Kinetic analysis of cellular internalization and expulsion of unstructured D-chirality cell penetrating peptides

Most cell penetrating peptides (CPPs) are unstructured and susceptible to proteolytic degradation. One alternative is to incorporate D-chirality amino acids into unstructured CPPs to allow for enhanced uptake and intracellular stability. This work investigates CPP internalization using a series of time, concentration, temperature, and energy dependent studies, resulting in a three-fold increase in uptake and 50-fold increase in stability of D-chirality peptides over L-chirality counterparts. CPP internalization occurred via a combination of direct penetration and endocytosis, with a percentage of internalized CPP expelling from cells in a time-dependent manner. Mechanistic studies identified that cells exported the intact internalized D-chirality CPPs via an exocytosis independent pathway, analogous to a direct penetration method out of the cells. These findings highlight the potential of a D-chirality CPP as bio-vector in therapeutic and biosensing applications, but also identify a new expulsion method suggesting a relationship between uptake kinetics, intracellular stability, and export kinetics.     

Magnesium technology 2002, part I: Primary production,environmental issues,high-temperature alloys

Conclusion It is expected that the research in high-temperature alloy development will reach a critical mass in about five years. This will not only raise the materials science of magnesium to a higher level but will also increase the confidence of the industry in magnesium as a structural performance material. Currently, applications are being actively pursued in United States Council for Automotive Research power train programs and European Council for Automotive R&D engine block programs to develop the technology for elevated-temperature magnesium. More research is still needed in this interesting materials field.     

Effectiveness of participatory ergonomic interventions on health outcomes: a systematic review

The objective of this study was to conduct a systematic review of the literature on the effectiveness of participatory ergonomic (PE) interventions for improving workers' health. The search strategy targeted six electronic databases and identified 442 potential articles. Each article was examined by pairs of reviewers for relevance (assessed a participative ergonomic workplace intervention, with at least one health outcome, published in English in peer reviewed literature). Twenty-three articles met relevance criteria and were then appraised for methodological strength. Using a best evidence synthesis approach, 12 studies that were rated as 'medium' or higher provided partial to moderate evidence that PE interventions have a positive impact on: musculoskeletal symptoms, reducing injuries and workers' compensation claims, and a reduction in lost days from work or sickness absence. However, the magnitude of the effect requires more precise definition.     

Controller design for flexible structure vibration suppression with robustness to contacts

Model-based feedback control of vibration in flexible structures can be complicated by the possibility that interaction with an external body occurs. If not accounted for, instability or poor performance may result. In this paper, a method is proposed for achieving robust vibration control of flexible structures under contact. The method uses robust linear state feedback, coupled with a state estimation scheme utilizing contact force measurement. Uncertain contact characteristics are modelled by a sector-bounded non-linear function, such that state feedback gains can be synthesized using a matrix inequality formulation of the Popov stability criterion. A separation theorem is used to establish a robust H₂ cost bound for the closed loop system. Experimental results from a multi-mode flexible structure testbed confirm that vibration attenuation and stability can be maintained over a broad range of contact characteristics, in terms of compliance and clearance.     

Purification, characterization, and high performance liquid chromatography assay of Salmonella glucose-1-phosphate cytidylyltransferase from the cloned rfbF gene

We report the purification and characterization of glucose-1-phosphate cytidylyltransferase, the first of five enzymes committed to biosynthesis of CDP-D-abequose from Salmonella enterica strain LT2. The purification was greatly facilitated by using a cloned rfbF gene encoding this enzyme. Pure enzyme was obtained by 64-fold enrichment in three chromatography steps. The NH2-terminal sequence of the purified enzyme was in agreement with the sequence predicted from the nucleotide sequence of the rfbF gene. The SDS-polyacrylamide gel electrophoresis estimated subunit M(r) of 31,000 agrees well with the M(r) of 29,035 calculated from the amino acid composition deduced from the nucleotide sequence of the rfbF gene. The glucose-1-phosphate cytidylyltransferase catalyzes a reversible bimolecular group transfer reaction and steady-state kinetic measurements, including product inhibition patterns, indicate that this reaction proceeds by a "ping-pong" type of mechanism. The Km values for CTP, alpha-D-glucose 1-phosphate, CDP-D-glucose, and pyrophosphate are 0.28, 0.64, 0.11, and 1.89 mM, respectively.     

