Shahid Sadoughi University of Medical Sciences International Journal of Medical Laboratory 2423-3706 4 1 2017 2 1 The Effect of Homeopathic Arnica on Macrophage Gene Expression 1 6 EN Salvatore Chirumbolo Evidence Based Complementary and Alternative Medicine (e-CAM Journal)-Editor -

Shahid Sadoughi University of Medical Sciences International Journal of Medical Laboratory 2423-3706 4 1 2017 2 1 Reference Interval of Cerebrospinal Fluid-Adenosine Deaminase Enzyme in Apparently Healthy Subjects- A Retrospective Study in India 7 14 EN Shalini N. Maksane Department of Biochemistry, Bhatia Hospital, Mumbai, Maharashtra, India. Rupali Parikh Department of Pathology, Bhatia Hospital, Mumbai, Maharashtra, India. Lakshmi Vaswani Department of Pathology, Bhatia Hospital, Mumbai, Maharashtra, India. Background and Aims: The aim of the present study was to re-evaluate the upper limit of normal range (ULN) for adenosine deaminase (ADA) level in cerebrospinal fluid (CSF) of healthy subjects in Indian population. Materials and Methods: Posteriori strategy was used for reference population selection. Individuals whose CSF samples had come to the laboratory of Bhatia Hospital, Mumbai (India) for ADA estimation during the period of January 2015-October 2016. After database search, 130 subjects were selected. Hexokinase method for CSF-glucose, turbidometric method for total protein and enzymatic method using the ADA assay kit BSBE for CSF-ADA was used. Non-parametric methodology recommended by IFCC/CLSI for reference intervals calculation was adopted. The 2.5 and 97.5 percentile formed the lower and upper limits of Reference Interval. Results: ULN range of CSF-ADA for the total study cohort was 10.7 U/L, which was quite higher compared to the ULN provided in the reagent kit insert (5.0 U/L). Upon gender wise comparison, female showed higher median CSF-ADA activity compared to male (Male-7.2; Female-8.2 p=0. 01), but upon application of robust methods, ULN of CSF-ADA did not show significant differences (ULN in Male-12.3 and in Female-12.5). Age wise analysis revealed that with increasing age; ULN of CSF-ADA showed increasing  till the age of 50 years and then showed decreasing trend with further increase in age. Conclusions: ADA level of 10.7 U/L in CSF should be adopted as the upper limit of normal range and gender based partitioning is not recommended. Age should be considered during the interpretation of CSF-ADA results for making this diagnostic test more clinically relevant.

Shahid Sadoughi University of Medical Sciences International Journal of Medical Laboratory 2423-3706 4 1 2017 2 1 Decreased Expression Levels of S100A12 and RAGE May Be Associated with Chronic HBV Infection 15 24 EN Yaser Eskandari Torbaghan Department of Immunology, International Campus, Shahid Sadoughi University of Medical Sciences, Yazd, Iran Mohammad Kazemi Arababadi Department of Laboratory Sciences, Faculty of Laboratory Sciences, Rafsanjan University of Medical Sciences, Rafsanjan, Iran. Ali Shams Department of Immunology, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. Background and Aims: Engagement of the receptor for advanced glycation end products (RAGE) and its ligand “S100A12 protein” induce a cascade of reactions that eventually might lead to develop an inflammatory response dependent on NF-κB. Although involvement of S100A12 and RAGE in some autoimmune disease have proved, in chronic hepatitis B (CHB) infection functions of the proteins are not clear thus far. Determining of expression of S100A12 and RAGE in peripheral blood cells of the CHB patients was the aim of the present study. Materials and Methods: In the case-control study the mRNA levels of S100A12 and RAGE genes of the sixty CHB patients and sixty healthy donors were measured by real-time polymerase chain reaction method. The patients and healthy donors were sex and age-matched. Results: The findings demonstrated expression of S100A12 and RAGE in the CHB patients significantly decreased compared to the healthy donor group (p=0.001). Expression levels of the genes were not altered among HBeAg-positive and HBeAg-negative CHB patients. The HBV-DNA copy number/ml did not affect the expression of S100A12 and RAGE in the patients (p>0.05). Conclusions: The results of the study suggested that down regulation of RAGE and S100A12, because of their role in inducing inflammation, might have a considerable role in chronicity of hepatitis B.

