ERK1/2 signaling induces skeletal muscle slow fiber-type switching and reduces muscular dystrophy disease severity

© 2019 American Society for Clinical Investigation. MAPK signaling consists of an array of successively acting kinases. ERK1 and -2 (ERK1/2) are major components of the greater MAPK cascade that transduce growth factor signaling at the cell membrane. Here, we investigated ERK1/2 signaling in skeletal muscle homeostasis and disease. Using mouse genetics, we observed that the muscle-specifc expression of a constitutively active MEK1 mutant promotes greater ERK1/2 signaling that mediates fber-type switching to a slow, oxidative phenotype with type I myosin heavy chain expression. Using a conditional and temporally regulated Cre strategy, as well as Mapk1 (ERK2) and Mapk3 (ERK1) genetically targeted mice, MEK1-ERK2 signaling was shown to underlie this fast-to-slow fber-type switching in adult skeletal muscle as well as during development. Physiologic assessment of these activated MEK1-ERK1/2 mice showed enhanced metabolic activity and oxygen consumption with greater muscle fatigue resistance. In addition, induction of MEK1-ERK1/2 signaling increased dystrophin and utrophin protein expression in a mouse model of limb-girdle muscle dystrophy and protected myofbers from damage. In summary, sustained MEK1-ERK1/2 activity in skeletal muscle produces a fast-to-slow fber-type switch that protects from muscular dystrophy, suggesting a therapeutic approach to enhance the metabolic effectiveness of muscle and protect from dystrophic disease

Rapid Pharmacokinetic and Biodistribution Studies Using Cholorotoxin-Conjugated Iron Oxide Nanoparticles: A Novel Non-Radioactive Method

Recent advances in nanotechnology have led to the development of biocompatible nanoparticles for in vivo molecular imaging and targeted therapy. Many nanoparticles have undesirable tissue distribution or unacceptably low serum half-lives. Pharmacokinetic (PK) and biodistribution studies can help inform decisions determining particle size, coatings, or other features early in nanoparticle development. Unfortunately, these studies are rarely done in a timely fashion because many nanotechnology labs lack the resources and expertise to synthesize radioactive nanoparticles and evaluate them in mice.To address this problem, we developed an economical, radioactivity-free method for assessing serum half-life and tissue distribution of nanoparticles in mice. Iron oxide nanoparticles coated with chitosan and polyethylene glycol that utilize chlorotoxin as a targeting molecule have a serum half-life of 7-8 hours and the particles remain stable for extended periods of time in physiologic fluids and in vivo. Nanoparticles preferentially distribute to spleen and liver, presumably due to reticuloendothelial uptake. Other organs have very low levels of nanoparticles, which is ideal for imaging most cancers in the future. No acute toxicity was attributed to the nanoparticles.We report here a simple near-infrared fluorescence based methodology to assess PK properties of nanoparticles in order to integrate pharmacokinetic data into early nanoparticle design and synthesis. The nanoparticles tested demonstrate properties that are excellent for future clinical imaging strategies and potentially suitable for targeted therapy

OXYGEN REACTIVE POLYMERS FOR TREATMENT OF TRAUMATIC BRAIN INJURY

Methods and compositions for treating traumatic brain injury . The methods and compositions utilize a multi - functional oxygen reactive polymer ( ORP ) that includes repeating units that include a reactive oxygen species ( ROS ) scavenging group and a polyalkylene oxide group . For theranostic applications , the oxygen reactive polymer fur ther includes a diagnostic group

Efficacy of transdermal 4% lidocaine patches for postoperative pain management after arthroscopic rotator cuff repair: a prospective trial.

Background: Postoperative pain management continues to be a challenging aspect of patient care. Lidocaine patches have shown efficacy in reducing pain in other surgical specialties and mixed results in orthopedic trials. We sought to determine the effectiveness of nonprescription lidocaine patches in reducing postoperative pain after arthroscopic rotator cuff repair. Methods: Patients undergoing primary arthroscopic rotator cuff repair were recruited from 3 surgeons at a single institution. All patients of each surgeon were randomized to a lidocaine patch or control group, with crossover occurring at the midpoint. Experimental group patients received 26 4% lidocaine gel-patches. They were provided written and visual instructions to begin wearing the lidocaine patches during daytime on postoperative day (POD) 2. They were to be switched every 8 hours and removed overnight. Control group patients received normal standard of care but did not receive a placebo control. Exclusion criteria included workmen\u27s compensation claims, ageyears, history of myocardial infarction, and history of lidocaine or adhesive allergies. The American Shoulder and Elbow Surgeons shoulder survey was completed preoperatively and 2-, 6-weeks, 3-, 4.5-, and 6-months postoperatively. A 14-day visual analog scale pain and medication log was completed three times daily following repair. All patients received interscalene nerve block with bupivacaine and general anesthesia. Results: 80 (40 control, 40 lidocaine) patients were enrolled, with 53 completing follow-up. Groups were demographically similar in age (P = .22), gender (P = .20), and body mass index (P = .77). They were similar in tear pattern (P = .95), concomitant acromioplasty (P = .44), concomitant biceps tenodesis (P = .07), and number of anchors used (P = .25). There was no difference in American Shoulder and Elbow Surgeons scores at any time points (range P = .28-P = .97). Reported 14-day pain logs were not different between study groups at any time points (range P = .07-P = .99). There was no difference in opioid consumption in the first 14 days after surgery (P = .38). The lidocaine group reported less satisfaction with their pain management beginning in the evening of POD 2 (P = .05). This continued until the afternoon of POD 8 (P = .03). Conclusion: Transdermal 4% lidocaine patches are not effective in reducing pain or opioid consumption after arthroscopic rotator cuff repair and were associated with reduced patient satisfaction

A protein domain interaction interface database: InterPare.

