Journal of Arak University of Medical Sciences

Expression of recombinant streptolysin O by Escherichia coli

Hamid Abtahi

Introduction: Streptolysin O (SLO) is an antigenic protein that is secreted by Streptococcus pyogenes. Streptococcal infections are diagnosed with anti streptolysin O. At present, streptolysin O is produced by vectors that have fusion protein. In this study streptolysin O without fusion protein vectors is produced. Materials and Methods: In this experimental study, Streptolysin O gene was amplified by Polymerase chain reaction (PCR) method and subcloned to prokaryotic expression vector pET28a. Escherichia coli BL21-DE3-plySs were transformed with pET28a-SLO and gene expression was induced by IPTG. Then it was purified by Ni-NTA kit. The concentration of SLO was assayed by Bradford method. To confirm recombinant SLO Western Blot was used. Results: The sequencing result was confirmed by Sanger method and was the same as SLO gene. Escherichia coli BL21 (DE3) pLysS was transformed with pET28a-SLO and gene expression was induced by IPTG. The expressed protein was purified by affinity chromatography with Ni-NTA resin. The concentration of purified protein was 100µg/ml. The integrity of product was confirmed by Western Blot analysis using a mouse anti streptolysin O. Conclusion: Data showed that recombinant SLO protein can be produced by pET28a in Escherichia coli. This protein maintains its antigenic effect very well. Therefore, recombinant SLO has same epitopes with natural form of this antigen.

The effects of exposure to simulation cell phone waves on gonads of male mouse

Javad Baharara

Introduction: The increasing users of microwave appliances such as cell phones caused an increasing interest on investigation of its adverse effects on human health and development of animals.This study is done to investigate the effects of simulating cell phone waves on gonads and sex cells of male Balb/C mouse. Materials and Methods: This is an experimental study in which with the use of generating apparatus of simulating cell phone waves, adult male mice were exposed to cell phone waves for 10 days (4 hours per day). Then structure and ultrastructure of testes and number of sex cells were examined by light and transmission electron microscope. Data was analyzed using t and Mann Whitney testes. Results: The results did not show any significant differences in the size and weight of testes in mice exposed to the simulating cell phone waves. Results also showed that the number of spermatogonia cells and primary spermatocytes and spermatids and sperms were significantly increased in exposed mice (p<0.05), but the number of sertoli cells were significantly decreased (p<0.05). The study of micrographs showed changes in ultrastructure of sexual cells, such as cleft and hetrochromatined nucleus and decrease of cell organelles and vacuolization of cytoplasm. Conclusion: Results indicate the effect of simulating cell phone waves on number and ultrastructure of sex cell in male Balb/C mouse.

Early identification of hearing impairment of neonates admitted to neonatal intensive care unit using otoacoustic emissions

Arash Bayat

Introduction: It is well recognized that hearing is critical to speech and language development, communication, and learning. Otoacoustic emission (OAE) is an efficient and sensitive method to identify subjects at risk for auditory impairment. Infants who require admission to neonatal intensive care unit are reported to be at 10-20 times greater risk for hearing impairment. The porpuse of this study was to investigate the incidence of hearing impairment in neonates screened by OAE. Materials and Methods: In a cross-sectional study, 148 newborns having risk factors for hearing impairment reffering to Taleghani hospital of Arak and Aboozar hospital of Ahwaz were evaluated. All clients had normal otoscopic findings. Transient evoked (TEOAE) and distortion-product otoacoustic emissions (DPOAEs) were measured in both ears. If the results of otoacoustic emissions were not normal, these tests were repeated one month later. Patients who did not pass the second stage were reffered for comprehensive auditory evaluations. Data analysis was performed using Kolmogrov-Smirnov and t-tests. Results: 41 and 28 cases could not pass the examining test at the first TEOAE and DPOAE examination, respectively. Also, 23 and 16 cases did not pass the examining test at the second TEOAE and DPOAE evaluations, respectively. These participants underwent auditory brainstem response evaluation and 11 of them had abnormal responses. In 5 cases of confirmed ones, hearing impairment was due to aminoglycoside side effects. Conclusion: OAE hearing screening of at-risk newborns is a clinically beneficial approach to early detection of hearing impairment. Regarding the high prevalence of hearing loss in our subjects (7.43%), prevention of its complications is highly recommended in this population.