Shahid Sadoughi University of Medical Sciences International Journal of Medical Laboratory 2423-3706 4 1 2017 2 1 Prevalence of qnr Genes in Extended-Spectrum β-Lactamase Producing Klebsiella pneumoniae Isolated from Clinical Urine Specimens in University Teaching Hospitals, Iran 25 33 EN Amin Dehghan Banadkouki Department of Medical Microbiology, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. Gilda Eslami Research Center for Food Hygiene and Safety, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. Hengameh Zandi Research Center for Food Hygiene and Safety, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. Ali Dehghan Banadkouki Laboratory of Imam Khomeini Hospital, Karaj, Iran. Background and Aims: Extended-spectrum β-lactamase (ESBL) producing gram negative bacteria are resistant to penicillins, narrow and extended-spectrum cephalosporins, and aztreonam; also they are frequently resistant to trimethoprim-sulfamethoxazole, aminoglycosides, and quinolones. This study aimed to investigate the prevalence of plasmid-mediated quinolone resistance (PMQR) determinants qnrA, qnrB and qnrS in ESBL producing Klebsiella pneumoniae isolates. Materials and Methods: In this descriptive-sectional study, 130 Klebsiella pneumoniae isolates were collected from urine specimens and identified by convectional biochemical tests from December 2013 to August 2014,. Antimicrobial susceptibility testing was performed by disk diffusion method (Kirby-Bauer). The presence of ESBLs was confirmed by combination disk tests. E.test method was used for determination of ceftazidime and ciprofloxacin minimum inhibitory concentration (MIC). qnr genes were investigated by multiplex polymerase chain reaction and after sequencing, indexed in Genbank database. Results: Out of 130 isolates, 46 (35.4%) isolates were identified as ESBL producers, considering all, the highest rate of resistance belonged to amoxicillin, cefotaxime and ceftriaxone (each one 100%) and the lowest rate of resistance was for meropenem and ertapenem (each one 4.3%). 45 (97.8%) isolates were resistant to ceftazidime (MIC ≤ 16) and 24 (52.2%) isolates were resistant to ciprofloxacin (MIC≤4). qnrB and qnrS genes were detected in 21 (45.7%) and 7 (15.2%) isolates, respectively. 7 (15.2%) isolates were positive for both qnrB and qnrS genes. qnrA was not detected. Conclusions: With respect to the high prevalence of qnr genes in ESBL producing Klebsiella pneumoniae isolates, quinolones and beta-lactam agents should be used with caution.

