BACKGROUND

The aim of our study is to determine the accuracy of magnetic resonance imaging (MRI) scan in relation to arthroscopic findings in patients presenting with chronic ankle pain and/or instability.

METHODS

All patients who underwent arthroscopy of the ankle as well as MRI from December 2005 to July 2008 in our institution were reviewed by the Orthopaedic surgeons. Twenty-four patients were identified and the results of MRI scans were compared with arthroscopic findings. This study specifically looked at anterior talofibular ligament (ATFL), calcaneofibular ligament (CFL) and osteochondral lesions (OCD). Arthroscopic findings were considered as a gold standard. There were 12 female and 12 male patients with an average age 39 years (11-65 years). Time interval between MRI scan and arthroscopy was 7.0 months (2-18 months).

RESULTS

In our study MRI showed 100% specificity for the diagnosis of ATFL and CFL tears and osteochondral lesions. However sensitivity was low particularly for CFL tears. Accuracy of MRI in detecting ATFL tear was 91.7%, CFL tear was 87.5% and osteochondral lesion was 83.3%.

CONCLUSIONS

We conclude that MRI scan has very high specificity and positive predictive value in diagnosing tears of ATFL, CFL and osteochondral lesions. However sensitivity was low with MRI. In a symptomatic patient negative results on MRI must be viewed with caution and an arthroscopy may still be required for a definitive diagnosis and treatment. However high resolution scans may differ in their ability to pick up these lesions and further research is required to assess their efficiency as evidence is not currently available.

The aims of this study were to measure the forces in the anterior talofibular ligament (ATFL) and calcaneofibular ligament (CFL) and the motion in the tibiotalar and subtalar joints during simulated weight-bearing in eight cadaver ankle specimens. An MTS test machine was used to apply compressive loads to specimens held in a specially designed testing apparatus in which the ankle position (dorsiflexion-plantarflexion and supination-pronation) could be varied in a controlled manner. The forces in the ATFL and CFL were measured with buckle transducers. Tibiotalar motion and total ankle joint motion were measured with an instrumented spatial linkage. The specimens were positioned sequentially at 10 degrees dorsiflexion, neutral, and 10 degrees and 20 degrees plantarflexion, and this sequence was repeated at 15 degrees supination, neutral pronation/supination, and 15 degrees pronation. Force and motion measurements were recorded in each of these positions with and without a 375 N compressive load simulating weight-bearing. From 10 degrees dorsiflexion to 20 degrees plantarflexion, all motion occurred in the tibiotalar joint. In contrast, the ratio of subtalar motion to tibiotalar motion was 3:1 for supination-pronation and 4:1 for internal-external rotation. Inverse loading patterns were observed for the ATFL and CFL from plantarflexion to dorsiflexion. Compressive loading did not affect CFL tension, but it magnified the pattern of increasing ATFL tension with plantarflexion. The largest increase in ATFL force was observed in supination and plantarflexion with a compressive load (76 +/- 23 N), whereas CFL tension mainly increased in supination and dorsiflexion with a compressive load (109 +/- 28 N). In conclusion, the results showed that the ATFL acted as a primary restraint in inversion, where injuries typically occur (combined plantarflexion, supination and internal rotation). Also, the subtalar joint was of primary importance for normal supination-pronation and internal-external rotation.

Coronatine (COR) is a plasmid-encoded phytotoxin synthesized by several pathovars of phytopathogenic Pseudomonas syringae. The COR biosynthetic gene cluster in P. syringae pv. glycinea PG4180 is encoded by a 32-kb region which contains both the structural and regulatory genes needed for COR synthesis. The regulatory region contains three genes: corP, corS, and corR. corS is thought to function as a histidine protein kinase, whereas corP and corR show relatedness to response regulators of the two-component regulatory paradigm. In the present study, we investigated whether CorR is a positive activator of COR gene expression. We also studied whether CorR specifically binds the DNA region located upstream of cfl, a gene located at the 5' end of the gene cluster encoding coronafacic acid, the polyketide portion of COR. Complementation analysis with a corR mutant, PG4180.P2, and transcriptional fusions to a promoterless glucuronidase gene (uidA) indicated that CorR functions as a positive regulator of COR gene expression. Deletion analysis of the 5' end of the cfl upstream region was used to define the minimal region required for COR gene expression. A 360-bp DNA fragment located over 500 bp upstream from the cfl transcriptional start site was used in DNase I protection assays to define the specific bases bound by CorR. An area extending from -704 to -650 with respect to the cfl transcriptional start site was protected by DNase I footprinting, indicating a rather large area of protection. This area was also conserved in the promoter region for cmaA, which encodes a transcript containing genes for coronamic acid synthesis, another intermediate in the COR biosynthetic pathway. The results obtained in the current study suggest that both the coronafacic acid and the coronamic acid structural genes are controlled by CorR, a positive activator of COR gene expression.

