Journal of Arak University of Medical Sciences

The Relation Between Accessory Gene Regulator (Agr) Types of S.aureus and Some Phenotypic Criteria

Meysam Hasan Nejad Bibalan

Background: Staphylococcus aureus is a gram-positive bacterium that has remained a persistent pathogen, causing infections such as endocarditis and toxic shock syndrome in humans. The accessory gene regulator (agr) system of Staphylococcus aureus is responsible for controlling the expression of many genes that code virulence factors and hemolysis.This study was carried out to determine the S.aureus agr group based on their source of isolation and any relation between agr speciﬁcity groups, pigmentation and hemolysis .

Materials and Methods: DNA of 194 S. aureus isolates were extracted by lysozym-phenol chloroform method, included 85clinical samples, 58 samples which isolated from nose of health care workers and 51 cases obtained from food product in Gorgan, North of Iran. PCR-based assays were used to evaluate agr locus nucleotide polymorphism for the identiﬁcation of agr specificity group. Pigmentation on nutrient agar medium and hemolysis on sheep Blood agar medium were assessed.

Results: The majority of isolates belonged to agr group I (43.3%), followed by agr group III (28.87%), agr group II (22.68%), and agr group IV (5.15%). The isolates belonged to agr group IV have greater ability to produce hemolysin (60%) whereas isolates belonged to agr group III have greater ability to produce pigment (60.5%).

Conclusion: agr group I was predominant among health care worker and food product specimens in Gorgan, North of Iran but in strains isolated from patient, agr group III was predominant. Investigation of the possible role of agr group III in Staphylococcus aureus infection in the next studies is recommended.

Determination of Extraction and Analysis Method for Imipenem in Hospital Sewage Samples Using High Performance Liquid Chromatography-UV Detector and with Solid Phase Extraction Column

Navid Safavy

Background: Drugs residual discharge into the environment through municipal and hospital wastewaters is one of the emergent environmental problems. Imipenem as a professional hospital antibiotic is widely used against gram- positive and negative bacteria and with entrance to the aquatic environments could prompt a lot of risks such as bacteria resistance, allergies, spoiling alga and daphnia and interrupting in wastewater treatment processes. Therefore there is a command to develop a method for extraction and determination of Imipenem from hospital sewage.

Materials and Methods: Solid phase extraction (SPE) was used to extract Imipenem from samples. Recovery percentage calculated at different pH of 3 and 7. The extracted samples analyzed by High Performance Liquid Chromatography (HPLC) equipped with UV detector. HPLC operated using borate buffer/methanol as mobile phase at flow rate of 0.7 ml/min, column temperature of 20 °C, and UV wavelength of 280-300 nm.

Results: Maximum recovery percentage was obtained 68% at pH=7. The best condition for HPLC was 80:20 ratio of borate buffer/methanol with pH=7.5 and at UV wavelength of 300 nm. Linearity calculated 0.9829, primary and intermediate precision both were more than 95%. Limit of detection (LOD) and limit of quantification (LOQ) were 3 and 10 µg/l respectively.

Conclusion: The method could simply and with significant reliability be applied to extract and determinate Imipenem in complex hospital wastewater matrixes.

The Inhibition of Nucleus Acumbens Cholinergic Receptors Decreased Memory Retention in Rat

Niloufar Darbandi

Background: It is well known that morphine influence learning and memory processes. The Nucleus accumbens (N.ac) which has an important role in reward participates in morphine-induced impairment of memory retention. Considering the cholinergic system is involved in the effects of morphine on learning and memory, in the present study, the effects of intra-N.ac injections of acetylcholine receptor antagonists alone or with morphine on memory retention and morphine-induced memory has been investigated in rats.

Materials and Methods: In this original research animals were bilaterally cannulated in the N.ac and a step-through passive avoidance task was used for the assessment of memory retention .

