Cloning and expression of codon-optimized recombinant darbepoetin alfa in Leishmania tarentolae T7-TR

Darbepoetin alfa is an engineered and hyperglycosylated analog of recombinant human erythropoietin (EPO) which is used as a drug in treating anemia in patients with chronic kidney failure and cancer. This study desribes the secretory expression of a codon-optimized recombinant form of darbepoetin alfa in Leishmania tarentolae T7-TR. Synthetic codon-optimized gene was amplified by PCR and cloned into the pLEXSY-I-blecherry3 vector. The resultant expression vector, pLEXSYDarbo, was purified, digested, and electroporated into the L. tarentolae. Expression of recombinant darbepoetin alfa was evaluated by ELISA, reverse-transcription PCR (RT-PCR), Western blotting, and biological activity. After codon optimization, codon adaptation index (CAI) of the gene raised from 0.50 to 0.99 and its GC content changed from 56 to 58. Expression analysis confirmed the presence of a protein band at 40 kDa. Furthermore, reticulocyte experiment results revealed that the activity of expressed darbepoetin alfa was similar to that of its equivalent expressed in Chinese hamster ovary (CHO) cells. These data suggested that the codon optimization and expression in L. tarentolae host provided an efficient approach for high level expression of darbepoetin alfa. © 2015 Elsevier Ltd. All rights reserved

Building resilient communities over time

Community resilience entails the community’s ongoing and developing capacity to account for its vulnerabilities and function amid and recover from disturbance. A holistic and systematic approach of the community on how it uses material and energy resources or how a society educates the members’ over time is required to learn from the past and adapt to the present and future opportunities and threads. Community resilience has a long history in the local communities, which is embedded in their culture and history around shared values and local knowledge based on a dedicated and robust collaboration among diverse groups of the community and the various actors from different backgrounds. The innovative partnership between various actors such as stakeholders, research entities, local communities, and third sector parties is required to overcome the complexities of resiliency building. Using local knowledge to understand the local needs better is crucial in developing local, sustainable solutions and building community resilience over time.Global Challenges (FSW

Comparison of antibacterial activities of Ircinia mutans extracts in two different seasons from Kish Island, Persian Gulf, Iran

Sponges, which constitute the phylum Porifera, are the most primitive of the multicellular animals, among all marine organisms screened. Marine sponges produce the largest number of structurally diversified natural products. In this study we investigated in vitro antimicrobial activity of Ircinia mutans collected from the Kish Island in the Persian Gulf against strains of bacteria Escherichia coli (ATCC 15224), Pseudomonas aeruginosa (ATCC 25619), Staphylococcus aureus (ATCC 1764), and Bacillus subtilis spizizenii (ATCC 6633). Diethyl etter, methanol and aqueous extracts of sponge were evaluated by using the Bacterial Broth Dilution Method. The results showed that the aqueous extracts didn’t have any antibacterial activity. Minimum Inhibitory Concentrations (MIC) of the winter diethyl etter extract was 2 mg/ml for E.coli and 20 mg/ml for P. aeruginosa, whereas the summer diethyl etter extract and both of methanol extracts did not show any activity. The MIC and MBC (Minimum Bacterial Concentration) of summer diethyl etter extracts were 2 mg/ml and 3mg/ml against S. aureus; and 5mg/ml and 10mg/ml when tested on B. subtilis. The MIC and MBC of winter diethyl etter extracts were measured as 1.5 mg/ml and 2mg/ml against S. aurous; and 5mg/ml and 10mg/ml when examined on B. subtilis. Summer and winter methanol and aqueous extracts of I. mutans did not show any activity against these bacteria. Therefore secondary metabolite solutions in diethyl etter contain components with antibacterial properties and can be used as antibiotics products

