Research/Original Articles

HPV16 E7-CT (gp96) fusion protein: Molecular cloning, expression and purification of a recombinant 6xHis-tagged protein in E. coli

Amin Daemi, Sahar Hosseinzadeh, Sima Rafati, Farnaz Zahedifard, Masoumeh Rajabi Bazl, Fatemeh Doustdari, Mehrdad Hashemi, Elnaz Agi, Azam Bolhassani

Archives of Advances in Biosciences, Vol. 3 No. 2 (2012), 17 June 2012

The development of a therapeutic vaccine against human papillomavirus (HPV) is important for the control of cervical cancer. E7 is the major transforming protein produced in cervical cancers, and therefore represents potential tumor-specific antigen that could be the target of immunotherapy for cervical cancer. Among different vaccine strategies, protein-based vaccines are capable of generating CD8+ T cell responses in vaccinated animals and humans. Recently, development of novel strategies that enhance protein vaccine potency is important for generation of effective cancer vaccines and immunotherapies. Heat shock proteins (HSPs) including Gp96 have been shown to act as potent immuno-adjuvant to enhance antigen-specific tumor immunity. Therefore, the HSP-based protein vaccines can be administered by fusing antigens to HSPs, in vitro. It has been known that the HSP fragments (e.g., N-/or C-terminal regions) as mini-chaperones are better choice for immunization. The most straightforward method to produce large amounts of recombinant protein suitable for a vaccine is to clone the gene into a prokaryotic expression vector and produce the protein in Escherichia coli. In current study, we describe cloning of the HPV16 E7 gene linked to C-terminal fragment of gp96, identification and purification of the resultant E7-CT (gp96) fusion protein for next usage as a potential vaccine candidate protein against HPV in a pre-clinical trial. The recombinant E7-CT (gp96) migrated as a 51 kDa protein in SDS-PAGE. In Western blot experiment, the existence of a 51 kDa band for rE7-CT (gp96) was confirmed by rabbit anti-His as well as mouse anti-HPV16 E7 monoclonal antibodies. The protein of interest was both in the insoluble and the soluble fraction; therefore, purification was performed under denaturating and native conditions by affinity chromatography on Ni-NTA resin using 6xHis-tag.

Volatile compounds of essential oil Centaurea behen L. grown in Iran

Akbar Esmaeili, Elham Khodadadi

Archives of Advances in Biosciences, Vol. 3 No. 2 (2012), 17 June 2012

      The essential oils from Compositae specie of Iran:Centaurea behen L. obtained by hydrodistillation was analyzed by GC and GC/MS.  β-Caryooyllane (24.5%), β-selinene (13.9%) and valencene (11.7%) were the main components among the sixteen constituents characterized in the oil of Centaurea behen representing  93.7% of the total components detected.

Antimicrobial activities of the essential oils of five Salvia species from

Mansoureh Paknejadi, Fatemeh Foroohi, Morteza Yousefzadi

Archives of Advances in Biosciences, Vol. 3 No. 2 (2012), 17 June 2012

      Essential oils are volatile, natural, complex compounds formed by aromatic plants as secondary metabolites. In nature, essential oils play an important role in protecting plants in form of antibacterials, antivirals, antifungals, insecticides properties and also against herbivores. They also may attract some insects to use them in dispersion of pollens and seeds, or repel undesirable others. Present study describes the results of our study on the chemical composition, and antimicrobial activity of the essential oils from aerial parts of aromatic plants which collected from province of Tehran. The aerial parts of plants were collected at full flowering stage. The essential oils were isolated by hydrodistillation and analyzed by combination of capillary GC and GC-MS. Also their antimicrobial activities were considered according to the disc diffusion method and MIC values. Finally, five plant species (Salvia sclarea, salvia multicaulis, salvia verticillata, salvia limbata, Salvia choloroleuca) which showed good significant antibacterial were presented. These five plants belong to labiates’ family. Among these proposed plants salvia multicaulis and  Salvia sclarea showed good antibacterial activity.

