1 1028-852X Pasteur Institute of Iran 344 Genetic Heterogeneity of PKD1 and PKD2 Genes in Iran and Determination of the Genotype/Phenotype Correlations in Several Families with Autosomal Dominant Polycystic Kidney Disease Radpour Ramin Rezaee Mina Haghighi Mahdi M. Ohadi Mina Najmabadi Hossein Hajibeigi Asghar 1 1 2006 10 1 1 8 14 08 2011 15 01 2006 Autosomal dominant polycystic kidney disease (ADPKD) is the most common genetic nephropathy, which is characterized by replacement of renal parenchyma with multiple cysts. In Iran, the disease prevalence within the chronic hemodialysis patient population is approximately 8-10%. So far, three genetic loci have been identified to be responsible for ADPKD. Little information is available concerning the pattern of linkage in Iranian population. In the present study, the linkage analysis was performed using three pairs of polymorphic microsatellite markers including 16AC2.5-CA (D16S291), SM7-CA (D16S283) and KG8-CA (intragenic marker at the 3' end of the gene). These markers are closely linked to the ADPKD1 locus and three pairs of the selected polymorphic microsatellite markers including YUNCA9 (D4S231), AFM155xe11 (D4S1534) and AFM224x6 (D4S423), which were closely linked to the ADPKD2 locus. In parallel, the genomic DNA of 150 unrelated healthy individuals was used to determine frequency, heterozygosity rate and polymorphic information content (PIC) for each marker. In our study, haplotypes were constructed in a number of ADPKD families using respective markers. Assignment of the disease gene loci was performed following phasing and haplotype construction, genotype/phenotype correlations were deduced from the constructed haplotypes. Analysis of clinical data confirms a milder ADPKD phenotype for PKD2 families in Iran. Our results showed relatively high heterozygosity rates and PIC values for some markers, while the most informative markers were KG8 (PIC: 0.772) and 16AC2.5 (PIC: 0.689) for PKD1 gene and AFM224x6 (PIC: 0.712) for PKD2 gene. We report here the first molecular genetic study of ADPKD and the existence of locus heterogeneity for ADPKD in Iranian population by performing linkage analysis on 15 affected families. Eleven families showed linkage to PKD1 and two families linked to PKD2 gene. In 2 families, PKD1 markers were common in all affected members but PKD2 markers were not informative.
345 Expression and in vitro Characterization of Herpes Simplex Virus 1 (HSV-1) ORF P Protein Karimi ali Nafisi mohammad reza 1 1 2006 10 1 9 13 14 08 2011 15 01 2006 Herpes simplex virus 1 (HSV-1) unspliced 8.3 latency associated transcript (LAT), which located in the long repeat sequences, has been shown to contain at least 16 open reading frames (ORF: A-P). One of these ORF, ORF P, maps almost entirely antisense to HSV-1 neurovirulence gene, ICP34.5. Both ORF P and ICP34.5 are located in the long repeat and are antisense overlapping genes. Therefore, in ORF P deletion mutants, ICP34.5 is also deleted and thus, the characterization of ORF P mutants is almost impossible. An alternative way to analyse its function is to determine those cellular and viral proteins which interact with ORF P. During these experiments, firstly, the expression of full length GlutationeS-transferase (GST)-ORF P fusion protein was optimised and then, using GST-pull down, it was shown that ORF P interacts with a viral and a few cellular proteins in vitro. Conclusively, ORF P might have some functions in HSV-1 replication cycle.  346 The Effect of Nucleus Tractus Solitarius Nitric Oxidergic Neurons on Blood Pressure in Diabetic Rats Kourosh Arami Massoomeh Sarihi Abdolrahman Malacoti Seyed Mansour Behzadi Gila Vahabian Mehrangiz Amiri Iraj 1 1 2006 10 1 15 19 14 08 2011 15 01 2006 It has been shown that nitric oxide is synthesized in the central nervous system as well as in vascular endothelial cells. Recently, it was reported that nitric oxide was involved in central cardiovascular regulation, baroreflex modulation, and involved in a reciprocal release with excitatory amino acids in the nucleus tractus solitarii of rats. The purpose of the present study was to investigate the possible interaction of nitric oxide and glucose in the nucleus tractus solitarii on blood pressure regulation. Male Wistar stereptozotocin induced diabetic rats were anesthetized with urethane. A cannula was inserted above the nucleus tractus solitarii and blood pressure was monitored intra-arterially. Unilateral microinjection of L-glutamate (2.3 nmol/60 nL) into the nucleus produced a decrease in blood pressure in diabetic rats. Microinjection of lidocaine (0.5 µl %2) increased blood pressure. Unilateral microinjection of sodium nitroprusside (100 mmol/60 nL) into the nucleus increased blood pressure in diabetic rats. After microinjection of sodium nitroprusside, the depressive responses to glutamate were significantly attenuated. These results demonstrated the probable role of glucose on blood pressure regulation in diabetic animals affecting on nitric oxidergic neurons and so it implicates