Results: Post-training subcutaneous administration of morphine dose dependently decreased the learning and induced amnesia. The administration of the same dose of morphine as pre-test treatment induced state-dependent learning. Pre-test intra- N.ac administration of atropine, scopolamine and mecamylamine in different doses alone cannot affect on memory retention. While, pretest intra- N.ac injection of these drugs before the administration of morphine dose dependently inhibited morphine state-dependent learning. The level of statistical significance was set at p<0.05 .

Conclusion: The processes of learning in animals can be affected by morphine and the opioids produce state-dependent learning. Moreover, it can be concluded that inactivation of the muscarinic and nicotinic acethylcoline receptors in the N.ac are involved in mediating morphine state-dependent learning.

Relationship Between White Blood Cell Count and Six Months Follow Up Outcomes in Patients with Acute Coronary Syndrome

Mahsa Ghasemi

Background: Inflammation has an important role in the pathogenesis of atherosclerosis. The aim of this study was to evaluate the role of WBC (White Blood Cell) count and incidence of morbidity and mortality in patients with ACS.

Materials and Methods: This prospective study was carried out on 101 patients with ACS who admitted in Bu-ali Sina hospital. All patients were stratified according to WBC categories in to 3 groups, (WBC 1 :<7000 mm3, WBC 2:7000 -10000 mm3, WBC 3:> 10000 mm3). Demographic and laboratory data such as acute reactive protein, cardiac biomarker and etc. were recorded. Adverse cardiac events and mortality were recorded to a phone or in person for six months of follow up period. The collected data were analyzed using SPSS software (Statistical Package for Social Sciences, version 17.0). The Fisher´s exact chi-square test and the student t-test were applied. P-values less than 0.05 were considered significant.

Results: In our study, 5 patients (31.25%) in third group had recurrent non fatal cardiac event and the same percent (31.25%) were died after 6 months follow up. Multivariate analysis showed WBC count >10000 mm3 was strongest predictor of outcome in our patients.

Conclusion: WBC count can be considered one of the strong independent predictor of mortality and cardiac event in patients with ACS.

Comparative Study on the Effects of Aloe Vera Extract in Clinical Strains of Staphylococcus Aureus, Klebsiella, Staphylococcus Epidermidis and Escherichia Coli Compared to Antibiotics of Choice

Mohammad Arjomandzadegan

Background: Nowadays, with the development of drug resistance, the use of herbs as an alternative to chemical drugs is considered by researchers. In this work, effects of Aloe vera extracts on clinical isolates was studied.

Materials and Methods: Aloe vera plant medicinal plants were obtained from a greenhouse. Three extracts including essential oils, extracts and no essential oils and essential oil extraction method also includes a complete extract of Aloe vera were prepared Percolation total. To investigate Microbiology extracts of two strains of Staphylococcus aureus and Staphylococcus epidermidis clinical strain of Gram-positive and Gram-negative isolates of Klebsiella pneumoniae and Escherichia coli strains Staphylococcus aureus ATCC25923 were used as well. Evaluate the effect of two methods: Kirby-Bauer disk with the minimum inhibitory concentration (MIC) was performed using microplate dilution. Turbidity was determined by an ELISA reader apparatus.

Results: All extracts of aloe vera on Klebsiella with a diameter of 32±2 mm mg/ml 285.7 concentration with microplate dilution method was 2.23 mg/ml. Staphylococcus aureus and MIC zone diameter of 30±2 mm and mg/ml 2.23, Staphylococcus epidermidis and Escherichia coli mg/ml4.46 mm 17.85 mm 30±5 mg/ml 17.85 respectively. Similar concentration of 17.85 mg ml Aloe Vera with a circle formed by the disk mc/ml 10 gentamicin was shown. This effect is similar to other bacteria antibiotics gentamicin, clindamycin, erythromycin, and Cefixime compared with Aloe Vera extract has been proven. Essential oils made from all parts of the same whole extract of aloe vera, but not essential extracts, bacteria studied were ineffective.

Conclusion: In this study the effects of similarity and some excess water Asrsarh Aloe Vera with common antibiotics on bacteria causing the infection was confirmed. Therefore, by production of appropriate pharmaceutical plant drugs with fewer side effects, bacterial infections couled be treated properly.

