DANESHVAR MEDICINE
Year:2006 | Volume:13 | Issue:62|Papers Count:14

Introduction: Monoclonal antibody has many applications against peroxidase among which peroxidase-anti peroxidase (PAP) complex formation is of great importance. This complex is used in many immunohistochemical and immunocytochemical staining techniques. The aim of this study was the production of a mouse antiperoxidase of hybridoma cell line to be used in peroxidase - anti peroxidase immunostaining.Materials & Methods: The Balblc mice were first immunized by repeated injections of horseradish peroxidase (HRP). After the confirmation of their immunization by ELISA, the spleen lymphocytes and SP2/0 myeloma cells were hybridized by using PEG. The hybridoma cells were then selected through the selective HAT medium. From seven rounds of cell fusion, 224 hybridoma clones were obtained, from which six clones produced monoclonal antibody against HRP. Once again from these six clones, two clones were selected and developed. Limiting dilution was performed twice and monospecific clones were developed.Results: Using the Isostrip kit revealed that both monoclonal anti peroxidase antibodies (P1F11D2, P2F6F3) were from lgG1 isotype class. In addition, ELISA test showed that the monoclonal antibodies don't affect the active site of the enzyme. Therefore it seems that the produced antibodies are suitable for formation of PAP immune complex.Conclusion: At present, this study is being carried out farther to produce PAP complex and investigate its practical usage in immunohistochemical and immunocyto chemical techniques.

Introduction: Epilepsy is neuronal disorder and mechanism of this disorder is not understood. In this study the role of adenosine A1 receptors of the CA1 region of hippocampus on piriform cortex-kindled seizures was investigated in rats.Material and methods: In full kindled animal by daily electrical stimulation drugs N6-cyclohexyladenosinc (CHA), an adenosine A1 receptor agonist at doses of 0.1, 1, 10 an 100 μm and 1,3 dimethy-8-cyclopenthylxanthin (CPT), and adenosine A1 receptors at doses 10 and 20 μm Bilaterally microinjected to CA1 region of hippocampus. 24h before drug injection, microinjected ACSF and data are as a control groups.Results: Obtained results showed that in kindled animals, (CHA), decreased the after discharge duration (ADD), stage 5 seizure duration (S5D) and seizure duration (SO), and increased the latency to stage 4 (S4L) significantly. CHA at the doses of 1 μM decreased only ADD and no changed were observed at the doses of 0.1 μM. Bilateral microinjection of (CPT), increased ADD at the dose of 20 μM significantly, but had no effect on seizure parameters at the dose of 10 μM Pretreatment of animals with CPT (10 μM) before CHA (10 and 100 μM), reduced the effect of CHA on seizure parameters significantly.Conclusion: Thus, it may be suggested that the hippocampal CA1 region may play an important role in relaying or spreading of seizure spikes elicited from the piriform cortex and activation of adenosine A1 receptors of this region can prevent these actions.

Introduction: This study was performed to investigate the rate of inner cell mass of blastocyst which was obtained from the two-cell embryos of mouse culture in the presence and absence of recombinant of human leukemia inhibitory factor.Materials and Methods: ICR female mice (6-8 weeks old) received intra peritoneal injection of 7.5 IU of pregnant mare serum gonadotropine for super ovulation. This was followed by intra peritoneal administration of 7.5 of hCG 46-48 hours later. The mice were then mated to mature ICR male mice and were checked for vaginal plug 20 hours later. Female mice were killed by cervical dislocation 48-50 hours after hCG administration. After washing flushing of the oviduct from the proximal end of the oviduct, two cell embryos were selected and collected. All two cell embryos were randomly divided into 4 groups (A, B C and D) and were cultured in special media for 120 hours in CO2 incubator. Group A: KSOM+AA, Group B: KSOM+AA 500 IU/ML LIF. Group C: KSOM+AA 1000 IU/ML. Group D: KSOM+AA 1500 IU/ML LIF media and were cultured in special media for 120 hours in CO2 incubator. Later, all blastocysts were collected and ICM was counted by differential staining technology.Results: The rates of ICM of blastocysts, which were obtained from different groups, were 19±2.6, 28±4.4, 24±2.1, and 26±2.2, respectively. This data indicated that the rate of ICM in B, C and D was statistically higher than A (p=0.02) Moreover, there was no statistical difference between the three groups (B, C and D).Conclusion: In brief, since leukemia inhibitory factor can improve the rate of ICM of blastocyst, it is suggested that this factor be added to blastocyst culture medium.    