Coronafacic acid (CFA), the polyketide component of the phytotoxin coronatine (COR), is activated and coupled to coronamic acid via amide bond formation, a biosynthetic step presumably catalyzed by the CFA ligase (cfl) gene product. The COR biosynthetic gene cluster in Pseudomonas syringae pv. glycinea PG4180 is located within a 32-kb region of a 90-kb plasmid designated p4180A. In the present study, a cloned region of p4180A complemented all CFA- mutants spanning an 18.8-kb region of the COR biosynthetic cluster. The genetic evidence presented in this study indicates that cfl and the CFA biosynthetic gene cluster are encoded by a single transcript and that transcription of all of the genes in this operon is directed by the cfl promoter. The cfl promoter was localized to a 0.37-kb region upstream of the transcriptional start site by progressive subcloning in pRG960sd, a vector containing a promoterless glucuronidase gene. Transcription of the cfl/CFA operon was temperature sensitive and showed maximal glucuronidase activity at 18 degrees C. Furthermore, transcription of the cfl/CFA operon was dependent on the functional activity of a modified two-component regulatory system located within the COR biosynthetic gene cluster. Thermoregulation of the cfl/CFA operon and the coronamic acid biosynthetic gene cluster via the modified two-component regulatory system is discussed.

The Controlled Oral Word Association (COWA) Test is a brief and sensitive measure of executive cognitive dysfunction. There are two commonly used forms of the test, one using the letters F, A, and S, and the other using C, F, and L. This study examines the relative difficulty of the two forms using a meta-analytic approach that includes multiple samples of normal individuals. The effects of age, education, gender composition, exclusion criteria, and age of study are also examined. Results indicate that the CFL form of the test is more difficult and that age, education, and the use of strict exclusion criteria influence performance. Performance is more variable for the FAS form, and age and age of study influence performance variability.

Polycystin-1 (Pc1) cleavage at the G protein-coupled receptor (GPCR) proteolytic site (GPS) is required for normal kidney morphology in humans and mice. We found a complex pattern of endogenous Pc1 forms by GPS cleavage. GPS cleavage generates not only the heterodimeric cleaved full-length Pc1 (Pc1(cFL)) in which the N-terminal fragment (NTF) remains noncovalently associated with the C-terminal fragment (CTF) but also a novel (Pc1) form (Pc1(deN)) in which NTF becomes detached from CTF. Uncleaved Pc1 (Pc1(U)) resides primarily in the endoplasmic reticulum (ER), whereas both Pc1(cFL) and Pc1(deN) traffic through the secretory pathway in vivo. GPS cleavage is not a prerequisite, however, for Pc1 trafficking in vivo. Importantly, Pc1(deN) is predominantly found at the plasma membrane of renal epithelial cells. By functional genetic complementation with five Pkd1 mouse models, we discovered that CTF plays a crucial role in Pc1(deN) trafficking. Our studies support GPS cleavage as a critical regulatory mechanism of Pc1 biogenesis and trafficking for proper kidney development and homeostasis.

BACKGROUND

Multiple techniques have been described for reconstruction of the lateral ligaments of the ankle. Most require extensive exposure and dissection, which may lead to potential problems with wound healing, higher risk of nerve injury, fibrosis, and stiffness. This study reports on the results of a minimally invasive method to reconstruct the ligaments using a semitendinosus tendon autograft and achieve a stable ankle while avoiding these problems.

METHODS

From September 2006 to May 2010, 25 patients (14 males, 11 females) with chronic ankle instability underwent lateral ligament reconstruction. The average age was 32.4 (range, 17 to 62) years old. A semitendinosus autograft was harvested through 2 small knee incisions. For the ankle reconstruction, 4 small incisions of 5 mm each were made at the medial and lateral side of the fibular tip, the talar neck, and the middle of the calcaneus. Anatomical reconstruction of the anterior talofibular ligament (ATFL) and calcaneofibular ligament (CFL) was then performed through these small incisions. The mean final follow-up was 32.3 (range, 12 to 56) months. AOFAS questionnaires were used to measure clinical outcomes and donor site morbidity and patient satisfaction are also reported. Preoperative and postoperative stress tests were performed and radiographic parameters were measured.

