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

Tularemia and Plague Survey in Rodents in Earthquake Zones in Southeastern Iran

OBJECTIVES: Earthquakes are one the most common natural disasters that lead to increased mortality and morbidity from transmissible diseases, partially because the rodents displaced by an earthquake can lead to an increased rate of disease transmission. The aim of this study was to evaluate the prevalence of plague and tularemia in rodents in the earthquake zones in southeastern Iran. METHODS: In April 2013, a research team was dispatched to explore the possible presence of diseases in rodents displaced by a recent earthquake magnitude 7.7 around the cities of Khash and Saravan in Sistan and Baluchestan Province. Rodents were trapped near and in the earthquake zone, in a location where an outbreak of tularemia was reported in 2007. Rodent serums were tested for a serological survey using an enzyme-linked immunosorbent assay. RESULTS: In the 13 areas that were studied, nine rodents were caught over a total of 200 trap-days. Forty-eight fleas and 10 ticks were obtained from the rodents. The ticks were from the Hyalomma genus and the fleas were from the Xenopsylla genus. All the trapped rodents were Tatera indica. Serological results were negative for plague, but the serum agglutination test was positive for tularemia in one of the rodents. Tatera indica has never been previously documented to be involved in the transmission of tularemia. CONCLUSIONS: No evidence of the plague cycle was found in the rodents of the area, but evidence was found of tularemia infection in rodents, as demonstrated by a positive serological test for tularemia in one rodent

Impact of Health Literacy, Self-efficacy, and Outcome Expectations on Adherence to Self-care Behaviors in Iranians with Type 2 Diabetes.

OBJECTIVES: Diabetic patients with higher health literacy (HL) may feel more confident in their ability to perform self-care behaviors and may have strong beliefs that diabetes-related behaviors will lead to specific outcomes. Our study aimed to document the relationships between HL, self-efficacy, outcome expectations, and diabetes self-care of patients with type 2 diabetes mellitus (T2DM) in Iran. METHODS: We conducted a cross-sectional observational study of 187 patients with T2DM. Participants completed the Functional Communicative and Critical Health Literacy scale, the Summary of Diabetes Self-Care Activities, the Diabetes Management Self-Efficacy Scale, Outcome Expectations Questionnaire, and a demographic questionnaire. RESULTS: Participants who received diabetes education (t = 5.79, p<0.001) and were married (F = 3.04, p<0.050) had better diabetes self-care behavior. There was a significant positive correlation between self-care behaviors and communicative HL (r = 0.455, p<0.010), critical HL (r = 0.297, p<0.010), self-efficacy (r = 0.512, p<0.010) and outcome expectations (r = 0.387, p<0.010). Diabetes education and marital status accounted for 16.9% of the variance in diabetes self-care. Self-efficacy, outcome expectations, communicative, and critical HL explained 28.0%, 1.5%, 3.7%, and 1.4% of the variance, respectively. CONCLUSIONS: This study revealed that the potential impact of self-efficacy, outcome expectations, communicative, and critical HL should be considered in the education program for patients with diabetes. We found self-efficacy to be the most important predictor of diabetes self-care. Therefore, the use of self-efficacy theory when designing patient education interventions could enhance diabetes self-care. It is essential that health care providers assess patient's HL levels to tailor health-related information specific to a domain of HL. This would fully inform patients and promote empowerment rather than simple compliance