Introduction: The aim of the present research was to determine the isolated proteus strains from urinary samples of out patients and hospitalized patients and to specify the antibiotic resistant pattern.Materials & Methods: In this study, four hundred urine samples were collected from various sections of Imam Khomeini hospital and Valie-Asr clinic in Tehran. Collected samples were assessed by bacteriology examinations. With respect to culture results and isolating one kind of pure coloni and also based on existing more than 100000 colony count in culture medium, typical infection related to proteus strains was identified.Determination of urinary typical infection was based on isolating 106 samples (26.5%) from 400 mentioned specimens, in which 11 samples (10.4%) were caused by proteus group.Results: Results showed that proteus strains were the most salient isolated bacterial agents after E.coli in 39 samples (36.9%).In this investigation from 11 cases proteus, (73%) 8 cases belonged to women and the rest to men. This phenomenon could be in part due to special physiology of females' urogenital tract. On the other hand, 9 (82%) isolated samples were related to hospitalized patients and others (18%) to out patients. Increased incidence of urinary infection could probably be due to long-term hospitalization, nonobservance of health in hospitals and related to, patient's physical weakness. Prevalence age was mainly 30-39.Conclusion: Drug susceptibility pattern showed that susceptible antibiotics for urogenital infections, caused by proteus strains, were Amikacin and Nalidixic acid (91%) and Ofloxacine, Ciprofloxacine and polymyxine B (82%) respectively. In contrast, the proteus strains were completely resistant (100%) to Clincamycin, Tetracylin, Ampicillin and Oxacillin. Thus, early detection and timely treatment can prevent from urinary infection complications.

Introduction: In this study, in vitro activities of two plants Allium sativum (garlic) and Allium cepa L. (onion) were tested against reference and native dermatophyte species and compared with the activity of a known antifungal drug; terbinafine (TBF).Material & Method: Agar dilution method was used for determining the antidermatophytic activities of above-mentioned compounds. The fungi were cultured on Sabouraud dextrose agar in the presence of different concentrations of selected compounds. All compounds were inhibited the growth of all examined fungi in a dose-dependent manner.Results: The minimal inhibitory concentrations (MICs) of TBF, onion and garlic were determined in the range of 0.005-0.165 (μg/ml), 62.5-500 (μl/ml) and 0.24-3.91 (μl/ml), for all tested dermatophytes, respectively. The MFC values of these compounds were measured as 0.330 (μg/ml), 1000 (μl/ml) and 7.81-31.25 (μl/ml), accordingly. TBF showed the highest antifungal activity against Microsporum canis MC-1 and M. gypsum PTCC 5069. M. canis strains MC-1 and PTCC 5069 were the most resistant dematophytes against onion and garlic extracts.Conclusion: On the whole, two compounds TBF and garlic showed suitable antifungal activities in very low concentrations against Trichophyton mentagrophytes, T. rubrum, M. gypseum and Epidermophyton fluccosum. With respect to the high anti-dermatophytic activity of onion and garlic extracts, finding of active antifungal fractions of these extracts and in vivo study of their effects in treatment of various types of dermatophytosis is recommended.

