Effects of Non-Aerobic Maximal Effort Exercise on Fatigue in Deconditioned Men and Women with Multiple Sclerosis

Multiple Sclerosis (MS) is a neurodegenerative disease of unknown etiology affecting women more frequently than men. Mental and physical fatigue complaints are often the most disabling symptoms for an MS patient. Both are multifactorial, potentially exacerbated by aerobic exercise, may prevent sustained physical functioning, and significantly interfere with activities of daily living1. A multi-center study was designed to investigate the effects of non-aerobic maximal effort exercise (MEE) for deconditioned persons with MS, with the expectation of minimizing fatigue. The IsoPUMP (Neuromuscular Engineering; Nashville, TN), is a specialized exercise and strength-sensing machine, designed to allow individuals to safely perform and record their non-aerobic MEE sessions. The Modified Fatigue Impact Scale (MFIS) and Multiple Sclerosis Functional Composite (MSFC) are common, accepted methods used to measure fatigue and function. The MFIS is a 21-item questionnaire which assesses the subjects’ perception of physical, cognitive, and psychosocial aspects of fatigue over a four-week period2. Each of the 21 items are scored on a scale from 0 (never) to 4 (almost always), and the total MFIS score is calculated by summing the circled number for each item. Total scores can range from 0 to 84; higher scores indicating a greater impact of fatigue on the person. The MFIS has three distinct subscales: (1) physical, (2) cognitive, and (3) psychosocial. These subscales can be scored independently by summing the questions that pertain to each subscale2. The MFIS physical subscale score can range from 0 – 36 and the MFIS cognitive subscale score can range from 0 – 40. The MSFC combines clinical measures used to assess lower limb function (Timed 25-Foot Walk [25-FW]), upper limb function (9-Hole Peg Test [9-HPT]), and cognition (Paced Auditory Serial Addition Test [PASAT-3”])3. The 25-FW is a quantitative measure of lower extremity function. The 9-HPT is a quantitative measure of arm and hand function where a subject inserts and then removes 9 pegs from a board, using one hand at a time. The time is recorded for each hand with the dominant hand trial first and the non-dominant hand trial second. The final score is recorded as the mean time for both hands. The PASAT-3” is a measure of cognitive function, specifically assessing auditory information processing speed, short-term memory, flexibility, and calculation ability. Cognitive dysfunction affects half of all MS patients; slowing ability to reason, concentrate, and recall5. In this test subjects listen to a series of 61 spoken numbers separated by 3 seconds and must add each number to the prior number. Their final PASAT-3” score is the number of correct additions in the series, with 60 reflecting a perfect score. The MSFC is then evaluated by creating Z-scores for each component, which compare each outcome with the average outcome of the study population. The three Z-scores are then averaged to create an overall composite score (the MSFC score) which represents change over time for that population of MS subjects3

The Effect of Progressive Non-Aerobic High-Intensity Maximal Effort Exercise (MEE) on the Health-Related Quality of Life in Patients with Multiple Sclerosis

Background: Studies indicate that Multiple Sclerosis (MS) patients are less satisfied with the quality of their lives than healthy individuals in similar circumstances. Common symptoms experienced include fatigue, cognitive dysfunction, pain, spasticity, depression, bladder/bowel dysfunction and sexual dysfunction. Several pharmacological and non-pharmacological methods have been employed for such symptoms to try to increase quality of life and reduce the mortality rate. Non-pharmacological methods recommended for MS patients include lifestyle modifications, exercise programs and physical therapy. MS patients easily fatigue during aerobic exercise but a non-aerobic progressive maximal effort exercise (MEE) protocol consisting of a few short, duration isometric and eccentric leg press and whole body lunges was previously seen to increase strength without increasing fatigue. The IsoPUMP® (Neuromuscular Engineering, Nashville TN) exercise system permitted safe conduct and measurement of muscle strength and duration during each exercise repetition

An Analysis of Functional Status in Multiple Sclerosis Patients after Progressive Non-Aerobic High-Intensity Maximal Effort Exercise (MEE)

Background: Multiple Sclerosis (MS) is a disease with a wide-ranging impact on functional status. MS patient function has been assessed using Multiple Sclerosis Functional Composite Score (MSFCS). The MSFCS includes the standardized scores (Z-score) of three functional tests: the Paced Auditory Serial Addition Test (PASAT-3”) for cognitive function, 9-Hole Peg Test (9-HPT) for upper extremity function, and timed 25-foot walk (25-TW) for lower extremity function. One of the most common symptoms experienced by MS patients is severe fatigue, often brought on suddenly by aerobic exercise. Non-aerobic maximal effort exercise (MEE) is thought to increase strength without increasing fatigue. The IsoPUMP® (Neuromuscular Engineering; Nashville, TN) is a stationary exercise device designed for patient use to safely perform MEE leg presses and whole body lunges using isometric and eccentric exercises. The progressive functional changes of the MS patients were tracked using the MSFCs at specific intervals during the study

