Nuclear Surveillance and Degradation of Hypomodified Initiator tRNA\u3csup\u3eMet\u3c/sup\u3e in \u3cem\u3eS. cerevisiae\u3c/em\u3e

The tRNA m1A58 methyltransferase is composed of two subunits encoded by the essential genes TRM6 and TRM61 (formerly GCD10 and GCD14). The trm6-504 mutation results in a defective m1A methyltransferase (Mtase) and a temperature-sensitive growth phenotype that is attributable to the absence of m1A58 and consequential tRNAiMet instability. We used a genetic approach to identify the genes responsible for tRNAiMet degradation in trm6 cells. Three recessive extragenic mutations that suppress trm6-504 mutant phenotypes and restore hypomodified tRNAiMet to near normal levels were identified. The wild-type allele of one suppressor, DIS3/RRP44, encodes a 3′-5′ exoribonuclease and a member of the multisubunit exosome complex. We provide evidence that a functional nuclear exosome is required for the degradation of tRNAiMet lacking m1A58. A second suppressor gene encodes Trf4p, a DNA polymerase (pol σ) with poly(A) polymerase activity. Whereas deletion of TRF4 leads to stabilization of tRNAiMet, overexpression of Trf4p destabilizes the hypomodified tRNAiMet in trm6 cells. The hypomodified, but not wild-type, pre-tRNAiMet accumulates as a polyadenylated species, whose abundance and length distribution both increase upon Trf4p overexpression. These data indicate that a tRNA surveillance pathway exists in yeast that requires Trf4p and the exosome for polyadenylation and degradation of hypomodified pre-tRNAiMet

Microbiological pattern of arterial catheters in the intensive care unit

<p>Abstract</p> <p>Background</p> <p>Intravascular catheter related infection (CRI) is one of the most serious nosocomial infections. Diagnostic criteria include a positive culture from the catheter tip along with blood, yet in many patients with signs of infection, current culture techniques fail to identify pathogens on catheter segments. We hypothesised that a molecular examination of the bacterial community on short term arterial catheters (ACs) would improve our understanding of the variety of organisms that are present in this niche environment and would help develop new methods for the diagnosis of CRI.</p> <p>Results</p> <p>The whole bacterial community presenting on all ACs was evaluated by molecular methods, i.e., a strategy of whole community DNA extraction, PCR amplification followed by cloning and 16S rDNA sequence analysis. Ten ACs were removed from patients suspected of CRI and 430 clones from 5 "colonised" and 5 "uncolonised" (semi-quantitative method) AC libraries were selected for sequencing and subsequent analysis. A total of 79 operational taxonomic units (OTUs) were identified at the level of 97% similarity belonging to six bacterial divisions. An average of 20 OTUs were present in each AC, irrespective of colonisation status. Conventional culture failed to reveal the majority of these bacteria.</p> <p>Conclusions</p> <p>There was no significant difference in the bacterial diversity between the 'uncolonised' and 'colonised' ACs. This suggests that vascular devices cultured conventionally and reported as non infective may at times potentially be a significant source of sepsis in critically ill patients. Alternative methods may be required for the accurate diagnosis of CRI in critically ill patients.</p

Normative EMG activation patterns of school-age children during gait

Gait analysis is widely used in clinics to study walking abnormalities for surgery planning, definition of rehabilitation protocols, and objective evaluation of clinical outcomes. Surface electromyography allows the study of muscle activity non-invasively and the evaluation of the timing of muscle activation during movement. The aim of this study was to present a normative dataset of muscle activation patterns obtained from a large number of strides in a population of 100 healthy children aged 6-11 years. The activity of Tibialis Anterior, Lateral head of Gastrocnemius, Vastus Medialis, Rectus Femoris and Lateral Hamstrings on both lower limbs was analyzed during a 2.5-min walk at free speed. More than 120 consecutive strides were analyzed for each child, resulting in approximately 28,000 strides. Onset and offset instants were reported for each observed muscle. The analysis of a high number of strides for each participant allowed us to obtain the most recurrent patterns of activation during gait, demonstrating that a subject uses a specific muscle with different activation modalities even in the same walk. The knowledge of the various activation patterns and of their statistics will be of help in clinical gait analysis and will serve as reference in the design of future gait studie