Purpose: Newcastle disease virus (NDV) is a paramyxovirus, which causes Newcastle disease. This disease brings about great economical losses in poultry industry. Fusion protein, which is an essential factor in paramyxovirus pathogenicity, consists of HR1, HR2 and a fusion peptide that plays an important role in the fusion mechanism of the virus. Mutation in these regions and the produced peptides can block virus fusion. In this study, we selected one of the native isolates of Newcastle disease virus (i.e. NR43) to study its genomic sequence as a paramyxovirus model with emphasizing on the important regions.Materials & Methods: After RNA extraction, two regions of genomic RNA with about 800 bp (A: 10-820 and B: 810-1636) were amplified by RT-PCR and the products were cloned into a T-vector, then the inserts were sequenced.Results and Discussion: Alignment of sequence A with other strains of Newcastle virus by using Blast software (www.ncbi.nlm.nih.gov/blast), showed maximum similarity, first to Russian strain Vol 95 (Y16169) and then to Chinese viruses. Cleavage site sequence (RRQRR) indicates that the virus is also Velogenic. Comparison of HR1 sequence of NR43 with other strains of NDV, showed a novel mutation amino acid number 82 (F instead of L). The study of B sequence, containing HR2 domain, showed highly conserved regions with other NDV HR2 domain.

Introduction: Experimental studies show that some cancer cells including colon cancer cells up regulate the expression of Intracellular Adhesion Molecule-1 (ICAM-1, CD54). In addition, these protein molecules shed from the cell surfaces to the environments that could be detected in culture medium or patient serums. Carcino-embryonic antigen (CEA) has been known as a tumor marker, and is commercially available for detecting and following up the gut malignancy, specially the colon cancer. To determine and compare the serum levels of these molecules in colon cancer patients before and after operation, the present study has been designed.Materials & Methods: a group of patients (66% male and 34%femaleswith age ranges of 56±11 years) who were diagnosed for colon cancer pre- & post-operation in a University hospital was investigated. Their peripheral bloods were taken and their serums were isolated by standard and routine method. Similar procedure was performed for a healthy matched group, and then the serums for the molecule assessment were obtained using photometric enzyme immunoassay (or, ELISA).Results: The pre-operation levels of serum soluble molecule of patients were as follows: sICAM-1=781±219 ng/ml, and CEA=53.6±22.7 μg/l. In addition, the data obtained from healthy subjects showed the basal levels of sICAM-1=179±39 ng/ml, and CEA=3, 6±1.6 μg/l. However, after operation the mean±SD of samples indicated the following: sICAM-1=585±48 ng/ml, and CEA=41.3±26.5±g/l.Conclusion: CEA concentration as a tumor marker significantly increased in all the patients, moreover, sICAM-1 was augmented in the patient group four times more than the physiologic levels in healthy control group, the mean assessment of both molecules compared to decrease gradually after operation. Thus, it can be concluded that similar to CEA, sICAM1 can be considered as tumor marker to evaluate and follow up the situation in colon cancer.

Objective: Symptoms are important for appropriate triage and beginning of rapid treatment. Classification of patients referred to the emergency departments into high, low or moderate risk group involves accurate recognition of risk factors and symptom predictors.Method and Material: This survey is a descriptive analytic study to determine symptom predictors and ACS risk factors in patients hospitalized in eight selected hospitals in coronary care units (Tehran). 446 patients with unstable angina or acute myocardial infarction were randomly selected.Exclusion criteria were CVA history, joint degenerative disease, trauma, asthma, pulmonary infection and renal failure. Data collecting instruments were a questionnaire consisting of demographic variables, risk factors, baseline variables, electrocardiography changes, cardiac enzymes and also ACS symptoms checklist.Results: Results showed that menopause and diabetes are the only effective variables on report of typical symptoms. The mostly reported symptom among diabetic patients was diaphoresis and chest symptom was dominant in non-diabetic patients. Chest symptom was also most common in the menopause. Results showed that there was no significant difference between typical symptoms of ACS with sex (p=0.98). Further more, chest symptoms (sub sternum pain, left chest pain with pressure, heaviness, tightness/squeezing sense) were the most common complaints. Typical symptoms were similar in both younger (<65 year old) and older (>65 year old) patients. Syncope (24.4%) and chest symptom (10.3%) were most common in older and younger patients, respectively. Regression analysis showed that we can successfully predict disease by symptom predictors.Prediction probability was 45% in younger patients and 27% in older in presence of all typical symptoms.Conclusion: typical symptoms are strong predictors of ACS in younger patients. However, they are not strong predictors of ACS in women, since prediction probability was 64% for men and 27% for women.  