Endonuclease-independent LINE-1 retrotransposition at mammalian telomeres

Long interspersed element-1 (LINE-1 or L1) elements are abundant, non-long-terminal-repeat (non-LTR) retrotransposons that comprise 17% of human DNA(1). The average human genome contains similar to 80-100 retrotransposition- competent L1s (ref. 2), and they mobilize by a process that uses both the L1 endonuclease and reverse transcriptase, termed target-site primed reverse transcription(3-5). We have previously reported an efficient, endonuclease-independent L1 retrotransposition pathway (ENi) in certain Chinese hamster ovary (CHO) cell lines that are defective in the non-homologous end-joining (NHEJ) pathway of DNA double-strand-break repair(6). Here we have characterized ENi retrotransposition events generated in V3 CHO cells, which are deficient in DNA-dependent protein kinase catalytic subunit (DNA-PKcs) activity and have both dysfunctional telomeres and an NHEJ defect. Notably, similar to 30% of ENi retrotransposition events insert in an orientation-specific manner adjacent to a perfect telomere repeat (5'-TTAGGG-3'). Similar insertions were not detected among ENi retrotransposition events generated in controls or in XR-1 CHO cells deficient for XRCC4, an NHEJ factor that is required for DNA ligation but has no known function in telomere maintenance. Furthermore, transient expression of a dominant-negative allele of human TRF2 ( also called TERF2) in XRCC4-deficient XR-1 cells, which disrupts telomere capping, enables telomere-associated ENi retrotransposition events. These data indicate that L1s containing a disabled endonuclease can use dysfunctional telomeres as an integration substrate. The findings highlight similarities between the mechanism of ENi retrotransposition and the action of telomerase, because both processes can use a 3' OH for priming reverse transcription at either internal DNA lesions or chromosome ends(7,8). Thus, we propose that ENi retrotransposition is an ancestral mechanism of RNA-mediated DNA repair associated with non-LTR retrotransposons that may have been used before the acquisition of an endonuclease domain.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/62964/1/nature05560.pd

Characterization of LINE-1 Ribonucleoprotein Particles

The average human genome contains a small cohort of active L1 retrotransposons that encode two proteins (ORF1p and ORF2p) required for their mobility (i.e., retrotransposition). Prior studies demonstrated that human ORF1p, L1 RNA, and an ORF2p-encoded reverse transcriptase activity are present in ribonucleoprotein (RNP) complexes. However, the inability to physically detect ORF2p from engineered human L1 constructs has remained a technical challenge in the field. Here, we have employed an epitope/RNA tagging strategy with engineered human L1 retrotransposons to identify ORF1p, ORF2p, and L1 RNA in a RNP complex. We next used this system to assess how mutations in ORF1p and/or ORF2p impact RNP formation. Importantly, we demonstrate that mutations in the coiled-coil domain and RNA recognition motif of ORF1p, as well as the cysteine-rich domain of ORF2p, reduce the levels of ORF1p and/or ORF2p in L1 RNPs. Finally, we used this tagging strategy to localize the L1–encoded proteins and L1 RNA to cytoplasmic foci that often were associated with stress granules. Thus, we conclude that a precise interplay among ORF1p, ORF2p, and L1 RNA is critical for L1 RNP assembly, function, and L1 retrotransposition

Interim Consequence Management Guidance for a Wide-Area Biological Attack

Abstract not provide

Emergence of 3D Printed Dosage Forms: Opportunities and Challenges

The recent introduction of the first FDA approved 3D-printed drug has fuelled interest in 3D printing technology, which is set to revolutionize healthcare. Since its initial use, this rapid prototyping (RP) technology has evolved to such as extent that it is currently being used in a wide range of applications including in tissue engineering, dentistry, construction, automotive and aerospace. However, in the pharmaceutical industry this technology is still in its infancy and its potential yet to be fully explored. This paper presents various 3D printing technologies such as stereolithographic, powder based, selective laser sintering, fused deposition modelling and semi-solid extrusion 3D printing. It also provides a comprehensive review of previous attempts at using 3D printing technologies on the manufacturing dosage forms with a particular focus on oral tablets. Their advantages particularly with adaptability in the pharmaceutical field have been highlighted, including design flexibility and control and manufacture which enables the preparation of dosage forms with complex designs and geometries, multiple actives and tailored release profiles. An insight into the technical challenges facing the different 3D printing technologies such as the formulation and processing parameters is provided. Light is also shed on the different regulatory challenges that need to be overcome for 3D printing to fulfil its real potential in the pharmaceutical industry

SIGffRid: A tool to search for sigma factor binding sites in bacterial genomes using comparative approach and biologically driven statistics