an interaction between nitric oxide and glucose in central cardiovascular regulation. Iran. Biomed. J. 10 (1): 15-19, 2006 347 Effect of Cuneiformis Nucleus Inactivation by Lidocaine Microinjection on the Analgesic Response of Morphine in Rats Sepehri Gholamreza Shafeiee Mohammad Naser 1 1 2006 10 1 21 26 14 08 2011 15 01 2006 The present study was performed to evaluate the analgesic effect of morphine microinjection into the cuneiformis nucleus (CnF) and the effect of inactivation of this area by lidocaine on pain modulation. Rats were anaesthetized by thiopental (45-60 mg/kg/i.p.) and placed in a stereotaxic instrument, and then a guide cannula was implanted just one mm above the CnF. Following surgery and recovery period, various doses of morphine (10, 20 and 40 µg/0.5 µl saline) and lidocaine 5% (0.5 µl) were microinjected into the CnF. Antinociceptive response was measured by tail flick latency (TFL) and maximal possible effect (% MPE) for 25 min at 5-min intervals, before and after any injection in control and experimental groups. The results of this study showed that morphine microinjection into the CnF increased TFL in a dose-dependent manner. TFL was also increased significantly after lidocaine microinjection. However, co-microinjection of morphine and lidocaine increased TFL which was less than morphine microinjection alone. The intravenous morphine injection with lidocaine microinjection increased TFL significantly, as compared to morphine microinjection. These effects were reversed by naloxone administration. In summary, the results of this study showed that morphine microinjection into the CnF caused a significant analgesic response which indicates that CnF may be involved in pain modulation. 348 Study of Correlation between Prostate Specific Antigen and Matrix Metalloproteinase-2 Activity in Benign and Malignant Prostate Hyperplasia Alizadeh Nastaran Safa Omid Khoramizadeh Mohammad reza Pezeshki Mohammad 1 1 2006 10 1 27 32 14 08 2011 15 01 2006 There are emerging data on novel tumor markers such as matrix metalloproteinase-2 (MMP-2 or gelatinase-A), which play a key role in tissue invasion and metastasis. We designed an investigation to assess the usefulness of MMP-2 activity as compared to prostate specific antigen (PSA) in cancer staging process. We have analyzed the circulating form of MMP-2 in serum samples of patients suffering from either benign prostate hyperplasia (BPH, n = 54) or prostate cancer (PC, n = 26), and prostatitis (n = 4) as compared to control normal individual (n = 26), respectively. Protein-content adjusted samples were separated by gelatin-embedded polyacrylamide gel electrophoresis then were subjected to densitometric analysis. Total PSA (tPSA) and free PSA (fPSA) were quantified using a standard ELISA technique. Correlation coefficient (r) between tPSA and MMP-2 activity in patient group was +0.938 and in controls group was 0.799 ( P <0.01). In addition, (r) between tPSA and MMP-2 activity in PC patients was 0.940 and in BPH patients was 0.962 ( P <0.01). (r) between fPSA and MMP2 activity in PC patients was 0.913, in BPH patients was 0.644, and in prostatitis patients was -0.994 (P <0.01). These results demonstrate that MMP-2, compared to PSA, might be considered as a better tumor marker in monitoring and screening patients with PC. 193 The Effects of Extremely Low Frequency Pulsed Electromagnetic Field on Collagen Synthesis of Rat Skin: a Biochemical and Histoligical Approach Ahmadian Shahin Rezaei Zarchi Saeed Bolouri Bahram 1 1 2006 10 1 33 38 03 08 2010 15 01 2006  The efficacious effects of pulsed electromagnetic field (PEMF) under the certain field parameters like frequency and the field intensity have been reported for various tissue and molecules. Since collagen is found abundantly in most tissue structures, this research was designed to further investigate the effects of extremely low frequency (ELF) PEMF on the synthesis of the epidermal collagen. To do the task, six groups of animals each consisting of eight mature male rats were selected randomly as one group for the control and five for the test. The field was generated by using a parallel set of Helmholtz coil. The first set of experiments was carried out at the peak intensity of 2 mT (milli Tesla) for different frequencies of 25, 50 and 100 Hz. Since the most effective frequency turned out to be 25 Hz, another set of experiment was conducted using this frequency and two different field intensities of 1 and 4 mT. The field was applied for 2.5 h/day lasting for 8 days, keeping the same procedure for the control group except for the field turned off. On the ninth day, the rats were sacrificed and the skin samples from the dorsal region were taken for biochemical assessment of collagen by measuring hydroxyproline content using Stegeman-Stalder method and histological assessment. The data indicated that pulsed electromagnetic field of 2 mT at 25 Hz increased the collagen synthesis (P<0.05). The other intensities and frequency setting did not have much distinguishable effect, however, at the frequency of 25 Hz and 4 mT, the field effect on the collagen increase was also noticeable. It was concluded that applying the field parameters of 25 Hz and 2 mT peak intensity for 2.5 h/day during eight days rendered a significant increase in collagen synthesis in rat skin. Histological observations were consistent with the biochemical findings.   