Background & Objectives: Today, various evidences indicate that Helicobacter pylorus is one of the significant factors in gastric ulcer and duodenal ulcers. This bacterium, which is seen in mucousal layer and in vicinity of epithelial cells causes, mucousa changes, cell disorders, tissue lesions, and inflammation. One of the important characteristics of this bacterium is producing large amounts of urease enzyme. The activity of urease enzyme can be an important virulence factor which plays a key role in causing damage to mucousa, and replacement of bacterium in mucousa of stomach. In this study, the role of urease enzyme in causing cell lesions was investigated. Methods: The study was carried out on 100 biopsy specimens. A speedy urea test was performed in endoscopy theatre, for all these samples. The biopsy samples were transferred to the laboratory in transport medium and were cultured in the selected medium. Then, the identification of bacteria was done by microscopic examination and biochemical tests. The cultures of 80 samples of biopsy were positive. The direct effect of supernatant containing urease enzyme extracts from 50 bacterium samples on HeLa cell. was investigated. Furthermore, urea with concentrations of 3.75 and 20 m.molar as well as enzyme containing supernatant were added to HeLa cell culture medium. To investigate the factors causing cell lesions, ammonia was used in PBS with concentrations of 40, 32, 22, 20, 15, 10, 8, 7 and 3 m.mol. Results: In the presence of urease enzyme and a concentration of 20 m.molar of urea, the colour of the culture medium turned into purple. This change was a sign of the production of large amounts of ammonia. Intracellular vacuoles were formed as well. By adding urease enzyme containing supernatant in the absence of urea, intercellular vacuole was not observed. Neither in concentrations of below 8 m.molar of ammonia was intercellular cell vacuole observed. While, in concentrations over 8 m. mol. the cytopathic effect was observed in the form of vacuoles, and in the concentration of 40 m. mol. of ammonia, the full erosion of cell layer occurred.Conclusion: Therefore, based on the results of this investigation, urease enzyme in not directly harmful. It is the ammonia produced as a result of hydrolyze of urea by urease enzyme which has toxic effects and causes inflammation and ulcers.

Introduction: With regard to the importance of QOL of patients with cancer, especially breast cancer, and conflicting researches on the quality and quantity of effect of counseling on QOL promotion in such patients, also, for determining the effect of group counseling on functional scales of QOL of patients with breast cancer treated with chemotherapy, this research was done in Cancer Institute of Imam Khomeini Hospital in 2003, Tehran.Material and Methods: This is a randomized clinical trial. 164 women (25- 55 years old) undergoing chemotherapy due to breast cancer and with mastectomy history, were studied in two groups: control group (Standard care n=82) and case group (counseling n=82). A counseling session was designed for the case group. QOL patients' were measured at the beginning and the end of study by two standard questionnaires (QLQ_Br 23, QLQ-C30). Results: Results revealed that nursing counseling improved QOL functional scales in the case group (p<0.001). General functional scales (Physical, role, emotional, cognitive and social functioning) and special functional scales (body image, sexual functioning and sexual enjoyment) were improved in the case group by means of counseling (p<0.001).Conclusion: Considering the results with which revealed the effect of group counseling program on promotion of QOL level in patients with breast cancer treated with chemotherapy, we suggest applying group counseling as an effective method in QOL promotion for such clients in their medical supportive centers.

Introduction: Myopia is becoming more of a visual problem in Asian countries.Purpose: To study tine changes in myopia with age among patients in Optometric clinic of Mashhad University of medical sciences.Methods: In this study, after a routine optometric examination, the prevalence rates and amount of myopia of the 2410 patients (1-97 years) from the optometry department, Mashhad University of Medical Sciences were analyzed by using X2 and ANOVA.Results: The results of this investigation showed that prevalence and amount of myopia (spherical equivalent) increased with increasing age from 1 to 20 years (p<0.0001) and had not worth considering until 30 years and then decreased.Conclusion and Discussion: These results indicated that 21 to 30 years is the best time for refractive surgery in myopic patients.