global decision support tool can be applied in any neighborhood in the world, and this study implements it at Taman Universiti neighborhood in Skudai, Malaysia. The PWA model has established the walkability score index which determines the grading rate of walkability accomplishment for each walkability variable of the under-survey neighborhood. Using the PWA grading index enables urban designers to manage properly the financial resource allocation for inspiring walkability in the targeted neighborhood

Genetic and antigenic analysis of type O and A FMD viruses isolated in Iran,

ABSTRACT FMD is one of the most highly contagious diseases of animals, caused by RNA virus belong to Picornaviridae family and Aphtovirus genus. A broad host range and occurrence of FMDV as seven serotypes and also intratypic antigenic variation without clear cut demarcations, which interferes with a concept of sub typing these factors make difficult conditions to diagnosis, control and eradication of disease. Therefore it is very important to characterize virus strains and monitoring the field virus to determine the relationship between field viruses and vaccine strains. The objective of this study was to characterize FMD type O, A, virus isolated from Iran between 2005 and 2006. 13 FMD type A and 6 type O viruses isolated from Iran between 2005 and 2006 were used in this study. All viruses adapted to IBRS2 cells and the clarified infected cell culture supernatants were used for typing by sandwich capture ELISA and extraction of viral RNA for RT-PCR reaction with the specific primer for each type. The PCR products were purified for sequencing. Sequence of 600 nucleotides at the 3` end of 1D gene of all samples subjected to phylogenic analysis and determine the antigenic relationship ("r" Value). All type A viruses that isolated from different province of Iran, sequenced in this study, were closely related to each other and A/iran/05 virus group. The sequencing results of type O isolated from Iran between 2005 and 2006 showed the close genetic relationship between field isolates and the Iranian vaccine strain. The result of average "r" Value detected by two dimensional virus neutralization test, for type A87IR was 0.46 (46%), type A05IR 0.78 (78%), type O Shabestar 0.81 (81%) and type O967 0.90 (90%)

Green driver: Travel behaviors revisited on fuel saving and less emission

Road transportation is the main energy consumer and major contributor of ever-increasing hazardous emissions. Transportation professionals have raised the idea of applying the green concept in various areas of transportation, including green highways, green vehicles and transit-oriented designs, to tackle the negative impact of road transportation. This research generated a new dimension called the green driver to remediate urgently the existing driving assessment models that have intensified emissions and energy consumption. In this regard, this study aimed to establish the green driver's behaviors related to fuel saving and emission reduction. The study has two phases. Phase one involves investigating the driving behaviors influencing fuel saving and emission reduction through a systematic literature review and content analysis, which identified twenty-one variables classified into four clusters. These clusters included the following: (i) FEf1, which is driving style; (ii) FEf2, which is driving behavior associated with vehicle transmission; (iii) FEf3, which is driving behavior associated with road design and traffic rules; and (iv) FEf4, which is driving behavior associated with vehicle operational characteristics. The second phase involves validating phase one findings by applying the Grounded Group Decision Making (GGDM) method. The results of GGDM have established seventeen green driving behaviors. The study conducted the Green Value (GV) analysis for each green behavior on fuel saving and emission reduction. The study found that aggressive driving (GV = 0.16) interferes with the association between fuel consumption, emission and driver's personalities. The research concludes that driver's personalities (including physical, psychological and psychosocial characteristics) have to be integrated for advanced in-vehicle driver assistance system and particularly, for green driving accreditation

A sustainable historic waterfront revitalization decision support tool for attracting tourists

Waterfront revitalization would be an effective strategy to preserve heritages, conserve the contaminated or abandoned site and inspire the identity and authenticity. However, there is no decision support tool to quantify and evaluate the sustainability accreditation of waterfronts in tourism attraction. This research aimed to identify the most potential waterfront typology in tourism attraction and develop the waterfront sustainable revitalization (SWR) index assessment model. The SWR index can assist policy makers and urban developers to analyze the heritage waterfronts using Analytical Hierarchy Process (AHP) method. The research found out the historic waterfront has the highest potential in tourism attraction among other typologies. And, pollution moderator is mostly important sub-criterion in tourism absorption (WC2.2 = 0.1294); followed by Identity (WC1.2 = 0.1272) and Safety and well-being (WC1.3 = 0.1043). The SWR index can be applied in any waterfronts in heritage cities around the world, while this research implemented it as a case study in Bandar Maharani, Muar, Malaysia. It resulted Bandar Maharani was ranked as grade C; means, usable waterfront to which extent environmental, social and physical revitalization are needed. The SWR index can be coupled with other decision-making methods in future, to reduce its inconsistencies and increasing accuracy