BACKGROUND: Most proteins function by interacting with other molecules. Their interaction interfaces are highly conserved throughout evolution to avoid undesirable interactions that lead to fatal disorders in cells. Rational drug discovery includes computational methods to identify the interaction sites of lead compounds to the target molecules. Identifying and classifying protein interaction interfaces on a large scale can help researchers discover drug targets more efficiently. DESCRIPTION: We introduce a large-scale protein domain interaction interface database called InterPare http://interpare.net. It contains both inter-chain (between chains) interfaces and intra-chain (within chain) interfaces. InterPare uses three methods to detect interfaces: 1) the geometric distance method for checking the distance between atoms that belong to different domains, 2) Accessible Surface Area (ASA), a method for detecting the buried region of a protein that is detached from a solvent when forming multimers or complexes, and 3) the Voronoi diagram, a computational geometry method that uses a mathematical definition of interface regions. InterPare includes visualization tools to display protein interior, surface, and interaction interfaces. It also provides statistics such as the amino acid propensities of queried protein according to its interior, surface, and interface region. The atom coordinates that belong to interface, surface, and interior regions can be downloaded from the website. CONCLUSION: InterPare is an open and public database server for protein interaction interface information. It contains the large-scale interface data for proteins whose 3D-structures are known. As of November 2004, there were 10,583 (Geometric distance), 10,431 (ASA), and 11,010 (Voronoi diagram) entries in the Protein Data Bank (PDB) containing interfaces, according to the above three methods. In the case of the geometric distance method, there are 31,620 inter-chain domain-domain interaction interfaces and 12,758 intra-chain domain-domain interfaces

The Seoul National University AGN Monitoring Project. IV. Hα Reverberation Mapping of Six AGNs and the Hα Size–Luminosity Relation

The broad-line region (BLR) size–luminosity relation has paramount importance for estimating the mass of black holes in active galactic nuclei (AGNs). Traditionally, the size of the Hβ BLR is often estimated from the optical continuum luminosity at 5100 Å, while the size of the Hα BLR and its correlation with the luminosity is much less constrained. As a part of the Seoul National University AGN Monitoring Project, which provides 6 yr photometric and spectroscopic monitoring data, we present our measurements of the Hα lags of high-luminosity AGNs. Combined with the measurements for 42 AGNs from the literature, we derive the size–luminosity relations of the Hα BLR against the broad Hα and 5100 Å continuum luminosities. We find the slope of the relations to be 0.61 ± 0.04 and 0.59 ± 0.04, respectively, which are consistent with the Hβ size–luminosity relation. Moreover, we find a linear relation between the 5100 Å continuum luminosity and the broad Hα luminosity across 7 orders of magnitude. Using these results, we propose a new virial mass estimator based on the Hα broad emission line, finding that the previous mass estimates based on scaling relations in the literature are overestimated by up to 0.7 dex at masses lower than 107M⊙

A Modifier Screen for Bazooka/PAR-3 Interacting Genes in the Drosophila Embryo Epithelium

The development and homeostasis of multicellular organisms depends on sheets of epithelial cells. Bazooka (Baz; PAR-3) localizes to the apical circumference of epithelial cells and is a key hub in the protein interaction network regulating epithelial structure. We sought to identify additional proteins that function with Baz to regulate epithelial structure in the Drosophila embryo.The baz zygotic mutant cuticle phenotype could be dominantly enhanced by loss of known interaction partners. To identify additional enhancers, we screened molecularly defined chromosome 2 and 3 deficiencies. 37 deficiencies acted as strong dominant enhancers. Using deficiency mapping, bioinformatics, and available single gene mutations, we identified 17 interacting genes encoding known and predicted polarity, cytoskeletal, transmembrane, trafficking and signaling proteins. For each gene, their loss of function enhanced adherens junction defects in zygotic baz mutants during early embryogenesis. To further evaluate involvement in epithelial polarity, we generated GFP fusion proteins for 15 of the genes which had not been found to localize to the apical domain previously. We found that GFP fusion proteins for Drosophila ASAP, Arf79F, CG11210, Septin 5 and Sds22 could be recruited to the apical circumference of epithelial cells. Nine of the other proteins showed various intracellular distributions, and one was not detected.Our enhancer screen identified 17 genes that function with Baz to regulate epithelial structure in the Drosophila embryo. Our secondary localization screen indicated that some of the proteins may affect epithelial cell polarity by acting at the apical cell cortex while others may act through intracellular processes. For 13 of the 17 genes, this is the first report of a link to baz or the regulation of epithelial structure