Journal of Arak University of Medical Sciences

Site-Directed Mutation, Cloning and Expression of Streptokinase for Producing a New Suitable Molecule for PEGylation

Reza Arabi Mianroodi

Background: Streptokinase is one of the most common and cost effective fibrinolytic drugs for treatment of heart attacks and vein thrombosis. Unlike many advantages over other thrombolytic drugs, administration of streptokinase can produce some complications such as immunologic reactions, hemorrhage and incomplete treatment due to relative short half life. Pegylation is one of the most common methods for improving of these shortcomings.

Materials and Methods: In this study, designing a proper candidate for specific pegylation with cysteine was done by means of SPDBviewer software. After a meaning ful mutation by SOEing PCR method, mutated (sk45cys) and intact SK (ski) genes were cloned in pET26-b vector and the structures were transformed in E.coli. Clones, Afrer growing, were expressed by IpTG and exptression of proteins was confirmed by SDS-PAGE and western blotting. The proteins were purified by affinity chromatography with NiNTA columns and amidolytic activity of purified proteins was assayed using chromogenic method and different concentrations of S2251 substrate.

Results: Results of activity assays showed that amidolytic activity of SK45cys had about 10% increase in comparison to Ski, after 30 minutes of complex formation with plasminogen.

Conclusion: Generally, it was concluded that, considering cys45 as a superficial aminoacid and also relative increase of activity, SK45cys can be considered a suitable protein for specific pegylation.

Personality Profile Investigation of Adoption Applicant Couples in Arak, 2014

Mehri Jamilian

Background: Adoption has a significant role for strengthening families without children and also resolution of psychological confilicts of unsupervised children. The goal of this study is to assess the psychological status of adoption applicant couples.

Materials and Methods: This is a descriptive- analytic study that is done with cooperation of welfare organization and Arak University of Medical Sciences in 2014. All adoption applicant couples referred to welfare organization for 1 year participated in the study. Data were gathered by using demographic questionnaire and MMPI-II. Data were showed by descriptive statistic and were analyzed by using mean, average and covariance analysis indices.

Results: Mean and standard deviation of all clinical scales and reliability of MMPI-II in all samples and also different subgroups of statistical population were under 65. All applicants had good mental health and psychologically were eligible for adoption. A group of applicants have pretended themselves better than what they are because of the fear of rejection that led to significant difference in k scale(p=0.019).

Conclusion: According to the findings of this study, applicants had good mental health and didn’t have any psychopathologic damage, but sometimes they have pretended themselves better than what they are. Investigation of other diagnostic tools in future studies is recommended.

Combined Effect of 5-Fluorouracil and Acriflavine on Mortality Rate of Colorectal Cancer Cell Lines

Ebrahim Eftekhar

Background: A small percent of patients with colorectal cancer (CRC) respond to 5-Fluorouracil (5-FU), as a first line of chemotherapy. In this study in, in order to design a new chemotherapy protocol, the effect of 5-FU and acriflavine (ACF) cotreatment on mortality rate of CRC cell lines was investigated.

Materials and Methods: Cytotoxicity of 5-FU and ACF against CRC cell lines (LS174T, SW480 and HCT116) was detected using MTT assay. Cells were treated with different concentrations of 5-FU (0.5-64 µM) or ACF (0.07-5 µM) for 72 hours and then cell viability and drugs IC50 was calculated. To assess the effect of ACF on anticancer activity of 5-FU, cells were cotreated with different concentrations of 5-FU and IC30 concentration of ACF.

Results: ACF and 5-FU suppress the viability of CRC cell lines in dose-dependent manner. 5-FU and ACF have most cytotoxic effect on LS174T and the lowest cytotoxic effect on SW480 cells. Cotreatment of ACF with 5-FU could not significantly change the sensitivity of cells against 5-FU (p>0.05).

Conclusion: In this study, the fatal and cytotoxic effect of ACF on three CRC cell lines was shown. However, cotreatment of ACF with 5-FU could not improve the anticancer activity of 5-FU.

Effectiveness of Self-Control Training on Quality of Life Dimensions in Migraine Patients

Roghayeh Nooripour Liavali

Background: Migraine is a chronic neurological disorder that leads patients to avoid any kind of activities. Since different factors are involved in migraine incidence and its triggers, so drugs are used to prevent or treat it are so variable. Also, combined medications are used to relieve migraine. This study examined the effectiveness of self-control training on quality of life in patients with migraine.

Materials and Methods: Statistic population of this study included all migraine patients in Ardabil in 2014(Estimation: N=1150) that 40 patients were selected by convenience sampling. Demographic and disease information questionnaire and quality of life questionnaire (SF-36) were used to collect data in clinical centers. Multivariate analysis of variance (MANCOVA) was used to analyze data, because present research was a experimental and clinical trial with pre-test and post-test with control group.