A critique of the application of sensory integration therapy to children with learning disabilities

Sensory integration (SI) therapy is a controversial--though popular--treatment for the remediation of motor and academic problems. It has been applied primarily to children with learning disabilities, under the assumption that such children (or at least a subgroup of them) have problems in sensory integration to which some or all of their learning difficulties can be ascribed. The present article critically examines the related issues of whether children with learning disabilities differentially exhibit concomitant problems in sensory integration, and whether such children are helped in any way by means specific to SI therapy. An overview of theoretical contentions and empirical findings pertaining to the first issue is presented, followed by a detailed review of recent studies in the SI therapy research literature, in an effort to resolve the second issue. Results of this critique raise serious doubts as to the validity or utility of SI therapy as an appropriate, indicated treatment for the clinical population in question--and, by extension, for any other groups diagnosed as having "sensory integrative dysfunction." It is concluded that the current fund of research findings may well be sufficient to declare SI therapy not merely an unproven, but a demonstrably ineffective, primary or adjunctive remedial treatment for learning disabilities and other disorders.     

Prosthetic valve endocarditis: superiority of surgical valve replacement versus medical therapy only

The objective of our study was to assess the long-term outcome of patients with prosthetic valve endocarditis. We used a multicenter, prospective, observational study design. Six university teaching hospitals with high volume cardiothoracic surgery participated. Seventy-four patients with prosthetic valve endocarditis as defined by explicit, objective criteria were selected for participation. All patients were followed up prospectively for 1 year. Thirty-one percent and 69% had development of endocarditis within 60 days of valve insertion ("early") and after 60 days ("late"), respectively. The most common causes were Staphylococcus epidermidis (40%), Staphylococcus aureus (20%), streptococcal species (18%), and aerobic gram-negative bacilli (11%). Physical signs of endocarditis (new or changing murmur, stigmata, emboli) were seen in 58%. At 6 months and 12 months, mortality was 46% and 47%, respectively. Surgical replacement of the infected valve led to significantly lower mortality (23%) as compared with medical therapy alone (56%), as assessed by both univariate and multivariate analyses (p < 0.05). Improved outcome was seen for the surgical group even when controlling for severity of illness at time of diagnosis. From these findings we conclude that accurate assessment of outcome in prosthetic valve endocarditis requires long-term follow-up of at least 6 months following diagnosis. Surgical therapy warrants greater scrutiny; evaluation in controlled clinical trials is appropriate.     

Electromyographic and neuromuscular variables in post-polio subjects

OBJECTIVE: Post-polio subjects experience functional deterioration many years after developing acute poliomyelitis and have been shown previously to have a deficit in strength recovery after isometric activity. This study characterized the size and stability of the motor units in a group of post-polio subjects with macro and single fiber electromyography (EMG) and correlated these variables with isometric strength, endurance, "work capacity," and strength recovery after fatiguing isometric exercise. DESIGN: A cohort of 12 post-polio subjects was tested for neuromuscular function. Electromyographic variables were determined on a separate day. SETTING: Volunteers were recruited from the community and tested in our neuromuscular research laboratory. SUBJECTS: A volunteer sample was obtained from advertisements. All subjects acknowledged post-polio syndrome symptoms. MAIN OUTCOME MEASURES: Neuromuscular variables were isometric knee extension peak torque, endurance (time to exhaustion) at 40% of maximal torque, tension time index, and recovery of torque at 10 minutes. Electromyographic variables were macro EMG and single fiber EMG (percent blocking and jitter). RESULTS: Macro EMG amplitude was ninefold the control value, and both jitter and blocking were greatly increased in comparison to control values. Isometric strength significantly (p < .05) correlated negatively with macro EMG amplitude. CONCLUSIONS: The weakest subjects had the greatest number of muscle fibers within the motor unit (as measured by macro EMG amplitude). Jitter and blocking did not correlate with neuromuscular function.     

'98 Escherichia coli SWISS-2DPAGE database update

The combination of two-dimensional polyacrylamide gel electrophoresis (2-D PAGE), computer image analysis and several protein identification techniques allowed the Escherichia coli SWISS-2DPAGE database to be established. This is part of the ExPASy molecular biology server accessible through the WWW at the URL address http://www.expasy.ch/ch2d/ch2d-top.html . Here we report recent progress in the development of the E. coli SWISS-2DPAGE database. Proteins were separated with immobilized pH gradients in the first dimension and sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the second dimension. To increase the resolution of the separation and thus the number of identified proteins, a variety of wide and narrow range immobilized pH gradients were used in the first dimension. Micropreparative gels were electroblotted onto polyvinylidene difluoride membranes and spots were visualized by amido black staining. Protein identification techniques such as amino acid composition analysis, gel comparison and microsequencing were used, as well as a recently described Edman "sequence tag" approach. Some of the above identification techniques were coupled with database searching tools. Currently 231 polypeptides are identified on the E. coli SWISS-2DPAGE map: 64 have been identified by N-terminal microsequencing, 39 by amino acid composition, and 82 by sequence tag. Of 153 proteins putatively identified by gel comparison, 65 have been confirmed. Many proteins have been identified using more than one technique. Faster progress in the E. coli proteome project will now be possible with advances in biochemical methodology and with the completion of the entire E. coli genome.