The Higher National Council of Medical Sciences Planning (HNCMSP) in Iran compile and approve educational programs by using expert academic staff, considering the country,s needs and studying the background of establishment of each major in other countries. Since the employment of graduates in occupations related to their majors is of utmost importance in educational planning, this survey has been done to investigate the graduates, employment and the factors affecting it, using a retrospective study.

This survey is done on 120 graduates of Laboratory Sciences (LS), Medical Records (MR) and Radiology Technology (RT), at Paramedical Faculty of Shahid Beheshti Medical University (1997-2000).The subjects were randomly selected and given a questionnaire and the data was statistically analyzed using one-way ANOVA,Chi-Square, and Exact Fishers test (SPSS 15). 63.5 percent of the samples were females and the rest were males with an average age of 35.The coordination between their aptitude and major was highest (61%) in LS graduates, but lowest (21%) in MR graduates. Also the coordination between majors and interest was highest in LS graduates (48.4%).The results indicate that 6% of the subjects were post graduate students, 91% were employed and 9% were jobless.75% of them believed that their jobs were quite related to their majors and interest. They also mentioned the same factors for finding a suitable job (P=0.328).

The results show that there is a direct relationship between the students, interest and aptitude on one hand, and their major and interest on the other. Moreover, because of the coordination between the majors and the countrys needs, the rate of unemployment has been found to be very low(9%).These findings can be helpful in reevaluation of educational planning, guidance of aptitudes towards the countrys economic priorities, and having a study of post graduation status.


Fecal carriage of Escherichia coli and Klebsiella spp. as major reservoirs of clinically important resistance markers

Maryam Razaghi, Parisa Torabi, Ehsan Nazemalhosseini Mojarad, Elahe Tajeddin, Somayeh Jahani Sherafat, Marjan Rashidan, Sima Sadat Seyedjavadi, Masoud Alebouyeh, Mohammad Reza Zali

Archives of Advances in Biosciences, Vol. 3 No. 2 (2012), 17 June 2012

Intestinal normal flora can become reservoirs of antibiotic resistance genes present among the strains responsible for nosocomial infections. It is suggested that gram negative intestinal bacterial flora have increased capacities to obtain antibiotic resistance genes and therefore can act as main reservoirs for transfer of resistance genes to other pathogenic bacteria. This study aimed to compare fecal carriage of clinically important resistance markers for more frequent members of enterobacteriacae between nondiarrheal and community associated diarrheal patients (control group) versus their counterparts from the patients with nosocomial infections (case group). 261 stool and 190 clinical samples were collected from outpatient and hospitalized patients from 6 hospitals in Tehran, Iran. The samples were cultured on MacConkey agar plates and colonies were identified by standard biochemical methods. Antibiotic sensitivity testing of the isolates against 13 antibiotics was performed according to the CLSI guideline using the disk diffusion method.   Among stool and clinical samples, more frequent identified enterobacteriaceae bacteria were included E. coli (58.99/ 3.15%), Klebsiella spp. (22.61/7.36%), and other members of enterobacteriaceae (8.86/1.06%), respectively. Overall, resistance against four of the main antibiotics (3th and 4th generation cephalosporins, gentamicin, imipenem, and ciprofloxacin) was significantly higher among the case group (50-75% versus 10-14%). Analysis of these results showed similar dissemination of resistance phenotypes among the isolates from the control group in ranges of 1.5-7.6% and 4.4% for E. coli and Klebsiella spp., respectively. Our results suggested that the fecal carriage of resistant phenotypes related to the β-lactam antibiotics in E. coli and Klebsiella spp. in compare to the clinical isolates is rapidly increasing. This may be caused by dissemination of β-lactamase producing E. coli in the community from the hospitals. There were no significant correlations between the two groups of the samples, as the clinical samples had shown 3 to 7 folds excess resistance phenotypes. Surveillance studies of the resistance patterns among the samples from different regions will provide awareness about dissemination of these bacteria within the community as reservoirs of main resistance markers.