Effect of the V364D mutation in membrane endocytosis of ROMK2 (Kir1.1b)

Saiid Hajihashemi

Introduction: Recent studies suggest that endocytosis of ROMK channels is important for regulation of K+ secretion in cortical collecting ducts. In this study the effect of V364D mutation is examined on the membrane turnover and stability of ROMK2 channel when expressing in Xenopus laevis oocytes. Materials and Methods: In this experimental study, oocytes were isolated by standard protocols using collagenase (Type 1A). Mutations of the cytoplasmic termini of ROMK2 were constructed using the quikchange approach for site-directed mutagensis. Xenopus oocytes were injected with cRNA encoding ROMK2 or V364D mutant three days prior to treatment with BFA solution (time 0). Brefeldin A (BFA) was added to the OR3 medium (+BFA) at concentrations of 5-25 μM (inhibit insertion of new proteins into the cell membrane) or ethanol as BFA vehicle (-BFA). Two-electrode voltage clamp (TEVC) was used to measure oocyte ROMK-dependent currents and membrane potential. Data was analysed using Student’s t-tests or ANOVA as appropriate. Results: Incubation of oocytes expressing ROMK2 channels in both 5µM and 25 µM BFA caused a reduction in the normalized steady state currents. The effect of BFA was dose dependent. In oocytes expressing the V364D mutant, there was no decay in current at any time point during incubation with BFA at either 5 M or 25 M. The fractional current for ROMK2 at 48h following treatment of oocytes with BFA was 0.24  0.05 (n=16) which was significantly different to V364D mutant (1.17  0.09). Conclusion: These results show that the V364D mutation increases the general stability of ROMK and renderes the protein resistant to endocytosis, consistent with the idea that there is an interaction between the C-terminal of ROMK2 and components of the endocytotic pathway. A functional PDZ domain (the S-E-V) plays a key role in determining stability of ROMK.

The survey of age at natural menopause according to determinants factors in Arak city in 2007

Mohammad Rafie

Introduction: Menopause is a natural period of life. It means the permanent cessation of menstruation and the end of reproductive period which occurs between 45-52 years old. It also serves as an indicator of population health as the occurrence of menopause indicates the increased risk for the onset of several chronic diseases. In this study, the mean age of natural menopause among women in Arak (Central Iran) was investigated, considering their reproductive and socio-economic-demographic factors and their health behaviors. Materials and Methods: This study was an observational cross-sectional study on a population base, carried out in 2007 year. Interviews and questionnaires were used for gathering data from 458 women resident in Arak city, who had undergone a natural menopause. Sampling was multistaged using postal zones and selecting samples in each zone. Data was analyzed using variance analysis, multiple regression, T and correlation coefficient tests. Results: Findings showed that the mean age of menopause in our sample was 48.2±2.34 years (95% CI: 47.98- 48.41) and its median was 49 years. The mean age of menopause in women who were never married was less than married women (P= 0.015). Low income level (P= 0.003), having physical activity (P= 0.004), number of pregnancies (P<0.001), age of last pregnancy (P=0.026) and place of birth (P=0.003) were meaningfull factors in age of menopause. Conclusion: This phenomenon must be documented in Iran because of its strong link with mortality and morbidity of the growing number of women who are reaching advanced ages whose one forth of life is after menopause. Therefore health education as the best way for changing their lifestyle is suggested.

Effect of Allium Sativum extract on acid and pepsin secretion in basal condition and stimulated with Pentagastrin in rat

Mehrdad Shahrani

Introduction: Allium Sativum (garlic) is used widely in the world, especially in Iran. This plant is used in cooking foods as an odorant. In Iran it is believed that it is useful for gastrointestinal disorders. So in this study the effect of Allium Sativum extract on acid and pepsin secretion in rat is investigated. Materials and Methods: This experimental study is performed on two groups of rats (12 in each group: control group and allium sativum group). After anesthesia with nesdonal (50 mg/kg, ip), rats had gone under surgical tracheotomy, laparatomy and gasterodeodenostomy. In garlic group Allium Sativum extract (100mg/kg) was introduced into the stomach by gasterodeodenostomy canula. In order to stimulation of acid and pepsin secretion, Pentagastrin was used (25mcg/kg, ip). Stomach secretion was washed out and acid was measured using titerimetery and pepsin using Anson method. Data was analyzed using t-test. Results: Allium Sativum caused a meaningful increase in acid and pepsin secretion in garlic group comparing to control group (p<0.001). Pentagastrin also increased acid and pepsin secretion in control group (p<0.001). Conclusion: Using garlic in nutritional regimen causes an increase in acid and pepsin secretion in those using this plant.