Comparison of the Efficacy and Safety of Insulin with Oral Hypoglycemic Agents in Management of Gestational Diabetes – an Evidence Based Medicine Approach

Habibollah Rahimi

Background: This study aimed to compare the efficacy and safety of insulin versus oral hypoglycemic drugs for gestational diabetes treatment, using evidence based medicine approach.

Materials and Methods: This secondary study was conducted by selecting a comprehensive systematic review through searching the Cochrane library. To calculate the number needed to treat (NNT), outcomes such as cesarean section, macrosomia, birth weight more than 90th percentile, operative vaginal delivery, labor induction, post partum hemorrhage, and shoulder dystocia has been considered while neonatal hypoglycemia, NICU hospitalization and fetal distress were considered for calculating number needed to harm (NNH). To calculate the likelihood of helped versus harmed (LHH), macrosomia and hypoglysmia were selected. Data were analyzed with Wilson score test.

Results: The NNT for above outcomes were 3, -14, -5, 16, -3, 31, and 32, respectively. Among them cesarean section was significance. Calculated NNH for side effects were 6, 15, and -17 which only was significance for neonatal hypoglycemia (p<0.05). The LHH was estimated -0.43.

Conclusion: Considering LHH, it seems that the benefit of insulin in management of gestational diabetes is more than metformin. But with respect to issues such as low sample size of trials, lack of assessment the maternal hypoglysemia as complication of insulin therapy, the findings should be considered cautiously.

Comparison of Efficacy of Doxycycline vs. Doxycycline with Omega- 3 in Treatment of Acne Vulgaris, a Triple-Blinded Clinical Trial

Hassan Faghani

Background: Acne vulgaris, is a chronic inflammatory disease of the pilosebaceous units. Genetic, environmental factors and nutrition influence on this disease. Recent studies indicate an essential role of nutrition in the development of acne, particularly with the effect on oxidative stress and inflammatory processes. Many studies have reported lower incidence of acne in individuals who receive diets containing large amounts of omega-3. The aim of this study was to determine the probable therapeutic effects of omega-3 for the treatment of acne.

Materials and Methods: In this study, 60 patients with moderate to severe facial acne were randomly divided into two treatment groups: 1) 100 mg doxycycline daily and one placebo capsule Omega -3 2) doxycycline 100 mg daily and one capsule 1000 mg Omega -3. Acne severity was determined based on GAGS system at the beginning of treatment and at the end of the sixth and twelfth weeks of treatment, with photographs obtained from each patient.

Results: There was a significant reduction in acne severity score (GAGS) at the end of sixth and twelfth weeks in the Omega-3 group in comparison to control group that received placebo.

Conclusion: Omega-3 fatty acids can improve moderate to severe inflammatory acne, probably due to their anti-inflammatory properties.

The Comparison Between Hydroethanolic Extraction of Pistacia Atlantica and Suzin Effects on Growth Inhibition of K562 Cell Line in Vitro Condition

Nooshin Naghsh

Background: Leukemiais a malignant and progressive disease of the Hematopoietic tissues of the body. The pistacia atlantica tree base, the geographic in large areas of the Mediterranean and the Middle East is growing. K562Cell class is considered as laboratory model of chronic phase of human CML. We compared the growth inhibitory effects SUZIN as a chemical compared with pistacia atlantica as a combined Zn plant antioxidant capacity in reducing cancer has been studied.

Materials and Methods: Pistachio nut was collected from around Kerman, then they were dried in room temperature and extraction was performed for 48 hours by maceration method. K562 cell class was incubated in medium RPMI-1640 fortified with 10%(v/v) FBS and 50% Streptomycin-Penicillin. Cytotoxic effect of hydro-ethanolic extract of pistacia atlantica against cancer cells K562 was evaluated in three interval by MTT method. Light absorbance by Eliza device was measured in wave length 540 nm. Statistical analysis was performed using SPSS15 software and ANOVA test.