The effect of illness perception on physical health-related quality of life promotion in multiple sclerosis (MS) patients attending peer support groups

Maryam Jadid Milani, Tahereh Ashktorab, Zhila Abed Saeedi, Hamid Alavi Majd

Archives of Advances in Biosciences, Vol. 3 No. 2 (2012), 17 June 2012

Illness perception influences patients’ decision to adopt effective behavior and achieve positive results such as adapting with the disease and improving functionality, and the interventions that increase illness perception can promote health. This study aimed to investigate the effect of illness perception on the physical health-related quality of life of MS patients attending peer support groups. This study with a quasi-experimental before-and-after design included 33 MS patients in three groups: male-only(n=10), female-only(n=11) and one with both males and females (mixed, n=12) that selected by convenience sampling Participants were required to attend 8 weekly sessions comprising 2 hours each. Instruments used to assess physical health related quality of life and illness perception were the physical health section of "Multiple Sclerosis Quality of Life Inventory (MSQLI)" and "Revised Illness Perception Questionnaire(IPQ-R)" respectively, which were completed by participants before and after attending the group sessions. The results showed that although illness perception of MS patients attending peer support groups did not show a significant increase, physical health significantly improved(p=0.001). Attending peer support group increased illness perception in the mixed group(p=0.01) and elevated physical health in men only and mixed group (p=0.03 for the mixed group and p=0.04 for men only group). Regression analysis showed a significant relationship between MS and physical health with efficacy of 0.54(p<0.001). The results showed that increased illness perception in MS patients improves their physical health. Therefore, we can improve MS patients’ physical health-related quality of life through peer support groups and hence promote patients’ quality of life.


Evaluation of Persian Academy approved genetics terms acceptance in upper graduate user population

Saeed Hesami Tackallou, Atefeh Ghanbari

Archives of Advances in Biosciences, Vol. 3 No. 2 (2012), 17 June 2012

Finding Persian equivalents for scientific terms is one of the aims of Academy of Persian language and literature, and more than 50 scientific committees are now working on this scope in terminology department of the academy. Genetics and biotechnology terminology committee is one of these teams that started his activity from 2009 and since then approved more than 500 of these terms for use in academic fields. In this research 101 questionnaires including 20 questioned term were given to more than 101 upper graduate users including MSc and PhD students and academic staff of randomly selected universities in Tehran and Semnan provinces. And then the evaluation of their acceptance was carried out by a model given by Cooper for the Study of Language Spread. All 20 studied terms were randomly selected from genetic approved terms and divided in to two groups: newly coined equivalents (those equivalents that was newly coined by the committee) and selected equivalents (those that have the selected equivalent before).

Results gathered and analyzed with statistical tests, and showed that most of accepted terms are among selected equivalents. By another words the newly coined terms has much less acceptability than the others. This research introduces a method for evaluation of approved Persian equivalents of scientific terms and besides show the state of some of these equivalents between user populations. The important point is that term selection for scientific terms including genetic terms; is not an obligatory rule, but is a proposal for meeting the researchers need to strength Persian language as a scientific language. And usage of these equivalents is completely on the part of researchers and students and their point of view to equivalents.


Review Article

Chronic myeloid leukemia as a stem cell-derived malignancy

Sadegh Babashah, Mostafa Rezaei-Tavirani, Mona Zamanian-Azodi, Najmaldin Saki

Archives of Advances in Biosciences, Vol. 3 No. 2 (2012), 17 June 2012

Chronic myeloid leukemia (CML) is a myeloproliferative disease of the hematopoietic stem cells, characterized by the presence of the Philadelphia (Ph) chromosome. Although imatinib inhibits the BCR-ABL kinase activity, clinical experiences confirm that imatinib may not target CML stem cells in vivo. The identification of signaling pathways responsible for the self-renewal properties of leukemic stem cells in CML will help in the discovery of novel therapeutic targets. Here we review signaling pathways including Wnt/β-catenin, Hedgehog, Alox5, and Foxo which play crucial roles in the maintenance of stem cell functions in CML. It is thought that inhibition of key genes that are part of self-renewal associated signaling pathways may provide an effective way to reduce aberrant stem cell renewal in CML.