Results: The results showed that there is a significant difference between mean in quality of life in migraine patients and control subjects. It means that physical health and mental health of quality of life was different between control and experimental groups after self- control training.

Conclusion: Self-control training can be used to enhance quality of life in migraine patients. These results have important and effective applications in the treatment of migraine patients. Generally, specialists of clinical centers can use this method alongside other treatment interventions.

Demodicosis and Associated Risk Factors in Young Adults in Khorramabad, 2015

Hamidreza Shokrani

Background: Demodex mites can infect the sebaceous glands and hair follicles from almost every area of human skin especially on the face. Demodicosis is considered to be a factor in the pathogenesis of some skin disorders. The aim of this study was to determine the prevalence of Demodex mites and to identify the association between risk factors and Demodex infection.

Materials and Methods: This cross-sectional study was conducted with 100 university students in Khorramabad, Iran. Samples were collected from three different regions of the face (nose, cheeks and forehead) using two different methods, including cellophane tape method (CTP) and skin pressurization. A questionnaire was filled for each individual and data were subjected to statistical analyses using chi-square test.

Results: The overall prevalence of Demodex mites was 55%. The detection rate of infection by cellophane tape method was much higher than that of the skin pressurization (p<0.01). The incidence of Demodex was found to occur more frequently in males and in people with oily skin (p<0.05). Furthermore, the infection rate among students with acne vulgaris was significantly higher than that of students without acne (p<0.05). We found a significant difference between acne vulgaris and density of Demodex mites (p<0.01).

Conclusion: The cellophane tape method (CTP) is an easy and suitable method for epidemiologic investigations. Skin types and gender were identified as risk factors. Our study suggests that Demodex infection could be associated with acne vulgaris.

Association between MMP9 Promoter Polymorphism and Breast Cancer Progression in Northwest of Iran

Farhad Mashayekhi

Background: MMP9, as a member of the MMPs family, codes a protein that is able to provide suitable infrastructures for the migration of cancer cells and angiogenesis within tumor. The aim of this study was to investigate the relationship between -1562 C> T polymorphism in MMP9 promoter and progression of breast cancer in northwest of Iran.

Materials and Methods: In this case-control study, 187 females from northwest of Iran were involved. Polymorphism of interest was determined by PCR-RFLP method using enzyme PaeI and statistical analysis was done by Med Calc software.

Results: Distribution of CC genotype in cancer and control groups was,44 and 62.5, tespectively and distribution of CT genotype in cancer and control groups was 56 and 37.5, respectively, In the statistical analysis, χ²and p value were respectively, 5.4 and 0.01. There is a significant association between this polymorphism and lymph nodes involvement and presence of tumor larger than 2cm³ (respectively p= 0.005 and p = 0.03). The results of this study showed -1562 C> T polymorphism in MMP9 promoter is associated with stage II and higher stages of breast cancer in our population. Furthermore, CT genotype may increase the risk of lymph node metastasis and presence of tumor larger than 2 cm³ (OR= 4).

Conclusion: According to the results of this study, MMP9 (-1562 C> T) polymorphism may be used as a biomarker for breast cancer prognosis. Although, to achieve more definitive results, it is necessary to examine a larger population.

Early Detection of Amyotrophic Lateral Sclerosis (ALS) using the Gait Motor Signal Frequency Analysis

Ataollah Abbasi

Background: ALS is a progressive neuro-muscular disease, which is characterized by motor neuron loss in the Central Nervous System (CNS) and Peripheral Nervous System (PNS). Up to now, no accurate clinical method for diagnosis of the disease have been provided. In most cases, ALS patients are unable to walk normally due to abnormalities in the nervous system. For this reason, one of the most appropriate methods in the diagnosis of ALS from other neurological diseases or from healthy volunteers is the gait motor signal analysis.

Materials and Methods: In this study, gait signals available in Physionet database have been used. The database consists of 13 patients with ALS (ALS1, ALS2, …, ALS13) and 16 normal subjects (CO1, CO2, …, CO16). The patients participating in this study had no history of any psychiatric disorders and did not use any assistive device for walking, like wheelchair. The power spectrum of stride, swing, and stance of normal subjects and patients was computed for both left and right legs. To provide appropriate inputs for the classifier, the frequency band of the power spectrum of all signals was divided into eight equal parts. The area of all regions was computed. Three frequency band of the lower range of power spectra selected as inputs of the classifier.

Results: In this study, power spectra, as frequency attributes, were used to explore probable differences of time series in both patients and healthy subjects.

Conclusion: Artificial Neural Network was used to classify normal and ALS groups with the accuracy of 83% for the test data set. It seems that the present algorithm can be used in discriminating patients from normal subjects in the early stages of the disease.