Induced spermiation of rainbow trout, Oncorhynchus mykiss, using a GnRh analogue

In this study, the benefits of using the first Iranian made GnRHa[D-Ala6 des Gly10] mGnRH ethylamide, to induce spermiation in male rainbow trout, Oncorhynchus mykiss, were evaluated, In addition, its effect on acceleration and synchronization, quality and quantity of milt and the plasma Testosterone (T) fluctuations were examined. For these purposes, 40 non-spermiating male rainbow trouts were injected with a mammalian gonadotropin releasing hormone analog (GnRHa) preparations of 0 (control), 30, 40 and 80 μg/kg B.W. or vehicle (propylene glycol). Spermiation was very synchronous and accelerated in treated groups. Six days after first injection, the cumulative spermiation rates reached respectively 40, 49 and 79% in injected groups (i.e. 30 to 80 μg/kg B.W.) while none of the control fish was spermiated. GnRHa injections advanced spermiation and reduced the average time to spermiation from 16 3.67 days for control group to 14.3± 2.2, 9.2 ±0.75 and 6.6± 0.29 days for treated groups, respectively (p0.05). Circulating levels of testosterone (T) prior to the GnRHa treatment were relatively low in all groups. Treatment with GnRHa induced significant increase in plasma T after 12 h, increasing it to 43.43±5.82, 38.66±5.63, 39.72±5.07 ng/ml in groups 2 to 4, respectively, which were higher than T levels for control (i.e. 24.58±7.13 ng/ml) p<0.05. These levels remained high up until 48 h in treated groups; but after this time, T levels reduced to the basal levels of time 0, except in 4 which had received its second GnRHa injection at time 48

Measurement of strain evolution in overloaded roller bearings using time-of-flight neutron diffraction

Neutron diffraction is an established method for non-destructively characterising residual stress or observing in situ strain during external stimuli. Neutron based stroboscopic techniques have previously been introduced for measuring strains undergoing cyclic processes but have not been used for tribological applications. This work presents a novel approach for measuring the evolution of radial strain in a rotating bearing through part of the component's lifetime. A cylindrical roller bearing was pre-overloaded to increase the probability of damage within a reasonable experimental time and to help develop further understanding of the influence such events have on bearing life, notably for the application of wind turbine gearbox bearing failure. The stroboscopic neutron diffraction technique was successful in measuring time-resolved contact strain, with a significant increase in compressive radial strain being observed after a suspected failure had been detected using condition monitoring techniques, implemented for validating damage propagation. Cyclic contact strains associated with rolling contact fatigue were also evaluated using neutron diffraction

Comparison of the tuberculin skin test and the QuantiFERON-TB Gold test in detecting latent tuberculosis in health care workers in Iran

OBJECTIVES: The tuberculin skin test (TST) and the QuantiFERON-TB Gold test (QFT) are used to identify latent tuberculosis infections (LTBIs). The aim of this study was to determine the agreement between these two tests among health care workers in Iran. METHODS: This cross-sectional study included 177 tuberculosis (TB) laboratory staff and 67 non-TB staff. TST indurations of 10 mm or more were considered positive. The Student's t-test and the chi-square test were used to compare the mean score and proportion of variables between the TB laboratory staff and the non-TB laboratory staff. Kappa statistics were used to evaluate the agreement between these tests, and logistic regression was used to assess the risk factors associated with positive results for each test. RESULTS: The prevalence of LTBIs according to both the QFT and the TST was 17 (95 confidence interval CI, 12% to 21%) and 16% (95% CI, 11% to 21%), respectively. The agreement between the QFT and the TST was 77.46%, with a kappa of 0.19 (95% CI, 0.04 to 0.34). CONCLUSIONS: Although the prevalence of LTBI based on the QFT and the TST was not significantly different, the kappa statistic was low between these two tests for the detection of LTBIs

Validating 3D two-parameter fracture mechanics models for structural integrity assessments