The relation of HbA1C Levels and major depression in patients with type 2 diabetes mellitus, referring to endocrine clinic of Arak

Bahman Salehi

Introduction: Depression is one of the most common affective disorders, and in addition to different neurotransmitters and life events, various internal stressors such as blood glucose, cholesterol, triglyceride and other factors are responsible for it. Considering hyperglycemia as a possible risk factor of this disorder and inconsistency in results of previous studies, we decided to assess the relationship between different levels of HbA1C and major depression. Materials and Methods: This is a cross-sectional analythical study that is carried out on 134 samples. They were chosen randomly among patients with type2 diabetes reffering to Vali-e-Asr hospital. The patients were divided into 2 groups on the basis of their HbA1C test results patients with HbA1C less than 7% (group A), consisting 55 persons and patients with HbA1C more than 7% and equal to it (group B), including 79 persons. Participants were assessed by Hamilton scale of depression. The relation between HbA1C levels and major depression in these 2 groups was assessed using Chi square and Mann Whitney U tests. Results: We found that 40% of depressed patients had HbA1C level lower than 7% and 60% of them had HbA1C level higher than 7%. No significant correlation was found between HbA1C and major depression. But the duration of disease, using Insulin and being hypertensive simultaneously was correlated to major depression in this study. Conclusion: This study indicates that uncontrolled diabetes and high level of blood glucose is cosidered of less importance as an undelying mechanism of depression in patients with type 2 diabetes.

Evaluation of the relation between Dehydroepiandrostrone level and Insulin sensitivity

Afsane Talali

Introduction: Dehydroepiandrostrone (DHEA) and its sulfate form is an adrenal hormone and is the most common steroidal hormone in body and despite of other adrenal hormones increases with age, so it is called youth fountain. Recently, DHEA is noticed increasingly because of its multiple effects. Its effect on insulin sensitivity is controversial. In this study the relation between Dehydroepiandrostrone (DHEA) level and Insulin sensitivity is investigated. Materials and Methods: In this cross-sectional analytical study, samples were selected among female relatives of diabetic patients referring to Isfahan Endocrine Research Center. They were divided into three groups including diabetic, impaired glucose tolerance and normal glucose tolerance, each 30 persons. In these groups blood sugar, lipid, CBC, Insulin and DHEA-S levels were evaluated and the rate of Insulin sensitivity was determined using Homeostatic Modle Assessment- Insulin Resistance (HOMA-IR) and the relation between insulin sensitivity rate and DHEA in the three groups was compared. Data was analyzed using one way analysis of variance and t-tests. Results: The mean of variables in the three groups of diabetic, impaired glucose tolerance and normal glucose tolerance for DHEA-S was 127, 113, 129 mcg/dl Insulin 7.7, 1.10, 8.3 mcg/ml HOMA-IR 1.6, 3.5, 1.8 and fasting blood sugar 86, 135, 89 mg/dl respectively. Comparison of mean of variables showed that the difference between DHEA-S in groups was not significant. Also there was no meaningful relationship between DHEA-S and blood sugar. In the group with normal glucose tolerance, DHEA-S with HOMA-IR and insulin had a partial meaningful relation (p<0.06). Conclusion: Overally DHEA has no meaningful relation with insulin sensitivity and HOMA-IR, but this relation is meaningful in the three groups and the relation between DHEA-S with HOMA-IR in the group with normal glucose tolerance is partially meaningful.

