Requirements for translation re-initiation in Escherichia coli: roles of initiator tRNA and initiation factors IF2 and IF3

Despite its importance in post-transcriptional regulation of polycistronic operons in Escherichia coli, little is known about the mechanism of translation re-initiation, which occurs when the same ribosome used to translate an upstream open reading frame (ORF) also translates a downstream ORF. To investigate translation re-initiation in Escherichia coli, we constructed a di-cistronic reporter in which a firefly luciferase gene was linked to a chloramphenicol acetyltransferase gene using a segment of the translationally coupled geneV–geneVII intercistronic region from M13 phage. With this reporter and mutant initiator tRNAs, we show that two of the unique properties of E. coli initiator tRNA – formylation of the amino acid attached to the tRNA and binding of the tRNA to the ribosomal P-site – are as important for re-initiation as for de novo initiation. Overexpression of IF2 or increasing the affinity of mutant initiator tRNA for IF2 enhanced re-initiation efficiency, suggesting that IF2 is required for efficient re-initiation. In contrast, overexpression of IF3 led to a marked decrease in re-initiation efficiency, suggesting that a 30S ribosome and not a 70S ribosome is used for translation re-initiation. Strikingly, overexpression of IF3 also blocked E. coli from acting as a host for propagation of M13 phage

Thermal conductivity, microstructure and hardened characteristics of foamed concrete composite reinforced with raffia fiber

Researchers have become enthralled with using natural fiber, which is a waste product from industrial processes, as an additive in cement-based materials. This is due to the fact that natural fiber is inexpensive, has principal carbon neutrality, and is obtainable in large quantities. Additionally, this fiber is made from a renewable resource. Hence it has a low density and is amenable to undergoing chemical alteration. The idea of this investigation is to discover the reactivity of raffia (raphia vinifera) fiber (RF) in low-density foamed concrete (FC). FC density of 950 kg/m3 was utilized. Workability, density, thermal conductivity, SEM analysis, compressive, bending, and tensile strengths were the parameters that were quantified and assessed. Based on the outcomes, it has been determined that the mechanical properties and thermal conductivity of FC-RF composites may be enhanced by using RF with an ideal reinforcing fraction content of 6%. Slump flow gradually decreased from 2% to 8% RF fraction content. The lowest slump flow was achieved by adding RF to the FC mixture at a fraction content of 8%. The density of FC-RF composites shows a developing tendency, likely because of the RF's comparatively high specific gravity and increasing fraction content. The addition of RF to FC considerably enhances the material's compressive, bending, and tensile strength. The optimal strength characteristics emerged when 6% RF was added to FC. Besides, the FC thermal conductivity improves as the weight percent of RF increases because the porous structure of FC with RF allows it to absorb heat

Progressive Rearrangement of Telomeric Sequences Added to Both the ITR Ends of the Yeast Linear pGKL Plasmid

Relocation into the nucleus of the yeast cytoplasmic linear plasmids was studied using a monitor plasmid pCLU1. In Saccharomyces cerevisiae, the nuclearly-relocated pCLU1 replicated in a linear form (termed pTLU-type plasmid) which carried the host telomeric repeats TG(1-3) of 300-350 bp at both ends. The telomere sequences mainly consisted of a major motif TGTGTGGGTGTGG which was complementary to part of the RNA template of yeast telomerase and were directly added to the very end of the pCLU1-terminal element ITR (inverted terminal repeat), suggesting that the ITR end played a role as a substrate of telomerase. The telomere sequences varied among isolated pTLU-type plasmids, but the TG(1-3) organization was symmetrically identical on both ends of any one plasmid. During cell growth under non-selective condition, the telomeric repeat sequences were progressively rearranged on one side, but not on the opposite side of pTLU plasmid ends. This indicates that the mode of telomeric DNA replication or repair differed between both ends. Clonal analysis showed that the intense rearrangement of telomeric DNA was closely associated with extreme instability of pTLU plasmids

Overexpression of DNA Polymerase Zeta Reduces the Mitochondrial Mutability Caused by Pathological Mutations in DNA Polymerase Gamma in Yeast