The Combined Effect of Regular Aerobic Exercise with Garlic Extract on Renal Apoptosis Regulatory Factors in Aged rats with Chronic Kidney Disease

masoumeh habibian

Background: Chronic kidney disease as an important risk factor is associated with some disorders which are key causes of death and disability in older people. Therefore, the aim of this study was to assess the combined effect of regular aerobic exercise with garlic extract on renal apoptosis regulatory factors in aged rats with chronic kidney disease.

Materials and Methods: In this experimental research, 42 aged male Wistar rats(48-52 weeks) were selected and randomly divided into 6 groups: control, doxorubicin, doxorubicin-salin, doxorubicin- garlic, doxorubicin - exercise, doxorubicin –garlic-exercise(combined). Chronic kidney disease was induced by a single subcutaneous injection 8.5 mg/kg of doxorubicin. Swimming training was programmed 3 days /week, 30 min/day for 8 weeks. Both the doxorubicin garlic and combined groups with garlic extract were administered by garlic gavage at a dose of 2.5 g/kg. The renal Bax and Bcl-2 levels were evaluated by ELIZA method. A one-way analysis of variance was used to data analysis (p<0.05).

Results: The results showed that induced chronic kidney disease was associated with a significant increase on Bax and a decrease on Bcl-2 in aged rats. Also, 8 weeks swimming training, garlic supplementation and the combined intervention significantly reversed these changes. Furthermore, no significant difference have been observed in the effect of these interventions on Bax and Bcl-2 in aged rats with chronic kidney disease.

Conclusion: It seems that the use of non-pharmacological treatment methods such as exercise training, garlic extract supplement, and combination of the both interventions may be effective in reducing apoptosis resulted from chronic kidney disease in aged rats.

Lack of Association between Tumor Necrosis Factor Alpha (TNFα) Gene -1031C/T Polymorphisms and Susceptibility to Inflammatory Bowel Disease (IBD)

Ali Mohammad Asgharian

Background: Inflammatory bowel disease (IBD) includes two basic categories ulcerative colitis (UC) and Crohn's disease (CD) that the etiology of which remains unclear. Tumor necrosis factor alpha (TNFα) promoter polymorphisms are a good candidate for susceptibility to IBD as there is a significant relationship between them. The main aim of this study was to assess TNFα gene polymorphisms with IBD susceptibility at positions -1031in Iranian patients.

Materials and Methods: In this case-control study, were studied 101 patients with IBD (86 ulcerative colitis, 15 Crohn's disease) and 100 healthy controls were studied. PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism) was used for determining of genotyping. In following, allele frequency and genotype distribution of polymorphism T> C in TNFα gene between the case and control groups were typed.

Results: The frequency of genotype TT, TC and CC among patients was 64.4%, 28.7% and 6.9% and in control group was 63%, 29% and 8%, respectively. Also, allele frequency T-1031 of TNFα gene in IBD patients was high, while there is no statistical significant(p>0.05).

Conclusion: There was no significant correlation between TNFα gene polymorphisms and susceptibility to IBD at position -1031. Our results showed that TNFα gene polymorphisms cannot be considered as a potential prognostic marker cause of IBD in Iranian population.

Studying the Association between TaqIA polymorphism in ANKK1 Gene and Heroin and Methamphetamine Addiction in Markazi Province

Rezvan Ghadbeigi

Background: ANKK1 (ankyrin repeat and kinase domain containing 1) gene is a member of the serine/threonine kinase family. This family involved in signal transduction pathways. This gene contains Taq1A (rs1800497) single nucleotide polymorphism. The A1 allele carriers of TaqIA polymorphism have shown reduced DRD2 (Dopamine Receptor D2) receptors. This decrease predisposes individuals to seek for addictive substances to compensate this deficiency in dopaminergic system. The present study investigated TaqIA (rs1800497) polymorphism in heroin and methamphetamine addiction.

Materials and Methods: In this case-control study, 91 male methadone-maintained heroin and methamphetamine addicts and 100 male healthy controls were studied. Genomic DNA extraction was carried out from peripheral blood through salting-out method and individuals were genotyped for TaqIA polymorphism by RFLP-PCR technique and TaqI enzyme was used for RFLP.

Results: This survey revealed the significantly higher frequency of the A1 allele of TaqIA polymorphism in patients than control individuals (p<0.001). The frequency of A1 allele in patient and control individuals was %51 and %22.5, respectively. The A1A1 genotype was detected in 25% of patients and 7% of controls (p<0.001, OR=9.7, 95% CI=3.64-25.85).

Conclusion: The results of this study revealed that the A1 allele of TaqIA polymorphism is significantly associated with heroin and methamphetamine addiction.