The effectiveness of Dexamethasone on pain, nausea and vomiting after hemorrhoidectomy

Shahin Fateh

Introduction: Post operative pain, nausea and vomiting are common complications after hemorrhoidectomy and cause unpleasant expriences for patients. Today, scientists are increasingly moving forward to find new drugs with fewer side effects and costs to control these complications. So this study was planned to evaluate the effect of Dexamethasone on post operative pain, nausea and vomiting. Materials and Methods: This study was a double blind randomized controlled clinical trial that carried out on 112 patients scheduled for hemorrhoidectomy. Patients were diveded into two groups. Case group received 8mg Dexamethasone and control group received 2 ml normal saline intramuscularly 30 minutes before the operation. The method of surgery and general anesthesia were the same in all patients. Pain and nausea score were recorded using visual analog scale (VAS) and vomiting was recorded by asking of them at 0, 2, 4, 6 hours after operation. The patients were followed for 6 hours after the end of the operation and data was analyzed using t and chi square tests. Results: There was a significant difference between the two groups regarding prevention of post operative pain and nausea (p=0.00001). But there was no significant difference between the two groups regarding prevention of post operative vomiting. Conclusion: The result of this study suggested that injection of Dexamethasone before hemorrhoidectomy can decrease incidence of postoperative pain and nausea but dose not have any effects on postoperative vomiting. Therefore Dexamethasone is recommended before hemorrhoidectomy.

The effect of Kegel exercise on pain relief post hemorrhoidetomy

Monireh Motevaselian

Introduction: Hemorrhoid is a common disorder causing pain, and its standard treatment is Hemorrhoidetomy. Also, pain is a common problem after surgery and control of it prevents physiologic and psychiatric complications. Kegel exercise is one of the relaxing and none invasive methods for reduction of pain. In this study the effect of kegel exercise on pain relief post hemorrhoidetomy is investigated. Materials and Methods: This study is a single blind randomized clinical trial. Samples were patients admitted for hemorrhoidetomy to Shahid Sadooghi hospital of Yazd. They were randomly divided in two experiment and control groups (each group 30 persons). Before operation, Kegel exercise was educated to experiment group and after surgery pain range were recorded. Data was analyzed using descriptive and analythic statistics (t and Pearson correlaton tests). Results: Findings showed that the mean score of post surgery pain in the first 48 hours after operation in experimental group was 26.50±5.82 and in control group 31.26±5.94, that have meaningful difference (P=0.003). Regarding the amount of analgesics, the mean use of analgesics after surgery in experimental group was 18.9±14.83 and in control group 52.83±29.89 which has meaningful difference (P=0.00001). Type of analgesics in the two groups 18, 24 and 48 hours after surgery with p=0.04, p=0.04 and p=0.05, had significant difference. There were not any correlations between pain intensity and numbers of Kegel exercise. Conclusion: Based on results, Kegel exercise decreased post hemorrhoidetomy pain and using analgesic drugs was decreased in experimental group. Preoperative education of this method is useful in pain management postoperatively.

The Relationship between diabetes and hearing impairment

Efat Mashhadi

Introduction: Diabetes mellitus is a chronic disease that has well known effects on most organs, but there have been controversies in few different studies about the effects of diabetes on hearing. So this study was carried out in 2003-2004 in Kermanshah Diabetes Research Center to assess the effect of diabetes on hearing. Materials and Methods: This is a case-control study. 82 diabetic cases (26 types I and 56 type 2 cases) were studied along with 82 controls matched both in age and sex. Cases were selected randomly, their range of age was 15 to 55 years and their duration of diabetes was more than 3 years. After completing questionnaires, all cases and controls undergone autoscopic examination. Those with other ear disease with hearing impairment were omitted. Audiometery including pure tone audiometery, speech audiometery, and impedance audiometrey with well tone AD19 formed was done on 164 (diabetic and control) persons. Data was analyzed using t-test. Results: Audiometric findings with different frequencies in the two groups revealed that there was a significant relationship between diabetes and hearing impairment (p<0.001). This relationship was more significant in high frequencie, older age and type 2 diabetes, but there were no relationship between control of blood sugar (HbA1c), duration of diabetes, sex, microvascular and macrovasculur complications with hearing loss in diabetic patients. Conclusion: These finding demonstrate that diabetes can cause mild to moderate hearing impairment especially in high frequencies. The mechanism of this effect is unknown. Complementary studies with more patients and with more accurate devices such as Otoacustic Emission (OAE) and Audiometric Brainstem Reflex (ABR) are recommended