RESULTS

The mean AOFAS score increased on average from 71.1 to 95.1 (P < .001). Two patients reported residual instability on uneven ground. No patient reported weakness or disability from the donor site. The satisfaction level was excellent in 20 patients and good in 5 patients. Significant improvement in stress radiographic parameters was noted for the talar tilt angle, with reduction from a mean of 14.0 to 3.8 degrees (P < .001); anterior talar displacement reduced from a mean of 12.3 to 4.6 mm (P < .001).

CONCLUSIONS

Reconstruction of the lateral ankle ligaments using a semitendinosus tendon autograft and a minimally invasive approach can achieve a stable ankle while avoiding extensive exposure and risk of nerve injury.

METHODS

Level IV, retrospective case series.

Cross-linked hyperbranched fluoropolymer (HBFP) and poly(ethylene glycol) (PEG) amphiphilic networks with PEG weight percentages of 14% (HBFP-PEG14), 29% (HBFP-PEG29), 45% (HBFP-PEG45), and 55% (HBFP-PEG55) were prepared on 3-aminopropyl)triethoxysilane (3-APS) functionalized microscope glass slides for marine antifouling and fouling-release applications. The surface-free energies (gamma(s)), polar (gamma(s)(p) and gamma(s)(AB)), and dispersion (gamma(s)(d) and gamma(s)(LW)) components were evaluated using advancing contact angles by two-liquid geometric-mean and three-liquid Lifshitz-van der Waals acid-base approaches. The HBFP coating exhibited a low surface energy of 22 mJ/m(2), while the gamma(s) and gamma(s)(p) of the cross-linked HBFP-PEG coatings increased proportionally with the PEG weight percentages in the networks. The adsorption of bovine serum albumin (BSA), lectin from Codium fragile (CFL), lipopolysaccharides from Escherichia coli (LPSE) and Salmonella minnesota (LPSS) upon glass, APS-glass, HBFP, PEG, and the cross-linked HBFP-PEG network coatings were investigated by fluorescence microscopy. The marine antifouling and fouling-release properties of the cross-linked HBFP-PEG coatings were evaluated by settlement and release assays involving zoospores of green fouling alga Ulva (syn. Enteromorpha; Hayden, H. S.; Blomster, J.; Maggs, C. A.; Silva, P. C.; Stanhope, M. J.; Waaland, J. R. Eur. J. Phycol. 2003, 38, 277). The growth and release of Ulva sporelings were also investigated upon the HBFP-PEG45 coating in comparison to a poly(dimethylsiloxane) elastomer (PDMSE) standard material. Of the heterogeneous cross-linked network coatings, the maximum resistances to protein, lipopolysaccharide, and Ulva zoospore adhesion, as well as the best zoospore and sporeling release properties, were recorded for the HBFP-PEG45 coating. This material also exhibited better performance than did a standard PDMSE coating, suggesting its unique applicability in fouling-resistance applications.

For operative reconstruction, precise anatomic information on the dimensions of the ankle ligaments is important and can help to optimize these procedures. The purpose of this study was to investigate the length and width dimensions of the ankle ligaments and to contrast the results with the published literature. Seventeen non-paired adult, formalin-fixed ankle specimen were dissected to expose the capsuloligamentous structures. The following ligaments were investigated: tibiofibular syndesmosis (anterior and posterior tibiofibular ligament/ATiFL and PTiFL), lateral ankle ligaments (anterior and posterior talofibular ligament, calcaneofibular ligament/ATFL, PTFL and CFL), medial ankle ligaments (deltoid ligament, anterior and posterior tibiotalar ligament/ATTL and PTTL). After identification of the ligaments, the dimensions were measured with a ruler and a sliding caliper. Additionally, the attachment area and the center of insertion (COI) were evaluated. The dimensions of the ligaments were recorded. Measurements were calculated and discussed according to the existing literature. The tibial COI of the ATiFL was situated 8.35 ± 2.05 mm from the inferior articular surface of the tibia and 5.04 ± 1.32 mm from the fibular notch. Its fibular COI was situated 25.45 ± 5.84 mm from the tip of the lateral malleolus and 3.12 ± 1.01 mm from the malleolar articular surface. The calcaneal COI of the CFL was situated 20.63 ± 3.56 mm anterior and 5.73 ± 1.89 mm plantar to the superior edge of the calcaneal. Its fibular attachment of the CFL was directly at the tip of the lateral malleolus, dorsal to the fibular attachment of the ATFL. Studies of the therapeutic options in severe ankle ligament injuries have shown better results in anatomical reconstructions compared to other operative treatments. To optimize these procedures, exact anatomical information on the dimensions of the ankle ligaments should be beneficial.