In-situ fracture tests were carried out on the I12 beamline at the Diamond Light Source. Four Al-Ti metal-matrix composites (MMCs), with two crack lengths, were studied to assess for the impact of in-plane constraint. Synchrotron X-ray computed tomography and synchrotron X-ray diffraction were used to measure total strain and elastic strain respectively. In this work, the measured elastic strains in the samples are detailed as a function of applied load and compared against those predicted from a 3D elastic-plastic finite element model. The modelled strains increased asymptotically towards the tip of the electro discharge machined notch. The experimental results do not highlight the same response, which is due to a combination of blunting and low experimental spatial resolution. Far field experimental and measured strain fields converged, notably in the test piece containing a long notch (a/W = 0.5) and higher levels of constraint

Reliability and validity of the persian version of compulsive eating scale (CES) in overweight or obese women and its relationship with some body composition and dietary intake variables

Objective: Compulsive or binge eating is a kind of disturbed eating behavior, which is mostly observed among dieting women, and is integrated with appetite disorder, and uncontrolled eating of plenty of junk food. The Compulsive Eating Scale (CES) created first by Kagan & Squires in 1984, is an eight-item self-reporting instrument that is made to measure the severity of binge eating disorder. The aim of this study was to provide the reliability and validity of the Persian version of Compulsive Eating Scale (CES) among overweight and obese women in Iran. Method: One hundred and twenty six (N = 126) overweight and obese women consented to participate in this study. We estimated the anthropometric indices, including body weight, height, waist and hip circumferences, a total body fat percentage, and visceral fat level with body analyzer all in standard situations. Then, the participants completed the CES. Next, to assess concurrent validity, Beck Depression Inventory, Spielberger anxiety scale, appetite visual analogue rating scale, Food Craving questionnaire, Three-Factor Eating Questionnaire-R18, and Restraint eating visual analogue rating scale were performed simultaneously. To assess test-retest reliability, CES was repeated for all the participants two weeks later. Moreover, we reported the internal consistency and factor analysis of this questionnaire. Furthermore, we estimated the concurrent correlation of CES with logically relevant questionnaires and body composition and anthropometric indices. Results: Based on the reliability analysis and factor analysis of the principal component by Varimax rotation, we extracted two factors: eating because of negative feelings, and overeating. Internal consistency (Cronbach's alpha) of the CES was 0.85 (Cronbach alpha of the factors was 0.85, and 0.74, respectively). The test-retest correlation of the CES was 0.89. Also, the split-half reliability of the questionnaire was established with the correlation coefficient between Sets I and II. The correlation was 0.85. Conclusion: This study provides preliminary support for the reliability and validity of the Persian version of the CES. This instrument would be helpful in measuring the clinical practice and research studies of obesity, appetite and eating disorders reliably and validly

Efficacy of tenofovir disoproxil fumarate therapy in nucleoside-analogue naive Iranian patients treated for chronic hepatitis B

Background: Tenofovir disoproxil fumarate (TDF) is a new effective treatment option for patients with chronic hepatitis B (CHB). Objectives: To evaluate TDF efficacy in nucleos(t)ide analogues (NAs)-naive Iranian patients with CHB. Patients and Methods: The NA-naive patients received TDF for at least six months. The primary endpoint was the proportion of patients achieving a complete virological response (CVR) during the treatment. Multivariate Cox regression analysis determined predictive factors independently associated with the time to CVR. The secondary endpoints were biochemical and serological responses, frequency of virological breakthrough, genotypic resistance development, safety and tolerability. Results: In all, 93 patients (64.5 hepatitis B e antigen HBeAg-negative) were eligible. Of these, 70 patients completed 24 months of treatment. The cumulative CVR rates in HBeAg-negative and HBeAg-positive patients were 87% versus 53% at 24 months, respectively. The multivariate Cox regression model showed only HBeAg positivity at baseline and a high baseline HBV DNA level were independent factors predicting a CVR. No patient achieved hepatitis B surface antigen (HBsAg) and HBeAg loss or seroconversion and no virologic breakthrough occurred. A new amino acid substitution (rtD263E) was observed to develop in 60% of patients with viremia. Conclusions: The cumulative CVR rates showed that patients with HBeAg-negative have better virologic respond than those with HBeAg-positive during the same period. The rtD263E mutation might be associated with partial resistance to TDF. © 2015, Kowsar Corp