Results: Pistacia atlantica in 24h and in concentration 100&mug/ml andSuzin in48h and in 12.5 &mug/ml,72 h and in 50 &mug/ml induced growth inhibition half of the cells were K562. Results obtained from changes in cell morphology influenced by hydro-ethanolic extract of pistacia atlantica and SUZIN suggest abnormal transformation of cells that probably represents apoptosis and necrosis.

Conclusion: Time and concentration against cytotoxic effect of Pistacia atlantica have the combined effect. Whileiron supplementation, alone time is due. Special concentration of pistacia atlantica having high antioxidant capacity with the Suzincan be considered as a potential target for inhibiting K562 cells in treatment of blood cancer.

Cytotoxicity of Zno and Ag/Zno Nano-Composites on Malignant Melanoma Cell Line (A375)

Nowruz Najafzadeh

Background: Melanoma is a malignant tumor of melanocytes, early-stage melanomas can be treated effectively with surgery alone, but more advanced cancers often incurable. The incidence of melanoma malignancy in most countries has risen faster than any other cancer types. It was the first time which we evaluated the cytotoxic effects of zno and Ag/zno nano-composites (NP) on melanoma cell line, A375, viability.

Materials and Methods: In this experimental study, A375 cell line was grown in RPMI-1640 supplemented with 10% FBS, penicillin/streptomycin (100 U/ml, 100 µg/ml) at 37◦C in 5% CO2, then the effects of different concentrations of zno and Ag/zno nano-composites on melanoma cell were evaluated by MTT, clonogenic survival assays, and acridine orange/ ethidium bromide staining.

Results: Herein, we demonstrated that Zno and Ag/zno nano-composites showed similar effects on cytotoxicity of melanoma cancer cells. In a dose dependent manner, a significant cytotoxicity was observed with increasing of zno and Ag/zno. The inhibitory concentration 50% (IC50) values of the nano-composites for A375 cell line after 24 hrs were 7.24±1.55 and 15.93±1.73 µg/ml for zno and Ag/zno, respectively.

Conclusion: The results showed that zno and Ag/zno has ability to induce cytotoxicity in the human melanoma cancer cell line in lower micromolar concentrations. In conclusion, these findings may introduce a new view on the mode of action and possible application of new nano-composites in the cancer chemotherapy.

The Role of Hippocampal Orexin-1 Receptors (OX1R) in Mediating the Effect of Morphine on CA1 Baseline Synaptic Response and Short Term Synaptic Plasticity

Narges Hosseinmardi

Background: Considering the increased activity of hypothalamic orexinergic neurons due to morphine administration, and its extensive projections to the hippocampus, it is probable that morphine effect on CA1 neuronal function is mediated by orexinergic system. So the effect of hippocampal orexin-1 receptors (OX1R) blockade on CA1 baseline synaptic response and short term synaptic plasticity was investigated.

Materials and Methods: In this experimental study, animals received morphine 10 mg/kg/12h/(SC) for 10 days. SB-334867-A, OX1R antagonist (0.5&mug/0.5 &mul), was microinjected intrahippcampally for OX1R inhibition before each morphine injection. Baseline synaptic response and short term synaptic plasticity were evaluated by field potential recording. fEPSP was recorded from CA1 following Schaffer collaterals stimulation. After Input/Output construction, short term synaptic plasticity was induced by paired pulse stimulations.

Results: Chronic use of morphine did not affect the baseline synaptic response (p>0.05). SB- 334867-A microinjection in CA1 did not have any effect on baseline synaptic response in morphine dependent rats. Morphine increased paired pulse index (PPI) at 80 ms inter pulse interval (IPI, p<0.05). SB-334867-A pretreatment did not affect this morphine induced PPI change.

Conclusion: The results suggest that orexin-1 receptors (OX1R) do not mediate the effect of morphine on baseline synaptic response and short term synaptic plasticity in CA1 area of the hippocampus.