OBJECTIVE

The purpose of this study was to determine the clinical utility of three bony tubercles: fibular obscure tubercle, talar obscure tubercle and tuberculum ligamenti calcaneofibularis, to serve as anatomical landmarks for defining the precise location of the origins and insertions of the anterior talofibular ligament (ATFL) and calcaneofibular ligament (CFL).

METHODS

Twelve lower extremity cadaveric specimens were procured. The detectability of the tubercles was tested using palpation and fluoroscopy with subsequent confirmation after dissection. If the tubercles were present, then distances from the identified tubercles to the footprint centres and the intersection of the ATFL and CFL were measured to allow precise localization of the ATFL and CFL origin and intersection sites. Further, if the tubercles were not detectable, then an attempt to provide an alternative means of localizing ATFL and CFL origin and insertion sites was made by measuring distances between alternative landmarks and other important structures. All the measurements were performed by two researchers, and the results were averaged.

RESULTS

The fibular obscure tubercle existed and was detectable in all specimens. It was located 1.3 mm proximal to the articular tip of the fibula, 2.7 mm to the intersection of the ATFL and CFL, 3.7 mm distal to the ATFL and 4.9 mm proximal to the CFL origins. The talar obscure tubercle existed 58 % of specimens and was detectable in 57 %. The talar obscure tubercle was located 1.4 mm to the ATFL. The ATFL insertion point was located 60 % of the distance from the inferolateral corner to the anterolateral corner of the of talar body along the anterior border of the talar lateral articular facet. The tuberculum ligamenti calcaneofibularis existed in 33 % of specimens and was detectable in 8 %. The CFL inserted 17 mm on a perpendicular projected line distal from the subtalar joint.

CONCLUSIONS

The fibular obscure tubercle was clinically relevant and reliable bony landmark of the ATFL and CFL origin location. However, the talar obscure tubercle was less reliable and the tuberculum ligamenti calcaneofibularis was rarely available and as such alternative landmarks for the ATFL and CFL insertion location should be utilized. The present study describes the utility of clinically relevant bony landmarks that may assist in identifying the origins and insertions of the ATFL and CFL to facilitate minimally invasive ankle stabilization surgery.

OBJECTIVE

A recent consensus statement has suggested ≥3 corner inflammatory lesions (CILs) or several corner fatty lesions (CFLs) as candidate criteria indicative of axial spondyloarthritis (SpA) on magnetic resonance imaging (MRI) of the spine. The aim of this study was to evaluate the diagnostic utility of these cutoffs in nonradiographic axial SpA and ankylosing spondylitis (AS).

METHODS

One hundred thirty consecutive patients with back pain who were ≤50 years of age and newly referred to 2 university clinics (cohorts A and B) were classified according to rheumatologist expert opinion based on results of clinical examination and pelvic radiography as having nonradiographic axial SpA (n = 50), AS (n = 33), or nonspecific back pain (n = 47). Cohort A also included 20 age-matched healthy controls. Four blinded readers assessed MRIs of the spine using the standardized Canada-Denmark module. Readers recorded CILs and CFLs in 23 discovertebral units. We tested the diagnostic utility (mean sensitivity and specificity over 4 readers) of the cutoff for the number of lesions on spinal MRI as proposed in the literature (≥2 or ≥3 CILs and ≥6 CFLs), and we tested for possible thresholds (from ≥1 CIL or CFL to ≥10 CILs or CFLs) for nonradiographic axial SpA and AS patients in both cohorts.

RESULTS

None of the spinal thresholds (≥2 or ≥3 CILs and ≥6 CFLs) showed clinically relevant diagnostic utility (positive likelihood ratio [LR] range 1.38-2.36) when comparing patients with nonradiographic axial SpA to patients with nonspecific back pain. A threshold of ≥6 CILs had moderate to substantial diagnostic utility (positive LR 13.26 and 6.74 in cohorts A and B, respectively) in nonradiographic axial SpA, while ≥4 CILs showed small diagnostic utility (positive LR 3.83 and 2.72 in cohorts A and B, respectively) but specificities of >0.90.

CONCLUSIONS

None of the previously proposed candidate criteria for a positive spinal MRI finding of axial SpA showed clinically relevant diagnostic utility in nonradiographic axial SpA. These results question the value of proposed definitions for a positive finding of SpA based on MRI of the spine alone.