Comparing the sacral ratio in children with various degrees of vesicoureteral reflux with children without reflux

Parsa Yousefi

Introduction: Children with reflux who fail prophylaxis management or reflux which do not resolve spontaneously are candidates for sugery. Delay in surgery increases the development of new renal scars. Finding a modal factor for prediction of reflux resolution likelihood can decrease complications of treatments. Regarding that sacral bone anomalies have a direct influence on the final function of urinary and bowel systems, this study is done to investigate the relationship between sacral ratio and reflux. Materials and Methods: This is a case-control study. The case group included 76 children less than 9 years old refered to Vali-e-Asr and Amir Kabir hospital clinics with urinary tract infection and according to voiding cystouretrograply (VCUG) results, children with III, IV and V grade vesicoureteral reflux (VUR) entered the study. The control group included 76 children with previous urinary tract infection and normal VCUG. Sacral ratio in the two groups was measured and was compared by student t, Chi square and Mann Whitney tests. Results: The case group included 24 males and 52 females with average age of 2.7 years old and the control group included 23 males and 53 females with the average age of 3 years. The percent of sacral ratio abnormality were 40% and 23.7% in the case and control groups respectively, which was significantly different (p=0.001). The difference between the mean sacral ratio in case group with the value of 80% and the control group with the value of 84% was not different. The ratio distribution pattern in the two groups was not different. Conclusion: The percent of sacral ratio abnormality in children with vesicoureteral reflux were more than children without reflux.

A case report of familial adenomatous polyposis

Mahmoud Amini

Introduction: Familial adenomatous polyposis (FAP) is a rare disease with hundreds to thousands adenomatous polyps in large intestine which inherits in the form of autosomal dominant, but 25% of patients have no positive family history. Rectorrhagia and rectal prolapsus are the most prevalent symptoms. Diagnosis is based on clinical findings and results of colon investigation, and prompt colectomy prevents carcinoma in patient. In this article a case report of familial adenomatous polyposis is presented. Case: The patient is a 17 years old teenager, a known case of familial adenomatous polyposis, who was refered with severe obstruction signs of gastrointestinal tract. The disease was diagnosied when he was 5 years old. In 6 years old he was treated by partital colectomy and in 11 years old by total colectomy and ileorectal anastomosis. The patient was investigated by endoscopy and small bowel transit. Because of many polyps in stomach, duodenum and jejunum polypectomy, gastrectomy and duodenotomy were done. Diffuse carcinoma in situ was reported in pathology. Conclusion: Rectal and colonic polyps are rarely obsereved below 10 years old, but rectorrhagia and rectal prolapsus suspects the diagnosis of FAP in children. In the case of dysplasia and carcinoma abdominal pain, anemia and weight loss will be presented. In children with suspecting FAP, screening of extracolonic manifestation and dysplasia should be considerd and elective surgery should be done.

A case report of hypomagnesemia hypercalciuria nephrocalcinosis in a 3 years old child

Abolhassan Seiedzade

Introduction: Hypomagnesemia hypercalciuria nephrocalcinosis (HHNC) is a rare syndrome due to mutation defect in CLDN-16 gene. Its product is paracellin, a protein in henle loop paracellur area which acts in magnesium and calaium reabsorption. In this article a case of HHNC is reported. Case: The patient is a 3 years old boy presented with tetany, polyuria and polydipsia. Imaging study revealed medullary nephrocalcinosis. Laboratory investigations showed hypomagnesemia, hypercalciuria, hypermagnesiuria and normal arterial blood gases. In 2 years follow up, no neurological diseases, growth failure, or rickets were seen. He was treated with Hydrochlorothiazide and Potassium polycitra for prevention of nephrocalcinosis and progressive renal failure. Conclusion: HHNC is a rare syndrome presenting with hypercalciuria, hypo- magnesemia nephrocalcalcinosis, tetany, polyuria and polydypsia which progresses to end stage renal failure in 2nd or 3rd decades. Early diagnosis and some treatment modalities like Hydrochlorothiazide or polycitra-k can delay this process.