Expression of bovine leukemia virus p24 protein in bacterial cell

Bovine leukemia virus (BLV) is a member of the family Retroviridae, genus Deltaretrovirus that has three important gene including gag, pol and env. This virus causes B-cell lymphocytosis and lymphosarcoma in cows. In the first step PCR product of gag gene of BLV isolated in different regions of Iran and BLV-FLK strain were cloned in to a pTZ57R/T vector, then insert were digested by BglII and XhoI restriction enzymes and cloned in to pET-28(a) as an expression vector. For the expression of p24 protein, the pET-28(a) recombinant vector was transformed in BL21(DE3) strain of E. coli competent cell and after induction of the protein having been expressed by IPTG, the presence of gag expressed protein was shown in immunoblotting and SDS-PAGE system. With respect to the remarkable frequency of infection to BLV in Iran and the necessity of controlling it through vaccination with recombinant vaccines of gp51, manufacturing and applying the recombinant p24 protein are vital goals in recognition and distinction between infection and responses caused by vaccine.H. Momtaz, F. Hemmatzadeh and H. Keyvanfa

Study of effector antigens of IBR virus on humoralimmunity

The objective of this study was to determine the protein pattern and antigenic structure of BHV-1. Ten viral isolates from cows with clinical signs of respiratory infections were selected for this study and confirmed by SN test. All samples were cultured on cell cultures and purified by ultracentrifugation. Purified isolates were electerophorsed using SDS-PAGE method and immunoblotted. Viral antigens were detected using anti BHV-1 bovine antiserum and molecular weight of the bands were determined. All samples showed identical band number and molecular weight except for one sample which revealed an additional 120-125 KDa band. Also immunoblotting data resulted in identified of twelve bands (150, 130, 115, 97, 77, 70, 55, 45, 40, 38, 32 and 25 KDa) suggesting involvement of these antigens in evoking humoral immune response in affected cattle.Hemmatzadeh, F., Keyvanfar, H., Ahmadi, M

Antigenic pattern of BVD viruses isolated in Iran

Processing of viral polyproteins of Bovine Viral Diarrhoea Virus (BVDV) by cellular and virus-encoded proteases gives rise to at least 11 structural and nonstructural proteins having molecular weights of 165, 125, 110, 93, 80, 70, 62, 53, 48, 44, 34 and 23 kDa. Some studies showed a small changes in molecular weight or number of the viral proteins. (Donis and Dubovi 1987). Cytopathic (CP) strains of BVDV expressing of NS3 (P80) and non-cytopathic (NCP) strains express the fusion protein NS2/3 (P125), but not NS3 (Qu et al., 2001), while in CP strains both NS2/NS3 and a discrete NS3 are observed (Agapov et al:, 2004, Balint et al., 2004). Proteinal pattern of various pestiviruses can be variable in different isolates and cleavage of viral proteins by viral and/or cellular proteases, would result in diversity in viral proteins (Becher et al., 196; Paton, 1995).F. Hemmatzadeh, H. Momtaz, H. Keyvanfar and E. Banihasanhttp://www.mendeley.com/research/antigenic-pattern-bvd-viruses-isolated-iran

PCR for detection of Ovine Herpesvirus-2 in cow and sheep of Iran

Malignant Catarrhal Fever (MCF) is one the most important viral diseases of livestock. Aetiologically, there are 2 forms of the disease, one is associated with Alcelaphine Herpes Virus-1 (AlHV-1) of the wildebeests and the other (SA-MCF) is associated with Ovine Herpes Virus-2 (OvHV-2). In order to detect the epidemiological condition of the disease (Ovine herpes virus-2 in cattle and sheep) in Iran, we collected 100 whole blood samples from clinically ill cattle with MCF, healthy cattle and sheep (more than one year-old). The specimens undergone PCR method in several stages. The 1st stage was performed by pair primers 556 and 775. The reviced a result a band of 422 DNA base pair in 100% of infected cattle and sheep and 75% in healthy cattle. In the 2nd stage, using pair primers 556 and 555, in the Semi-nested PCR, 3 bands of base pairs 238, 340 and 420 of DNA were traced in the specimens. It is noteworthy, that the specimens colleted from sheep and clinically infected cattle with MCF, had all three bands and the healthy cattle specimens had only the 2 DNA band of 420 and 340, which were belonged to OvHV-2 genome.Hassan Momtaz, Farhid Hemmatzadeh, Hadi Keyvanfar and Behnam Abbasia