The antibacterial working range of six lysozymes was tested under ambient and high pressure, on a panel of five gram-positive (Enterococcus faecalis, Bacillus subtilis, Listeria innocua, Staphylococcus aureus and Micrococcus lysodeikticus) and five gram-negative bacteria (Yersinia enterocolitica, Shigella flexneri, Escherichia coli O157:H7, Pseudomonas aeruginosa and Salmonella typhimurium). The lysozymes included two that are commercially available (hen egg white lysozyme or HEWL, and mutanolysin from Streptomyces globisporus or M1L), and four that were chromatographically purified (bacteriophage lambda lysozyme or LaL, bacteriophage T4 lysozyme or T4L, goose egg white lysozyme or GEWL, and cauliflower lysozyme or CFL). T4L, LaL and GEWL were highly pure as evaluated by silver staining of SDS-PAGE gels and zymogram analysis while CFL was only partially pure. At ambient pressure each gram-positive test organism displayed a specific pattern of sensitivity to the six lysozymes, but none of the gram-negative bacteria was sensitive to any of the lysozymes. High pressure treatment (130-300 MPa, 25 degrees C, 15 min) sensitised several gram-positive and gram-negative bacteria for one or more lysozymes. M. lysodeikticus and P. aeruginosa became sensitive to all lysozymes under high pressure, S. typhimurium remained completely insensitive to all lysozymes, and the other bacteria showed sensitisation to some of the lysozymes. The possible applications of the different lysozymes as biopreservatives, and the possible reasons for the observed differences in bactericidal specificity are discussed.

Microvascular leakage due to endothelial barrier dysfunction is a prominent feature of T helper 2 (Th2) cytokine mediated allergic inflammation. Interleukin-4 (IL-4) is a potent Th2 cytokine, known to impair the barrier function of endothelial cells. However, the effectors mediating IL-4 induced cytoskeleton remodeling and consequent endothelial barrier dysfunction remain poorly defined. Here we have used whole genome transcriptome profiling and gene ontology analyses to identify the genes and processes regulated by IL-4 signaling in human coronary artery endothelial cells (HCAEC). The study revealed Wnt5A as an effector that can mediate actin cytoskeleton remodeling in IL-4 activated HCAEC through the regulation of LIM kinase (LIMK) and Cofilin (CFL). Following IL-4 treatment, LIMK and CFL were phosphorylated, thereby indicating the possibility of actin stress fiber formation. Imaging of actin showed the formation of stress fibers in IL-4 treated live HCAEC. Stress fiber formation was notably decreased in the presence of Wnt inhibitory factor 1 (WIF1). Non-invasive impedance measurements demonstrated that IL-4 increased the permeability and impaired the barrier function of HCAEC monolayers. Silencing Wnt5A significantly reduced permeability and improved the barrier function of HCAEC monolayers upon IL-4 treatment. Our study identifies Wnt5A as a novel marker of IL-4 activated vascular endothelium and demonstrates a critical role for Wnt5A in mediating IL-4 induced endothelial barrier dysfunction. Wnt5A could be a potential therapeutic target for reducing microvascular leakage and edema formation in Th2 driven inflammatory diseases.

OBJECTIVE

To gain a better understanding of the precise anatomy of the lateral ligaments of the ankle through a systematic review of published cadaveric studies in order to improve anatomical minimally invasive surgery (MIS) for treatment of chronic ankle instability (CAI).

METHODS

A systematic review of the literature was performed using the PubMed, EMBASE, Cochrane databases and Web of Science on June 2015 with the two search concepts: "lateral ligament of the ankle" and "anatomy". Anatomical studies that reported gross anatomy of the anterior talar fibular ligament (ATFL) and calcaneal fibular ligament (CFL) in English were included to assess the morphology and origins and insertions of the ligaments. All records found in the literature search were screened by title and abstract. Potentially relevant articles were selected for full-text review. Each of the identified articles was reviewed and included in qualitative synthesis. The following data were abstracted from the included articles: authors, date of publication, sample size, mean age, the length and the width of the each ligament, number of bundle of the ATFL and the location and the footprint of the origins and insertions for the ATFL and CFL.

