Molecular analysis of the clavulanic acid regulatory gene isolated from an Iranian strain of Streptomyces clavuligerus, PTCC 1709

Objective: The clavulanic acid regulatory gene (claR) is in the clavulanic acid biosynthetic gene cluster that encodes ClaR. This protein is a putative regulator of the late steps of clavulanic acid biosynthesis. The aim of this research is the molecular cloning of claR, isolated from the Iranian strain of Streptomyces clavuligerus (S. clavuligerus). Materials and Methods: In this experimental study, two different strains of S. clavuligerus were used (PTCC 1705 and DSM 738), of which there is no claR sequence record for strain PTCC 1705 in all three main gene banks. The specific designed primers were subjected to a few base modifications for introduction of the recognition sites of BamHI and ClaI. The claR gene was amplified by polymerase chain reaction (PCR) using DNA isolated from S. clavuligerus PTCC 1705. Nested-PCR, restriction fragment length polymorphism (PCR-RFLP), and sequencing were used for molecular analysis of the claR gene. The confirmed claR was subjected to double digestion with BamHI and ClaI. The cut claR was ligated into a pBluescript (pBs) vector and transformed into E. coli. Results: The entire sequence of the isolated claR (Iranian strain) was identified. The presence of the recombinant vector in the transformed colonies was confirmed by the colony-PCR procedure. The correct structure of the recombinant vector, isolated from the transformed E. coli, was confirmed using gel electrophoresis, PCR, and double digestion with restriction enzymes. Conclusion: The constructed recombinant cassette, named pZSclaR, can be regarded as an appropriate tool for site directed mutagenesis and sub-cloning. At this time, claR has been cloned accompanied with its precisely selected promoter so it could be used in expression vectors. Hence the ClaR is known as a putative regulatory protein. The overproduced protein could also be used for other related investigations, such as a mobility shift assa

The Role of Urine Polymerase Chain Reaction Test in Diagnosis of Genitourinary Tuberculosis

Abstract BACKGROUND: The genitourinary system is one of he most common sites of infection in non-pulmonary tuberculosis (TB). The clinical symptoms and radiologic findings of urinary TB are nonspecific. Current diagnostic tests are of low sensitivity and labor-intensive. Therefore, this study was aimed to evaluate diagnostic value of urine PCR in genitourinary tuberculosis (GNTB). METHODS: This was a descriptive study on 33 patients with confirmed genitourinary TB. Demographic data, clinical symptoms, laboratory and radiologic findings were collected. For each patient, three consecutive early morning urine specimens were examined by PCR. The diagnostic value of PCR in mycobacterium tuberculosis (MTB) in comparison with standard microbiological methods was assessed. FINDINGS: There were 33 patients with a mean age of 47.27 16.1 years. The most common presenting symptoms were irritative voiding symptoms (51.5%), flanks pain (27.2%), gross hematuria (9%) and suprapubic pain (9%). Laboratory findings in U/A were hematuria (75.8%) and pyuria (60.6%). IVU was abnormal in 61.5% of patients. Most common abnormalities were pyelocalyceal dilation (44%), ureteral stricture and hydroureter (37%) and multiple small calyceal deformities (25%). Of the 33 patients PCR for MTB was positive in 16 cases (48.5%). In patients with abnormal IVU, PCR was positive in 62.5%. CONCLUSION: A high index of clinical suspicion is necessary for diagnosis of GUTB. PCR is recommended for instant diagnosis and screening before further examination, it cannot be the only method in identification of GUTB

Assessing the capability of satellite data for soil mapping

The purpose of this research is to evaluate the potential of using landsat ETM+ data for soil mapping. The study area is located in center of Iran and covers about 1300 ha. The database of studied area was created by introducing topographic map (1:25,000), soil map (1:20,000) and reports and satellite data. After pre-possessing stage, selection of the best informative bands was carried out using optimum index factor (OIF) calculation and principle component analysis (PCA). Results showed that bands TM1, TM4 and TM6-2 contain the highest information and the lowest redundancy. Besides the mentioned bands, TM5 and TM7 were considered for digital image classification. The images were classified using maximum likelihood classifier into seven mapping units. Separability of mapping units examined at 95% confidence level. Comparison of the prepared soil map from satellite data and ground truth showed a relatively high accuracy of 80%. Also, comparison of prepared soil map from satellite data and detailed soil maps prepared using conventional methods showed imagery data could increase the classification and interpretative purity percentage up to 50% and 85%, respectively. The results indicated high potential of imagery data for inventory and increasing the precision of existing soil maps. Therefore, incorporation of high-resolution satellite data for soil survey especially in arid and semi-arid regions is highly recommended