343 Detection of Survivin Gene Expression in Formalin-Fixed Paraffin-Embedded Tissues of Human Osteosarcoma: Its Potential Usefulness in Diagnosis and Prognosis of Bone Tumors Babaei Esmaeil Mowla seyed javad Shariat Torbaghan Shams Emadi Bayegi Mojtaba 1 1 2006 10 1 39 45 13 08 2011 15 01 2006 Osteosarcoma is a relatively uncommon malignancy however, it is the most common form of primary malignant bone tumors in human. Diagnosis and prognosis of patients with osteosarcoma is limited to clinico-radiopathological parameters, whereas molecular markers of tumor aggression have been poorly identified. Survivin, an inhibitor of apoptosis (IAP), is unique for its expression in human tumors and fetal tissues but not in non-dividing normal adult cells. It mediates suppression of apoptosis in many cancers including bone tumors, and plays a role in tumor progression and chemotherapy resistance. In the present study, the expression of survivin was evaluated by heminested RT-PCR for amplifiable mRNA extracted from 23 formalin-fixed, paraffin-embedded (FFPE) specimens of high-grade osteosarcoma as well as 8 non-tumoral bone tissues. ß2-microglobulin (ß2m) gene expression was also evaluated, and used as an internal control. Survivin gene expression was detected in 82.6% (19/23) of high-grade osteosarcomas. In contrast, there was no gene expression in non-tumoral bone samples as well as the normal tissues obtained from the margin of some osteosarcoma samples. In conclusion, our data revealed that the expression of survivin is limited to osteosarcoma cells and associated with high-grade malignancies. Therefore, evaluating surviving gene expression might have a potential usefulness in diagnosis and prognosis of bone tumors. 349 Comparison of PCR and Culture Methods for Diagnosis of Enteropathogenic Campylobacter in Fowl Feces Havaei Seyed Asghar Salehi Rasol Bokaeina Mohammad Fazeli Seyed Ali 1 1 2006 10 1 47 50 14 08 2011 15 01 2006 Enteritis due to Campylobacter is the most common cause of acute bacterial diarrhea worldwide. In most cases, infection occurs as a result of consuming contaminated water or food, especially raw meat of fowls. Campylobacters are saccharolytic and fastidious bacteria. These traits limit the number of available biochemical tests by which isolates may be differentiated. These limitations might, in principle, be overcome by the use of PCR techniques, which is the aim of the present study. To compare the culture technique with PCR assay, a total of 116 fecal samples from fowls were tested using these two techniques for the presence of Campylobacters . Campylobacter strains were isolated from 11 (9.4%) out of 116 fecal cultures from fowls (8 C. jejuni and 3 C. coli ). Using PCR assays, the number of positive Campylobacter s increased to 27 (23%). Of these 27 positive samples, 18 were C. jejuni and 9 were C. coli . The sensitivity and specificity of PCR in comparison to the culture method were found to be 100 and 84.7%, respectively. According to the present study, it is proposed that the PCR is a reliable and sensitive method which can be used as a diagnostic technique for the detection of Campylobacter in fowls’ samples. 350 Protective Immunity in Mice with Whole Body of Echinococcus granulosus Hashemitabar Gholam Reza Razmi Gholam Reza Naghibi Aboulghasem 1 1 2006 10 1 51 55 14 08 2011 23 11 2015 Despite the establishment of extensive and successful control programmes in some countries or regions, Echinococcus granulosus still has a very wide geographical distribution. Mature E. granulosus (n = 120) obtained from Parasitology Department, School of Veterinary Medicine, Ferdowsi University of Mashhad (Iran). Soluble protein of whole body of parasite was prepared by freeze-thawing in liquid nitrogen and at 42ºC for three times. The sample was homogenized in a blender, sonicated at 110 V, 170 W for 3 times for 15 s each and then centrifuged at 10,000 ×g for 15 min. Final yield was kept in -20ºC. Ten mice were randomly divided into 2 groups of 5. Each mouse in test group was vaccinated subcutaneously with 100 µg (100 µl) of whole body of E. granulosus plus 100 µl of Freund’s complete adjuvant (FCA), respectively. Mice in control group were vaccinated with adjuvant in PBS. Second vaccination was conducted after four weeks with the same preparation except that FCA was replaced by Freund’s incomplete adjuvant (FIA). Three weeks after the second vaccination, each mouse was challenged with 2000 protoscolices, intraperitoneally. Mouse autopsy was carried out eight months post challenge. Our results show that none of the vaccinated mice with the whole body of E. granulosus had cysts that indicate 100% protective immunity.