RESULTS

Sixteen studies were identified indicating the length of the ATFL and CFL was 12-24.8 and 18.5-35.8 mm, respectively, while the width was 5-11.1 and 4.6-7.6 mm, respectively. Fibular origins of the ATFL and CFL were located on the anterior border of distal fibula at a distance of 10-13.8 and 5.3-8.5 mm proximal to the tip of the fibula, respectively. The talar insertion of the ATFL was located 14.2-18.1 mm to the subtalar joint or 11.3-14.8 mm to the anterolateral corner of the talar body. The calcaneal insertion of the CFL was located 12.1-13 mm to the subtalar joint or 13.2-27.1 mm to the peroneal tubercle on the lateral wall of calcaneus.

CONCLUSIONS

Systematic review of the literature of the research for the ATFL and CFL has identified the morphology of the ligaments and their location of origins and insertions. This is the best available data about the ATFL and CFL which will facilitate more precise anatomical MIS for treatment of CAI.

METHODS

Systematic review, Level IV.

Identification of actin-depolymerizing factor homology (ADF-H) domains in the structures of several related proteins led first to the formation of the ADF/cofilin family, which then expanded to the ADF/cofilin superfamily. This superfamily includes the well-studied cofilin-1 (Cfl-1) and about a dozen different human proteins that interact directly or indirectly with the actin cytoskeleton, provide its remodeling, and alter cell motility. According to some data, Cfl-1 is contained in various human malignant cells (HMCs) and is involved in the formation of malignant properties, including invasiveness, metastatic potential, and resistance to chemotherapeutic drugs. The presence of other ADF/cofilin superfamily proteins in HMCs and their involvement in the regulation of cell motility were discovered with the use of various OMICS technologies. In our review, we discuss the results of the study of Cfl-1 and other ADF/cofilin superfamily proteins, which may be of interest for solving different problems of molecular oncology, as well as for the prospects of further investigations of these proteins in HMCs.

By exploiting the thermoplastic and photosensitive nature of SU-8 photoresists, different types of hierarchical pillar arrays with variable aspect ratios are fabricated through capillary force lithography (CFL), followed by photopatterning. The thermoplastic nature of SU-8 enables the imprinting of micropillar arrays with variable aspect ratios by CFL using a single poly(dimethylsiloxane) mold, simply by tuning the initial film thickness of SU-8 on a substrate. The pillar array is subsequently photopatterned through a photomask, followed by post-exposure baking above the glass transition temperature (T(g)) of SU-8. The pillars in the exposed region become highly crosslinked and, therefore, neither soluble nor able to reflow above T(g), whereas the pillars in the unexposed regions can reflow and flatten out. Two developing strategies are investigated after UV exposure of the SU-8 pillar arrays including i) solvent development and drying and ii) thermal reflow to create bilevel hierarchical structures with short pillars and single-level, dual-scaled, high-aspect-ratio (up to 7.7) pillars in a microdot array, respectively.

BACKGROUND

This was the second study in a Phase 3 program treating crow's feet lines (CFL) with onabotulinumtoxinA.

OBJECTIVE

To evaluate the efficacy and safety of onabotulinumtoxinA treatment of CFL alone or with glabellar lines (GL).

METHODS

This multicenter, double-blind, placebo-controlled, repeat treatment, 7-month study randomized subjects with moderate-to-severe CFL and GL (maximum contraction) to onabotulinumtoxinA 44 U (CFL: 24 U, GL: 20 U; n = 305), onabotulinumtoxinA 24 U (CFL: 24 U, GL: placebo; n = 306), or placebo (n = 306). Coprimary end points were investigator-assessed and subject-assessed proportion of subjects achieving a CFL Facial Wrinkle Scale Grade of 0 or 1 (maximum smile; Day 30, Cycle 1). Additional efficacy end points and safety/adverse events (AEs) were evaluated.

RESULTS

All primary and secondary end points were achieved; statistically significant differences favored onabotulinumtoxinA (p < .001, all comparisons vs placebo). Investigator and subject responder rates were: CFL, 54.9% and 45.8%; CFL + GL, 59.0% and 48.5%; and placebo, 3.3% (both), respectively. Responder rates on other end points also significantly favored onabotulinumtoxinA treatments. Most AEs were mild or moderate. Two subjects discontinued: 1 serious AE unrelated to treatment (myocardial infarction) and 1 treatment-related AE (injection site pain).

CONCLUSIONS

OnabotulinumtoxinA was effective and well tolerated for treating moderate-to-severe CFL alone or in combination with GL.