Shahid Sadoughi University of Medical Sciences International Journal of Medical Laboratory 2423-3706 4 1 2017 2 1 Prevalence of Virulence Factors and Antimicrobial Resistance of Streptococcus agalactiae and Streptococcus uberis in Ruminant Sub-clinical Mastitic Milk in Iran 34 47 FA Hassan Momtaz Department of Microbiology, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran. Mahvash eliyasi Department of Microbiology, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran. Reza Soleimani Faculty of Veterinary Medicine, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran Ali Jazayeri Faculty of Veterinary Medicine, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran Background and Aims: The aim of this study was to determine the virulence and antibiotic resistance properties of Streptococcus agalactiae (S. agalactiae) and Streptococcus uberis (S. uberis) isolated from the ruminant’s sub-clinical mastitic milk in Iran. Materials and Methods: Positive samples in the CMT test were cultured and S. agalactiae and S. uberis have been identified using biochemical tests and polymerase chain reaction (PCR). Virulence and antibiotic resistance genes were studied using PCR. A total of 350 milk samples were collected randomly from 4 major provinces of Iran. We chose 55 bovine, 43 ovine, and 32 caprine herds from these regions. Results: Totally, 34.52% of bovine, 25% of ovine, and 25% of caprine milk samples were found to be contaminated with S. agalactiae and also, 27.38% of bovine, 25% of ovine, and 25% of caprine milk samples were found to be contaminated with S. uberis. The most commonly detected virulence genes of S. agalactiae were cfb (68.75%), scpB and pavA (43.75%). Also, the most commonly detected pathogenic genes of S. uberis were sua and hasB (83.33%), cfu (75.00%) and skc (70.83%). Totally, tetO (89.28%), ermA (82.14%), mefA (73.21%) and tetM (69.64%) were the most common antibiotic resistance genes in Streptococcus isolates. The most resistance were detected against tetracycline (87.5%) and erythromycin (83.92%), respectively. Conclusions: Our results showed that animal milk samples should be controlled, especially in colder seasons of the year in order to study the presences of S. agalactiae and S. uberis.

Shahid Sadoughi University of Medical Sciences International Journal of Medical Laboratory 2423-3706 4 1 2017 2 1 The Relation of Serum Bilirubin Level with the Severity and Complexity of Coronary Artery Disease and Long-term Outcomes in the Patients Undergoing Primary Percutaneous Coronary Intervention 48 55 EN Arsalan Salari Department of Cardiology, Guilan Interventional Cardiovascular Research Center, Heshmat Hospital, Guilan University of Medical Sciences, Rasht, Iran. Fatemeh Moaddab Department of Nursing, Guilan Interventional Cardiovascular Research Center, Heshmat Hospital, Guilan University of Medical Sciences, Rasht, Iran. Mahsa Pourabdollah Tootkaboni Department of Cardiology, Guilan Interventional Cardiovascular Research Center, Heshmat Hospital, Guilan University of Medical Sciences, Rasht, Iran. Maryam Shakiba Department of Epidemiology, Guilan Interventional Cardiovascular Research Center, Heshmat Hospital, Guilan University of Medical Sciences, Rasht, Iran. Mohadeseh Poursadeghi Department of Nursing, Guilan Interventional Cardiovascular Research Center, Heshmat Hospital, Guilan University of Medical Sciences, Rasht, Iran. Background and Aims: Bilirubin has been considered an antioxidant that protects against atherosclerosis. The aim of this study was to evaluate the relationship of serum bilirubin level with the severity and complexity of coronary artery disease (CAD) and long-term outcome in the patients undergoing primary percutaneous coronary intervention (PCI). Materials and Methods: This prospective cohort study was performed on 136 patients with STEMI undergoing PCI in Heshmat Hospital, Rasht, Iran during 2 years. The complexity was calculated by using dedicated software of the syntax score and the severity of CAD was determined based on the number of vessel involvement. Also, long-term outcomes were evaluated by major adverse cardiac events (MACE ) after one year. Results: The mean total bilirubin level was 0.95 ± 0.69 mg/dl; 60.3% of the patients were in the low range of syntax score (0-22) and 38.2 % had two vessels involved. In examining the relationship between serum bilirubin level with the severity and complexity of CAD and MACE, There was no statistically significant association (p>0.05); but, the majority of the patients with high bilirubin levels had a lower number of vessels involved and the complexity of the vessels; Also, MACE was observed in the patients with low bilirubin levels. Conclusions: According to the results, attention to serum bilirubin level could be important in CAD and the protective effect of bilirubin can be used for new therapeutic decisions. Further studies on a larger sample of patients have been recommended to confirm and complete these findings.