The AURORA Gigabit Testbed

AURORA is one of five U.S. networking testbeds charged with exploring applications of, and technologies necessary for, networks operating at gigabit per second or higher bandwidths. The emphasis of the AURORA testbed, distinct from the other four testbeds, BLANCA, CASA, NECTAR, and VISTANET, is research into the supporting technologies for gigabit networking. Like the other testbeds, AURORA itself is an experiment in collaboration, where government initiative (in the form of the Corporation for National Research Initiatives, which is funded by DARPA and the National Science Foundation) has spurred interaction among pre-existing centers of excellence in industry, academia, and government. AURORA has been charged with research into networking technologies that will underpin future high-speed networks. This paper provides an overview of the goals and methodologies employed in AURORA, and points to some preliminary results from our first year of research, ranging from analytic results to experimental prototype hardware. This paper enunciates our targets, which include new software architectures, network abstractions, and hardware technologies, as well as applications for our work

An Overview of the AURORA Gigabit Testbed

AURORA is one of five U.S. testbeds charged with exploring applications of, and technologies necessary for, networks operating at gigabit per second or higher bandwidths. AURORA is also an experiment in collaboration, where government support (through the Corporation for National Research Initiatives, which is in turn funded by DARPA and the NSF) has spurred interaction among centers of excellence in industry, academia, and government. The emphasis of the AURORA testbed, distinct from the other four testbeds, is research into the supporting technologies for gigabit networking. Our targets include new software architectures, network abstractions, hardware technologies, and applications. This paper provides an overview of the goals and methodologies employed in AURORA, and reports preliminary results from our first year of research

Evaluation of range of motion restriction within the hip joint

In Total Hip Arthroplasty, determining the impingement free range of motion requirement is a complex task. This is because in the native hip, motion is restricted by both impingement as well as soft tissue restraint. The aim of this study is to determine a range of motion benchmark which can identify motions which are at risk from impingement and those which are constrained due to soft tissue. Two experimental methodologies were used to determine motions which were limited by impingement and those motions which were limited by both impingement and soft tissue restraint. By comparing these two experimental results, motions which were limited by impingement were able to be separated from those motions which were limited by soft tissue restraint. The results show motions in extension as well as flexion combined with adduction are limited by soft tissue restraint. Motions in flexion, flexion combined with abduction and adduction are at risk from osseous impingement. Consequently, these motions represent where the maximum likely damage will occur in femoroacetabular impingement or at most risk of prosthetic impingement in Total Hip Arthroplasty

Requirements for a Robust Animal Model to Investigate the Disease Mechanism of Autoimmune Complications Associated With ARF/RHD

The pathogenesis of Acute Rheumatic Fever/Rheumatic Heart Disease (ARF/RHD) and associated neurobehavioral complications including Sydenham's chorea (SC) is complex. Disease complications triggered by Group A streptococcal (GAS) infection are confined to human and determining the early events leading to pathology requires a robust animal model that reflects the hallmark features of the disease. However, modeling these conditions in a laboratory animal, of a uniquely human disease is challenging. Animal models including cattle, sheep, pig, dog, cat, guinea pigs rats and mice have been used extensively to dissect molecular mechanisms of the autoimmune inflammatory responses in ARF/RHD. Despite the characteristic limitations of some animal models, several rodent models have significantly contributed to better understanding of the fundamental mechanisms underpinning features of ARF/RHD. In the Lewis rat autoimmune valvulitis model the development of myocarditis and valvulitis with the infiltration of mononuclear cells along with generation of antibodies that cross-react with cardiac tissue proteins following exposure to GAS antigens were found to be similar to ARF/RHD. We have recently shown that Lewis rats injected with recombinant GAS antigens simultaneously developed cardiac and neurobehavioral changes. Since ARF/RHD is multifactorial in origin, an animal model which exhibit the characteristics of several of the cardinal diagnostic criteria observed in ARF/RHD, would be advantageous to determine the early immune responses to facilitate biomarker discovery as well as provide a suitable model to evaluate treatment options, safety and efficacy of vaccine candidates. This review focuses on some of the common small animals and their advantages and limitations

The conventional gait model - success and limitations

The Conventional Gait Model (CGM) is a generic name for a family of closely related and very widely used biomechanical models for gait analysis. After describing its history, the core attributes of the model are described followed by evaluation of its strengths and weaknesses. An analysis of the current and future requirements for practical biomechanical models for clinical and other gait analysis purposes which have been rigorously calibrated suggests that the CGM is better suited for this purpose than any other currently available model. Modifications are required, however, and a number are proposed

Are mice good models for human neuromuscular disease? Comparing muscle excursions in walking between mice and humans

The mouse is one of the most widely used animal models to study neuromuscular diseases and test new therapeutic strategies. However, findings from successful pre-clinical studies using mouse models frequently fail to translate to humans due to various factors. Differences in muscle function between the two species could be crucial but often have been overlooked. The purpose of this study was to evaluate and compare muscle excursions in walking between mice and humans