Toxin-based identification procedures are useful for differentiating Pseudomonas syringae pathovars. A biological test on peptone-glucose-NaCl agar in which the yeast Rhodotorula pilimanae was used proved to be more reliable for detecting lipodepsipeptide-producing strains of P. syringae than the more usual test on potato dextrose agar in which Geotrichum candidum is used. A PCR test performed with primers designed to amplify a 1, 040-bp fragment in the coding sequence of the syrD gene, which was assumed to be involved in syringomycin and syringopeptin secretion, efficiently detected the gene in pathovars that produce the lipodepsipeptides. Comparable results were obtained in both tests performed with strains of the syringomycin-producing organisms P. syringae pv. syringae, P. syringae pv. atrofaciens, and P. syringae pv. aptata, but the PCR test failed with a syringotoxin-producing Pseudomonas fuscovaginae strain. The specificity of the test was verified by obtaining negative PCR test results for related pathovars or species that do not produce the toxic lipodepsipeptides. P. syringae pv. syringae was detected repeatedly in liquid medium inoculated with diseased vegetative tissue and assayed by the PCR test. Our procedure was also adapted to detect P. syringae pv. morsprunorum with a cfl gene-based PCR test.

Convenience foods enable the consumer to save time and effort in food activities, related to shopping, meal preparation and cooking, consumption and post-meal activities. The objective of this paper is to report on the attitudes and reported behaviour of food consumers in Great Britain based on a review of their convenience food lifestyle (CFLs). The paper also reports the development and application of a segmentation technique that can supply information on consumer attitudes towards convenience foods. The convenience food market in Great Britain is examined and the key drivers of growth in this market are highlighted. A survey was applied to a nationally representative sample of 1000 consumers (defined as the persons primarily responsible for food shopping and cooking in the household) in Great Britain in 2002. Segmentation analysis, based on the identification of 20 convenience lifestyle factors, identified four CFL segments of consumers: the 'food connoisseurs' (26%), the 'home meal preparers' (25%), the 'kitchen evaders' (16%) and the 'convenience-seeking grazers' (33%). In particular, the 'kitchen evaders' and the 'convenience-seeking grazers' are identified as convenience-seeking segments. Implications for food producers, in particular, convenience food manufacturers are discussed. The study provides an understanding of the lifestyles of food consumers in Great Britain, and provides food manufacturers with an insight into what motivates individuals to purchase convenience foods.

Tropolone emerged from the screening of a chelator fragment library (CFL) as an inhibitor of the Zn(2+)-dependent virulence factor, Pseudomonas aeruginosa elastase (LasB). Based on this initial hit, a series of substituted tropolone-based LasB inhibitors was prepared, and a compound displaying potent activity in vitro and in a bacterial swarming assay was identified. Importantly, this inhibitor was found to be specific for LasB over other metalloenzymes, validating the usage of tropolone as a viable scaffold for identifying first-in-class LasB inhibitors.

The management of chronic lateral instability of the ankle remains controversial. In general, the anterior talofibular ligament (ATFL) must be reconstructed in all patients. Some will also need reconstruction of the calcaneofibular ligament (CFL) (or its function) to regain stability of both the ankle and the subtalar joints, and to avoid recurrence of instability. After reconstruction, most authors report good to excellent results in 80% to 85% of patients. We describe the augmented reconstruction technique of ATFL and CFL with a semitendinosus tendon allograft through a peroneal bone tunnel fixed with biodegradable anchors, and advocate this procedure as a safe, effective method to manage lateral ankle instability.

Cardiovascular disease remains the leading cause of death worldwide(1). Cardiac tissue engineering holds much promise to deliver groundbreaking medical discoveries with the aims of developing functional tissues for cardiac regeneration as well as in vitro screening assays. However, the ability to create high-fidelity models of heart tissue has proven difficult. The heart's extracellular matrix (ECM) is a complex structure consisting of both biochemical and biomechanical signals ranging from the micro- to the nanometer scale(2). Local mechanical loading conditions and cell-ECM interactions have recently been recognized as vital components in cardiac tissue engineering(3-5). A large portion of the cardiac ECM is composed of aligned collagen fibers with nano-scale diameters that significantly influences tissue architecture and electromechanical coupling(2). Unfortunately, few methods have been able to mimic the organization of ECM fibers down to the nanometer scale. Recent advancements in nanofabrication techniques, however, have enabled the design and fabrication of scalable scaffolds that mimic the in vivo structural and substrate stiffness cues of the ECM in the heart(6-9). Here we present the development of two reproducible, cost-effective, and scalable nanopatterning processes for the functional alignment of cardiac cells using the biocompatible polymer poly(lactide-co-glycolide) (PLGA)(8) and a polyurethane (PU) based polymer. These anisotropically nanofabricated substrata (ANFS) mimic the underlying ECM of well-organized, aligned tissues and can be used to investigate the role of nanotopography on cell morphology and function(10-14). Using a nanopatterned (NP) silicon master as a template, a polyurethane acrylate (PUA) mold is fabricated. This PUA mold is then used to pattern the PU or PLGA hydrogel via UV-assisted or solvent-mediated capillary force lithography (CFL), respectively(15,16). Briefly, PU or PLGA pre-polymer is drop dispensed onto a glass coverslip and the PUA mold is placed on top. For UV-assisted CFL, the PU is then exposed to UV radiation (λ = 250-400 nm) for curing. For solvent-mediated CFL, the PLGA is embossed using heat (120 °C) and pressure (100 kPa). After curing, the PUA mold is peeled off, leaving behind an ANFS for cell culture. Primary cells, such as neonatal rat ventricular myocytes, as well as human pluripotent stem cell-derived cardiomyocytes, can be maintained on the ANFS(2).