Mutations of Dual Oxidase 2 (DUOX2) Gene among patients with Permanent and Transient Congenital Hypothyroidism

Objective: The prevalence of congenital hypothyroidism (CH) is high in Isfahan, Iran. In addition, it has different etiologies compared with other countries. The rate of parental consanguinity is also high in the city. Moreover, DUOX2 gene is effective in transient CH and permanent CH due to dyshormonogenesis. Therefore, the aim of this research was to investigate the mutations of DUOX2 gene in patients with transient CH and permanent CH due to dyshormonogenesis. Methodology: In this descriptive, prospective study, patients diagnosed with transient and permanent CH due to dyshormonogenesis during CH screening program were selected. Venous blood samples were obtained to determine the 3 mutations (Q36H, R376W, and D506N) of DUOX2 gene using polymerase chain reaction (PCR) method by specific primers and complementary methods such as restriction fragment length polymorphism (RFLP) and singlestrand conformation polymorphism (SSCP). Results: In this study, 25 patients with transient CH and 33 subjects with permanent CH due to dyshormonogenesis were studied. In addition, 30 children were studied as the control group. We did not find any mutations of the 3 mentioned mutations of DUOX2 gene. Conclusion: Considering the findings of the current study, further studies with other methods are required to evaluate other gene mutations such as pendrin, sodium-iodide symporter (NIS) and thyroglobuli

Coincidence of Compound Odontoma and Cemento Ossifying Fibroma; A Rare Case Report

Objective: Cemento-ossifying fibroma defines as a relative rare osteogenic neoplasm of the jaw. This tumor includes fibrous and osseous components. Odontoma is the most common odontogenic tumor containing enamel ,dentin ,cementum and pulp tissue. in this paper we report a  rare case of ossifying fibroma associated with compound odontoma in the mandible. Case: A 37-years-old woman was referred to Oral Medicine department , Shahid Beheshti Dental School with complaint of swelling in the anterior part of the mandible, over 6 years period. Clinical examination revealed mandibular enlargement in right –anterior region with labial and lingual expansion and canine missing . panoramic view showed a large mixed radiolucent -  radiopaque lesion associated with impacted canine.The differential diagnosis include calcifying odontogenic cyst (COC) and cemento-ossifying fibroma(COF). Histopathologic examination established diagnosis of COF with multiple compound odontoma .Conclusion: The relationship between the occurrence of these two lesions is not clear and more studies are needed to establish the relationship between them

Comparison of Vibration Amplitude in Isfahan Subway Due to Track Structure- An Experimental Study

Increasing the stability of structures and reducing the maintenance cost of slab track superstructures compared to ballasted tracks are among the reasons for the tendency to use this category of superstructures in the railway industry. Vibration reduction methods can be divided into three categories, source, propagation path, and receiver. In general, the slab track structures in Iran are divided into three categories: direct fixation track (DFT), floating slab track (FST), and high resilient fastener (HRF). Although railway tracks are a safe, economical and fast transportation system and can lead to the strengthening of the tourism industry, in the long term, vibrations can damage many historical structures in the city of Isfahan. FST and HRF systems are used in the structure of Isfahan subway track. In this paper, the accelerations (longitudinal, lateral, and vertical) of the Isfahan subway vehicle were measured in 30 stations (15 go stations and 15 return stations). The results showed that the HRF system compared to the FST has a significant effect in reducing the range of vibrations and ultimately the safety of the train and the ride comfort. For example, in the area between Si-O-Se-Pol and Imam Hossein Square, due to the track structure type (HRF), the maximum acceleration and RMS acceleration are in the range of 1.5 and 0.3 m/s2, respectively, while in other stations these values were extracted up to 4 and 0.7 m/s2, respectively