Shahid Sadoughi University of Medical Sciences International Journal of Medical Laboratory 2423-3706 4 1 2017 2 1 The Relaxant Effect of Carvacrol on Acetylcholine-induced Contraction in Isolated Rat's Ileum 56 62 FA Hamed Faghihi Department of Physiology, International Campus, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. Abolghasem Abbasi Sarcheshme Department of Anatomy, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran Seyed Hassan Hejazian Department of Physiology, Herbal Medicine Research Center, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran Background and Aims: The antioxidant and anti-inflammatory effects of carvacrol were reported. The aim of the current study was an investigation the relaxant effects of carvacrol on acetylcholine-induced contraction in isolated rat's ileum. Materials and Methods: In this study, the tissues separated from ileum were fixed on the organ bath containing Tyrode's solution. The amplitude of contractions has been recorded using isotonic transducer. Before and after subjecting the tissues with saline and different concentrations of carvacrol,the procedure was elicited by cumulative logarithmic concentrations of acetylcholine (Ach). Results: Our results indicated that carvacrol at 10-2, 10-3 and 10-4, but not 10-5 M decreased acetylcholine (10-4 M)-induced contraction (10-2 M at p<0.001; 10-3 and 10-4 at p<0.05 vs. Ach+saline). The antispasmodic effect of carvacrol was examined and showed that in concentrations of 10-5, 10-4 and 10-3 M, but not 10-6 M, the contractive effect of acetylcholine10-3 M was prevented significantly. The concentrations of 1, 2 and 4 µM of verapamil were also tested and indicated that 4 µM of verapamil reduced contraction by 22.1±.5% (p<0.01 vs. saline). In samples treated with combined carvacrol (10-5 M) and verapamil (4 µM) contraction percentage was decreased by 35±14%, which was significantly different compared to the Ach+saline group as well as the single treated groups (p<0.01 and p<0.05, respectively). Conclusions: According to our findings, due to the effect of carvacrol and verapamil, the relaxant mechanism of carvacrol may be mediated by Ca+2 release from the sarcoplasmic reticulum and the response of the contractile system.

Shahid Sadoughi University of Medical Sciences International Journal of Medical Laboratory 2423-3706 4 1 2017 2 1 Determination of Helicobacter pylori Antigen in Stool Samples and Comparison with Rapid Urease Test in Patients Suspected of Helicobacter Infection 63 68 EN Reza Marvinam Department of Immunology, Reproductive Immunology Research Center, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. Hossein Hadinedoushan Department of Immunology, Reproductive Immunology Research Center, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. Mahdi Dehghanmanshadi Department of Immunology, Reproductive Immunology Research Center, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. Fateme Zare Department of Immunology, Reproductive Immunology Research Center, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. Background and Aims: Helicobacter pylori (H. Pylori) is the microorganism that infects nearly half of word's population. There are several invasive and non-invasive methods for diagnosis of infection. The main objective of this study was to evaluate antigen of H. Pylori in feces with H. pylori stool antigen (HpSA) test and comparison with rapid urease test (RUT) in the patients suspected to be infected. Material and Methods: one hundred thirty-seven subjects (56 males, 81 females) were recruited from those patients undergoing a gastrointestinal endoscopic examination in the endoscopy units of Shahid Sadoughi university of medical sciences. One biopsy specimen was obtained from the stomach and each biopsy specimen per subject was tested for the presence of urease using the commercially available CLO test. Stool specimens were taken concurrently with the endoscopic examination and tested by Enzyme-linked immunosorbent assay (ELISA) method for presence of HpSA. In this study, RUT was considered as a gold standard test. Results: The mean age was 40.4±1.12 years. 13.3% of samples were shown HpSA-positive and negative RUT and 12.9% were shown HpSA-negative and positive RUT. Sensitivity and specificity of HpSA test was 86.6% and 87.1%, respectively. Positive and negative predictive values and accuracy were 89%, 84.3%, 86.2%, respectively. Conclusions: Our findings showed that the stool enzyme immunoassay for H. pylori is a useful method for the primary diagnosis of H. pylori in the patients suspected to be infected.