Hepatocellular carcinoma (HCC) is the most common type of liver cancer. Despite the availability of several treatment strategies, resistance to chemotherapeutic agents, which limits the effectiveness of anticancer drugs, is a major problem in cancer therapy. In this study, we used a histone deacetylases inhibitor (HDACi) to establish drug-resistant HCC cells and further analyzed the molecular mechanisms underlying the development of resistance in HCC cells. Compared with the parental cells, HDACi-resistant cells showed high metastatic and pro-survival abilities. Two-dimensional electrophoresis data showed that the cofilin-1 (CFL-1) protein was altered in HDACi-resistant cells and was highly expressed in resistant cells compared with parental cells. The molecular function of CFL-1 is actin depolymerization, and it is involved in tumor metastasis. In this study, we showed that CFL-1 inhibition decreased cell migration and increased cell apoptosis in HDACi-resistant cells. We observed that HDACi induced ROS accumulation in cells and apoptosis via promotion of the CFL-1 interaction with Bax and CFL-1 translocation to the mitochondria, resulting in cytochrome C release. Importantly, phosphorylation of CFL-1 by activated extracellular signal-regulated kinases 1 and 2 (ERK1/2) confers strong protection against HDAC inhibitor-induced cell injury. p-CFL-1 shows a loss of affinity with Bax and will not translocate to mitochondria, stably remaining in the cytoplasm. These results indicate that phosphorylation to inactivate CFL-1 decreased the chemosensitivity to HDAC inhibitors and resulting in drug resistance of HCC cells.

Rapid nitric oxide (NO) production via endothelial NO synthase (eNOS) activation represents a major signaling pathway for the cardiovascular protective effects of estrogens; however, the pathways after NO biosynthesis that estrogens use to function remain largely unknown. Covalent adduction of a NO moiety to cysteines, termed S-nitrosylation (SNO), has emerged as a key route for NO to directly regulate protein function. Cofilin-1 (<em>CFL</em>1) is a small actin-binding protein essential for actin dynamics and cytoskeleton remodeling. Despite being identified as a major SNO protein in endothelial cells, whether SNO regulates <em>CFL</em>-1 function is unknown. We hypothesized that estradiol-17β (E2β) stimulates SNO of <em>CFL</em>1 via eNOS-derived NO and that E2β-induced SNO-<em>CFL</em>1 mediates cytoskeleton remodeling in endothelial cells. Point mutation studies determined Cys80 as the primary SNO site among the 4 cysteines (Cys39/80/139/147) in <em>CFL</em>1. Substitutions of Cys80 with Ala or Ser were used to prepare the SNO-mimetic/deficient (C80A/S) <em>CFL</em>1 mutants. Recombinant wild-type (wt) and mutant <em>CFL</em>1 proteins were prepared; their actin-severing activity was determined by real-time fluorescence imaging analysis. The activity of C80A <em>CFL</em>1 was enhanced to that of the constitutively active S3/A <em>CFL</em>1, whereas the other mutants had no effects. C80A/S mutations lowered Ser3 phosphorylation. Treatment with E2β increased filamentous (F)-actin and filopodium formation in endothelial cells, which were significantly reduced in cells overexpressing wt-<em>CFL</em>. Overexpression of C80A, but not C80S, <em>CFL</em>1 decreased basal F-actin and further suppressed E2β-induced F-actin and filopodium formation compared with wt-<em>CFL</em>1 overexpression. Thus, SNO(Cys80) of cofilin-1 via eNOS-derived NO provides a novel pathway for mediating estrogen-induced endothelial cell cytoskeleton remodeling.