Entropic forces drive self-organization and membrane fusion by SNARE proteins

SNARE proteins are the core of the cell’s fusion machinery and mediate virtually all known intracellular membrane fusion reactions on which exocytosis and trafficking depend. Fusion is catalyzed when vesicle-associated v-SNAREs form trans-SNARE complexes (“SNAREpins”) with target membrane-associated t-SNAREs, a zippering-like process releasing ∼65 kT per SNAREpin. Fusion requires several SNAREpins, but how they cooperate is unknown and reports of the number required vary widely. To capture the collective behavior on the long timescales of fusion, we developed a highly coarse-grained model that retains key biophysical SNARE properties such as the zippering energy landscape and the surface charge distribution. In simulations the ∼65-kT zippering energy was almost entirely dissipated, with fully assembled SNARE motifs but uncomplexed linker domains. The SNAREpins self-organized into a circular cluster at the fusion site, driven by entropic forces that originate in steric–electrostatic interactions among SNAREpins and membranes. Cooperative entropic forces expanded the cluster and pulled the membranes together at the center point with high force. We find that there is no critical number of SNAREs required for fusion, but instead the fusion rate increases rapidly with the number of SNAREpins due to increasing entropic forces. We hypothesize that this principle finds physiological use to boost fusion rates to meet the demanding timescales of neurotransmission, exploiting the large number of v-SNAREs available in synaptic vesicles. Once in an unfettered cluster, we estimate ≥15 SNAREpins are required for fusion within the ∼1-ms timescale of neurotransmitter release

Particle acceleration and reconnection in the solar wind

An emerging paradigm for the dissipation of magnetic turbulence in the supersonic solar wind is via localized quasi-2D small-scale magnetic island reconnection processes. An advection-diffusion transport equation for a nearly isotropic particle distribution describes particle transport and energization in a region of interacting magnetic islands [1; 2]. The dominant charged particle energization processes are 1) the electric field induced by quasi-2D magnetic island merging, and 2) magnetic island contraction. The acceleration of charged particles in a “sea of magnetic islands” in a super-Alfvénic flow, and the energization of particles by combined diffusive shock acceleration (DSA) and downstream magnetic island reconnection processes are discussed

Irradiation-induced Ag nanocluster nucleation in silicate glasses: analogy with photography

The synthesis of Ag nanoclusters in sodalime silicate glasses and silica was studied by optical absorption (OA) and electron spin resonance (ESR) experiments under both low (gamma-ray) and high (MeV ion) deposited energy density irradiation conditions. Both types of irradiation create electrons and holes whose density and thermal evolution - notably via their interaction with defects - are shown to determine the clustering and growth rates of Ag nanocrystals. We thus establish the influence of redox interactions of defects and silver (poly)ions. The mechanisms are similar to the latent image formation in photography: irradiation-induced photoelectrons are trapped within the glass matrix, notably on dissolved noble metal ions and defects, which are thus neutralized (reverse oxidation reactions are also shown to exist). Annealing promotes metal atom diffusion, which in turn leads to cluster nuclei formation. The cluster density depends not only on the irradiation fluence, but also - and primarily - on the density of deposited energy and the redox properties of the glass. Ion irradiation (i.e., large deposited energy density) is far more effective in cluster formation, despite its lower neutralization efficiency (from Ag+ to Ag0) as compared to gamma photon irradiation.Comment: 48 pages, 18 figures, revised version publ. in Phys. Rev. B, pdf fil

Diffusive Shock Acceleration and Reconnection Acceleration Processes

Shock waves, as shown by simulations and observations, can generate high levels of downstream vortical turbulence, including magnetic islands. We consider a combination of diffusive shock acceleration (DSA) and downstream magnetic-island-reconnection-related processes as an energization mechanism for charged particles. Observations of electron and ion distributions downstream of interplanetary shocks and the heliospheric termination shock (HTS) are frequently inconsistent with the predictions of classical DSA. We utilize a recently developed transport theory for charged particles propagating diffusively in a turbulent region filled with contracting and reconnecting plasmoids and small-scale current sheets. Particle energization associated with the anti-reconnection electric field, a consequence of magnetic island merging, and magnetic island contraction, are considered. For the former only, we find that (i) the spectrum is a hard power law in particle speed, and (ii) the downstream solution is constant. For downstream plasmoid contraction only, (i) the accelerated spectrum is a hard power law in particle speed; (ii) the particle intensity for a given energy peaks downstream of the shock, and the distance to the peak location increases with increasing particle energy, and (iii) the particle intensity amplification for a particular particle energy, f(x,c/c_0)/f(0,c/c_0), is not 1, as predicted by DSA, but increases with increasing particle energy. The general solution combines both the reconnection-induced electric field and plasmoid contraction. The observed energetic particle intensity profile observed by Voyager 2 downstream of the HTS appears to support a particle acceleration mechanism that combines both DSA and magnetic-island-reconnection-related processes