Mental imagery can improve performance in a visuomotor task: a pilot study

This experiment assessed the effectiveness of two interventions; mental imagery and physical training as compared to a control group, on their ability to improve visuomotor accuracy, measured by scores achieved on a visuomotor task. For mental imagery participants had to imagine throwing a dart and for physical training participants physically practised dart throwing. Measurements were recorded pre-intervention, after two weeks of training (Mid-intervention), after five weeks of training (Outcome Measure 1) and two weeks after training (Outcome Measure 2). Comparison of Mid-intervention, Outcome Measure 1 and Outcome Measure 2 with baseline showed both interventions to significantly increase performance on dart throwing compared to the Control group. Our findings show that, as well as traditional physical practise, mental imagery can effectively improve performance on a fine visuomotor task. This is an important finding highlighting possible applications of mental imagery in those with limited motor abilities to maintain or enhance motor movement

Protective effect of crocin against reperfusion-induced cardiac arrhythmias in anaesthetized rats

The aim of the present study was to investigate the effects of crocin - a natural antioxidant derived from saffron - on cardiac reperfusion-induced arrhythmia and antioxidant systems such as catalase and superoxide dismutase (SOD) enzyme activities, glutathione (GSH) and malondialdehyde (MDA, as a marker of lipid peroxidation) levels. Rats in 4 experimental groups were administered crocin (20 mg/kg/day) or vehicle (i.p.) for 21 days with or without cardiac ischemia-reperfusion (IR). At the end of this period, hearts of anaesthetized animals in IR and “Cr + IR” groups were subjected to 10 min occlusion of the left anterior descending coronary artery and thereafter reperfused for 30 min. The results suggest that crocin is partially capable of suppressing reperfusion-induced arrhythmias. Compared to control group, ischemic-reperfusion injury significantly decreased SOD activity and GSH level and increased MDA level of heart muscle. “Cr + IR” group showed remarkably increased catalase activity in heart tissue (28.7 ± 6.6 vs. 23.6 ± 4.1 U/mg protein, P < 0.05) compared to the IR group. The level of cardiac tissue SOD activity in the “Cr + IR” group animals did not decline significantly compared to rats that were administered crocin alone with no ischemia. The results suggest a protective role of crocin on cardiac reperfusion arrhythmias which may at least partially be related to stability or even amplification of antioxidant systems. Crocin may potentially be useful for treatment or prevention of arrhythmias in patients with ischemic heart disease and this issue remains to be investigated in future clinical studies

Detection of Nocardia, Streptomyces and Rhodococcus from bronchoalveolar lavage specimens of patients with HIV by Multiplex PCR Assay

Background: Nocardia, Streptomyces and Rhodococcus are life threatening opportunistic pathogens under immunodeficiency conditions, particularly among patients infected with HIV. Rapid and accurate detection of these infections can improve immune health quality, patient management and appropriate treatment. The aim of this study was to design a novel multiplex-PCR assay for rapid diagnosis of these three organisms directly from bronchoalveolar lavage (BAL) specimens of patients infected with HIV.Methods: The genus specific primers were designed for directdetection of Nocardia, Streptomyces and Rhodococcus in a single tube multiplex PCR. This PCR specifically amplified the target genes from pure cultures. It subsequently was applied on BAL specimens of 29 HIV positive patients that had previously been culture negative for actinomycete bacteria, of which Nocardia, Streptomyces and Rhodococcus are members.Results: Of 29 respiratory clinical specimens, there were positive for Nocardia spp. and one was positive for Streptomyces spp using the multiplex PCR assay. The sequencing of the PCR products identified the species as Nocardia cyriacigeorgica (n=2), Nocardia farcinica and Streptomyces albus.Conclusion: This novel multiplex PCR assay yielded reliable results for accurate identification of Nocardia, Streptomyces and Rhodococcus from BAL while the results of bacterial culture were negative.