<p>Abstract</p> <p>Background</p> <p>Many programs have been developed to identify transcription factor binding sites. However, most of them are not able to infer two-word motifs with variable spacer lengths. This case is encountered for RNA polymerase Sigma (<it>σ</it>) Factor Binding Sites (SFBSs) usually composed of two boxes, called -35 and -10 in reference to the transcription initiation point. Our goal is to design an algorithm detecting SFBS by using combinational and statistical constraints deduced from biological observations.</p> <p>Results</p> <p>We describe a new approach to identify SFBSs by comparing two related bacterial genomes. The method, named SIGffRid (SIGma Factor binding sites Finder using R'MES to select Input Data), performs a simultaneous analysis of pairs of promoter regions of orthologous genes. SIGffRid uses a prior identification of over-represented patterns in whole genomes as selection criteria for potential -35 and -10 boxes. These patterns are then grouped using pairs of short seeds (of which one is possibly gapped), allowing a variable-length spacer between them. Next, the motifs are extended guided by statistical considerations, a feature that ensures a selection of motifs with statistically relevant properties. We applied our method to the pair of related bacterial genomes of <it>Streptomyces coelicolor </it>and <it>Streptomyces avermitilis</it>. Cross-check with the well-defined SFBSs of the SigR regulon in <it>S. coelicolor </it>is detailed, validating the algorithm. SFBSs for HrdB and BldN were also found; and the results suggested some new targets for these <it>σ </it>factors. In addition, consensus motifs for BldD and new SFBSs binding sites were defined, overlapping previously proposed consensuses. Relevant tests were carried out also on bacteria with moderate GC content (i.e. <it>Escherichia coli</it>/<it>Salmonella typhimurium </it>and <it>Bacillus subtilis</it>/<it>Bacillus licheniformis </it>pairs). Motifs of house-keeping <it>σ </it>factors were found as well as other SFBSs such as that of SigW in <it>Bacillus </it>strains.</p> <p>Conclusion</p> <p>We demonstrate that our approach combining statistical and biological criteria was successful to predict SFBSs. The method versatility autorizes the recognition of other kinds of two-box regulatory sites.</p

SIGffRid: A tool to search for sigma factor binding sites in bacterial genomes using comparative approach and biologically driven statistics

<p>Abstract</p> <p>Background</p> <p>Many programs have been developed to identify transcription factor binding sites. However, most of them are not able to infer two-word motifs with variable spacer lengths. This case is encountered for RNA polymerase Sigma (<it>σ</it>) Factor Binding Sites (SFBSs) usually composed of two boxes, called -35 and -10 in reference to the transcription initiation point. Our goal is to design an algorithm detecting SFBS by using combinational and statistical constraints deduced from biological observations.</p> <p>Results</p> <p>We describe a new approach to identify SFBSs by comparing two related bacterial genomes. The method, named SIGffRid (SIGma Factor binding sites Finder using R'MES to select Input Data), performs a simultaneous analysis of pairs of promoter regions of orthologous genes. SIGffRid uses a prior identification of over-represented patterns in whole genomes as selection criteria for potential -35 and -10 boxes. These patterns are then grouped using pairs of short seeds (of which one is possibly gapped), allowing a variable-length spacer between them. Next, the motifs are extended guided by statistical considerations, a feature that ensures a selection of motifs with statistically relevant properties. We applied our method to the pair of related bacterial genomes of <it>Streptomyces coelicolor </it>and <it>Streptomyces avermitilis</it>. Cross-check with the well-defined SFBSs of the SigR regulon in <it>S. coelicolor </it>is detailed, validating the algorithm. SFBSs for HrdB and BldN were also found; and the results suggested some new targets for these <it>σ </it>factors. In addition, consensus motifs for BldD and new SFBSs binding sites were defined, overlapping previously proposed consensuses. Relevant tests were carried out also on bacteria with moderate GC content (i.e. <it>Escherichia coli</it>/<it>Salmonella typhimurium </it>and <it>Bacillus subtilis</it>/<it>Bacillus licheniformis </it>pairs). Motifs of house-keeping <it>σ </it>factors were found as well as other SFBSs such as that of SigW in <it>Bacillus </it>strains.</p> <p>Conclusion</p> <p>We demonstrate that our approach combining statistical and biological criteria was successful to predict SFBSs. The method versatility autorizes the recognition of other kinds of two-box regulatory sites.</p

Queen mandibular pheromone: questions that remain to be resolved

The discovery of ‘queen substance’, and the subsequent identification and synthesis of keycomponents of queen mandibular pheromone, has been of significant importance to beekeepers and to thebeekeeping industry. Fifty years on, there is greater appreciation of the importance and complexity of queenpheromones, but many mysteries remain about the mechanisms through which pheromones operate. Thediscovery of sex pheromone communication in moths occurred within the same time period, but in this case,intense pressure to find better means of pest management resulted in a remarkable focusing of research activityon understanding pheromone detection mechanisms and the central processing of pheromone signals in themoth. We can benefit from this work and here, studies on moths are used to highlight some of the gaps in ourknowledge of pheromone communication in bees. A better understanding of pheromone communication inhoney bees promises improved strategies for the successful management of these extraordinary animals