In yeast, DNA polymerase zeta (Rev3 and Rev7) and Rev1, involved in the error-prone translesion synthesis during replication of nuclear DNA, localize also in mitochondria. We show that overexpression of Rev3 reduced the mtDNA extended mutability caused by a subclass of pathological mutations in Mip1, the yeast mitochondrial DNA polymerase orthologous to human Pol gamma. This beneficial effect was synergistic with the effect achieved by increasing the dNTPs pools. Since overexpression of Rev3 is detrimental for nuclear DNA mutability, we constructed a mutant Rev3 isoform unable to migrate into the nucleus: its overexpression reduced mtDNA mutability without increasing the nuclear one

Electromechanical analysis of an adaptive piezoelectric energy harvester controlled by two segmented electrodes with shunt circuit networks

This paper presents an adaptive power harvester using a shunted piezoelectric control system with segmented electrodes. This technique has spurred new capability for widening the three simultaneous resonance frequency peaks using only a single piezoelectric laminated beam where normally previous works only provide a single peak for the resonance at the first mode. The benefit of the proposed techniques is that it provides effective and robust broadband power generation for application in self-powered wireless sensor devices. The smart structure beam with proof mass offset is considered to have simultaneous combination between vibration-based power harvesting and shunt circuit control-based electrode segments. As a result, the system spurs new development of the two mathematical methods using electromechanical closed-boundary value techniques and Ritz method-based weak-form analytical approach. The two methods have been used for comparison giving accurate results. For different electrode lengths using certain parametric tuning and harvesting circuit systems, the technique enables the prediction of the power harvesting that can be further proved to identify the performance of the system using the effect of varying circuit parameters so as to visualize the frequency and time waveform responses

Observational study of chronic myeloid leukemia Italian patients who discontinued tyrosine kinase inhibitors in clinical practice.

It is judged safe to discontinue treatment with tyrosine kinase inhibitors for chronic myeloid leukemia in experimental trials on treatment free remission. We collected a total of 293 Italian patients with chronic phase chronic myeloid leukemia who discontinued tyrosine kinase inhibitors in deep molecular response. 72% of patients were on treatment with imatinib, 28% with second generation tyrosine kinase inhibitors at the time of discontinuation. Median duration of treatment with the last tyrosine kinase inhibitor was 77 months (IQR 54;111), median duration of deep molecular response was 46 months (IQR 31;74). Duration of treatment with tyrosine kinase inhibitors and duration of deep molecular response were shorter with 2nd generation tyrosine kinase inhibitors than with imatinib (p<0.001). 88% of the Italian patients discontinued per clinical practice and reasons for stopping treatment were: toxicity for 20% of patients, pregnancy for 6% patients and shared decision between treating physician and patient for 62% of cases. After a median follow-up of 34 months (Min-Max 12-161) overall estimated treatment free remission was 62% (95% CI 56;68). At 12 months treatment free remission was 68% (95% CI 62;74) for imatinib, 73% (95% CI 64;83) for 2nd generation tyrosine kinase inhibitors. Overall median time to restart treatment was 6 months (IQR 4;11). No progressions occurred. Although our study has the limitation of a retrospective study, our experience within the Italian population confirms that discontinuation of imatinib and 2nd generation tyrosine kinase inhibitors is feasible and safe in the clinical practic

Systemic importance of financial institutions: regulations, research, open issues, proposals

In the field of risk management, scholars began to bring together the quantitative methodologies with the banking management issues about 30 years ago, with a special focus on market, credit and operational risks. After the systemic effects of banks defaults during the recent financial crisis, and despite a huge amount of literature in the last years concerning the systemic risk, no standard methodologies have been set up to now. Even the new Basel 3 regulation has adopted a heuristic indicator-based approach, quite far from an effective quantitative tool. In this paper, we refer to the different pieces of the puzzle: definition of systemic risk, a set of coherent and useful measures, the computability of these measures, the data set structure. In this challenging field, we aim to build a comprehensive picture of the state of the art, to illustrate the open issues, and to outline some paths for a more successful future research. This work appropriately integrates other useful surveys and it is directed to both academic researchers and practitioners