Clade identification of symbiotic zooxanthellae of dominant sclerectinian coral species of intertidal pools in Hengam Island

Zooxanthellae of reef-building corals are unicellular dinoflagellates of the Symbiodinium genus, which has an important role in bleaching phenomenon. Symbiodinium and their coral hosts are sensitive to environmental stresses that include salinity, high temperatures, low temperatures, extreme light levels and turbidity. Tidal pools have harsh conditions due to lack of nutrients, food and pronounced changes in physical conditions such as pH, salinity and temperature, hence the study of symbiotic zooxanthellae on coral reefs of tidal pool seems to be necessary. Samples of five coral species that include Siderastrea savignyana, Coscinaraea columna, Anomastrea irregulariis, Cyphastrea serailia, Psammocora superficialis were collected at intertidal pool of Hengam Island in the northern Persian Gulf. Partial 28S nuclear ribosomal (nr) DNA of Symbiodinium were amplified by polymerase chain reaction (PCR) and then PCR products were analyzed by the phylogenetic analyses of the LSU DNA sequences based on PAUP and Clustal X software. The results showed that there are at least two clades of Symbiodinium from Hengam Island. Clade D was detected from 3 of the coral species whileclade C was found in 2 species only. This study showed dominance of clade D at intertidal pool in Hengam Island and the dominace of clade D might be explained by the high environmental stresses for the Persian Gulf.Key words: Persian Gulf, clade D, tides, Symbiodinium and Hengam Island

Hypermethylation in the ZBTB20 gene is associated with major depressive disorder.

This is the final version of the article. Available from BioMed Central via the DOI in this record.BACKGROUND: Although genetic variation is believed to contribute to an individual's susceptibility to major depressive disorder, genome-wide association studies have not yet identified associations that could explain the full etiology of the disease. Epigenetics is increasingly believed to play a major role in the development of common clinical phenotypes, including major depressive disorder. RESULTS: Genome-wide MeDIP-Sequencing was carried out on a total of 50 monozygotic twin pairs from the UK and Australia that are discordant for depression. We show that major depressive disorder is associated with significant hypermethylation within the coding region of ZBTB20, and is replicated in an independent cohort of 356 unrelated case-control individuals. The twins with major depressive disorder also show increased global variation in methylation in comparison with their unaffected co-twins. ZBTB20 plays an essential role in the specification of the Cornu Ammonis-1 field identity in the developing hippocampus, a region previously implicated in the development of major depressive disorder. CONCLUSIONS: Our results suggest that aberrant methylation profiles affecting the hippocampus are associated with major depressive disorder and show the potential of the epigenetic twin model in neuro-psychiatric disease.The study was funded by the Wellcome Trust; European Community’s Seventh Framework Programme (FP7/2007-2013). The study also receives support from the National Institute for Health Research (NIHR) Clinical Research Facility at Guy’s & St Thomas’ Davies et al. Genome Biology 2014, 15:R56 Page 9 of 12 http://genomebiology.com/2014/15/4/R56 NHS Foundation Trust and NIHR Biomedical Research Centre based at Guy's and St Thomas' NHS Foundation Trust and King's College London. Matthew Davies is supported by the EU FP7 grant EuroBATS (No. 259749). Tim Spector is an NIHR senior Investigator and is holder of an ERC Advanced Principal Investigator award. Further funding support for this project was obtained from the European Research Council (project number 250157). The members of the UK Brain Expression Consortium (UKBEC) are: (1) Department of Molecular Neuroscience, UCL Institute of Neurology, London, UK: John A Hardy, Mina Ryten, and Daniah Trabzuni; (2) Department of Medical and Molecular Genetics, King's College London, UK: Michael E Weale, Adaikalavan Ramasamy and Paola Forabosco; (3) Department of Pathology, The University of Edinburgh, Wilkie Building, Teviot Place, Edinburgh, UK: Colin Smith and Robert Walker. Australia: funding for phenotype and blood collection was from NHMRC grants to Nick Martin and NIH grants to Andrew Heath and Pamela Madden. We thank David Smyth for database management, Lisa Bowdler for sample preparation, and the twins for their cooperation