Brain Cortical Mapping by Simultaneous Recording of Functional Near Infrared Spectroscopy and Electroencephalograms from the Whole Brain During Right Median Nerve Stimulation

To investigate relationships between hemodynamic responses and neural activities in the somatosensory cortices, hemodynamic responses by near infrared spectroscopy (NIRS) and electroencephalograms (EEGs) were recorded simultaneously while subjects received electrical stimulation in the right median nerve. The statistical significance of the hemodynamic responses was evaluated by a general linear model (GLM) with the boxcar design matrix convoluted with Gaussian function. The resulting NIRS and EEGs data were stereotaxically superimposed on the reconstructed brain of each subject. The NIRS data indicated that changes in oxy-hemoglobin concentration increased at the contralateral primary somatosensory (SI) area; responses then spread to the more posterior and ipsilateral somatosensory areas. The EEG data indicated that positive somatosensory evoked potentials peaking at 22 ms latency (P22) were recorded from the contralateral SI area. Comparison of these two sets of data indicated that the distance between the dipoles of P22 and NIRS channels with maximum hemodynamic responses was less than 10 mm, and that the two topographical maps of hemodynamic responses and current source density of P22 were significantly correlated. Furthermore, when onset of the boxcar function was delayed 5–15 s (onset delay), hemodynamic responses in the bilateral parietal association cortices posterior to the SI were more strongly correlated to electrical stimulation. This suggests that GLM analysis with onset delay could reveal the temporal ordering of neural activation in the hierarchical somatosensory pathway, consistent with the neurophysiological data. The present results suggest that simultaneous NIRS and EEG recording is useful for correlating hemodynamic responses to neural activity

Analysis of the mitochondrial and nuclear genomes of two basidiomycetes, Coprinus cinereus and Coprinus stercorarius

The mitochondrial and nuclear genomes of Coprinus stercorarius and C. cinereus were compared to assess their evolutionary relatedness and to characterize at the molecular level changes that have occurred since they diverged from a common ancestor. The mitochondrial genome of C. stercorarius (91.1 kb) is approximately twice as large as that of C. cinereus (43.3 kb). The pattern of restriction enzyme recognition sites shows both genomes to be circular, but reveals no clear homologies; furthermore, the order of structural genes is different in each species. The C. stercorarius mitochondrial genome contains a region homologous to a probe derived from the yeast mitochondrial var1 gene, whereas its nuclear genome does not. By contrast, the C. cinereus nuclear, but not mitochondrial, genome contains a region homologous to the var1 probe. Only a small fraction of either the nuclear or mitochondrial genomes, perhaps corresponding to the coding sequences, is capable of forming duplexes in interspecies solution reassociations, as measured by binding to hydroxylapatite. Those sequences capable of reassociating were found to have approximately 15% divergence for the mitochondrial genomes and 7%–15% divergence for the nuclear genomes, depending on the conditions of reassociation.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/46959/1/294_2004_Article_BF00447385.pd

Achlya mitochondrial DNA: gene localization and analysis of inverted repeats

Mitochondrial DNA from four strains of the oomycete Achlya has been compared and nine gene loci mapped, including that of the ribosomal protein gene, var1 . Examination of the restriction enzyme site maps showed the presence of four insertions relative to a map common to all four strains. All the insertions were found in close proximity to genic regions. The four strains also cotained the inverted repeat first observed in A. ambisexualis (Hudspeth et al. 1983), allowing an examination by analysis of retained restriction sites of the evolutionary stability of repeated DNA sequences relative to single copy sequences. Although the inverted repeat is significantly more stable than single copy sequences, more detailed analysis indicated that this stability is limited to the portion encoding the ribosomal RNA genes. Thus, the apparent evolutionary stability of the repeat does not appear to derive from the inverted repeat structure per se.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/47563/1/438_2004_Article_BF00330510.pd